38 - pc_1_1_Jul_Sep_2011

Pharmacognosy Communications An Official Publication of Pharmacognosy Network Worldwide [Phcog.Net]

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Editor-in-Chief

Dr Ian Cock Biomolecular and Physical Sciences Griffith University, Nathan campus,

170 Kessels Rd, Nathan, Queensland 4111 Australia

Editorial Board Members

Dr. William N. Setzer, Professor and Chair Department of Chemistry

The University of Alabama in Huntsville Huntsville, AL 35899, USA

Dr. Khuraman MUSTAFAYEVA Pharmaceutical faculty

Azerbaijan Medical University

Dr David Ruebhart HydroTox Services Melbourne Australia

Dr. Omayma A. El Dahshan, Ph D Pharmacognosy Dept., Faculty of pharmacy, Ain shams University,

Cairo, Egypt

Dr. Michał Tomczyk Medical University of Białystok, Faculty of Pharmacy,

Department of Pharmacognosy, ul. Mickiewicza 2a, 15-089 Białystok, Poland

Prof. Ameenah Gurib-Fakim, CEPHYR Ltd (Centre for Phytotherapy and Research)

7th Floor, Cyber Tower 2 Ebene, Mauritius

Dr. Philip G. Kerr PhD School of Biomedical Sciences

Charles Sturt University Wagga Wagga NSW 2678

Australia

Prof. Dr. Rimantas Venskutonis Department of Food Technology Kaunas University of Technology

Radvilenu pl. 19, Kaunas LT-50254, Lithuania

Editor - Publications

Dr. Mueen Ahmed KK

Aim and Scope Phcog Commn. is aimed at a broad readership, publishing articles on all aspects of pharmacognosy, and related fields. The journal aims to increase understanding of pharmacognosy as well as to direct and foster further research through the dissemination of scientific information by the publication of manuscripts.

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Contents

Pharmacognosy Communications www.phcogcommn.org

Volume 1 | Issue 1 | Jul-Sep 2011

Editorial

Pharmacognosy Communications: The Scope of Pharmacognosy 1I.E. Cock

Invited Review

Plant Drugs Used to Combat Menace of Anxiety Disorders 4Reecha Madaan, Suresh Kumar, Gundeep Bansal, Anupam Sharma

Review Article

Problems of Reproducibility and Efficacy of Bioassays Using Crude Extracts, with Reference to Aloe vera 52I.E. Cock

Research Article

Cassane-type diterpenoids from the genus Caesalpinia 63R. A. Dickson, T. C. Fleischer, P. J. Houghton

Azadirachtolide: An anti-diabetic and hypolipidemic effects from Azadirachta indica leaves 78Dineshkumar B, Analava Mitra, Manjunatha M

Research Letter

Antimicrobial and anti-inflammatory activities of the leaves of Clerodendrum splendens leaves 85Fleischer, TC, Mensah, AY, Oppong, AB, Mensah, MLK, Dickson, RA, Annan, K

Chemical Examination and Hair Growth studies on the Rhizomes of Hedychium spicatum Buch.-ham 90G. Venkateswara Rao, T. Mukhopadhyay, M. S. L. Madhavi, S. Lavakumar

World Wide Web

Inside Pharmacognosy: A Blog [Pharmanocognosy.in] 94

Medicinal Plant Images

Eucalyptus ficifolia and Chondrodendron tomentosum 95

Department Profile

Biomolecular and Physical Sciences, Griffith University, Australia. 96

Upcoming Events 99

Instructions 100

(c) Copyright 2011 EManuscript Publishing Services, India 1

Editorial



*Correspondence: Tel.: +61 7 37357637; fax: +61 7 37355282E-mail: [email protected] (I. E. Cock).DOI: 10.5530/pc.2011.1.1

Pharmacognosy Communications: The Scope of PharmacognosyI. E. Cocka,b*aBiomolecular and Physical Sciences, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Brisbane, Queensland 4111, Australia. bEnvironmental Futures Centre, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Brisbane, Queensland 4111, Australia

Pharmacognosy is the branch of pharmacology that studies drugs in their crude and/or natural states.[1] In general, when we describe pharmacognosy, we are usually referring to plant based medicinal systems. However, it is important to note that medicinal preparations may also be derived from animal sources as well as from fungi and microorganisms. Indeed, the discovery of the fungal antibiotic agent penicillin (from Penicillinum spp.) [2] is one of the most important medicinal findings to date. Many other useful medicinal products are also derived from fungi including the immunosuppressant mycophenolic acid (also from Penicillinum spp.)[3] and purgative anthraquinone emodin (from Penicillium islandicum).[4] Also, numerous hallucinogenic substances (eg. psilocin and psilocybin) are produced by Psilocybe spp. (family Tricholometaceae) of fungi.[5]

Similarly, numerous medicinal agents are produced by bacteria, especially further antibiotic agents. Very early studies demonstrated the antibiotic potential of bacteria towards other bacterial species. In 1887 it was accidently discovered that prior injection of Streptococcus erysipelatis protected guinea pigs from developing cholera when injected with Vibrio cholera.[6] Furthermore, it was also shown that previous injection of either Streptococcus erysipelatis or Pseudomonas aeruginosa also prevented the development of anthrax in experimental animals injected with Bacillus anthracis[6] and that pre-injection of sterilised cultures of the protective bacteria have the same protective effect as live bacteria.[7] This discovery stimulated further studies into the antibiotic activity of bacteria, resulting in the discovery of streptomycin, chloramphenicol, chlortetracycline, tetracycline, erythromycin, neomycin and numerous other antibiotics, especially from Streptomyces spp. (family Streptomycetaceae). Other bacteria, particularly Bacillus spp., are noted for their production of antibiotic polypeptides such as actinomycin,[8] bacitracin,[9] tyrothrycin[10] and polymixin.[10] These antibiotic polypeptides were initially not widely used as they also display strong cytotoxic

properties. More recently, there is renewed interest in their use due to their antitumor potential. Indeed, the bacterial antibiotic polypeptides doxorubicin, daunorubicin and actinomycin D are now routinely used in the treatment of a variety of cancers.[11,12]

Although the number of animal derived pharmacognostical agents is small when compared to fungi, bacteria and plants, there has recently been an increase in interest in marine creatures as a source of new drugs. Marine invertebrates in particular, account for much of the recent publications describing animal pharmacognosy. Some species of sponges have been found to have antibacterial, antifungal, antimalarial, cytotoxic and anticancer bioactivities.[13] Furthermore, sponges produce interesting metabolites including bromophenols, cyclic peroxides, peroxyketals and modified sesquiterpenes which warrant further investigation. [13] The soft coral Sarcophyton glaucum produces the diterpenoids sarcophytol A and sarcophytol A, which have tumour inhibiting bioactivity.[14]

Whilst marine animals are receiving much recent interest, there are also many examples of pharmacognostical agents derived from terrestrial animals. For examples, bees (Apis mellifica) provide us with multiple useful medicinal properties. The antimicrobial activity of honey produced by bees feeding on some plant species is known to be exceptionally good. Manuka honey (made by bees feeding on the Eastern Australian/New Zealand plant Leptospermum scoparium) is an especially good antimicrobial agent. [15] Additionally, beeswax and royal jelly are also reported to have therapeutic properties.[16] Toad skins contain cardioactive agents and were used to treat oedema prior to the development of more effective agents.[17] Pharmacognostic agents produced by vertebrates include lanolin from wool, gelatine and musk. In my own region of the world (Australia) there is also much interest in oils obtained from emu for its many therapeutic properties. [18]

Inorganic chemicals may also have important medicinal properties. Silver is particularly well known for its antibacterial activity[19] and has been used since the times of ancient Greece. Silver nanoparticles have also been shown to have a potent cytoprotective bioactivity towards HIV infected cells.[20] Gold thiolates have been

2

Cock: The Scope of Pharmacognosy

• Natural product discovery and evaluation• Mechanistic studies• Method and technique development and evaluation• Isolation, identification and structural elucidation of natural

products• Synthesis and transformation studies

We look forward to receiving your valuable pharmacognosy communications.

REfEREnCES1. The American Heritage Medical Dictionary, 2007, Houghton Mifflin Company, USA.

2. Fleming A, 1928, On the antibacterial action of cultures of a Penicillium with special reference to their use in the isolation of B. Influenza. British Journal of Experimental Pathology, 10, 216-226.

3. Florey HW, Gilliver K, Jennings MA, Sanders AG, 1946, Mycophenolic acid, an antibiotic from Penicillium brevi-campactum Dierckx. Lancet, 1, 46-49.

4. Ghosh AC, Manmade A, Demain AL, 1977, Toxins from Penicillium islandicum Sopp. In Mycotoxins in Human and Animal Health, Edited by Rodricks JV, Hesseltine CW, Mehlman MA, Pathotox, Chicago, USA, 625-638.

5. Hofmann A, Heim R, Brack A, Kobel H, 1958, Psilocybin ein psychotroper Wirkstoff aus dem moscikanischen Rauschpilz Psilocybe mexicana Heim. Experientia, 14, 107.

6. Bouchard C, 1889, Influence qu’exerce sur la maladie charbonneuse l’inoculation du bacilli pyocyanique, Comptes Rendus de l’Académie des Sciences, 108, 713-714.

7. Woodhead GS, Wood C, 1889, De l’action antidotique exercée par les liquids pyocyaniques sur le cours de la maladie charbonneuse. Comptes Rendus de l’Académie des Sciences, 109, 985-988.

8. Waksman SA, Woodruff HB, 1940, Bacteriostatic and bactericidal substances produced by soil actinomycetes. Proceedings of the Society for Experimental Biology and Medicine, 45, 609-614.

9. Johnson BA, Anker H, Meleney FL, 1945, Bacitracin: a new antibiotic produced by a member of the B. Subtilise group. Science, 102, 376-377.

10. Dubos RJ, Hotchkiss RD, 1941, The production of bactericidal substances by aerobic sporulating Bacilli. Journal of Experimental Medicine, 73, 5, 629-640.

11. Lasek W, Giermasz A, Kuc K, Wańkowicz A, Feleszko W, Golab J, Zagozdzon R, Stoklosa T, Jakobisiak M, 1996, Potential of the anti-tumor effect of actinomycin D by tumor necrosis factor α in mice: Correlation between in vitro and in vivo results. International Journal of Cancer, 66, 374-379.

12. Weiss RB, 1992, The anthracyclines: will we ever find a better doxorubicin? Seminars in Oncology, 19, 6, 670-686.

13. Fusetani N, Matsunaga S, 1993, Bioactive sponge peptides. Chemistry Reviews, 93, 1793-1806.

14. Wei H, Frenkel K, 1992, Suppression of tumor promoter-induced oxidative events and DNA damage in vivo by sarcophytol A: A possible mechanism of antipromotion. Cancer Research, 52, 2298-2303.

15. Brophy JJ, Goldsack RJ, Bean AR, Forster PI, Lepschi BJ, 1991, Leaf essential oils of the genus Leptospermun (Mytaceae) in Eastern Australia. Part 5, Leptospermum continentale and its allies. Flavour and Fragrance Journal, 14, 98-104.

16. Fujii A, 1995, Pharmacological effect of royal jelly. Honeybee Science, 16, 97-104.

17. Chen KK, Kovariková A, 1967, Pharmacology and toxicology of toad venom. Journal of Pharmaceutical Sciences, 56, 12, 1535-1541.

18. Whitehouse MW, Turner Ag, Davis CKC, Roberts MS, 1998, Emu oil(s): A source of non-toxic transdermal anti-inflammatory agents in Aboriginal medicine, Inflammopharmacology, 6, 1-8.

19. Feng QL, Wu J, Chen GQ, Cui FZ, Kim TN, Kim JO, 2000, A mechanistic study of the antibacterial effect of silver ions on Escherichia coli and Staphylococcus aureus, Journal of Biomedical Materials Research, 52, 662-668.

20. Sun RWY, Chen R, Chung NPY, Ho CM, Lin CLS, Che CM, 2005, Silver nanoparticles fabricated in Hepes buffer exhibit cytoprotective activities towards HIV-1 infected cells. Chemistry Communications, 40, 5059-5061.

21. Parish RV, Cottrill SM, 1987, Medicinal gold compounds, Gold Bulletin, 20, 3-12.

22. Easmon J, Pürstinger G, Heinisch G, Roth T, Fiebig HH, Holzer W, Jäger W, Jenny M, Hofmann J, 2001, Synthesis, cytotoxicity, and antitumor activity of copper(II)

used in the treatment of rheumatoid arthritis and as anti-tumour agents (as reviewed in Parish and Cottrill).[21] A variety of copper and iron complexes demonstrate potent cytotoxic activities against human cancer cells.[22] Recent studies have highlighted the importance of selenium in blocking the production of reactive oxygen species (ROS) and thus blocking oxidative stress and its associated disease states and medical conditions. [23] A variety of other inorganic molecules and ions also have medicinal promise, possibly also through their maintenance of cellular redox state.

Despite the importance of pharmacognostic agents from fungi, microorganisms and animals, plants provide us with the greatest variety of medicinal agents and arguably hold the most promise for future drug discovery. Asian medicinal botany in particular has been especially well documented. Traditional Chinese Medicinal (TCM) systems and Indian Ayuverda are widely practiced with approximately 85% of Indians regularly using crude plant formulations for the treatment of various diseases and ailments.[24] Similarly, African and Middle Eastern medicinal ethnobotanies are also widely practiced well documented. Even allopathic/Western medicine practiced in developed countries owes much to our understanding of plant based remedies. Indeed, it has been estimated that approximately 25% of all prescription drugs currently in use are originally derived from plants.[26,27] Furthermore, approximately 75% of new anticancer drugs marketed between 1981 and 2006 are derived from plant compounds.[26] Recently, there has been an increase in interest in pharmacognosy and natural therapies due to the perception that natural therapeutics offer a safer alternative than synthetic formulations due to their organic origin. This is reflected in the dramatic increase in publications in pharmacognosy journals over the period 2005-2010.[28] It is evident that a further publication outlet is required to accommodate this expanding field.

Pharmacognosy Communications is a new journal published by Pharmacognosy Network Worldwide [www.phcog.net]. We aim to publish high quality original research articles, methods, techniques and evaluation reports, critical reviews, short communications, commentaries and editorials of all aspects of pharmacognosy research. The journal is aimed at a broad readership, publishing articles on all aspects of pharmacognosy, and related fields. The journal aims to increase understanding of pharmacognosy as well as to direct and foster further research through the dissemination of scientific information by the publication of manuscripts. The submission of original contributions in all areas of pharmacognosy are welcomed.

The journal aims to cater the latest outstanding developments in the field of pharmacognosy and natural products and drug design covering but not limited to the following topics:

• Pharmacognosy and pharmacognistic investigations• Research based ethnopharmacological evaluations• Biological evaluation of crude extracts, essential oils and pure

isolates

3

Cock: The Scope of Pharmacognosy

25. Newman DJ, Cragg GM, Snader KM, 2000, The influence of natural products on drug discovery. Natural Product Reports, 17, 215-234.

26. Hostettmann K, Hamburger M, 1993, Search for new lead compounds of natural origin. In Perspectives in Medical Chemistry, Testa B, Kyburz E, Fuhrer W, Giger R (eds), Verlag Helvitica Acta, Basel.

27. Ahmed MKK, 2011, New challenges in the new year for Pharmacog Mag.: 5 years of quality publication. Pharmacognosy Magazine, 7, 25, 1-3.

and iron(II) complexes of 4N-azabiclo[3.2.2]nonane thiosemicarbazones derived from acyl diazines, Journal of Medicinal Chemistry, 44, 13, 2164-2171.

23. Venardos K, Harrison G, Headrick J, Perkins A, 2004, Effects of dietary selenium on glutathione peroxidise and thioredoxin reductase activity and recovery from cardiac ischemia-reperfusion, Journal of Trace Elements in Medicine and Biology, 18, 1, 81-88.

24. Kamboj VP, 2000, Herbal medicine. Current Science, 78, 35-39.

AbouT jouRnAl

P h a r m a c o g n o s y Communications [Phcog Commn.] www.phcogcommn.org is a new journal published by Pharmacognosy Network Worldwide [www.phcog.net]. It is a peer reviewed journal aiming to publish high quality original research articles, methods, techniques and evaluation reports, critical reviews, short communications, commentaries and editorials of all aspects of medicinal plant research. The journal is

aimed at a broad readership, publishing articles on all aspects of pharmacognosy, and related fields. The journal aims to increase understanding of pharmacognosy as well as to direct and foster

further research through the dissemination of scientific information by the publication of manuscripts. The submission of original contributions in all areas of pharmacognosy are welcome.

The journal aims to cater the latest outstanding developments in the field of pharmacognosy and natural products and drug design covering but not limited to the following topics:


isolates• Natural product discovery and evaluation• Mechanistic studies• Method and technique development and evaluation• Isolation, identification and structural elucidation of natural


4 (c) Copyright 2011 EManuscript Publishing Services, India

Invited Review



*Correspondence: [email protected]; [email protected].: +91-9872981142, +91-9815916142DOI: 10.5530/pc.2011.1.2

Plant Drugs Used to Combat Menace of Anxiety DisordersReecha Madaan*1, Suresh Kumar2, Gundeep Bansal2, Anupam Sharma3

1Chitkara College of Pharmacy, Chitkara University, Rajpura, Punjab, India ([email protected]). 2Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala- 147 002, Punjab, India ([email protected]). 3Pharmacognosy Division, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh-160 014, India ([email protected])

INTRODUCTION

Anxiety Disorders: An OverviewGlobal scenario of persons afflicted by mental disorders is alarming.[1] About 500 million people suffer from neurotic, stress related and somatoform problems, 200 million from mood disorders, 83 million from mental retardation, 30 million from epilepsy, 22 million from dementia, and 16 million from schizophrenia. Anxiety disorders are serious medical illnesses that have affected 1/8th of total population worldwide irrespective of gender, age, religion, nationality and profession.[2] Anxiety Disorders Association of America (ADAA) described anxiety disorders as the most common mental illness in the US, that have affected 19.1 million (13.3%) of the adult (18-54 years) US population.[3] A study commissioned by ADAA on ‘The Economic Burden of Anxiety Disorders’ revealed that anxiety disorders cost the US more than $42 billion a year, almost one-third of the $148 billion total mental health bill for the US. In India, prevalence rate for all mental disorders is 65.4 per 1000 population, and that for anxiety neurosis is 18.5 per 1000 population.[4] The Global Research on Anxiety and Depression (GRAD) network, a consortium of world’s leading psychiatric epidemiologists and clinical researchers, during the 154th annual meeting of ‘American

Psychiatric Association’ (APA) has observed that, “a significant number of world’s population is plagued by chronic and excessive anxiety, also known as generalized anxiety disorder (GAD), which is more serious than those of lung disease, sleep disorders and major depression, and affects more than 5% of the world population”.[5] Following is the categories of anxiety disorders. [3,6]

1. Panic disorder (PD) is characterized by panic attacks, sudden feeling of terror that strike repeatedly and without warning. Physical symptoms include chest pain, heart palpitations, sweating, trembling, shortness of breath, dizziness, abdominal discomfort, fear of losing control, fear of dying, tingling sensations, and hot flushes. Panic disorders have affected 6 million (2.7%) adult US population. Women are twice more likely to be afflicted than men.

2. Obsessive–compulsive disorder (OCD) is characterized by uncontrollable obsessions (recurring thoughts or impulses that are intrusive or inappropriate and cause the sufferer anxiety) and compulsions (repetitive behaviours or rituals). It has affected 2.2 million (1%) adult US population. It is equally common among men and women.

3. Post-traumatic stress disorder (PTSD) is characterized by persistent symptoms (nightmares, flashbacks, numbing of emotions, depression, feeling angry and irritable) that occur after experiencing a traumatic event such as war, rape, child abuse and natural disaster. It has affected 7.7 million (3.5%) adult US population. Women are more likely to be afflicted by this disorder.

ABSTRACT: In present era, a sudden holocaust of mental disorders, and recognition of severe side effects and addiction liabilities associated with long term administration of widely prescribed synthetic drugs have aroused the attention of researchers towards natural resources. This review includes 351 references, and emphasizes pharmacological reports on anxiolytic plant products and formulations. Various chemical constituents (with structures), isolated from different plants, responsible for antianxiety activity, and their possible mechanism of actions have been incorporated in this review.The review has been compiled using references from major databases like Chemical Abstracts, Medicinal and Aromatic Plants Abstracts, PubMed, Scirus, Science Direct and Online Journals. It has been concluded that preliminary antianxiety activity studies have been carried out on crude extracts of most of traditonally used and clinically potential plants. Such plantsneed to be explored properly with a view to isolate anxiolytic constituents, and to evaluate their possible mode of actions.

KEY WORDS: Antianxiety activity, chemical constituents, mechanism of action, pharmacology

Invited Review

5

Madaan, et. al.: Plant Drugs Used to Combat Menace of Anxiety Disorders

Thought patternsNegative thoughts can actually create physical symptoms of anxiety.

Management of anxiety disordersSuch a horrid emergence of mental disorders has attracted the attention of researchers towards various pharmacotherapeutic approaches for the management of these ‘modernization borne diseases’.[10] Barbiturates, benzodiazepines (BZDs), azaspirones, norepinephrine and serotonin-reuptake inhibitors, monoamine oxidase inhibitors and phenothiazines are some of the commonly used psychotropic drugs.[10] Among these, BZDs are the most widely prescribed synthetic chemical drugs for the treatment of anxiety, insomnia, epilepsy, and stress. Regular use of BZDs causes deterioration of cognitive functioning, addiction, physical dependence and tolerance.[10-12] Abrupt cessation of chronic treatment with BZDs causes the appearance of withdrawal effects comprising re-bound anxiety, restlessness, epilepsy, and motor agitation.[13,14] In the light of adverse effects associated with the synthetic drugs, researchers have been exploring natural resources to find out safer and effective drugs. Investigating plants, based on their use in traditional systems of medicine, is a sound, viable and cost effective strategy to develop new drugs.[15] Plants like Valeriana officinalis, Nardostachys jatamansi, Withania somnifera and Panax ginseng have been used extensively in various traditional systems of therapy because of their adaptogenic and psychotropic properties. Inclusion of these well-established CNS affecting plants in the arsenal of modern therapeutics has revived the faith of researchers in the plants.[16]

Targets for Treatment of AnxietyWith anxiety, various brain neurotransmitters and hormones levels change immediately. In particular, monoamines, such as norepinephrine, serotonin and dopamine, are involved in mood, stress and other physical homeostasis.[17] Serotonin and norepinephrine mainly regulate stress and negative mood in the mammalian brain, and their dysfunctions cause various mood disorders, such as social anxiety disorder and depression.[18] Dopamine also regulates mood and emotion-related behaviors and has a motivation/reward function and conditional fear responses.[19,20] Various anxiolytics and antidepressants aim at monoamine neurocircuitry, such as their receptors and transporters.[21]

The 5-hydroxytryptamine 1A (5-HT1A) receptor is viewed as a relevant target for the treatment of psychiatric disorders, notably anxiety and depression.[22] 5-HT1A receptors are located at the presynaptic and postsynaptic sites.[23] The somatodendritic autoreceptor, when activated by systemic stimulation, is believed to exert anxiolytic-like effects and to reduce 5-HT release both in the cell body and in the terminal regions of the serotonergic neurons.[24] The other 5-HT1A receptor is localized postsynaptically to the serotonergic neurons in the hippocampus, septum, amygdala, and cortex, where it increases signal transfer, which leads to an inhibition of the firing activity.[25]

4. Social phobia or Social anxiety disorder (SAD) is characterized by an intense fear of situations where embarrassment may occur. Physical symptoms include palpitations, tremors, sweating, diarrhoea, confusion and blushing. It has affected 15 million (6.8%) US adult population. It is equally common among men and women.

5. Specific phobia (SP) is characterized by the excessive fear of an object or a situation, exposure to which causes an anxious response. Specific phobias affect an estimated 19 million (8.7%) US adult population and are twice as common in women as in men.

6. Generalized anxiety disorders (GAD) are characterized by chronic, exaggerated worry about everyday routine life events and activities, lasting at least six months. Physical symptoms include fatigue, trembling, muscle tension, headache or nausea. It has affected an estimated 6.8 million (3.1%) US adult population and is twice as common in women as in men. Though, GAD is the most frequent anxiety disorder, yet only 20% of patients receive proper treatment.[7] GAD results loss of 6 for every 30 work-impairment days.

Causes of Anxiety DisordersVarious factors causing anxiety disorders are described below. [8-9]

Heredity/Genetic factorsAnxiety disorders (PD and OCD) tend to run in families. Studies have shown that if one of the twins has an anxiety disorder, the second is more likely to have an anxiety disorder.

Brain chemistryThe symptoms of long term social anxiety disorder can be attributed to the improper chemical balance in the brain. Several neurotransmitters namely serotonin, norepinephrine, gamma-amino butyric acid (GABA), which are produced in the brain, directly affect one’s feelings about a given situation. Thus brain, too, appears to play a role in the onset of anxiety disorders because symptoms of anxiety disorders are often relieved by medications that alter the level of chemicals in the brain.

PersonalityPeople with low self-esteem and poor coping skills are more prone to anxiety disorders. Conversely, an anxiety disorder that begins in childhood may itself contribute to the development of low self-esteem.

Life experiencesLong term exposure to abuse, violence, poverty or stressful experiences (the early death of a parent, bad marital or family relationships, or traumatic experiences) may affect individual’s susceptibility to anxiety disorders.

Stress overload/Lifestyle factorsExcessive stress over time, and poor lifestyle habits such as overwork, lack of sleep, poor diet and lack of regular exercise promote anxiety.

6


In brain, Nitric oxide synthase (NOS) has been localized in regions involved with anxiety, such as hypothalamus, amygdala and hippocampus.[36,37] Inhibition of NOS by nonselective or by relatively selective inhibitors of nNOS produced antianxiety-like effect. Neurosteroids can rapidly alter the excitability of central nervous system by modulating neurotransmitter-gated ion channels such as GABAA and N-methyl-D-aspartate receptors. [38] Anxiolytic, anticonvulsant and anaesthetic effects of neuroactive steroids are mediated by their capacity to positively modulate GABAA receptor. 5-alpha reductase, the enzyme that converts into 5-alpha-reduced metabolites like the GABAA positive neuroactive steroid 3-alpha-hydroxy-5-alpha-pregnan-20-one, thus, few drugs exhibits anxiolytic action via an indirect activation of the GABA-ergic system through neuroactive steroids.[39]

PLANTS HAVING ANTIANXIETY ACTIVITY

Antianxiety activity reports of various plants, and plant constituents and formulations have been presented in tables 1 and 2. Various patented formulations of anxiolytic plant drugs have been depicted in table 3. Various review articles published on anxiolytic plants are shown in table 4.

GABA is a major inhibitory transmitter in the central nervous system. The γ-aminobutyric acid type A (GABAA) receptor, the chloride ion channel complex and the central benzodiazepine receptors located on the neuronal membranes within this complex have been suggested to play an important role in the regulation of the stress and anxiety states.[26,27] The benzodiazepine binding site and GABAA receptor are structurally and functionally coupled. [28] Benzodiazepines (BZDs) have become the primary pharmacological treatment for generalized anxiety disorder. However, BZDs are often associated with tolerance development and withdrawal symptoms, which pose a risk of relapse upon discontinuation.[29,30]

Monoamine oxidase (MAO) catalyzes the oxidative deamination of a variety of monoamines such as dopamine, norepinephrine and serotonin. The MAO reaction yields aldehydes and hydrogen peroxide (H2O2), which induces apoptosis.[31] Increased endogenous MAO inhibitory activity (tribulin activity) is associated with conditions associated with stress and anxiety, both in animals and in man. [32] Rat brain tribulin activity is significantly augmented by anxiogenic agents like pentylenetetrazole, and this effect can be prevented by anxiolytic agents.[33] Inhibition of MAO and subsequent H2O2 generation effectively prevents depression and various oxidative stresses in the brain.[34] The presence of plant-derived MAO inhibitors suggests that such plant extracts could be useful as potential neuroprotectants in the treatment or prevention of depression.[35]

OH

O

(1)

O

OO

O

O

(2)

N

H

O R 1

R 2

R 1 R 2

(3) OH

(4) H

OH

H

O O

(5)

7


OHO

OOH

OH

OH

(6)

O

OH

OH

HO

O

OH

OH

(7)

NHO

O

NH2

O

(8)

H

OH

HO

H

(9)

O

O OH

HO

(10)

CH3

OCH3

OH

OHOH

O

OCH2OH

OH

OHO

HO

HOH2C CH3

H3C

CHO

CH3

O

OCH2

OH OH

OH

(11)

CHO

CH2OOCCH2CH(CH3)2

(12)

8


OR O

R

O

O

(13), R = β-Gentiobiosyl

O

(14)

OOH

HO

OH

OH

OH

(15)

HO

CH3

H3CO

(16)

OH

HO

O

OH

OCH3H3CO

(17)

O(18)

N

RO

R'

CH3O

CH3O

R R’(19) CH3 OH(20) H H(21) H OH

N

RO

H3CO

H3CO

R

(22) H

(23) CH3

N

N

N

N

O

NH2

OH

HO

HO

(24)

9


O

R 3O

H H

O

OH

OR 2R 1

H3CO 2C

R1 R2 R3

(25)

(26)

H H H

H OH H

OOH

OH

(27)

O

O

O

O

OH

OO

OH O

R 1

R 2

H3C

C(CH 3)3

R 1 R 2

(28) H H

N

OCH3

OCH3

(29)

OOH

CH3HOR

O

OH

HO

OH OH

(30), R=CH3(31), R =CH2OH

O

OH

HO

O

OHO

HO

HOHO

CH2OH

(32)

O

OH

HO

O

OH

OH

R 1

R 2

R2

(33) -Glc

(34)

H

H -Glc

R1

O

O

O

O

O

HO

CH3O

(35)

β

β

10


OH

OH

(36)

O

OH

CH2

H2C

OH

(37)

O

OH

HO

OH O

(38)

N

O

N

HHO

CH3

H3C

HOCH3

OCH3

OCH3

H3CO

(39)

O

OO

OH

HOH

OH

OH

OH

OH OH

OH

OH

OH

HO

HO

HOHO

HO

H

O

OO

O

O

H

H

H

H

(40)

11


O O

HO

OH

OR

HO

OHHO

R

(41) D-glucose

(42) H

OHO

OOH

(43)

O

OH

HO

O

OH

OH

(44)

O

OOH

HO

O OH

OH

OH

(45)

CH COOH

OH

OH

CH

(46)

O2

3

45

6O

7

8

9

10

1

11

1213

14

OCH 3

R 1

R 2

R 3

R 4

R 5

R1 R2 R3 R4 R5 C5-C6

(47) H OCH 2OH H =

(48) H OCH 2OH H

(49) H H H H H H

(50) H H H H H =

(51) H H H H H = H

(52) H OCH 3 H H = HH

C7-C8

O

O O

N

O

O

(53)

12


O2

3

45

6O

7

8

9

10

1

11

1213

14

OCH3

R1

R2

R3

R4

R5

R3

(54)(55)(56)(57)(58)(59)(60)

R1 R2 R4 R5 C5-C6 C7-C8

OCH2OOCH2OOCH2OOCH2OOCH2OOCH2O

H H H =H H =H H =H H H = =H H = =H H H = =

OCH3O CH3

OCH3

OCH3

OCH3

OCH3H H H =

H

H

H

HO

(61)

H

H

H

HO

(62)

OHO

OH

OH

OH

OH

HOOC

(63)

O

O

O

O

O

O

O

O

H H

H

(64)

O

O

(65)

OHO

OOH

OCH3

(66)

OH

H

OHH

OH

OH

HH

OO

COOH

HOOH O

(67)

13


O

HOOH O

HO

(68)

HO

(69)

O

OH

HO

OH O

H3C

(70)

O OO O O

OHHO OOH

OH

OH

OHOHHO

H3C

OCH3

(71)

O

O

OH

HO

OO O

O

OH OH

OH

HO

HOH3C

OCH3

(72)

CH3

COOH

H

CH3

CH3

(73)

NH

O

HN NH

N

O O

O

CH3

CH3

(74)

O

(75)

14


O

CH3

CH3

CH3

(76)

O

(77)

O

(78)

CH3

H2CHH

CH3

CH3

(79)

OH

(80)

OH

(81)

OH

(82)

N

N

O

O

O

H

H

H

(83)

CONCLUSION

In present era, a sudden holocaust of mental disorders, and recognition of severe side effects and addiction liabilities associated with long term administration of widely prescribed synthetic drugs have aroused the attention of researchers towards natural resources. Plants like Valeriana officinalis, Nardostachys jatamansi, Withania somnifera and Panax ginseng have been used extensively in various traditional systems of therapy because of their adaptogenic and psychotropic properties. Inclusion of these well-established CNS affecting plants in the arsenal of

modern therapeutics has revived the faith of researchers in the plants.

In present review article, amongst 143 plants reported to possess antianxiety activity (Table 1):

(a) only 07 plants have been tested clinically, (b) preliminary antianxiety activity screening on crude extracts

has been carried out on 90 plants. Such plants need to be explored with a view to isolate active constituents and their mode of actions,

15

Madaan, et. al.: Plant Drugs Used to Combat Menace of Anxiety DisordersTa

ble

1: L

ist

of

vari

ou

s p

lan

ts r

epo

rted

to

po

sses

s an

tian

xiet

y ac

tivi

ty.

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

01A

bies

pin

drow

Roy

le

(Pin

acea

e)

Talis

pat

ra, S

ilver

F

ir, P

ind

row

Fir

Eth

anol

ext

ract

of

leav

es50

and

100

m

g/kg

, ora

lly

once

dai

ly fo

r 3

days

Wis

tar

rats

Ele

vate

d pl

us m

aze

(EP

M),

Ope

n fie

ld te

st (

OF

T),

Ele

vate

d ze

ro m

aze

(EZ

M)

—A

nxio

lytic

[40]

02A

chill

ea m

illef

oliu

m

Linn

. (C

ompo

sita

e)

Yarr

ow

, Milf

oil

Aqu

eous

ext

ract

of

flow

ers

12 m

g/kg

, p.o

.Fe

mal

e W

ista

r ra

tsC

onfli

ct b

ehav

iour

—A

nxio

lytic

[41]

03A

coru

s ca

lam

us

Linn

. (A

race

ae)

Bac

h/B

acop

a m

onni

eri L

inn.

(S

crop

hula

riace

ae)

Bra

hm

i

Pow

der

of w

hole

pla

nt50

0 m

g T

DS

fo

r 6

wee

ks81

Pat

ient

s su

fferin

g fr

om

anxi

ety

diso

rder

Ele

ctro

phys

iolo

gica

l par

amet

ers

like

EE

G, E

CG

—Im

prov

emen

t in

ne

rvou

snes

s,

rest

less

ness

, ir

ritab

ility

, poo

r co

ncen

trat

ion,

sl

eep

and

loss

of

app

etite

[42]

04A

ctae

a sp

icat

a Li

nn.

(Api

acea

e)

Ban

eber

ry,

Gra

pew

ort

Fla

vono

idal

moi

ety

2 m

g/kg

, p.o

.La

ca m

ice

EP

M—

Anx

ioly

tic[4

3]

(a)

Met

hano

l ext

ract

(b

) P

olyp

heno

l fra

ctio

n(a

) 10

0 m

g/kg

, p.

o.

(b)

50 m

g/kg

, p.

o.

Laca

mic

eE

PM

—A

nxio

lytic

[44]

05A

dian

tum

te

trap

hyllu

m H

umb.

&

Bon

pl. e

x W

illd.

(A

dian

tace

ae)

Fo

url

eaf

mai

den

hai

r

Eth

anol

ext

ract

(95

%)

of

leav

es20

0 m

g/kg

, p.

o.M

ale

Spr

ague

Daw

ley

rats

OF

T, E

PM

, Aco

ustic

sta

rtle

re

spon

se te

st—

Anx

ioly

tic[4

5]

06A

ethu

sa c

ynap

ium

Li

nn. (

Api

acea

e)

Fo

ol’s

Par

sley

Fatty

aci

d: tr

idec

a-7,

9,11

-trie

noic

aci

d(1)

is

olat

ed fr

om m

etha

nol

extr

act o

f ae

rial p

arts

20 m

g/kg

, p.o

.S

wis

s al

bino

mic

e[1

-(3-

chlo

rphe

nyl)p

iper

azin

e]

indu

ced

hypo

loco

mot

ion

test

—A

nxio

lytic

[46]

07A

lbiz

zia

julib

rissi

n D

uraz

z. (

Faba

ceae

)S

ilktr

ee, M

imo

sa,

Nem

un

oki

Aqu

eous

ext

ract

of

stem

ba

rk10

0 an

d 20

0 m

g/kg

, p.o

.M

ale

SD

rat

sE

PM

Ser

oton

ergi

c sy

stem

Anx

ioly

tic[4

7]

Aqu

eous

ext

ract

of

bark

200

mg/

kg,

p.o.

for

seve

n da

ys

Mal

e S

D r

ats

EP

MIn

tera

ctio

n w

ith

5-H

T1A

rece

ptor

Anx

ioly

tic[4

8]

08A

lbiz

zia

lebb

eck

Ben

th. (

Faba

ceae

)S

iris

tre

e, A

lbiz

ia

Sap

onin

s ric

h n-

buta

nolic

frac

tion

of

petr

oleu

m e

ther

ext

ract

fr

om le

aves

25 o

r 50

mg/

kg, p

.o.

Alb

ino

Sw

iss

mic

eE

PM

Inhi

bitio

n of

G

AB

Aer

gic

tran

smis

sion

Anx

ioly

tic a

nd

noot

ropi

c[4

9]

16


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

09A

loys

ia p

olys

tach

ya

Gris

eb.

(Ver

bena

ceae

) B

urr

ito

Hyd

ro-a

lcoh

olic

ext

ract

(6

0% e

than

ol)

of le

aves

1.56

to 5

0 m

g/kg

, i.p

.Fe

mal

e S

prag

ue D

awle

y ra

tsE

PM

, For

ced

Sw

imm

ing

Test

(F

ST

)—

Anx

ioly

tic a

nd

antid

epre

ssan

t[5

0]

Eth

anol

ext

ract

of

aeria

l pa

rts

1.0,

10.0

and

10

0.0

mg/

kg,

p.o.

Sw

iss

albi

no m

ale

mic

eE

PM

Oth

er m

echa

nism

th

an B

ZD

-bs

mod

ulat

ion

at th

e G

AB

AA r

ecep

tors

Anx

ioly

tic

with

out

seda

tive

effe

cts

[51]

10A

lpin

ia

zeru

mbe

t(P

ers.

) B

urtt

& R

M

(Zin

gibe

race

ae)

Sh

ell f

low

er, P

ink

po

rcel

ain

lily

Ess

entia

l oil

from

leav

esIn

hala

tion

3.5

mg/

L ai

rM

ale

ICR

mic

eLi

ght/D

ark

mod

el (

LDM

) , O

FT,

E

PM

—A

nxio

lytic

[52]

11A

ngel

ica

Ess

entia

l oil

30.0

mg/

kg,

p.o.

Mal

e S

wis

s m

ice

EP

M, L

DM

—A

nxio

lytic

[53]

Ess

entia

l oil

21 m

g/kg

, p.o

.M

ale

Wis

tar

rats

Soc

ial i

nter

actio

n in

rat

s (S

I),

Hol

e B

oard

Tes

t (H

BT

)—

Anx

ioly

tic[5

4]

12A

ngel

ica

dahu

rica

(Fis

ch. e

x H

offm

.)

Ben

th. (

Api

acea

e)

Dah

uri

an a

ngel

ica

Fur

anoc

oum

arin

–

Phe

llopt

erin

(2)

isol

ated

fr

om m

etha

nol e

xtra

ct o

f ro

ots

IC50

= 0

.36

mic

roM

In v

itro

—B

ZD

rec

epto

rs

agon

ist

Anx

ioly

tic[5

5]

13A

niba

rip

aria

(N

ees)

M

ez (

Laur

acea

e)

Ro

sew

oo

d

Rip

arin

III(

3) is

olat

ed

from

unr

ipe

frui

ts25

and

50

mg/

kg, i

.p.

Mal

e S

wis

s m

ice

EP

M, F

ST

—A

nxio

lytic

, an

tidep

ress

ant

[56]

Rip

arin

I (4

) is

olat

ed

from

unr

ipe

frui

ts25

and

50

mg/

kg, i

.p.

Mal

e S

wis

s m

ice

EP

M, O

FT,

HB

T—

Anx

ioly

tic[5

7]

Rip

arin

- III

(3)

isol

ated

fr

om u

nrip

e fr

uits

25 a

nd 5

0 m

g/kg

, p.o

.M

ale

Sw

iss

mic

eO

FT,

EP

M, H

BT

—A

nxio

lytic

but

de

void

of

seda

tive

activ

ity

[58]

14A

nnon

a ch

erim

olia

M

ill. (

Ann

onac

eae)

C

her

imoy

a,

Cu

star

d a

pp

le

Hex

ane

extr

act o

f le

aves

6.25

, 12.

5,

25.0

and

50.

0 m

g/kg

, p.o

.

Alb

ino

mic

eM

ouse

avo

idan

ce e

xplo

rato

ry

beha

vior

, Mar

ble

bury

ing

test

(M

BT

)

GA

BA

/BZ

D

rece

ptor

com

plex

Anx

ioly

tic[5

9]

15A

nnon

a di

vers

ifolia

S

aff.

(Ann

onac

eae)

L

lam

a, A

no

na

bla

nca

Pal

mito

ne(5

) is

olat

ed

from

hex

ane

extr

act o

f le

aves

0.3,

1, 3

, 10

and

30 m

g/kg

i.p

.

Alb

ino

mic

eE

PM

—A

nxio

lytic

[60]

17

Madaan, et. al.: Plant Drugs Used to Combat Menace of Anxiety Disorders16

Apo

cynu

m v

enet

um

Linn

. (A

pocy

nace

ae)

Do

gb

ane

Eth

anol

ext

ract

of

leav

es30

and

125

m

g/kg

, p.o

.M

ale

C75

BL/

6 m

ice

EP

MIn

volv

emen

t of

GA

BA

ergi

c sy

stem

Anx

ioly

tic[6

1]

Kae

mpf

erol

(6)

isol

ated

fr

om h

ydro

-alc

ohol

ic

extr

act (

70%

eth

anol

) of

le

aves

>0.0

2 m

g/kg

, p.

o.M

ale

BL6

/C57

J m

ice

EP

MB

ZD

rec

epto

r in

tera

ctio

nA

nxio

lytic

[62]

17A

roni

a m

elan

ocar

pa

Mic

hx. (

Ros

acea

e)

Bla

ck c

ho

keb

erry

Frui

t jui

ce5

and

10 m

l/kg

, p.o

.W

ista

r ra

tsS

I, O

FT

—A

nxio

lytic

[63]

18A

zadi

rach

ta in

dica

A

. Jus

s. (

Mel

iace

ae)

Nee

m t

ree

Aqu

eous

ext

ract

from

le

aves

10, 2

0, 5

0, 1

00

and

200

mg/

kg, p

.o.

Wis

tar

rats

EP

M, O

FT

—A

nxio

lytic

[64]

Aqu

eous

ext

ract

from

le

aves

500

mg/

kg/

day

× 15

day

sM

ale

Cha

rles-

Fost

er a

lbin

o ra

tsO

FT

and

Mor

ris w

ater

maz

eIn

crea

se in

as

corb

ic a

cid

leve

l of

brai

n w

hich

falls

dur

ing

brai

n is

chem

ia

Anx

ioly

tic[6

5]

19B

aphi

a ni

tida

Lodd

. (F

abac

eae)

A

fric

an

san

dal

wo

od

, B

arw

oo

d

Eth

yl a

ceta

te e

xtra

ct o

f le

aves

100-

400

mg/

kg, p

.o.

Adu

lt al

bino

mic

e of

eith

er

sex

EP

M, Y

maz

e—

Anx

ioly

tic[6

6]

20B

yrso

carp

us

cocc

ineu

s S

chur

n.

and

Tho

nn.

(Con

nara

ceae

) K

imb

ar m

ahal

ba

Aqu

eous

ext

ract

of

leav

es20

0 an

d 40

0 m

g/kg

, p.o

.A

lbin

o m

ice

of e

ither

sex

Hex

obar

bito

ne in

duce

d sl

eepi

ng

time,

Y-m

aze,

EP

M, H

BT

—A

nxio

lytic

and

se

dativ

e[6

7]

21C

allu

na v

ulga

ris

Linn

. (H

ull)

(Eric

acea

e)

Hea

ther

Que

rcet

in(7

) is

olat

ed

from

met

hano

l ext

ract

of

aeria

l par

ts

41µg

/mg

—In

vitr

oIn

hibi

tion

of

MA

O-A

Anx

ioly

tic[6

8]

22C

alot

ropi

s gi

gant

ea

(L.)

Dry

and.

(A

pocy

nace

ae)

Gia

nt

Milk

wee

d,

Cro

wn

Flo

wer

, Aak

Alc

ohol

ic e

xtra

ct o

f pe

eled

roo

ts25

0 an

d 50

0 m

g/kg

, p.o

. A

lbin

o ra

ts o

f ei

ther

sex

EP

M, H

ot p

late

met

hod,

Ace

tic

acid

indu

ced

writ

hing

, A

sses

smen

t of

loco

mot

or

activ

ity, r

ota

rod

and

PT

Z-

indu

ced

conv

ulsi

ons

—A

nxio

lytic

, an

ticon

vuls

ant,

anal

gesi

c an

d se

dativ

e

[69]

23C

amel

lia s

inen

sis

(L.)

O. K

untz

e (T

heac

eae)

G

reen

tea

L-th

eani

ne(8

)10

mg/

kg, p

.o.

Spr

ague

Daw

ley

rats

EP

MIn

crea

se in

do

pam

ine

leve

ls b

ut n

ot

GA

BA

A r

ecep

tor

inte

ract

ion

Anx

ioly

tic[7

0]

18


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

24C

anna

bis

sativ

a Li

nn.

(Can

naba

ceae

) B

han

g

Can

nabi

diol

(9)

15, 3

0 an

d 60

nm

ol,

intr

a-dl

PAG

(D

orso

late

ral

peri

aque

duct

al

gray

)

Mal

e W

ista

r ra

tsE

PM

, Vog

el c

onfli

ct te

stC

anna

bidi

ol

inte

ract

ion

with

5H

T1A

rec

epto

rs

in d

IPA

G in

bra

in

Anx

ioly

tic[7

1]

Can

nabi

diol

(9)

15, 3

0 an

d 60

nm

ol,

intr

a-B

NS

T

bila

tera

l in

ject

ions

Mal

e W

ista

r ra

tsE

PM

, Vog

el c

onfli

ct te

stFa

cilit

ates

lo

cal 5

-HT

1A

rece

ptor

-m

edia

ted

neur

o-tr

ansm

issi

on

Anx

ioly

tic[7

2]

25C

asim

iroa

edul

is

Llav

e &

Lex

. (R

utac

eae)

W

hit

e S

apo

te,

Zap

ote

bla

nco

Aqu

eous

ext

ract

of

Leav

es25

and

35

mg/

kg, i

.p.

Wis

tar

rats

EP

M, O

FT

—A

nxio

lytic

[73]

Hyd

ro-a

lcoh

olic

(60

%

etha

nol)

extr

act o

f le

aves

40, 8

0, 1

60,

and

320

mg/

kg, p

.o. i

n m

ice,

or

1.56

, 3.

12,

6.25

,12.

5 an

d 50

mg/

kg,

i.p. i

n ra

ts

Mal

e an

d fe

mal

e S

prag

ue-

Daw

ley

rats

Spo

ntan

eous

mot

or a

ctiv

ity,

EP

M, F

ST,

HB

T, M

BT

—A

nxio

lytic

, an

tidep

ress

ant

and

seda

tive

[74]

26C

asim

iroa

prin

glei

(S

. Wat

son)

Eng

l. (R

utac

eae)

P

rin

gle

’s Z

apo

te

Ess

entia

l oil

from

leav

es79

5 an

d 10

00

mg/

kg, p

.o.

Wis

tar

rats

EP

M, O

FT,

HB

T

—A

nxio

lytic

and

se

dativ

e[7

5]

27C

assi

a si

amea

Lam

. (F

abac

eae)

K

aso

d, S

iam

ese

cass

ia

Bar

akol

(10

)10

mg/

kg, i

.p.

Mal

e w

ista

r ra

tsE

PM

—A

nxio

lytic

[76,

77

]

28C

ecro

pia

glaz

ioui

S

neth

(U

rtic

acea

e)

Em

bau

ba,

Yar

um

o

(a)

Aqu

eous

ext

ract

of

leav

es

(b)

But

anol

ic fr

actio

n of

aq

ueou

s ex

trac

t of

leav

es

(a)

0.5

and

1.0

g/kg

, p.o

. (b

) 25

-100

m

g/kg

, p.o

.

Mal

e ad

ult S

wis

s m

ice

EP

M—

Anx

ioly

tic[7

8]

29C

elas

trus

pa

nicu

latu

s W

illd.

(Cel

astr

acea

e)

Jyo

tish

mat

i, M

aak

kan

gn

i

Pet

role

um e

ther

ext

ract

of

see

ds3.

2 g/

kg/d

ay

for

5 da

ysA

lbin

o m

ice

Beh

avio

ural

dis

inhi

bitio

n m

odel

—A

nxio

lytic

[79]

Oil

of s

eeds

1 an

d 1.

5 g/

kg,

i.p.

Wis

tar

rats

OF

T, E

PM

, Thi

rsty

rat

con

flict

pa

radi

gmS

erot

oner

gic

mec

hani

smA

nxio

lytic

[80]

19


Cen

tella

asi

atic

a (L

.)

Urb

. (U

mbe

llife

rae)

G

otu

Ko

la

Pow

dere

d dr

ug12

g/d

ay, p

.o.

Dou

ble-

blin

d, p

lace

bo-

cont

rolle

d st

udy

in 2

0 su

bjec

ts

Sig

nific

antly

atte

nuat

ed th

e pe

ak o

f ac

oust

ic s

tart

le

resp

onse

am

plitu

de

—A

nxio

lytic

[81]

(a)

Mar

kete

d fo

rmul

atio

ns

(b)

Met

hano

l ext

ract

(c

) E

thyl

ace

tate

ext

ract

(d

) A

siat

icos

ide(

11)

(a)

500

mg/

kg,

p.o.

(b

) 30

47 m

g/kg

, p.o

. (c

) 11

1 m

g/kg

, p.

o.

(d)

3 m

g/kg

, p.

o.

Mal

e S

prag

ue-D

awle

y (S

D)

rats

EP

M, O

FT,

SI,

loco

mot

or a

ctiv

ity,

puni

shed

drin

king

, nov

el c

age

test

—A

nxio

lytic

[82]

31C

entr

anth

us r

uber

(L

.) D

C

(Val

eria

nace

ae)

Red

val

eria

n

Val

epot

riate

–

valtr

ate(

12)

5 m

g/kg

, p.o

.W

ista

r ra

tsIn

hibi

tion

of o

rient

atio

n re

flexe

s an

d m

otor

act

ivity

—A

nxio

lytic

[83]

32C

erat

onia

sili

qua

Linn

. (Fa

bace

ae)

Car

ob

tre

e

Met

hano

l ext

ract

of

leav

es a

nd p

ods

Pod

s -

12.1

7 ng

and

Lea

ves

- 18

.7 n

g di

azep

am

equi

vale

nt

In v

itro

—B

ZD

rec

epto

r in

tera

ctio

nA

nxio

lytic

[84]

33C

inna

mom

um

cass

ia B

lum

e.

(Lau

race

ae)

Cas

sia

Bar

k,

Ch

ines

e ci

nn

amo

n

50%

Eth

anol

ext

ract

fr

om s

tem

bar

ks75

0 m

g/kg

, p.

o.M

ale

ICR

mic

eE

PM

Reg

ulat

ion

of

5-H

T1A

and

GA

BA

re

cept

or s

yste

m

Anx

ioly

tic[8

5]

34C

issu

s si

cyoi

des

Linn

. (V

itace

ae)

Po

ssu

m g

rap

e vi

ne,

Pri

nce

ss v

ine

Hyd

ro-a

lcoh

olic

ext

ract

(7

0% e

than

ol)

of le

aves

300,

600

and

10

00 m

g/kg

, i.p

.

Mal

e an

d fe

mal

e S

wis

s al

bino

mic

e E

PM

, HB

T, M

BT,

Sod

ium

P

ento

barb

ital-i

nduc

ed s

leep

ing

time,

PT

Z-in

duce

d co

nvul

sion

—A

nxio

lytic

, an

ticon

vuls

ant

[86]

35C

itrus

aur

antiu

m

Linn

. (R

utac

eae)

B

itte

r O

ran

ge

Ess

entia

l oil

from

pee

l (E

OP

) of

leav

es1

g/kg

, p.o

.M

ale

Sw

iss

mic

eE

PM

, OF

T—

Anx

ioly

tic[8

7]

Ess

entia

l oil

from

frui

ts

0.5

and

1.0

g/kg

, p.o

.M

ale

Sw

iss

mic

eLD

M, M

BT

—A

nxio

lytic

[88]

36C

itrus

sin

esis

Lin

n.

(Rut

acea

e)

Sw

eet

Ora

ng

e,

Blo

od

Ora

ng

e

Ess

entia

l oil

100,

200

and

40

0 µl

Wis

tar

mal

e ra

tsE

PM

, LD

M—

Anx

ioly

tic[8

9]

37C

litor

ia te

rnat

ea

Linn

. (P

apili

onac

eae)

B

utt

erfly

pea

Met

hano

l ext

ract

of

root

s10

0-40

0 m

g/kg

, p.o

.M

ale

Sw

iss

albi

no m

ice

and

Wis

tar

rats

EP

M, L

DM

—A

nxio

lytic

[90]

38C

onvu

lvul

us

plur

icau

lis C

hois

y. (C

onvo

lvul

acea

e)

Sh

ankh

pu

spi

Eth

yl a

ceta

te fr

actio

n of

et

hano

l ext

ract

of

the

aeria

l par

ts

100

mg/

kg, p

.o.

Spr

ague

-Daw

ley

rats

and

S

wis

s al

bino

mic

eE

PM

, OF

T a

nd r

otar

od

perf

orm

ance

—

Anx

ioly

tic

[91]

39C

opai

fera

ret

icul

ata

Duc

ke

(Leg

umin

osae

) B

razi

lian

co

pai

ba

Ess

entia

l oil

100,

400

and

80

0 m

g/kg

, i.p

.W

ista

r ra

tsE

PM

—A

nxio

lytic

[92]

20


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

40C

oria

ndru

m s

ativ

um

Linn

. (U

mbe

llife

rae)

C

ori

and

er, D

han

iya

Aqu

eous

ext

ract

of

seed

s10

0 m

g/kg

, p.

o.M

ale

albi

no m

ice

EP

M—

Anx

ioly

tic[9

3]

41C

rocu

s sa

tivus

Lin

n.

(Lili

acea

e)

Saf

fro

n, A

utu

mn

cr

ocu

s

Cro

cin(

13)

isol

ated

from

aq

ueou

s ex

trac

t of

red

drie

d st

igm

as

50 m

g/kg

, i.p

. W

ista

r ra

tsLD

M—

Anx

ioly

tic[9

4]

(a)

Aqu

eous

ext

ract

of

stig

mas

(b

) C

roci

n(13

)(c

) S

afra

nal (

14)

(a)

56, 8

0, 3

20

and

560

mg/

kg, i

.p.

(b)

50, 2

00

and

600

mg/

kg, i

.p.

(c)

0.05

, 0.1

5 an

d 0.

35 m

l/kg

, i.p

.

Raz

i mal

e m

ice

EP

M, O

FT,

Pen

toba

rbita

l sl

eepi

ng ti

me,

Rot

arod

test

—A

nxio

lytic

(A

t lo

wer

dos

e),

hypn

otic

(A

t hi

gher

dos

e)

[95]

42C

roto

n ce

ltidi

foliu

s B

aill.

(E

upho

rbia

ceae

) S

ang

ue-

de-

adav

e

Pro

anth

ocya

nidi

n(15

) ric

h fr

actio

n is

olat

ed

from

aqu

eous

ext

ract

of

bark

3 m

g/kg

, i.p

.W

ista

r ra

tsE

PM

—A

nxio

lytic

[96]

43C

roto

n ze

hntn

eri

Pax

& H

offm

an

(Eup

horb

iace

ae)

Can

ela

de

Cu

nh

a

Met

hyl e

ugen

ol(1

6) fr

om

esse

ntia

l oil

1, 3

and

10

µl/1

00 g

, p.o

.M

ale

Wis

tar

rats

OF

T, S

I, E

PM

, HB

T, F

ST

—

Ant

idep

ress

ant

and

mild

an

xiol

ytic

[97]

44C

urcu

ma

long

a Li

nn.

(Zin

gibe

race

ae)

Cu

rcu

ma,

Tu

rmer

ic

Cur

cum

in(1

7)20

mg/

kg, i

.p.

Sw

iss

albi

no m

ice

EP

M, O

FT,

LD

M, S

IIn

volv

emen

t of

indu

cibl

e N

OS

Anx

ioly

tic[9

8]

45C

ymbo

pogo

n ci

trat

us (

DC

.) S

tapf

(P

oace

ae)

Lem

on

gra

ss,

Gin

ger

gra

ss

Citr

al (

18)

or te

a ab

afad

o20

0 m

g/kg

, i.p

.M

ale

albi

no S

wis

s m

ice

OF

T, R

ota-

rod

test

, Spo

ntan

eous

m

otor

act

ivity

, Bar

bitu

rate

sl

eepi

ng-t

ime,

Tra

nsco

rnea

l el

ectr

osho

ck, P

TZ

-indu

ced

conv

ulsi

ons,

Pun

ishe

d re

spon

se

test

—C

entr

al

Ner

vous

de

pres

sant

[99]

Ess

entia

l oil

0.5

and

1.0

g/kg

, i.p

.S

wis

s m

ale

mic

eE

PM

, LD

M—

Anx

ioly

tic[1

00]

46D

avill

a ru

gosa

P

oire

t (D

illen

iace

ae)

Cip

o-C

abo

clo

, Fir

e vi

ne

Hyd

ro-a

lcoh

olic

ext

ract

(7

0% e

than

ol)

of s

tem

s 15

mg/

kg, p

.o.

Mal

e W

ista

r ra

tsE

PM

, OF

T—

Anx

ioly

tic[1

01]

47D

rym

aria

cor

data

(L

.) W

illd.

ex

Roe

m.

& S

chul

t. (C

aryo

phyl

lace

ae)

Tro

pic

al c

hic

kwee

d

Hyd

ro-a

lcoh

olic

ext

ract

(5

0% e

than

ol)

of le

aves

100

mg/

kg,

p.o.

Sw

iss

albi

no m

ice

EP

M, L

DM

, OF

T, H

BT

—A

nxio

lytic

[102

]

21


Duc

rosi

a an

ethi

folia

B

oiss

. (A

piac

eae)

H

azza

, Haz

zaz

Ess

entia

l oil

25, 5

0, 1

00,

200

and

400

mg/

kg, p

.o.

Sw

iss

albi

no m

ice

EP

M, S

pont

aneo

us m

otor

ac

tivity

, Ket

amin

e-in

duce

d sl

eep

time

—A

nxio

lytic

but

no

t sed

ativ

e[1

03]

49E

chin

acea

pur

pure

a (L

.) M

oenc

h.

(Ast

erac

eae)

C

on

e fl

ow

er

(a)

E. p

urpu

rea

root

ex

trac

t (et

hano

l 4%

v/v

; E

chin

acos

ide

4%)

(b)

E. p

urpu

rea

herb

ex

trac

t (et

hano

l 60%

m

/m; t

otal

phe

nols

4%

) (c

) E

. ang

ustif

olia

roo

t ex

trac

t (et

hano

l 85%

v/v

; E

chin

acos

ide

4%)

(d)

E. p

urpu

rea

root

ex

trac

t (et

hano

l 70%

v/v

)

3-7

mg/

kg, p

.o.

Mal

e W

ista

r ra

tsE

PM

, SI,

shoc

k in

duce

d so

cial

av

oida

nce

test

, OF

T—

Onl

y ex

trac

t (d

) sh

owed

an

xiol

ytic

ac

tivity

[104

]

50E

chiu

m a

moe

num

F

isch

. Et M

ey.

(Bor

agin

acea

e)

Vip

er’s

bu

glo

ss,

Red

fea

ther

s

Aqu

eous

ext

ract

of

flow

ers

5, 1

0, 3

0, 6

2.5,

80

and

125

m

g/kg

, i.p

.

Mal

e N

MR

I alb

ino

mic

eE

PM

—A

nxio

lytic

[105

]

Hyd

ro-e

than

ol e

xtra

ct

(80%

) of

the

plan

t flo

wer

s

50 m

g/kg

, i.p

.M

ale

TO m

ice

EP

M—

Anx

ioly

tic[1

06]

51E

clip

ta a

lba

Linn

. (A

ster

acea

e)

Bh

rin

gar

aj, F

alse

d

aisy

(a)

Aqu

eous

, hyd

ro-

alco

holic

ext

ract

s (b

) H

ydro

lyze

d fr

actio

n ob

tain

ed fr

om w

hole

pl

ant

(a)

150

and

300

mg/

kg,

p.o.

(b

) 30

mg/

kg,

p.o.

Wis

tar

rats

Lo

com

otor

act

ivity

, EP

M, H

BT,

C

old

rest

rain

t ind

uced

gas

tric

ul

cer

and

whi

te b

lood

cel

l cou

nt

in th

e m

ilk in

duce

d le

ukoc

ytos

is

chal

leng

e

Noo

trop

ic,

seda

tive,

an

xiol

ytic

and

an

tistr

ess

[107

]

52E

ryth

rina

mul

ungu

M

art.

(Pap

ilion

acea

e)

Mu

lun

gu

, C

ort

icei

ra

Hyd

ro-a

lcoh

olic

ext

ract

(7

0% e

than

ol)

from

the

inflo

resc

ence

Acu

te (

200

mg/

kg, p

.o.)

ch

roni

c (5

0 m

g/kg

, p.o

. for

7

days

)

Mal

e W

ista

r ra

tsE

leva

ted

T m

aze

(ET

M),

LDM

, C

at o

dor

test

—A

nxio

lytic

[108

, 10

9]

Wat

er :

Alc

ohol

(7:

3)

extr

act o

f in

flore

scen

ceA

cute

stu

dy

200

and

400

mg/

kg, p

.o.

and

chro

nic

stud

y fo

r 21

da

ys, 5

0 an

d 20

0 m

g/kg

, p.

o.

Mal

e W

ista

r ra

tsE

TM

—A

nxio

lytic

[110

]

Ery

thrin

ian

alka

loid

s i.e

(+

)-α–

hydr

oxye

rsot

rine(

19),

eryt

hrav

ine(

20)

and

(+)-

11-α

–hyd

roxy

er

ythr

avin

e(21

) is

olat

ed

from

hyd

ro-a

lcoh

olic

ex

trac

t of

flow

ers

3 an

d 10

mg/

kg, p

.o.

Mal

e S

wis

s M

ice

EP

M, L

DM

—A

nxio

lytic

[111

]

22


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

Cru

de e

xtra

ct (

CE

), E

ryth

rinia

n al

kalo

ids:

(+

)-α–

hydr

oxye

rsot

rine(

19),

eryt

hrav

ine(

20)

and

(+)-

11-α

–hyd

roxy

er

ythr

avin

e(21

) is

olat

ed

from

hyd

ro-a

lcoh

olic

ex

trac

t of

flow

ers

3-10

mg/

kg,

p.o.

C

E (

50, 1

00,

200

and

400

mg/

kg, p

.o.)

Mal

e S

wis

s m

ice

T-m

aze,

Loc

omot

or a

ctiv

ity te

st—

Anx

ioly

tic[1

12]

53E

ryth

rina

sube

rosa

R

oxb.

(Fa

bace

ae)

Co

ral t

ree

Alk

aloi

ds –

E

ryso

dine

(22)

and

er

ysot

hrin

e(23

) is

olat

ed

from

hyd

ro-a

lcoh

olic

ex

trac

t of

flow

ers

3 an

d 10

mg/

kg, p

.o.

Mal

e al

bino

mic

eE

PM

, LD

M—

Anx

ioly

tic[1

13]

54E

ryth

rina

velu

tina

Will

d. (

Faba

ceae

) B

ico

-De-

Pap

agai

o

Wat

er :

Alc

ohol

(7:

3)

extr

act o

f st

em b

ark

Acu

te s

tudy

-

200

and

400

mg/

kg, p

.o.,

and

chro

nic

stud

y -

50 a

nd

200

mg/

kg,

p.o.

Mal

e W

ista

r ra

tsE

TM

—A

nxio

lytic

[110

]

Hyd

ro-e

than

ol e

xtra

ct o

f st

em b

ark

50 a

nd 1

00

mg/

kg, p

.o. f

or

23-2

6 da

ys

Adu

lt m

ale

Sw

iss

albi

no

mic

eE

PM

—A

nxio

lytic

[114

]

55E

schs

chol

zia

calif

orni

ca C

ham

. (P

apav

erac

eae)

C

alifo

rnia

po

pp

y,

Go

ld p

op

py

Hyd

ro-a

lcoh

olic

ext

ract

(6

0% e

than

ol)

of a

eria

l pa

rts

25 m

g/kg

, i.p

.M

ale

Sw

iss

mic

eLD

MB

ZD

rec

epto

r in

tera

ctio

nA

nxio

lytic

[115

]

70%

eth

anol

ext

ract

of

aeria

l par

ts10

0 to

300

m

g/kg

, i.p

.M

ale

Wis

tar

rats

CC

l 4 in

duce

d ne

urop

athi

c pa

in,

hot p

late

and

car

rage

enan

in

duce

d pa

in

—A

nxio

lytic

and

an

ti-

neur

opat

hic

pain

[116

]

56E

upho

rbia

hir

ta

Linn

. (E

upho

rbia

ceae

) A

sth

ma

wee

d

Aqu

eous

ext

ract

of

who

le p

lant

12.5

and

25

mg/

kg, i

.p.

Sw

iss

albi

no m

ice

Sta

ir ca

se te

st, L

DM

—A

nxio

lytic

[117

]

57E

upho

ria lo

ngan

a La

mar

ck

(Sap

inda

ceae

) L

on

gan

Ari

llus

(a)

Met

hano

l ext

ract

(b

) ad

enos

ine(

24)

isol

ated

from

pul

p or

fle

sh

(a)

2 g/

kg, s

.c.

(b)

30 m

g/kg

, s.

c.

Mal

e dd

Y m

ice

Vog

el ty

pe a

nti-c

onfli

ct m

etho

d—

Anx

ioly

tic[1

18]

58E

upho

rbia

ner

rifol

ia

Linn

. (E

upho

rbia

ceae

) In

dia

n s

pu

rge

tree

, O

lean

der

sp

urg

e

Hyd

ro-a

lcoh

olic

(50

%

etha

nol)

extr

act o

f le

aves

400

mg/

kg,

p.o.

Sw

iss

albi

no m

ice

EP

M—

Anx

ioly

tic[1

19]

23


Eur

ycom

a lo

ngifo

lia

Jack

(S

imar

ouba

ceae

) To

ng

kat

ali,

Pen

awar

bia

s

Chl

orof

orm

, n-b

utyl

al

coho

l and

wat

er

frac

tions

obt

aine

d fr

om

met

hano

l ext

ract

of

root

s

0.3

g/kg

, p.o

. fo

r 5

days

tw

ice

daily

Alb

ino

mic

eE

PM

, OF

T, F

oot s

hock

indu

ced

fligh

ting

beha

viou

r—

Anx

ioly

tic[1

20]

60E

volv

ulus

als

inoi

des

Linn

. (C

onvo

lvul

acea

e)

Sh

ankh

pu

shp

i

Eth

yl a

ceta

te fr

actio

n of

et

hano

l ext

ract

of

the

aeria

l par

ts

100

mg/

kg, p

.o.

Spr

ague

-Daw

ley

rats

and

S

wis

s al

bino

mic

eE

PM

, OF

T a

nd r

otar

od

perf

orm

ance

—

Anx

ioly

tic,

neur

omus

cula

r co

ordi

natio

n an

d an

tioxi

dant

[91]

61G

alph

imia

gla

uca

Cav

. (M

alpi

ghia

ceae

) C

ald

ero

na

amar

illa

Gal

phim

ine

B(2

5),

galp

him

ine

A(2

6) a

nd

galp

him

ine

rich

frac

tions

(G

RF

s) o

btai

ned

from

m

etha

nol e

xtra

ct o

f ae

rial p

arts

15 m

g/kg

, i.p

.M

ale

ICR

mic

eE

PM

—A

nxio

lytic

[121

]

Met

hano

l ext

ract

of

aeria

l par

ts12

5, 2

50, 5

00,

1000

and

20

00 m

g/kg

, p.

o.

ICR

alb

ino

mic

eE

PM

, LD

M, F

ST

—A

nxio

lytic

and

an

tidep

ress

ant

[122

]

Cap

sule

s co

ntai

ning

310

m

g of

aqu

eous

ext

ract

of

aer

ial p

arts

310

mg

twic

e da

ily fo

r 4

wee

ks

A c

ontr

olle

d ra

ndom

ized

do

uble

blin

d cl

inic

al tr

ial

HA

MA

sca

le, t

he c

linic

al g

loba

l im

pres

sion

sca

le a

nd p

atie

nt

glob

al e

valu

atio

n

—A

nxio

lytic

[123

]

62G

arde

nia

jasm

inoi

des

Elli

s (R

ubia

ceae

) C

ape

jasm

ine

Kam

isho

yosa

n50

-200

mg/

kg,

p.o.

M

ale

ddY

mic

eS

I—

Anx

ioly

tic[1

24]

63G

astr

odia

ela

ta

Blu

me

(Orc

hida

ceae

) T

ian

ma

(Ch

ina)

; G

astr

od

ia

Tub

er(E

ng

lish

n

ame)

(a)

Aqu

eous

ext

ract

of

rhiz

omes

(b

) Phe

nolic

con

stitu

ents

: 4-

hydr

oxyl

-ben

zyl

alco

hol,

and

benz

alde

hyde

and

its

phen

olic

con

stitu

ents

(a)

400

mg/

kg,

p.o.

(b

) 50

and

100

m

g/kg

, i.p

.

Mal

e IC

R m

ice

EP

MIn

tera

ctio

n w

ith

5-H

T(1

A) r

ecep

tor

Anx

ioly

tic[1

25]

64G

else

miu

m

sem

perv

irens

(L.

) A

it. (

Loga

niac

eae)

C

aro

lina

yello

w

Jasm

ine

Met

hano

l ext

ract

of

root

s an

d rh

izom

es15

0 m

g/kg

, p.

o.S

wis

s al

bino

mic

e E

PM

—A

nxio

lytic

[126

]

Cen

tesi

mal

dilu

tions

of

hydr

o-al

coho

lic e

xtra

ct

of p

lant

as

in

hom

eopa

thic

sys

tem

5C, 9

C a

nd

30C

dilu

tions

ICR

-CD

1 m

ale

mic

eLD

M, O

FT

—A

nxio

lytic

[127

]

24


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

65G

inkg

o bi

loba

Lin

n.

(Gin

kgoa

ceae

) G

inkg

o,

Mai

den

hai

r tr

ee

Aqu

eous

and

eth

anol

ex

trac

ts o

f le

aves

5

and

10 m

g eq

uiva

lent

In v

itro

usin

g ra

t bra

in

mito

cond

rial e

xtra

ct—

Inhi

bitio

n of

m

onoa

min

e ox

idas

e (M

AO

A

and

B)

Anx

ioly

tic[1

28]

Gin

kgo

bilo

ba e

xtra

ct

(EG

b-76

1)8-

16 m

g/kg

, i.p

.W

ista

r A

F r

ats

SI

GA

BA

/ BZ

D/

Cl-

chan

nel

rece

ptor

in

tera

ctio

n

Anx

ioly

tic[1

29]

Gin

kgol

ic a

cid(

27)

conj

ugat

es (

GA

C)

isol

ated

from

chl

orof

orm

: m

etha

nol e

xtra

ct (

2:1)

of

the

leav

es

0.6

mg/

kg, p

.o.

Cha

rles

Fost

er r

ats

EP

M, O

FT,

nov

elty

-indu

ced

feed

ing

late

ncy

and

SI

—A

nxio

lytic

[130

]

G. b

iloba

ext

ract

(G

BE

), st

anda

rdiz

ed to

con

tain

24

% g

inkg

o-fla

vogl

ycos

ides

and

6%

gi

nkgo

-ter

peno

id

lact

ones

or

gink

golid

e A

(28)

0.5

and

1.0

g/kg

, p.o

. for

7

days

; 1

and

2 m

g/kg

, p.

o. fo

r fiv

e da

ys

Mal

e dd

Y m

ice

EP

MO

ther

mec

hani

sm

but n

ot th

roug

h G

AB

A/ B

ZD

/C

l- ch

anne

l re

cept

or

inte

ract

ion

Anx

ioly

tic[1

31]

66G

lycy

rrhi

za g

labr

a Li

nn. (

Legu

min

osae

) L

ico

rice

, Mu

leth

i

Hyd

ro-a

lcoh

olic

ext

ract

of

roo

ts a

nd r

hizo

mes

10-3

00 m

g/kg

, i.p

.S

wis

s al

bino

mic

eE

PM

, foo

t sho

ck in

duce

d ag

gres

sion

—A

nxio

lytic

[132

]

67H

edyo

smum

br

asili

ense

Mar

t. (C

hlor

anth

acea

e)

Ch

a d

e bu

gre

Eth

anol

ext

ract

of

aeria

l pa

rts

100

mg/

kg, i

.p.

Mal

e S

wis

s al

bino

mic

eE

PM

, OF

T, B

arbi

tura

te-in

duce

d sl

eepi

ng ti

me

test

—A

nxio

lytic

and

se

dativ

e[1

33]

68H

eter

opte

rys

glab

ra

Hoo

k. &

Arn

. (M

alpi

ghia

cae)

R

edw

ing

Eth

anol

ext

ract

of

frui

ts35

0 m

g/kg

, p.

o.D

BA

/2J

mic

eS

leep

wak

eful

ness

cyc

le,

elec

troe

ncep

halo

gram

(E

EG

) an

d vi

sual

evo

ked

pote

ntia

ls

(VE

P)

—A

nxio

lytic

and

se

dativ

e[1

34]

69H

ibis

cus

sabd

ariff

a Li

nn. (

Mal

vace

ae)

Jam

aica

so

rrel

, R

ed s

orr

el

Aqu

eous

, hyd

ro-

alco

holic

, and

eth

anol

ex

trac

t of

caly

xes

of

plan

t

300

mg/

kg,

p.o.

Wis

tar

rats

EP

M, k

etam

ine-

indu

ced

slee

p—

Anx

ioly

tic a

nd

seda

tive

(at

mul

tiple

do

ses)

[135

]

70H

ippe

astr

um

vitta

tum

(L’

Her

it)

Her

bert

(A

mar

yllid

acea

e)

Am

aryl

lis

Isoq

uino

line

alka

loid

: M

onta

nine

(29)

isol

ated

fr

om e

than

ol e

xtra

ct o

f bu

lbs

Anx

ioly

tic a

nd

seda

tive

(1-1

0 m

g/kg

, i.p

.),

antic

onvu

lsan

t (3

0 an

d 60

, m

g/kg

, i.p

.)

Sw

iss

albi

no m

ice

EP

M, S

odiu

m p

ento

barb

ital-

indu

ced

slee

p, P

TZ

-pro

voke

d co

nvul

sion

s, F

ST

—A

nxio

lytic

, mild

se

dativ

e an

d an

ticon

vuls

ant

but n

ot

antid

epre

ssan

t

[136

]

25


Hyp

eric

um

perfo

ratu

m L

inn.

(G

uttif

erae

) S

t Jo

hn

’s w

ort

H. p

erfo

ratu

m e

xtra

ct

LI60

—In

vitr

o—

β re

cept

or

activ

atio

nA

nxio

lytic

[137

]

Sta

ndar

dize

d ex

trac

t of

the

who

le p

lant

, co

ntai

ning

0.5

4% to

tal

hype

ricin

s [0

.11%

hy

peric

in(3

0) a

nd 0

.43%

ps

eudo

hype

ricin

(31)

]and

0.

09%

pro

tofo

rms

2778

and

18

52 m

g/kg

, p.

o.

Mal

e S

prag

ue–D

awle

y ra

tsO

FT,

LD

MIn

hibi

tory

in

fluen

ce o

n gl

utam

ater

gic

tran

smis

sion

m

edia

ted

by

NM

DA

rec

epto

rs

Anx

ioly

tic[1

38]

Lyop

hiliz

ed a

queo

us

extr

act

5 m

g/kg

, p.o

.M

ale

albi

no S

wis

s m

ice

EP

M—

Anx

ioly

tic[1

39]

Hyd

ro-a

lcoh

olic

ext

ract

of

who

le p

lant

100

or 2

00

mg/

kg, p

.o. O

D

for

3 da

ys

Wis

tar

rats

EP

M, O

FT,

EZ

M, n

ovel

ty-in

duce

d su

ppre

ssed

feed

ing

late

ncy,

SI

Affe

ct

mon

oam

ines

co

ncen

trat

ion

in

rats

’ bra

in

Anx

ioly

tic[1

40]

H. p

erfo

ratu

m e

xtra

ct L

I 16

0 30

0 m

g/kg

, p.

o. fo

r 21

da

ys

Mal

e al

bino

Sw

iss

mic

eM

ouse

def

ense

test

bat

tery

—A

nxio

lytic

[141

]

H. p

erfo

ratu

m e

xtra

ct L

I 16

030

0 m

g/kg

, p.o

Mal

e al

bino

Sw

iss

mic

eE

TM

—A

nxio

lytic

[142

]

H. p

erfo

ratu

m e

xtra

ct L

I 16

038

0 m

g/kg

/da

y ch

roni

c ad

min

istr

atio

n

C57

BL/

6J M

ice

OF

T, L

DM

, FS

T—

Anx

ioly

tic a

nd

antid

epre

ssan

t[1

43]

H. p

erfo

ratu

m e

xtra

ct L

I 16

015

0 an

d 30

0 m

g/kg

, p.o

.S

wis

s al

bino

mic

eM

BT,

FS

T—

Anx

ioly

tic a

nd

antid

epre

ssan

t[1

44]

Hyd

ro-a

lcoh

olic

ext

ract

of

who

le p

lant

200-

400

mg/

kg, p

.o.

Mal

e La

ca m

ice

Mir

rore

d ch

ambe

r, E

PM

, EZ

M—

Anx

ioly

tic[1

45]

72Ja

trop

ha c

iliat

a M

. A

rg.

(Eup

horb

iace

ae)

Hu

anar

po

Vite

xin(

32),

iso-

orie

ntin

(33)

and

or

ient

in(3

4) fr

om

met

hano

l ext

ract

of

Ste

ms

40 m

g/kg

, s.c

. M

ale

ddY

mic

eV

ogel

type

Ant

icon

flict

effe

ct in

m

ice

—A

nxio

lytic

[146

]

73K

ielm

eyer

a co

riace

a M

art.

(Clu

siac

eae)

P

áu s

anto

Eth

anol

ext

ract

of

leav

es12

0 m

g/kg

/da

y, p.

o.M

ale

Wis

tar

rats

EP

M—

Anx

ioly

tic[1

47]

26


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

74La

vand

ula

angu

stifo

lia M

iller

(L

amia

ceae

) E

ng

lish

Lav

end

er

Ess

entia

l oil

from

leav

esIn

hala

tion

0.1-

1.0

ml

Adu

lt m

ale

Spr

ague

-D

awle

y al

bino

rat

sO

pen

field

beh

avio

r te

stLa

vend

er o

il po

tent

iate

s th

e re

spon

ses

of

GA

BA

rec

epto

rs

at lo

w

conc

entr

atio

ns

and

inhi

bits

re

spon

ses

of

GA

BA

rec

epto

rs

at h

igh

conc

entr

atio

ns in

vi

tro

Anx

ioly

tic[1

48,

149]

Lave

nder

oil

I ml/1

00 g

, i.p

.M

ale

ICR

Mic

eG

alle

r ty

pe c

onfli

ct te

st—

Anx

ioly

tic[1

50]

Lave

nder

odo

ur—

Mat

ure

mal

e an

d fe

mal

e ge

rbils

EP

M—

Anx

ioly

tic[1

51]

75Le

ptos

perm

um

scop

ariu

m J

.R. e

t G.

Fors

t. (M

yrta

ceae

) M

anu

ka o

r Tea

tre

e

(a)

Hyd

ro-a

lcoh

olic

ex

trac

t (70

% e

than

ol)

(b)

5,7-

dim

etho

xyfla

vone

(1

), 5,

7-di

met

hoxy

-6-

met

hylfl

avon

e (2

), 5-

hydr

oxy-

7-m

etho

xy-6

-m

ethy

lflav

one

(3)

and

5-hy

drox

y-7-

met

hoxy

-6,

8-di

met

hylfl

avon

e (4

)

(a)

250

mg/

kg,

p.o.

(b

) IC

50-

valu

es o

f 2.

1 m

icro

M (

1), 4

5 m

icro

M (

2),

3.3

mic

roM

(3)

an

d 40

m

icro

M (

4)

(a)

Rat

s (b

) In

vitr

o ra

dio

rece

ptor

as

say

with

[3H

] Flu

nitr

aze-

pam

Loco

mot

ion

stud

yIn

tera

ctio

n w

ith

GA

BA

A/B

ZD

re

cept

or

Anx

ioly

tic[1

52,

153]

76Li

ppia

alb

a (M

ill.)

N

.E. B

row

n (V

erbe

nace

ae)

Cid

reir

a, B

ush

y m

atg

rass

Thr

ee c

hem

otyp

es o

f es

sent

ial o

il (E

O1,

EO

2,

EO

3) fr

om le

aves

EO

1 an

d E

O3

(100

mg/

kg,

i.p.)

and

EO

2 (2

5 m

g/kg

, i.p

.)

Mal

e S

wis

s M

ice

EP

M, O

FT

and

rot

arod

—A

nxio

lytic

and

m

yore

laxa

nt[1

54]

77Lo

esel

ia m

exic

ana

Bra

nd

(Pol

emon

iace

ae)

Mex

ican

fal

se

calic

o, E

spin

osi

lla

Dap

hnor

etin

(35)

isol

ated

fr

om h

ydro

-alc

ohol

ic

extr

act (

60%

eth

anol

) of

w

hole

pla

nt

1.8,

3.7

, 7.5

an

d 15

.0 m

g/kg

, i.p

.

Mal

e IC

R m

ice

OF

T, E

PM

—A

nxio

lytic

[155

]

78M

agno

lia d

ealb

ata

Zuc

c.

(Mag

nolia

ceae

) E

loxo

chit

i

Eth

anol

ext

ract

of

leav

es10

0 an

d 30

0 m

g/kg

, p.o

.M

ale

Sw

iss

albi

no m

ice

EP

M, H

BT,

exp

lora

tory

rea

rings

, S

odiu

m p

ento

barb

ital-i

nduc

ed

hypn

osis

, PT

Z-in

duce

d se

izur

es

—A

nxio

lytic

, se

dativ

e an

d an

ticon

vuls

ant

[156

]

27


79M

agno

lia O

bava

ta

Thu

nb.

(Mag

nolia

ceae

) Ja

pan

ese

big

leaf

m

agn

olia

Hon

okio

l (36

)0.

2-1

mg/

kg,

p.o.

for

seve

n da

ys

Mal

e m

ice

of th

e dd

Y

stra

inE

PM

—A

nxio

lytic

[157

]

Hon

okio

l (36

)0.

2 m

g/kg

, se

ven

days

, p.

o.

Mal

e m

ice

of th

e dd

Y

stra

inE

PM

—A

nxio

lytic

[158

]

Obo

vato

l (37

)isol

ated

fr

om le

aves

0.2,

0.5

and

1.

0 m

g/kg

, p.o

.IC

R m

ale

mic

eE

PM

, HB

TG

AB

A-B

ZD

- re

cept

ors

/ C

l- cha

nnel

ac

tivat

ion

Anx

ioly

tic[1

59]

80M

artic

aria

ch

amom

ila L

inn.

or

Mat

ricar

ia r

ecut

ita

(Ast

erac

eae)

G

erm

an

cham

om

ile,

Am

eral

e

Api

geni

n(38

) is

olat

ed

from

aqu

eous

ext

ract

of

bran

chle

ts w

ith fl

ower

s

3 m

g/kg

, i.p

.M

ale

C F

l mic

eE

PM

, HB

T, L

ocom

otor

act

ivity

te

st, H

oriz

onta

l-wire

test

, S

eizu

re te

stin

g

Inte

ract

ion

with

G

AB

AA/B

ZD

re

cept

or

Anx

ioly

tic a

nd

mild

sed

ativ

e at

10

times

do

se

[160

]

Api

geni

n(38

)30

mM

In v

itro,

Rat

s—

Inte

ract

ion

with

G

AB

AA/B

ZD

re

cept

or

Anx

ioly

tic[1

61-

163]

81M

elis

sa o

ffici

nalis

Li

nn. (

Lam

iace

ae)

Lem

on

bal

m,

Co

mm

on

bal

m

Cyr

acos

: hy

dro-

alco

holic

(3

0% e

than

ol)

extr

act o

f ae

rial p

arts

120,

240

and

36

0 m

g/kg

, p.

o. fo

r 15

da

ys

C57

Bl/6

Jico

mic

eE

PM

In

hibi

ts G

AB

A-T

(t

rans

amin

ase)

ac

tivity

and

in

crea

se G

AB

A

leve

l in

brai

n

Anx

ioly

tic[1

64]

82M

itrag

yna

parv

ifolia

R

oxb.

(R

ubia

ceae

) K

aim

, Gu

likad

am

Met

hano

l, et

hyl a

ceta

te

and

alka

loid

ric

h fr

actio

n of

ste

m b

ark

200

and

400

mg/

kg, p

.o.

Sw

iss

albi

no m

ice

EP

M, M

BT

Inte

ract

ion

with

G

AB

A r

ecep

tors

Anx

ioly

tic[1

65]

83M

orin

da c

itrifo

lia

Linn

. (R

ubia

ceae

) N

on

i, In

dia

n

mu

lber

ry

(a)

Met

hano

l ext

ract

of

frui

ts

(b)

But

anol

ic fr

actio

n

(a)

IC50

– 2

2.8

µg/m

l (b

) IC

50 –

27.

2 µg

/ml

In v

itro

—G

AB

AA a

goni

stA

nxio

lytic

[166

]

84N

aucl

ea la

tifol

ia

J.E

.Sm

ith

(Rub

iace

ae)

Neg

ro p

each

, A

fric

an p

each

Dec

octio

n fr

om b

ark

of

the

root

s 80

and

160

m

g/kg

, i.p

.A

dult

Sw

iss

mal

e m

ice

EP

M, D

iaze

pam

-indu

ced

slee

p,

ME

S-,

Str

ychn

ine-

, PT

Z-in

duce

d co

nvul

sion

s te

st, O

FT

—A

nxio

lytic

[167

]

85N

elum

bo n

ucife

ra

Gae

rtne

r (N

ymph

yaea

ceae

) S

acre

d w

ater

lilly

, P

ink

lotu

s

Nef

erin

e(39

) is

olat

ed

from

met

hano

l ext

ract

of

embr

yos

of th

e se

eds

100

mg/

kg, i

.p.

Mal

e IC

R m

ice

EP

M—

Anx

ioly

tic[1

68]

86N

epet

a pe

rsic

a B

oiss

. (La

mia

ceae

) C

atm

int

Hyd

ro-a

lcoh

olic

ext

ract

(8

0% e

than

ol)

of a

eria

l pa

rts

50 m

g/kg

, i.p

.M

ale

NM

RI m

ice

EP

M—

Anx

ioly

tic[1

69]

87O

cim

um

grat

issi

mum

Lin

n.

(Lam

iace

ae)

Van

a Tu

lsi

Met

hano

l ext

ract

of

leav

es

200

and

400

mg/

kg, p

.o.

Sw

iss

albi

no m

ice

OF

T, P

TZ

-indu

ced

seiz

ure

test

—A

nxio

lytic

and

an

ticon

vuls

ant

[170

]

28


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

88O

cim

um s

anct

um

Linn

. (La

mia

ceae

) Tu

lsi,

Ho

ly b

asil

Aqu

eous

ext

ract

of

who

le p

lant

200

mg/

kg,

p.o.

Sw

iss

Mic

eE

PM

, Pas

sive

avo

idan

ce

para

digm

, Sco

pola

min

e an

d di

azep

am in

duce

d am

nesi

a

—A

nxio

lytic

and

no

otro

pic

effe

cts

[171

]

Aqu

eous

ext

ract

of

who

le p

lant

500

mg/

caps

ule

twic

e da

ily a

fter

mea

l

35 m

ale

and

fem

ale

hum

an b

eing

sH

amilt

on’s

brie

f ps

ychi

atric

ra

ting

scal

e (B

PR

S)

—A

nxio

lytic

[172

]

89P

achy

rhiz

us e

rosu

s Li

nn. (

Legu

min

osae

) B

ang

kwan

g,

Jica

ma

Eth

anol

ext

ract

of

seed

s15

0 m

g/kg

, p.

o.S

wis

s al

bino

mic

eS

tairc

ase

test

, EP

M, a

ggre

ssiv

e be

havi

or, P

ento

barb

itone

in

duce

d sl

eepi

ng ti

me,

lo

com

otor

act

ivity

, rot

orod

test

—S

edat

ive,

an

tianx

iety

m

uscl

e re

laxa

nt a

nd

antia

ggre

ssiv

e ac

tivity

[173

]

90P

anax

gin

seng

C

.A.M

eyer

(A

ralia

ceae

) C

hin

ese,

Ja

pan

ese,

Ko

rean

g

inse

ng

, Nin

jin

Gin

seng

ext

ract

G-1

1510

0 m

g/kg

, p.

o.W

ista

r ra

tsV

ogel

con

flict

pro

cedu

re—

Anx

ioly

tic[1

74]

Aqu

eous

ext

ract

of

whi

te

and

red

root

s po

wde

r20

and

50

mg/

kg, p

.o. t

wic

e da

ily fo

r 5

days

Mal

e W

ista

r st

rain

alb

ino

rats

and

alb

ino

mic

eE

PM

, OF

T, c

onfli

ct b

ehav

ior

in

thirs

ty r

ats,

foot

sho

ck in

duce

d fig

htin

g in

pai

red

mic

e

Dec

reas

e M

AO

ac

tivity

in b

rain

Anx

ioly

tic[1

75]

But

anol

frac

tions

of

root

s of

red

(R

G)

and

sun

gins

eng

(SG

)

RG

(10

0 m

g/kg

, p.o

.) a

nd

SG

(25

and

50

mg/

kg, p

.o.)

Alb

ino

mic

eE

PM

—A

nxio

lytic

[176

]

(a)

Gin

seng

roo

t pow

der

(b)

Cru

de s

apon

in

gins

eng

frac

tion

(c)

Gin

seno

side

Rb1

(40)

(a)

300,

600

an

d 12

00 m

g/kg

, p.o

(b

) 50

, 100

, an

d 20

0 m

g/kg

, p.o

(c

) 2.

5, 5

and

10

mg/

kg, i

.p

Mal

e IC

R a

lbin

o m

ice

EP

M—

Anx

ioly

tic[1

77]

(a)

Gin

seng

aqu

eous

ex

trac

t (b

) G

inse

nosi

des

Rg3

(41)

and

Rh2

(42)

fr

om r

oots

(a)

50 a

nd 1

00

mg/

kg, p

.o.

(b)

5, 1

0 an

d 25

mg/

kg, p

.o.

once

dai

ly fo

r 3

days

Mal

e IC

R m

ice

EP

MIn

tera

ctio

n w

ith

GA

BA

/BZ

D

rece

ptor

s

Anx

ioly

tic[1

78]

91P

anax

qui

nque

foliu

s Li

nn. (

Ara

liace

ae)

Am

eric

an g

inse

ng

Sap

onin

s50

and

100

m

g/kg

, p.o

.M

ale

Sw

iss

albi

no m

ice

EP

M, L

DM

, HB

T—

Anx

ioly

tic[1

79]

29


Pas

siflo

ra a

ctin

ia

Hoo

ker

(Pas

siflo

race

ae)

Wild

bel

l ap

ple

, m

arac

ujá

-do

-mat

o

Hyd

ro-e

than

ol (

HE

) an

d M

etha

nol (

ME

) E

xtra

ct fr

om le

aves

HE

(30

0 an

d 60

0 m

g/kg

, p.

o.)

ME

(10

0 an

d 30

0 m

g/kg

, p.

o.)

Mal

e al

bino

-Sw

iss

mic

eE

PM

G

AB

AA r

ecep

tor

inte

ract

ion

Anx

ioly

tic[1

80]

93P

assi

flora

ala

ta

Dry

ande

r (P

assi

flora

ceae

) W

ing

ed-s

tem

P

assi

on

flo

wer

Hyd

ro-a

lcoh

olic

ext

ract

(4

0% e

than

ol)

of le

aves

50,1

00 o

r 15

0 m

g/kg

, i.p

.A

dult

fem

ale

Wis

tar

rats

EP

M—

Anx

ioly

tic[1

81]

Spr

ay d

ried

pow

der

of

aque

ous

extr

act o

f le

aves

400

and

800

mg/

kg,

p.o

.A

dult

mal

e W

ista

r ra

tsE

PM

—A

nxio

lytic

[182

]

Aqu

eous

ext

ract

of

leav

es50

and

100

m

g/kg

, i.p

.W

ista

r ra

tsE

PM

—A

nxio

lytic

[183

, 18

4]

94P

assi

flora

coe

rule

a Li

nn.

(Pas

siflo

race

ae)

Blu

e P

assi

on

fl

ow

er

Chr

ysin

(43

)1

mg/

kg, i

.p.

Mal

e C

F1

mic

eE

PM

, HB

TIn

tera

ctio

n w

ith

BZ

D r

ecep

tors

Anx

ioly

tic[1

85,

186]

95P

assi

flora

edu

lis

Sim

s (P

assi

flora

ceae

) B

at-L

eave

d

Pas

sio

n f

low

er

Hyd

ro-a

lcoh

olic

ext

ract

(4

0% e

than

ol)

of le

aves

50, 1

00 a

nd

150

mg/

kg, i

.p.

Adu

lt fe

mal

e W

ista

r ra

tsE

PM

—A

nxio

lytic

[181

]

(a)

Aqu

eous

ext

ract

(b

) Tot

al fl

avon

oid

frac

tion

(c)

Lute

olin

-7-O

-(2-

rh

amno

syl g

luco

side

)(4

4) fr

om to

tal f

lavo

noid

fr

actio

n of

aqu

eous

ex

trac

t of

leav

es

(a)

230

mg/

kg,

p.o.

(b

) 10

0 m

g/kg

, p.

o.

(c)

30 m

g/kg

, p.

o.

Adu

lt m

ale

Sw

iss

mic

eE

PM

, MB

T—

Anx

ioly

tic[1

87]

Spr

ay d

ried

pow

der

of

aque

ous

extr

act o

f le

aves

400

and

800

mg/

kg,

p.o

.A

dult

mal

e W

ista

r ra

tsE

PM

—A

nxio

lytic

[182

]

Aqu

eous

ext

ract

50

, 100

and

15

0 m

g/kg

, i.p

.W

ista

r ra

tsE

PM

—A

nxio

lytic

[183

, 18

4]

Aqu

eous

ext

ract

of

mat

ure

frui

ts a

nd it

s bu

tano

lic fr

actio

n

100

and

300

mg/

kg, p

.o.

Adu

lt m

ale

Sw

iss

mic

eLD

M, E

thyl

eth

er–i

nduc

ed

hypn

osis

, PT

Z-in

duce

d co

nvul

sion

s

—A

nxio

lytic

and

se

dativ

e bu

t no

t an

ticon

vuls

ant

[188

]

96P

assi

flora

inca

rnat

a Li

nn.

(Pas

siflo

race

ae)

Pas

sio

n f

low

er,

May

po

p

Aqu

eous

ext

ract

(P

assi

payT

M, I

ran

Dar

ouk)

45 d

rops

/day

fo

r 4

wee

ksA

dou

ble

blin

d ra

ndom

ized

tr

ial o

n 36

pat

ient

with

G

AD

HA

MA

sco

res

—A

nxio

lytic

[189

]

Met

hano

l ext

ract

of

aeria

l par

ts12

5 m

g/kg

, p.

o.S

wis

s A

lbin

o m

ice

EP

M—

Anx

ioly

tic[1

90-

193]

Hom

oeop

athi

c fo

rmul

atio

ns10

0 m

g/kg

, p.

o.S

wis

s A

lbin

o m

ice

EP

M—

Anx

ioly

tic[1

94]

Ben

zofla

vone

nuc

leus

as

basi

c m

oiet

y co

mpo

und

from

met

hano

l ext

ract

10 m

g/kg

, p.o

.S

wis

s A

lbin

o m

ice

EP

M

—A

nxio

lytic

[195

]

30


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

Met

hano

l ext

ract

of

aeria

l par

ts a

nd C

hrys

in

2 m

g/kg

, i.p

.M

ale

Spr

ague

-Daw

ley

rats

EP

MIn

tera

ctio

n w

ith

GA

BA

/BZ

D-

rece

ptor

s

Anx

ioly

tic[1

96,

197]

Tabl

et c

onta

inin

g 1.

01

mg

benz

ofla

vone

(B

ZF

) 50

0 m

g, p

.o.

A d

oubl

e bl

ind

plac

ebo-

cont

rolle

d st

udy

on 6

0 pa

tient

s w

ith a

nxie

ty

Num

eric

al r

atin

g sc

ale,

Trie

ger

dot t

est a

nd th

e di

git-

sym

bol

subs

titut

ion

test

—A

nxio

lytic

[198

]

Hyd

ro-e

than

ol e

xtra

ct

(50%

eth

anol

) of

aer

ial

part

s

375

mg/

kg,

p.o.

Mal

e B

L6/C

57 J

mic

eE

PM

Inte

ract

ion

with

G

AB

A r

ecep

tors

Anx

ioly

tic[1

99,

200]

(a)

Hyd

ro-a

lcoh

olic

ex

trac

t (50

% e

than

ol)

of

aeria

l par

ts

(b)

But

anol

frac

tion

(c)

Chl

orof

orm

ext

ract

(a)

150

and

300

mg/

kg,

p.o.

(b

) 2.

1 an

d 4.

2 m

g/kg

, p.o

. (c

) 0.

17 a

nd

0.34

mg/

kg,

p.o.

Mal

e C

57B

L/6J

mic

eE

PM

—A

nxio

lytic

[201

]

97P

assi

flora

qu

adra

ngul

aris

Lin

n.

(Pas

siflo

race

ae)

Gia

nt

gra

nad

illa

Aqu

eous

and

eth

anol

ex

trac

t of

leav

es25

0 an

d 50

0 m

g/kg

, p.o

.A

dult

mal

e W

ista

r ra

ts a

nd

Sw

iss

mic

eE

PM

, OF

T, H

BT

—A

nxio

lytic

[202

]

98P

erill

a fr

utes

cens

(L

.) B

ritto

n (L

amia

ceae

) P

urp

le P

erill

a, W

ild

red

bas

il

Ros

mar

inic

aci

d (4

5)an

d ca

ffeic

aci

d(46

) is

olat

ed

from

hyd

ro-a

lcoh

olic

ex

trac

t of

leav

es

10 m

g/kg

, p.o

.A

lbin

o m

ice

FS

T

Mod

ulat

ion

of th

e α 1A

- ad

reno

cept

or-

med

iate

d si

gnal

tr

ansd

uctio

ns a

nd

also

atte

nuat

es

the

dow

n re

gula

tion

of

BD

NF

tr

ansc

riptio

n

Anx

ioly

tic[2

03]

99P

etiv

eria

alli

acea

Li

nn.

(Phy

tola

ccac

eae)

G

uin

ea h

en w

eed

Hex

ane,

hyd

ro-a

lcoh

olic

, an

d pr

ecip

itate

d hy

dro-

alco

holic

ext

ract

(5

0%)

of r

oots

100

and

200

mg/

kg, i

.p. a

nd

p.o.

Fem

ale

Sw

iss

mic

eE

PM

, OF

T—

Anx

ioly

tic[2

04]

Who

le p

lant

ext

ract

300

and

900

mg/

kg, p

.o.

Mal

e al

bino

Sw

iss

mic

eE

PM

, OF

T—

Anx

ioly

tic[2

05]

31


0P

iper

met

hyst

icum

Fo

rst.

(Pip

erac

eae)

K

ava,

Kaw

a

WS

149

0 ex

trac

t 50

mg,

p.o

.25

-wee

k m

ultic

ente

r ra

ndom

ized

pla

cebo

-co

ntro

lled

doub

le-b

lind

tria

l on

121

out

patie

nts

suffe

ring

from

anx

iety

of

non-

psyc

hotic

orig

in

HA

MA

, som

atic

and

psy

chic

an

xiet

y, C

linic

al G

loba

l Im

pres

sion

(C

GI)

, Sel

f-R

epor

t S

ympt

om In

vent

ory-

90 It

ems

revi

sed,

and

Adj

ectiv

e M

ood

Sca

le

—A

nxio

lytic

[206

]

Kav

a ex

trac

t LI 1

503

tabl

ets

daily

eq

uiva

lent

to

135

mg

kava

py

rone

s da

ily

for

12 w

eeks

Con

trol

led

clin

ical

tria

l on

a 37

-yea

r-ol

d fe

mal

e ou

tpat

ient

with

GA

D, S

P

and

SA

D

CG

I sca

le, A

MD

P –

mod

ule,

H

AM

A, H

amilt

on d

epre

ssio

n sc

ale,

Bec

k an

xiet

y in

vent

ory,

Spe

ilber

ger

trai

t anx

iety

in

vent

ory

—A

nxio

lytic

[207

]

Kav

a ex

trac

t st

anda

rdiz

ed to

30%

ka

vala

cton

es

280

mg/

day

for

4 w

eeks

Pat

ient

s su

fferin

g w

ith

GA

DB

aror

efle

x co

ntro

l of

hear

t rat

e (B

RC

) an

d re

spira

tory

sin

us

arrh

ythm

ia (

RS

A)

—A

nxio

lytic

[208

]

Kav

a-K

ava

spec

ial

extr

act W

S 1

490

50 m

g/da

y fo

r 4

wee

ksA

ran

dom

ized

dou

ble-

blin

d pl

aceb

o-co

ntro

lled

clin

ical

tr

ial o

n 37

pat

ient

with

D

SM

-IV

GA

D

HA

MA

Sca

le, H

ospi

tal A

nxie

ty

and

Dep

ress

ion

Sca

le (

HA

DS

), S

elf-

Ass

essm

ent o

f R

esili

ence

an

d A

nxie

ty (

SA

RA

)

—A

nxio

lytic

[209

]

Hyd

ro-a

lcoh

olic

ext

ract

of

roo

ts12

0-24

0 m

g/kg

, p.o

.W

ista

r ra

tsE

PM

—A

nxio

lytic

[210

]

Kav

a K

ava

LI15

0 ex

trac

t40

0 m

g/da

yA

ran

dom

ized

dou

ble-

blin

d pl

aceb

o-co

ntro

lled

clin

ical

tr

ial o

n 12

9 pa

tient

s su

fferin

g fr

om G

AD

HA

MA

Sca

le a

nd B

oern

er

Anx

iety

Sca

le (

BoE

AS

), C

GI,

a sl

eep

ques

tionn

aire

(sf

-13)

, and

a

qual

ity o

f lif

e qu

estio

nnai

re

—A

nxio

lytic

[211

]

Sam

ples

con

tain

ing

12.8

-100

% to

tal

kava

lact

ones

, and

fr

actio

ns c

onta

inin

g ka

vala

cton

es 1

-6(4

7-52

) in

var

ying

con

cent

ratio

n (0

.1-6

7.5%

)

Coc

kere

ls

(Gal

lus

gallu

s;

stra

in W

36)

i.p. i

njec

tions

of

diffe

rent

co

ncen

trat

ions

Chi

ck s

ocia

l sep

arat

ion

proc

edur

e —

Anx

ioly

tic[2

12]

Eth

anol

ext

ract

of

the

aeria

l par

ts12

5 m

g/kg

an

d 88

mg/

kg,

i.p.

Sw

iss

albi

no m

ice

Mir

rore

d ch

ambe

r av

oida

nce

assa

y an

d E

PM

—

Anx

ioly

tic[2

13]

Kav

a-K

ava

spec

ial

extr

act W

S 1

490

50 m

g/da

y fo

r 4

wee

ksA

ran

dom

ized

dou

ble-

blin

d pl

aceb

o-co

ntro

lled

clin

ical

tr

ial o

n 14

1 pa

tient

s su

fferin

g fr

om n

euro

tic

anxi

ety

The

tota

l sco

re o

f th

e A

nxie

ty

Sta

tus

Inve

ntor

y (A

SI)

obs

erve

r ra

ting

scal

e, s

truc

ture

d w

ell-b

eing

sel

f-ra

ting

scal

e (B

f-S

) an

d C

GI

—A

nxio

lytic

[214

]

Kav

a-K

ava

spec

ial

extr

act W

S 1

490

50-3

00 m

g/da

y fo

r 4

wee

ks

A r

ando

miz

ed d

oubl

e-bl

ind

plac

ebo-

cont

rolle

d cl

inic

al

tria

l on

230

patie

nts

suffe

ring

from

neu

rotic

an

xiet

y

HA

MA

Sca

le, s

ubje

ctiv

e w

ell-

bein

g sc

ale

(Bf-

s), E

rlang

er

Anx

iety

, Ten

sion

, Agg

ress

ion

Sca

le (

EA

AS

), C

GI,

The

Brie

f P

erso

nalit

y S

truc

ture

Sca

le a

nd

The

Adj

ectiv

e C

heck

list

—A

nxio

lytic

[215

, 21

6]

101

Pip

er s

olm

sian

um C

. D

C. (

Pip

erac

eae)

P

arip

aro

ba

Em

ulsi

on o

f th

e es

sent

ial o

il fr

om a

eria

l pa

rts

5 or

10%

v/v

Sw

iss

mal

e m

ice

EP

M—

Anx

ioly

tic[2

17]

32


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

102

Pip

er tu

berc

ulat

um

Jacq

. (P

iper

acea

e)

Pim

enta

dar

ta a

nd

P

imen

ta L

on

ga

Pip

lart

ine

(53)

am

ide

alka

loid

isol

ated

from

ro

ots

50 a

nd 1

00

mg/

kg, i

.pS

wis

s m

ale

mic

e E

PM

, OF

T—

Anx

ioly

tic[2

18]

103

Pol

ygal

a sa

bulo

sa

A.W

. Ben

nett

(Pol

ygal

acea

e)

Tim

utu

-pin

hei

rin

ho

Thr

ee d

ihyd

rost

yryl

-2-

pyro

nes

I-III

(54

-56)

and

fo

ur s

tyry

l-2-p

yron

es I-

IV

(57-

60)

isol

ated

from

et

hyl a

ceta

te fr

actio

n of

hy

dro-

etha

nol (

HE

) ex

trac

t of

who

le p

lant

HE

, fra

ctio

ns

(250

, 500

and

10

00 m

g/kg

), p.

o.,

Dih

ydro

styr

yl-

2-py

rone

s an

d st

yryl

-2-

pyro

nes

(0.3

fm

ol–2

5 pm

ol,

i.c.v

.)

Mal

e ad

ult S

wis

s m

ice

EP

M, P

ento

barb

ital-a

nd e

thyl

et

her-

indu

ced

hypn

osis

, P

TZ

-indu

ced

conv

ulsi

ons,

R

ota-

rod

test

—H

ypno

tic,

antic

onvu

lsan

t an

d an

xiol

ytic

[219

]

Dih

ydro

styr

yl-2

-py

rone

s(54

-56)

and

st

yryl

-2-

pyro

nes(

57-6

0)is

olat

ed fr

om e

thyl

ac

etat

e fr

actio

n of

hy

dro-

etha

nol (

HE

) ex

trac

t of

who

le p

lant

0.3

fmol

–25

pmol

, i.c

.v.

Mal

e ad

ult S

wis

s m

ice

EP

MB

ZD

rec

epto

r in

tera

ctio

nA

nxio

lytic

[220

]

104

Pol

ygal

a te

nuifo

lia

Will

d.

(Pol

ygal

acea

e)

Yuan

Zh

i

Pol

ygal

a sa

poni

ns

40, 8

0 an

d 16

0 m

g/kg

, p.

o.

Mal

e ad

ult S

wis

s m

ice

EP

M, O

FT,

HB

T—

Anx

ioly

tic[2

21]

105

Pro

tium

he

ptap

hyllu

m (

Aub

l.)

Mar

ch.

(Bur

sera

ceae

) B

rasi

l res

intr

ee

α an

d β

amyr

in (

61-6

2)

pent

acyc

lic tr

iterp

enes

is

olat

ed fr

om s

tem

bar

k re

sin

10, 2

5 an

d 50

m

g/kg

i.p.

or

p.o.

Mal

e S

wis

s m

ice

EP

M, O

FT

BZ

D r

ecep

tor

inte

ract

ion

Anx

ioly

tic[2

22]

106

Pru

nus

dom

estic

a Li

nn.

(Ple

uron

ectid

ae)

Mir

abel

le, P

lum

, A

lu b

ukh

ara

Chl

orog

enic

aci

d(63

)is

olat

ed fr

om fr

uits

20

mg/

kg, i

.p.

Sw

iss

albi

no m

ale

mic

eE

PM

, LD

M, f

ree

expl

orat

ory

test

—A

nxio

lytic

[223

]

107

Pul

satil

la n

igric

ans

Sto

erck

(R

anun

cula

ceae

) P

asq

uef

low

er,

Win

dfl

ow

er,

Mea

do

w a

nem

on

e

Met

hano

l ext

ract

of

aeria

l par

ts20

0 m

g/kg

, p.

o.La

ca m

ice

EP

M—

Anx

ioly

tic[2

24]

33


8P

unic

a gr

anat

um

Linn

. (P

unic

acea

e)

Po

meg

ran

ate,

G

ran

ada

Eth

anol

ext

ract

of

seed

s10

0, 2

50, a

nd

500

mg/

kg,

p.o.

Youn

g an

d ol

d m

ale

Sw

iss

albi

no m

ice

EP

M, P

ento

barb

ital-i

nduc

ed

slee

ping

tim

e, F

ST,

tail

flick

and

ho

t pla

te te

st

—A

nxio

lytic

, an

tidep

ress

ant

and

anti-

noce

cept

ive

[225

]

109

Rho

diol

a ro

sea

Linn

.(R

hizo

phor

acea

e)

Arc

tic

roo

t, G

old

en

roo

t, R

ose

roo

t

Hyd

ro-a

lcoh

olic

ext

ract

(c

onta

ins

3% r

osav

in

and

1% s

alid

rosi

de)

15 m

g/kg

, p.o

.M

ale

CD

1 m

ice

LDM

—A

nxio

lytic

[226

]

110

Rol

linia

muc

osa

(Jac

q.)

Bai

ll.

(Ann

onac

eae)

W

ild s

ug

ar a

pp

le

Hex

ane

extr

act o

f le

aves

1.62

to 6

.25

mg/

kg, p

.o.

Alb

ino

mic

eA

void

ance

exp

lora

tory

beh

avio

r pa

radi

gmG

AB

A/B

ZD

re

cept

ors

inte

ract

ion

Anx

ioly

tic[2

27]

111

Rub

us b

rasi

liens

is

Mar

tius

(Ros

acea

e)

Am

ora

bra

nca

Eth

anol

ext

ract

of

leav

es15

0 m

g/kg

, pe

r ga

vage

Mal

e W

ista

r ra

ts a

nd S

wis

s m

ice

EP

MIn

tera

ctio

n w

ith

GA

BA

A r

ecep

tor

Anx

ioly

tic[2

28,

229]

112

Rut

a ch

alep

ensi

s Li

nn. (

Rut

acea

e)

Fri

ng

ed r

ue,

h

erb

-of-

gra

ce

Eth

anol

ext

ract

of

aeria

l pa

rts

300

mg/

kg,

p.o.

Mal

e S

wis

s al

bino

mic

eP

TZ

-indu

ced

seiz

ures

, sod

ium

pe

ntob

arbi

tal-i

nduc

ed h

ypno

sis,

ex

plor

ator

y ac

tivity

, anx

iety

by

unfa

mili

ar e

nviro

nmen

t and

no

cice

ptio

n

—A

nxio

lytic

, an

ticon

vuls

ant,

seda

tive,

an

tinoc

icep

tive

[230

]

113

Sal

ix a

egyp

tiaca

Li

nn. (

Sal

icac

eae)

E

gyp

tian

m

usk

will

ow

Aqu

eous

ext

ract

of

flow

ers

100

mg/

kg, i

.p.

Mal

e N

MR

I mic

eE

PM

—A

nxio

lytic

[231

]

114

Sal

via

cinn

abar

ina

M.M

arte

ns &

G

aleo

tti (L

amia

ceae

) S

age,

Wild

Ku

s

A d

iterp

enoi

d C

MP

I10

mg/

kg, p

.o.

Alb

ino

mic

eE

PM

, FS

T—

Anx

ioly

tic[2

32]

115

Sal

via

divi

noru

m

Epl

ing

& J

átiv

a (L

amia

ceae

) D

ivin

er’s

sag

e

Sal

vino

rin-A

(64)

0.00

1-10

00

µg/k

g, s

.c.

Adu

lt m

ale

Spr

ague

-Daw

ley

rats

EP

M, F

ST,

Spo

ntan

eous

mot

or

activ

ity in

mic

e, T

ail s

uspe

nsio

n te

st

k-op

ioid

and

en

doca

nnab

inoi

d sy

stem

s

Anx

ioly

tic,

antid

epre

ssan

t[2

33]

116

Sal

via

eleg

ans

Vah

l. (L

amia

ceae

) S

carl

et p

inea

pp

le

Hyd

ro-a

lcoh

olic

(60

%

etha

nol)

extr

act o

f le

aves

and

flow

er

125,

250

, 500

, 10

00 a

nd

2000

mg/

kg,

p.o.

Mal

e IC

R m

ice

EP

M, L

DM

, OF

T—

Anx

ioly

tic[2

34]

60%

eth

anol

ext

ract

of

leav

es12

.5 m

g/kg

, i.p

.S

prag

ue D

awle

y ra

tsE

PM

, FS

T—

Psy

chot

ropi

c[2

35]

117

Sal

via

milt

iorr

hiza

B

ge. (

Lam

iace

ae)

Red

sag

e

Dite

rpen

e qu

inin

e –

Milt

irone

(65)

isol

ated

fr

om e

ther

eal e

xtra

ct o

f ro

ots

10-6

0 m

g/kg

, p.

o.A

lbin

o m

ice

Four

pla

te te

stB

ZD

rec

epto

r in

tera

ctio

nA

nxio

lytic

[236

]

118

Sal

via

reut

eran

a B

oiss

. (La

mia

ceae

) S

age

Hyd

ro-a

lcoh

olic

ext

ract

(8

0% e

than

ol)

of a

eria

l pa

rts

100

mg/

kg,

i.p

.M

ale

Syr

ian

mic

eE

PM

, Spo

ntan

eous

loco

mot

or

activ

ity—

Anx

ioly

tic[2

37]

34


ble

1: C

on

tin

ued

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

119

Sap

indu

s m

ukor

ossi

G

aert

n.

(Sap

inda

ceae

) S

oap

ber

ry,R

ith

a

Met

hano

l ext

ract

of

seed

s an

d fr

uits

200

and

400

mg/

kg, p

.o.

Alb

ino

mic

eE

PM

, Y-m

aze,

HB

T, A

ctop

hoto

-m

eter

, MB

TG

AB

Aer

gic

tran

smis

sion

Anx

ioly

tic[2

38]

120

Sau

ssur

ea la

ppa

C.B

. Cla

rke

(Ast

erac

eae)

K

uth

, Ku

sth

a

Ess

entia

l oil

Inha

latio

nA

nxie

ty in

a w

oman

in

labo

urB

ehav

iour

al r

espo

nses

—A

nxio

lytic

[239

]

121

Scu

tella

ria

baic

alen

sis

Geo

rgi

(Lab

iata

e)

Hu

ang

qin

A m

onof

lavo

noid

W

ogon

in(6

6), i

sola

ted

from

dic

hlor

omet

hane

ex

trac

t of

root

s

7.5,

15

and

30

mg/

kg, p

.o.

Mal

e IC

R m

ice

EP

MIn

tera

ctio

n w

ith

GA

BA

/ BZ

D

rece

ptor

Anx

ioly

tic[2

40]

Onl

y 2’

-OH

flav

ones

is

olat

ed fr

om

dich

loro

met

hane

, wat

er

extr

acts

of

root

s

IC50

val

ues

0.00

8 to

100

µM

In v

itro,

For

ebra

ins

of

Spr

ague

-Daw

ley

rats

R

adio

rec

epto

r B

ZD

-S a

ssay

Inte

ract

ion

with

G

AB

AA

/ B

ZD

re

cept

or

Anx

ioly

tic[2

41]

(a)

5,7,

2’-t

rihyd

roxy

-6,8

-di

met

hoxy

flav

ones

(b

) 5,

7-di

hydr

oxy-

6-m

etho

xyfla

vone

isol

ated

fr

om d

ichl

orom

etha

ne

extr

act o

f ro

ots

6.05

mM

In v

itro

Rad

io r

ecep

tor

BZ

D-S

ass

ay(a

) In

tera

ctio

n w

ith G

AB

AA

/ B

ZD

re

cept

or (

agon

ist)

(b

) In

tera

ctio

n w

ith G

AB

AA

/ B

ZD

re

cept

or

(sel

ectiv

e an

tago

nist

)

Anx

ioly

tic[2

42,

243]

Fla

vono

id b

aica

lin(6

7)

and

its a

glyc

one

baic

alei

n(68

)

Bai

cale

in (

10

mg/

kg, i

.p.)

an

d ba

ical

in

(20

mg/

kg, i

.p.)

Mal

e IC

R m

ice

Vog

el s

hock

con

flict

test

Inte

ract

ion

with

B

ZD

bin

ding

site

of

GA

BA

A

rece

ptor

s

Anx

ioly

tic[2

44]

122

Scu

tella

ria la

terif

lora

Li

nn. (

Labi

atae

) B

lue

sku

llcap

, H

oo

dw

ort

Aqu

eous

ext

ract

of

root

s10

0 m

g/m

l, or

ally

Mal

e S

prag

ue-D

awle

y ra

tsE

PM

Inte

ract

ion

with

G

AB

AA

/ B

ZD

re

cept

or

Anx

ioly

tic[2

45]

(a)

Fla

vono

id

baic

alin

(67)

in e

than

ol

extr

act o

f ro

ots

(b)

baic

alei

n(68

) in

et

hano

l ext

ract

of

root

s (c

) E

than

ol e

xtra

ct o

f ro

ots

(d)

glut

amin

e in

wat

er

extr

act o

f ro

ots

(a)

40 m

g/g,

p.

o.

(b)

33 m

g/g,

p.

o.

(c)

1.6

mg/

g,

p.o.

(d

) 31

mg/

g,

p.o.

Adu

lt m

ale

Spr

ague

-D

awle

y ra

tsE

PM

, SI

Inte

ract

ion

with

G

AB

AA

/ B

ZD

re

cept

or

Anx

ioly

tic[2

45]

123

Sec

urid

aca

long

eped

uncu

lata

Fr

esen

(P

olyg

alac

eae)

V

iole

t tr

ee

Aqu

eous

roo

ts e

xtra

ct10

0-40

0 m

g/kg

, p.o

.A

lbin

o m

ice

of e

ither

sex

EP

M, Y

-maz

e, S

tryc

hnin

e- a

nd

Pic

roto

xin-

indu

ced

seiz

ure,

H

exob

arbi

tone

- in

duce

d sl

eep

test

, Exp

lora

tory

act

ivity

—A

nxio

lytic

, an

ticon

vuls

ant,

seda

tive

[246

]

35


4S

esba

nia

gran

diflo

ra

(L.)

Poi

r. (F

abac

eae)

A

gat

i

Ben

zene

:eth

yl a

ceta

te

(BE

) fr

actio

n of

the

acet

one

solu

ble

part

of

petr

oleu

m e

ther

ext

ract

of

leav

es

100

and

200

mg/

kg, p

.o.

Mal

e S

prag

ue-D

awle

y ra

ts

and

mal

e m

ice

(NIN

str

ain)

EP

M, M

ES

-, P

TZ

-, S

tryc

hnin

e-,

Lith

ium

-pilo

carp

ine-

and

elec

tric

ally

indu

ced

seiz

ures

, P

ento

barb

ital-i

nduc

ed s

leep

, A

mph

etam

ine

anta

goni

sm

Incr

ease

in th

e br

ain

cont

ent o

f G

AB

A a

nd 5

-HT

Anx

ioly

tic a

nd

antic

onvu

lsan

t[2

47]

125

Son

chus

ole

race

us

Linn

. (A

ster

acea

e)

So

w t

his

tle,

Milk

y ta

ssel

Hyd

ro-a

lcoh

olic

(5

0% e

than

ol)

and

dich

loro

met

hane

ext

ract

fr

om a

eria

l par

ts

30-3

00 m

g/kg

, p.

o.A

dult

mal

e S

wis

s m

ice

EP

M—

Anx

ioly

tic[2

48]

126

Sph

aera

nthu

s in

dicu

s Li

nn.

(Ast

erac

eae)

M

un

di

Pet

role

um e

ther

ext

ract

(P

E),

90%

eth

anol

ex

trac

t (E

E),

Wat

er

extr

act (

WE

) of

flow

ers

PE

(10

mg/

kg,

i.p.)

, EE

(10

m

g/kg

, i.p

.),

WE

(30

mg/

kg,

i.p.)

Sw

iss

albi

no m

ale

mic

eE

PM

, OF

T, F

oot s

hock

indu

ced

aggr

essi

on

—A

nxio

lytic

[249

]

Hyd

ro-a

lcoh

olic

E

xtra

ct (

50%

eth

anol

)fr

om fu

lly g

row

n flo

wer

ing

herb

100

mg/

kg,

p.o.

Alb

ino

Wis

tar

mic

e an

d ra

ts o

f ei

ther

sex

EP

M—

Anx

ioly

tic[2

50]

127

Spo

ndia

s m

ombi

n Li

nn.

(Ana

card

iace

ae)

Ho

g p

lum

, Jo

bo

, Ye

llow

mo

mb

in

Eth

anol

ext

ract

of

leav

es12

.5 to

100

m

g/kg

, i.p

.A

lbin

o w

ista

r ra

ts a

nd

Sw

iss

mic

eM

uric

idal

act

ion

of r

ats,

Por

solt’

s F

ST

GA

BA

A r

ecep

tor

Anx

ioly

tic a

nd

antid

epre

ssan

t[2

51]

128

Sta

chys

la

vand

ulifo

lia V

ahl.

(Lam

iace

ae)

Lav

end

elb

laet

rig

e an

d W

oo

d b

eto

ny

Hyd

ro-a

lcoh

olic

ext

ract

(8

0% e

than

ol)

and

esse

ntia

l oil

of a

eria

l pa

rts

100

mg/

kg, i

.p.

Mal

e TO

mic

eE

PM

—A

nxio

lytic

[252

]

Pet

role

uum

eth

er (

PF

), et

hyl a

ceta

te (

EF

) an

d w

ater

(A

F)

frac

tions

of

hydr

o-al

coho

lic e

xtra

ct

(80%

eth

anol

) of

aer

ial

part

s

PF

(25

and

50

mg/

kg, i

.p.)

E

F (

25 a

nd 5

0 m

g/kg

, i.p

.)

AF

(50

mg/

kg,

i.p.)

Mal

e S

yria

n m

ice

EP

M—

Anx

ioly

tic[2

53]

129

The

obro

ma

caca

o Li

nn. (

Ste

rcul

iace

ae)

Cac

ao, C

ho

cola

te

tree

, Kak

ao

Mas

s or

cak

e10

0 m

g/10

0g,

o.s.

Mal

e W

ista

r st

rain

rat

sE

TM

Con

ditio

nal f

ear

rela

ting

beha

viou

r, bu

t did

not

affe

ct

the

conc

entr

atio

n of

bra

in

mon

oam

ines

su

ch a

s no

r-ep

inep

hrin

e,

sero

toni

n an

d do

pam

ine

Anx

ioly

tic[2

54]

36


Tab

le 1

: Co

nti

nu

ed

S.

No

. B

iolo

gic

al s

ou

rce

Ext

ract

/Fra

ctio

n/Is

ola

teD

ose

An

imal

/ H

um

an b

ein

g

Exp

erim

enta

l mo

del

/ A

sses

smen

t o

f cl

inic

al

par

amet

ers

Mec

han

ism

o

f ac

tio

nA

ctiv

ity

Ref

.

130

Tili

a am

eric

ana

Linn

.var

. mex

ican

a (M

alva

ceae

) A

mer

ican

b

assw

oo

d,

Am

eric

an li

nd

en

β-si

tost

erol

(69)

isol

ated

fr

om h

exan

e ex

trac

t of

inflo

resc

ence

s

1 to

10

mg/

kg,

i.p.

Mal

e S

wis

s al

bino

mic

eE

PM

, HB

T, s

odiu

m

pent

obar

bita

l-ind

uced

hyp

nosi

s an

d am

bula

tory

act

ivity

—A

nxio

lytic

and

S

edat

ive

at

high

er d

ose

(30

mg/

kg, i

.p.)

[255

, 25

6]

Met

hano

l ext

ract

from

br

acts

and

flow

ers

[Tili

rosi

de(7

0) m

ain

cons

titue

nt]

25-1

00 m

g/kg

, p.

o.A

lbin

o IC

R m

ice

EP

M—

Anx

ioly

tic[2

57]

Aqu

eous

ext

ract

of

inflo

resc

ence

s [Q

uerc

etin

(7)

and

kaem

pfer

ol(6

) m

ay b

e ac

tive

cons

titue

nts]

10-3

00 m

g/kg

, p.

o.M

ale

Sw

iss

albi

no m

ice

EP

M, H

BT,

sod

ium

pen

toba

rbita

l (S

P)-

indu

ced

hypn

osis

po

tent

iatio

n, a

mbu

lato

ry

Act

ivity

—A

nxio

lytic

and

se

dativ

e[2

58]

131

Tili

a to

men

tosa

M

oenc

h (M

alva

ceae

) S

ilver

Lim

e

Hyd

ro-a

lcoh

olic

ext

ract

(7

0% e

than

ol)

of

Inflo

resc

ence

s an

d bu

tano

l fra

ctio

n

Dos

e eq

uiva

lent

to 1

g

of p

lant

m

ater

ial

Sw

iss

albi

no M

ice

EP

M, H

BT

Inte

ract

ion

with

B

ZD

rec

epto

rs

Anx

ioly

tic[2

59]

132

Trifo

lium

pra

tens

e Li

nn. (

Faba

ceae

) R

ed c

love

r

Isof

avon

es (

MF

11R

CE

) 80

mg

Wom

en w

ith p

ostm

eno-

paus

al a

nxie

tyH

AD

S,

Zun

g’s

self-

rat

ing

depr

essi

on

scal

e

—A

nxio

lytic

and

an

tidep

ress

ant

[260

]

133

Turn

era

aphr

odis

iaca

War

d (T

urne

race

ae)

Dam

ian

a

Met

hano

l ext

ract

of

aeria

l par

ts25

mg/

kg, p

.o.

Laca

mic

eE

PM

—A

nxio

lytic

[261

]

Hom

oeop

athi

c fo

rmul

atio

ns50

mg/

kg, p

.o.

Laca

mic

eE

PM

—A

nxio

lytic

[262

]

Api

geni

n(38

) is

olat

ed

from

met

hano

l ext

ract

of

aeria

l par

ts

2 m

g/kg

, p.o

.La

ca m

ice

EP

M—

Anx

ioly

tic[2

63]

134

Unc

aria

rh

ynch

ophy

lla (

Miq

.)

Jack

s (R

ubia

ceae

) C

at’s

Cla

w h

erb

Aqu

eous

ext

ract

of

root

s20

0 m

g/kg

/da

y p.

o fo

r 7

days

Mal

e S

D r

ats

and

mal

e IC

R m

ice

EP

M, H

BT

Ser

oton

ergi

c ne

rvou

s sy

stem

Anx

ioly

tic[2

64]

135

Vac

cini

um a

shei

R

eade

(E

ricac

eae)

B

lueb

erry

Ant

hocy

anin

frac

tion

from

96%

eth

anol

ext

ract

of

ber

ries

0.6–

1.0

and

2.6–

3.2

mg/

kg/d

ay, p

.o.

Adu

lt m

ale

Sw

iss

mic

e (a

ged

3 m

onth

s)E

PM

, OF

T, In

hibi

tory

avo

idan

ce—

Mem

ory-

enha

ncin

g,

anxi

olyt

ic a

nd

loco

mot

ion

incr

easi

ng

effe

cts

[265

]

136

Val

eria

na e

dulis

ssp.

P

roce

ra N

utt.

ex T

orr.

(Val

eria

nace

ae)

Tob

acco

ro

ot

Hyd

ro-a

lcoh

olic

(70

%

etha

nol)

extr

act o

f ro

ots

100,

300

and

10

00 m

g/kg

, p.

o.

Mal

e IC

R m

ice

PT

Z-in

duce

d se

izur

es,

Exp

lora

tory

rea

ring,

Rot

arod

—A

nxio

lytic

, an

ticon

vuls

ant

and

myo

rela

xant

[266

]

37


7V

aler

iana

gl

echo

mifo

lia M

eyer

(V

aler

iana

ceae

) V

aler

ian

Val

epot

riate

frac

tion

10 m

g/kg

, p.o

.M

ale

albi

no S

wis

s m

ice

EP

M, O

FT

—A

nxio

lytic

[267

]

138

Val

eria

na o

ffici

nalis

Li

nn.

(Val

eria

nace

ae)

Val

eria

n

Hyd

ro-a

lcoh

olic

ext

ract

ca

lled

vald

an d

rops

Val

dan

drop

s o.

s. d

aily

for

15 d

ays

Fem

ale

albi

no m

ice

Beh

avio

ural

stu

dies

in

loco

mot

ion

test

and

FS

T—

Anx

ioly

tic[2

68]

Val

epot

riate

s83

.1 m

g pe

r da

y36

pat

ient

s w

ith G

AD

DS

M

III-R

Psy

chic

fact

or o

f H

AM

AS

igni

fican

t re

duct

ion

in th

e ps

ychi

c fa

ctor

of

HA

MA

Anx

ioly

tic[2

69]

6-m

ethy

lapi

geni

n (7

0)an

d he

sper

idin

(71)

, is

olat

ed fr

om r

oots

and

rh

izom

es

Hes

perid

in (

4 m

g/kg

, i.p

.),

6-m

ethy

l-ap

igen

in (

1 m

g/kg

, i.p

.)

Adu

lt m

ale

Wis

tar

rats

EP

M ,

HB

TIn

tera

ctio

n w

ith

GA

BA

A /

BZ

D

rece

ptor

Anx

ioly

tic[2

70,

271]

Fla

vono

id li

narin

(72)

4 an

d 7

mg/

kg,

i.p.

Alb

ino

mic

eH

BT

—A

nxio

lytic

[272

]

Dic

hlor

omet

hane

ext

ract

of

roo

ts0.

2 g/

kg, p

.o.

Mal

e W

ista

r ra

tsE

PM

—A

nxio

lytic

[273

]

Val

eren

ic a

cid(

73)

isol

ated

from

hyd

ro-

alco

holic

ext

ract

of

root

s

3 m

g/kg

, i.p

.Fe

mal

e ho

oded

rat

sE

PM

GA

BA

(A)

–erg

ic

syst

emA

nxio

lytic

[274

, 27

5]

139

Vite

x ne

gund

u Li

nn.

(Ver

bena

ceae

) F

ive-

leav

ed c

has

te

tree

Eth

anol

ext

ract

of

root

s10

0 an

d 20

0 m

g/kg

, p.o

.S

wis

s al

bino

mic

eE

PM

, LD

M—

Anx

ioly

tic[2

76]

140

With

ania

som

nife

ra

(Lin

n.)

Dun

al

(Sol

anac

eae)

A

shw

agan

dh

a

Gly

cow

ithan

olid

es

isol

ated

from

the

root

s20

and

50

mg/

kg, p

.o. f

or 5

da

ys

Wis

tar

rats

SI,

Nov

elty

-indu

ced

supp

ress

ed

feed

ing

late

ncy,

EP

M

—A

nxio

lytic

[277

]

Aqu

eous

ext

ract

of

root

s50

, 200

and

50

0 m

g/kg

Wis

tar

rats

EP

M—

Anx

ioly

tic[2

78]

141

Zin

gibe

r of

ficin

ale

Linn

. (Z

ingi

bera

ceae

) G

ing

er

Ben

zene

frac

tion

of

acet

one

solu

ble

part

of

petr

oleu

m e

ther

ext

ract

of

drie

d rh

izom

es

15 a

nd 3

0 m

g/kg

, i.p

.M

ale

Spr

ague

-Daw

ley

rats

EP

M—

Anx

ioly

tic[2

79]

142

Ziz

iphu

s ju

juba

M

iller

(R

ham

nace

ae)

Des

i Ber

Eth

anol

ext

ract

of

seed

s0.

5 g/

kg, p

.o.

Mal

e IC

R m

ice

Bla

ck/w

hite

test

, EP

M a

nd

ambu

lato

ry b

ehav

iour

test

—A

nxio

lytic

[280

]

Alc

ohol

ic e

xtra

ct o

f se

eds

320

mg/

kg,

p.o.

thyr

oid

tabl

et fo

r ni

ne

days

Yin

def

icie

ncy

mic

eE

PM

, LD

MIn

crea

se th

e G

AB

A a

nd

expr

essi

on o

f G

AB

AA

Anx

ioly

tic[2

81]

San

join

ine-

A(7

4) fr

om

alka

loid

al fr

actio

n of

se

eds

2.0

mg/

kg, p

.o.

Mal

e IC

R m

ice

EP

M, O

FT,

HB

TG

AB

Aer

gic

tran

smis

sion

Anx

ioly

tic[2

82]

38


Tab

le 2

: Lis

t o

f va

rio

us

anxi

oly

tic

form

ula

tio

ns

and

co

mp

ou

nd

s.

S.

No

. F

orm

ula

tio

n/E

xtra

ct/F

ract

ion

/Iso

late

Do

seA

nim

al/H

um

an

bei

ng

sE

xper

imen

tal m

od

el/C

linic

al

stu

die

s p

aram

eter

sM

ech

anis

m o

f ac

tio

nA

ctiv

ity

Ref

.

01O

ils o

f ro

se, y

lang

-yla

ng, a

nd C

ham

omile

ex

trac

ted

from

flow

ers

of R

osa

sp.,

Can

anga

od

orat

a, a

nd A

nthe

nis

nobi

lis (

or M

atric

aria

ch

amom

illa)

, res

pect

ivel

y. or

ange

oil

extr

acte

d fr

om th

e rin

d of

Citr

us s

p..

200–

1600

mg/

kg,

i.p.

Mal

e IC

R m

ice

Gel

ler

conf

lict t

est a

nd V

ogel

’s

conf

lict t

est

Oth

er m

echa

nism

bu

t not

thro

ugh

GA

BA

/BZ

D

Onl

y ro

se o

il ex

hibi

ted

anxi

olyt

ic

activ

ity

[283

]

Lave

nder

oil

1600

mg/

kg, i

.p.

Mal

e IC

R m

ice

Gel

ler

type

con

flict

test

—A

nxio

lytic

[284

]

Ess

entia

l oils

:

(a)

Lave

nder

oil

– 2-

phen

ethy

l alc

ohol

, ci

tron

ella

l(75)

(b)

Ros

e oi

l – 1

,8 c

ineo

le(7

6), m

enth

one(

77),

pule

gone

(78)

, met

hyl a

lcoh

ol,

cary

ophy

llene

(79)

(c)

Pep

perm

int -

men

thol

(80)

1 m

l/100

g, i

.p.

Mal

e IC

R m

ice

(a)

Ant

icon

flict

(b

) In

crea

sed

ambu

lato

ry e

ffect

(c

) In

crea

sed

ambu

lato

ry e

ffect

Dop

amin

e re

cept

or

invo

lvem

ent

Anx

ioly

tic[2

85]

Ros

e oi

l1

ml/1

00 g

, i.p

.M

ale

ICR

mic

eG

elle

r co

nflic

t tes

t and

Vog

el’s

co

nflic

t tes

t—

Anx

ioly

tic[2

86]

Ros

e oi

lIn

hala

tion

(1.0

, 2.

5 or

5.0

% w

/w)

Adu

lt W

ista

r m

ale

rats

EP

M—

Anx

ioly

tic[2

87]

Lem

on o

ilIn

hala

tion

1 m

lIC

R s

trai

n m

ice

EP

M, F

ST,

OF

T5-

HT

nerg

ic

path

way

and

the

supp

ress

ion

of D

A

activ

ity r

elat

ed to

en

hanc

ed

5-H

Tne

rgic

ne

uron

s

Anx

ioly

tic a

nd

antid

epre

ssan

t[2

88]

Ner

oli e

ssen

tial o

ilIn

hala

tion

100

µlG

erbi

lsLo

com

otor

act

ivity

, FS

T—

Anx

ioly

tic[2

89]

02M

onot

erpe

nic

phen

ol –

Car

vacr

ol(8

1) fr

om

esse

ntia

l oil

frac

tion

of O

rega

no a

nd T

hym

e12

.5, 2

5, 5

0 m

g/kg

, p.o

.M

ale

Sw

iss

mic

eE

PM

GA

BA

ergi

c tr

ansm

issi

onA

nxio

lytic

[290

]

03M

onot

erpe

ne a

lcoh

ol –

Isop

uleg

one(

82)

25, 5

0 m

g/kg

, i.p

.M

ale

Sw

iss

mic

eO

FT,

EP

M, H

BT,

Tai

l sus

pens

ion

and

FS

T—

Anx

ioly

tic a

nd

antid

epre

ssan

t[2

91]

04In

dole

alk

aloi

d al

ston

ine(

83)

1 m

g/kg

, i.p

.M

ale

Sw

iss

mic

eLD

M—

Anx

ioly

tic

[292

]

05A

swal

100-

250

mg/

l, i.p

.G

uine

a pi

gs a

nd

Long

-Eva

ns r

ats

Ext

race

llula

r an

d w

hole

cel

l pa

tch

clam

p re

cord

ings

on

CA

1 py

ram

idal

neu

rons

Het

ero-

gene

ous

with

cal

cium

an

tago

nism

Anx

ioly

tic a

nd

antie

pile

ptic

[293

]

06Ir

idol

con

tain

ing

com

poun

ds ir

idoi

ds10

mg/

kg, p

.o.

Mal

e ra

ts a

nd

patie

nts

with

ar

teria

l hy

pert

ensi

on I

- II

degr

ee,

acco

mpa

nied

by

psyc

ho-e

mot

iona

l di

stur

banc

es

Beh

avio

ural

par

amet

ers

Anx

ioly

tic[2

94]

39


Kav

ospo

ral f

orte

(S

tand

ardi

zed

extr

act o

f K

ava)

150

mg

for

a w

eek

Clin

ical

tria

l (20

pa

tient

s w

ith

situ

atio

nally

in

duce

d an

xiet

y)

Two

self-

rate

d sc

ale

and

one

obse

rver

rat

ed s

cale

—A

mel

iora

tion

of

anxi

ety

aris

es in

co

nnec

tion

with

m

amm

ary

biop

sy

[295

]

08S

tand

ardi

zed

prod

uct c

onta

inin

g M

elis

sa

offic

inal

is a

nd V

aler

iana

offi

cina

lis60

0 m

gD

oubl

e bl

ind,

pl

aceb

o co

ntro

lled,

ra

ndom

ized

cro

ss

over

, 24

heal

thy

volu

ntee

rs

Def

ined

Inte

nsity

Str

esso

r S

imul

atio

n (D

ISS

) an

d C

ogni

tive

perf

orm

ance

—A

mel

iora

ted

the

nega

tive

effe

cts

of

the

DIS

S o

n ra

tings

of

anxi

ety

[296

]

09Z

ingi

com

b, a

pre

para

tion

cons

istin

g of

Z

. offi

cina

le a

nd G

. bilo

ba e

xtra

cts

0.5,

1, 1

0 or

100

m

g/kg

, in

trag

astr

ical

ly

Mal

e W

ista

r ra

tsO

ne-t

rial s

tep-

thro

ugh

avoi

danc

e ta

sk—

Anx

ioly

tic[2

97]

10P

olyh

erba

l for

mul

atio

n50

, 100

, 300

mg/

kg, p

.o.

Mal

e S

wis

s m

ice

EP

M a

nd F

ST

—A

ntia

nxie

ty a

nd

antid

epre

ssan

t[2

98]

11S

uanz

aore

ntan

g, C

hine

se m

edic

ine

(Sem

en

Ziz

iphi

Spi

nosa

e, R

hizo

ma

Chu

anxi

ong,

P

oria

, Rhi

zom

a A

nem

arrh

enae

, Rad

ix e

t R

hizo

ma

Gly

cyrr

hiza

e)

5 g

Hum

an b

eing

s—

Dec

reas

e se

roto

nerg

ic

activ

ity

Anx

ioly

tic[2

99,

300]

12S

ho-ju

-sen

(S

K),

a Ja

pane

se h

erba

l m

edic

ine,

con

tain

s a

wat

er e

xtra

ct o

f S

asa

kurin

ensi

s M

akin

o et

Sib

ata

(Kum

azas

a;

Poa

ceae

) le

aves

(S

S),

etha

nol e

xtra

ct o

f P

inus

den

siflo

ra S

iebo

ld e

t Zuc

earin

i (J

apan

ese

red

pine

; Pin

acea

e) (

PN

) an

d P

anax

gin

seng

C.A

. Mey

er (

Gin

seng

; A

ralia

ceae

) (P

X)

in th

e ra

tio o

f 8:

1:1

SK

(10

%

solu

tions

for

7 da

ys)

Mal

e m

ice

of th

e dd

Y s

trai

nE

PM

—A

nxio

lytic

[301

]

13K

ami-S

hoya

-San

(T

J-24

) is

one

of

the

trad

ition

al C

hine

se h

erba

l med

icin

e:

Bup

leur

um s

corz

oner

aefo

llium

Will

d.

(Bup

leur

i Rad

ix; B

uple

urac

eae)

, Pae

onia

la

ctifl

ora

Pal

las

(Pae

onia

e R

adix

; P

aeon

acea

e), A

trac

tylo

des

lanc

era

(Thu

nb.)

D

C. (

Act

ract

ylod

is L

ance

ae R

hizo

ma;

C

ompo

sita

e), A

rcha

ngel

ica

offic

inal

is H

offm

. (A

ngel

icae

Rad

ix; U

mbe

llife

rae)

, Por

ia c

ocos

(S

chw

.) W

olf

(Hoe

len;

Pol

ypor

acea

e),

Gar

deni

a ja

smin

oide

s E

llis

(Gar

deni

ae

Fruc

tus;

Rub

iace

ae),

Pae

onia

suf

frut

icos

a A

ndr.

(Mou

tan

Cor

tex;

Pae

onac

eae)

, G

. gla

bra

(Gly

cyrr

hiza

e R

adix

; Le

gum

inos

ae),

Z. o

ffici

nale

(Z

ingi

beris

R

hizo

ma;

Zin

gibe

race

ae),

Men

tha

arve

nsis

M

alin

vaud

(M

enth

ae H

erba

; Lab

iata

e)

25-1

00 m

g/kg

, p.

o.A

lbin

o m

ice

SI

5α-r

educ

tase

in

hibi

tor,

invo

lvem

ent o

f ne

uros

tero

id

synt

hesi

s fo

llow

ed

by G

AB

A r

ecep

tor

stim

ulat

ion

Anx

ioly

tic[3

02]

40


ble

2: C

on

tin

ued

S.

No

. F

orm

ula

tio

n/E

xtra

ct/F

ract

ion

/Iso

late

Do

seA

nim

al/H

um

an

bei

ng

sE

xper

imen

tal m

od

el/C

linic

al

stu

die

s p

aram

eter

sM

ech

anis

m o

f ac

tio

nA

ctiv

ity

Ref

.

14B

otan

ical

ext

ract

s

(a)

Aqu

eous

ext

ract

of

the

Rut

acea

e fa

mily

(b

) hy

dro-

alco

holic

ext

ract

of

Alc

hem

illa

eryt

hrop

oda

Juz.

(La

dies

Man

tle; R

osac

eae)

(a)

28 a

nd 5

6 m

g/kg

, i.p

. (b

) 12

.5 a

nd 2

5 m

g/kg

, i.p

.

Chi

ckC

hick

soc

ial s

epar

atio

n-st

ress

pr

oced

ure

—A

nxio

lytic

[303

]

(a)

Aq.

Ext

ract

of

Mel

issa

offi

cina

lis(b

) A

q. e

xtra

cts

of C

ente

lla a

siat

ica

and

Val

eria

na o

ffici

nalis

(c

) A

q. e

xtra

cts

of M

aric

aria

rec

utita

and

H

umul

us lu

pulu

s

(a)

IC50

– 0

.35

mg/

ml

(b)

1 m

g/m

l (c

) 0.

11-0

.65

mg/

ml

In v

itro

(a)

inhi

bit G

AB

A

tran

sam

inas

e (b

) st

imul

ate

glut

amic

aci

d de

carb

oxyl

ase

(c)

inhi

bit g

luta

mic

ac

id d

ecar

boxy

lase

Anx

ioly

tic[3

04]

15D

ieta

ry p

rodu

cts:

Die

tary

soy

ph

ytoe

stro

gens

Phy

to-

estr

ogen

rich

Phy

to-6

00 d

iet

Long

–Eva

ns

mal

es a

nd

fem

ales

rat

s

EP

M—

Anx

ioly

tic[3

05]

Tab

le 3

: Lis

t o

f an

xio

lyti

c p

aten

ted

form

ula

tio

ns.

S. N

o.

Co

mp

osi

tio

n o

f fo

rmu

lati

on

Act

ivit

yR

ef.

01L-

tryp

toph

an, l

inse

ed o

il, th

yme

oil,

aque

ous

extr

acts

of

St-

John

’s w

ort,

Are

naria

bl

osso

m, V

aler

ian,

Ele

cam

pane

Man

agem

ent o

f st

ress

incl

udin

g sl

eep

dist

urba

nces

, agg

resi

vene

ss,

inst

abili

ty o

f te

mpe

r an

d st

ate

of a

nxie

ty[3

06]

02T

hean

ine,

gre

en te

a, r

ed g

inse

ng, S

asam

orph

a pu

rpur

asce

ns e

xtra

cts

Man

agem

ent o

f an

xiet

y an

d st

ress

[307

]03

Wat

er, a

q.-a

lcoh

olic

and

CO

2 ex

trac

t of

Forg

et-m

e-no

t (M

yoso

tis)

Anx

ioly

tic, n

ootr

opic

, ant

icon

vuls

ant a

nd c

ereb

ropr

otec

tive

activ

ity[3

08]

04Ta

blet

, pill

or

gran

ule

com

pris

ing

Bup

leur

i Rad

ix, R

hizo

ma

Ane

mar

rhen

ae, s

apon

in

com

pone

nt is

olat

ed fr

om S

emen

Ziz

iphi

spi

nosa

eTr

eatm

ent o

f ac

ute

anxi

ety

(Pan

ic a

nxie

ty)

and

chro

nic

anxi

ety

(Gen

eral

ized

an

xiet

y)[3

09]

05E

xtra

cts

of p

lant

s co

ntai

ning

bet

ulin

ic a

cid

and

its d

eriv

ativ

esA

nxio

lytic

[310

]06

Cas

sia

tora

aq.

Ext

ract

Anx

ioly

tic[3

11]

07R

osam

arin

ic a

cid

isol

ated

from

Per

illa

extr

act

Ant

ianx

iety

and

ant

idep

ress

ant

[312

]08

Hyd

ro-a

lcoh

olic

ext

ract

of

Pip

er m

ethy

stic

um le

aves

Anx

ioly

tic, a

ntic

onvu

lsan

t, m

uscl

e re

laxa

nt, a

nalg

esic

, sle

ep in

duci

ng,

antii

nfla

mm

ator

y an

d ne

urop

rote

ctiv

e[3

13]

09H

omoe

opat

hic

com

plex

com

pris

ing

Aco

nite

, Ave

na s

ativ

a, P

assi

flora

inca

rnat

a,

Scu

tella

ria la

terif

olia

, Str

amon

ium

and

Val

eria

naA

ntia

nxie

ty[3

14]

10Ta

blet

, gra

nule

, cap

sule

s or

ora

l liq

uid

of m

etha

nol e

xtra

ct o

f R

umex

mad

aio

Ant

ianx

iety

[315

]11

Loze

nges

con

tain

ing

Citr

us p

ectin

, tris

odiu

m c

itrat

e, r

aw s

ugar

, wat

er, g

luco

se fr

ucto

se

syru

p, a

nd m

ixtu

re o

f la

vend

er o

il, e

xtra

cts

of M

elis

sa, h

op a

nd o

atA

ntia

nxie

ty[3

16]

12P

heno

lic c

ompo

unds

hav

ing

phen

olic

mol

ecul

e co

vale

ntly

link

ed a

n ox

ygen

con

tain

ing

grou

p, a

nito

gen

or a

noth

er o

xyge

n co

ntai

ng g

roup

and

C1-

C4

alko

xy g

roup

obt

aine

d fr

om m

onoc

otyl

edon

pla

nts

like

corn

Ant

ianx

iety

[317

]

41


ble

4: L

ist

of

revi

ew a

rtic

les

pu

blis

hed

on

an

xio

lyti

c p

lan

ts, a

nd

th

eir

con

stit

uen

ts a

nd

form

ula

tio

ns.

S. N

o.

Info

rmat

ion

ava

ilab

leP

lan

t d

rug

sP

hyto

con

stit

uen

ts

rep

ort

edT

her

apeu

tic

acti

vity

re

po

rted

Mo

de

of

acti

on

rep

ort

edR

ef.

01P

harm

acol

ogic

al r

epor

tsV

aler

iana

offi

cina

lis, M

elis

sa

offic

inal

is, P

assi

flora

inca

rnat

a,

Hum

ulus

lupu

lus,

Lav

endu

la

offic

inal

is, P

iper

met

hyst

icum

, Tili

a pl

atyp

hyllo

s, L

eonu

rus

cord

iaca

, H

yper

icum

per

fora

tum

—A

nti-a

nxie

ty a

nd

antid

epre

ssan

t—

[318

]

02Tr

aditi

onal

use

s, c

hem

ical

co

nstit

uent

s, p

harm

acol

ogic

al r

epor

tsC

atha

edu

lis, C

ola

spec

ies,

Dat

ura

spec

ies,

Pau

siny

stal

ia y

ohim

be,

Tabe

rnan

the

ibog

a

—P

sych

oact

ive

—[3

19]

03P

harm

acol

ogic

al r

epor

ts(a

) E

phed

ra s

peci

es, P

aulli

nia

spec

ies,

Cat

ha e

dulis

(b)

Can

nabi

s sa

tiva,

Tab

erna

nthe

ib

oga,

Psy

chot

ria v

iridi

s,

Ban

iste

riops

is

(c)

Pas

siflo

ra in

carn

ata,

Val

eria

na,

Pip

er m

ethy

stic

um

—(a

) A

dapt

ogen

(b

) H

allu

cino

geni

c (c

) A

nalg

esic

and

an

xiol

yitc

—[3

20]

04C

linic

al r

epor

tsP

iper

met

hyst

icum

, Gin

kgo

bilo

ba,

Gal

phim

ia g

lauc

a, M

atric

aria

re

cutit

a, P

assi

flora

inca

rnat

a,

Val

eria

na o

ffici

nalis

—A

nxio

lytic

, Gen

eral

ized

an

xiet

y di

sord

ers

—[3

21]

05P

harm

acol

ogic

al r

epor

ts, M

ode

of

actio

nG

inkg

o bi

loba

, Hyp

eric

um

perfo

ratu

m, V

aler

iana

offi

cina

lis,

Pan

ax g

inse

ng

—A

nxio

lytic

Inte

ract

ion

with

rec

epto

rs o

f C

NS

(γ-

amin

obut

yric

aci

d,

glut

amat

e, d

opam

ine,

m

usca

rinic

and

ade

nosi

ne

rece

ptor

s)

[322

]

06P

harm

acol

ogic

al r

epor

tsK

ava,

Sku

llcap

, Lem

on b

alm

(M

elis

sa

offic

inal

is),

Val

eria

na o

ffici

nalis

, P

assi

flora

, Die

tary

sup

plem

ents

—A

nxio

lytic

—[3

23]

Pha

rmac

olog

ical

rep

orts

and

Clin

ical

re

port

sH

awth

orn

and

Cal

iforn

ia P

oppy

, Im

mat

ure

Oat

See

d, P

assi

onflo

wer

, Le

mon

balm

, Ver

vain

, Lav

ende

r an

d Li

nden

—N

ervi

ne, a

nxio

lytic

—[3

24]

07C

linic

al r

epor

tsP

iper

met

hyst

icum

, Bac

opa

mon

nier

a, K

ava

—A

nxio

lytic

—[3

25]

08P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s an

d M

echa

nism

of

actio

nH

erba

l dru

gs /

Her

bal c

onst

ituen

ts—

Psy

chia

tric

dis

orde

rs—

[326

]

09P

hyto

chem

ical

rep

orts

, P

harm

acol

ogic

al r

epor

tsB

razi

lian

plan

ts (

39-a

nxio

lytic

and

28

- hy

pnot

ic)

Fla

vono

ids,

ess

entia

l oi

ls, p

heno

lic a

cids

, al

kalo

ids

Anx

ioly

tic a

nd h

ypno

tic—

[327

]

10P

harm

acol

ogic

al r

epor

tsFo

ods

Anx

ioly

tic a

nd

antid

epre

ssan

t—

[328

]

42


ble

4: C

on

tin

ued

S. N

o.

Info

rmat

ion

ava

ilab

leP

lan

t d

rug

sP

hyto

con

stit

uen

ts

rep

ort

edT

her

apeu

tic

acti

vity

re

po

rted

Mo

de

of

acti

on

rep

ort

edR

ef.

11P

hyto

chem

ical

rep

orts

, P

harm

acol

ogic

al r

epor

tsB

razi

lian

plan

ts(a

) F

lavo

noid

s (b

) A

lkal

oids

(c

) E

ssen

tial o

il (d

) Li

gnan

s (e

) Tan

nins

(f

) Trit

erpe

ne a

nd

sapo

nins

(a)

Ana

lges

ic, a

ntip

yret

ic,

antia

nxie

ty, h

ypno

tic

(b)

Hal

luci

noge

n,

stim

ulan

t (c

) A

ntip

yret

ic, a

ntia

nxie

ty

(d)

Hal

luci

noge

n (e

) A

ntia

nxie

ty

(f)

Hyp

notic

—[3

29]

12P

harm

acol

ogic

al r

epor

tsE

ight

y fiv

e he

rbal

dru

gs—

Anx

ioly

tic, a

ntid

epre

ssan

t, ne

urol

eptic

, ant

idem

entia

—[3

30]

13P

harm

acol

ogic

al r

epor

ts, M

ode

of

actio

nM

etha

nol e

xtra

cts

of tr

aditi

onal

pl

ants

—P

sych

othe

rape

utic

ac

tivity

In v

itro

radi

olig

and

rece

ptor

bi

ndin

g an

d en

zym

e as

says

su

ch a

s ac

etyl

chol

ine

este

rase

, cho

line

acet

yl

tran

sfer

ase,

mon

oam

ine

oxid

ase

A a

nd B

.

Sel

ectiv

ely

on G

AB

AA, N

MD

A

and

MA

O r

ecep

tors

[331

]

14P

harm

acol

ogic

al r

epor

ts, M

ode

of

actio

nE

than

ol e

xtra

cts

of 3

1 tr

aditi

onal

pl

ants

—A

nxio

lytic

and

an

tiepi

lept

icG

AB

AA –

BZ

D r

ecep

tor,

Inhi

bitio

n of

GA

BA

tr

ansa

min

ase

[332

]

15C

linic

al r

epor

tsN

atur

al r

emed

ies

such

as

St J

ohn’

s W

art,

Kav

a K

ava,

Pas

sion

flow

er,

Inos

itol,

Val

eria

n ro

ot, M

elat

onin

, O

meg

a-3-

fatty

aci

ds, s

-ade

nosy

l-L-

met

hion

ine

—A

nxio

lytic

—[3

33]

16P

harm

acol

ogic

al r

epor

ts(a

) K

ava

kava

roo

ts

(b)

Gin

kgo

extr

act

—(a

) A

nxio

lytic

(b

) N

ootr

opic

—[3

34]

17P

harm

acol

ogic

al r

epor

ts, C

linic

al

repo

rts

Hyp

eric

um p

erfo

ratu

m—

Ant

idep

ress

ant,

anxi

olyt

ic, n

ootr

opic

, se

dativ

e, a

nalg

esic

, an

ticon

vuls

ant,

antis

chiz

ophr

enic

, al

coho

l, ni

cotin

e an

d ca

ffein

e de

addi

ctio

n

—[3

35]

18E

ffic

acy,

Saf

ety

Pro

file,

P

harm

acol

ogic

al r

epor

tsK

ava

—A

nxio

lytic

—[3

36]

19S

afet

y pr

ofile

Kav

a—

Anx

ioly

tic—

[337

]

20C

linic

al r

epor

tsK

ava

—A

nxio

lytic

—[3

38-

341]

21P

harm

acol

ogic

al r

epor

ts, S

ide

effe

cts,

M

ode

of a

ctio

nK

ava

lact

ones

—(a

) A

nalg

esic

, ane

sthe

tic

(b)

Anx

ioly

tic

Sid

e ef

fect

s- S

kin

rash

an

d ka

va d

erm

opat

hy

(a)

Non

-opi

ate

path

way

(b

) G

AB

A r

ecep

tor

bind

ing

[342

]

43


Phy

toch

emic

al r

epor

ts,

Pha

rmac

olog

ical

rep

orts

Mat

ricar

ia r

ecut

itaF

lavo

noid

s, p

heno

lic

com

poun

ds, e

ssen

tial o

ilA

ntio

xida

nt, a

ntim

icro

bial

, an

tipla

tele

t, an

tiinf

lam

mat

ory,

antim

utag

enic

, an

tispa

smod

ic, a

nxio

lytic

, ch

oles

tero

l low

erin

g

—[3

43]

23P

harm

acol

ogic

al r

epor

tsZ

izyp

hus

juju

ba—

Anx

ioly

tic, s

edat

ive,

hy

pnot

ic, a

phro

disi

ac,

antic

ance

r, hy

pote

nsiv

e,

antii

nfla

mm

ator

y

—[3

44]

24P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s—

Alk

aloi

ds in

Sce

letiu

m

and

Mes

embr

yant

ham

acea

e

Nar

cotic

-anx

ioly

tic,

hallu

cino

geni

c—

[345

]

25P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s—

Can

nabi

noid

s, δ

9 -te

trah

ydro

cann

abin

ol,

cann

abid

iol

Sed

ativ

e, h

ypno

tic,

anxi

olyt

ic, a

ntid

epre

ssan

t, an

tipsy

chot

ic,

antic

onvu

lsan

t

—[3

46]

26P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s—

Fla

vono

ids

– ch

rysi

n,

apig

enin

and

se

mis

ynth

etic

de

rivat

ives

of

flavo

ne

Anx

ioly

tic—

[347

]

27P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s, M

ode

of a

ctio

n—

Fla

vono

ids

Anx

ioly

ticB

ZD

site

on

GA

BA

A[3

48]

28P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s—

Food

pro

tein

s –

δ-op

ioid

pe

ptid

es, g

lute

n,

exor

phin

s, r

ubis

colin

s

Ant

inoc

icep

tive,

mem

ory

enha

ncin

g, a

nxio

lytic

—[3

49]

29P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s, M

ode

of a

ctio

n—

Terp

enoi

ds:

Mon

oter

peno

ids

(lina

lool

, α-t

hujo

ne,

born

eol,

vale

potr

iate

s);

Ses

quite

rpen

oids

(v

aler

enic

aci

d,

arte

mis

inin

); D

iterp

enoi

ds

(gin

kgol

ides

, for

skol

in,

salv

inor

ine

A);

Trite

rpen

oids

(g

inse

nosi

des)

; M

erot

erpe

noid

s (c

anna

bino

ids)

Sed

ativ

e, a

nxio

lytic

, an

tinoc

icep

tive,

an

ticon

vuls

ant,

hallu

cino

geni

c

—[3

50]

30P

hyto

cons

titue

nts,

Pha

rmac

olog

ical

re

port

s, M

ode

of a

ctio

n—

Wog

onin

(F

lavo

noid

)A

nxio

lytic

B

ZD

bin

ding

site

of

GA

BA

A

and

mod

ulat

ion

of r

ecep

tor

activ

ity

[351

]

44


(c) chemical constituents responsible for antianxiety activity have been reported in 53 plants and

(d) possible mechanism of action has been reported in 41 plants.

Seven formulations of plant drugs, 02 well known classes of phytoconstitunts and 03 pure constituents present in various plants, and nutraceuticals reported to possess antianxiety activity in battery of experimental models of anxiety have been compiled in present work (Table 2). Twelve anxiolytic formulations containing plants have been patented (Table 3). A survey of literature revealed that 30 review articles have been published on anxiolytic plant formulations and specific plant covering broad aspects as phytochemistry, pharmacology, clinical studies, toxicology and safety profiles (Table 4).

This review article would of immense help to natural product researchers to select traditionally used and clinically potential plants for their future research work.

AKNOWLEDGEMENT

Authors are grateful to Mr Rakesh Chawla, Lecturer, S.D. College of Pharmacy, Barnala for providing necessary full research articles for compilation of this review.

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ABBREVIATIONS

ADAA American Psychiatric AssociationAPA Anxiety Disorders Association of AmericaASI Anxiety Status InventoryBRC Baroreflex control of heart rateBZD BenzodiazepinesBoEAS Boerner Anxiety ScaleCNS Central nervous systemCGI Clinical Global ImpressiondlPAG Dorsolateral peri aqueductalECG ElectrocardiogramEEG ElectroencephalographicEPM Elevated plus mazeEZM Elevated zero mazeEAAS Erlanger Anxiety, Tension, Aggression ScaleFRA Federal Regulatory AuthoritiesFST Forced swimming testGABA Gamma-amino butyric acidGAD Generalized anxiety disorderGRAD Global Research on Anxiety and DepressionHAMA Hamilton Anxiety Scale

HBT Hole board testHADS Hospital Anxiety and Depression Scale5-HT1A 5-hydroxytryptamine 1Ai.p. IntraperitoneallyLDM Light / Dark modelMBT Marble burying testMAO Monoamine oxidaseNOS Nitric oxide synthaseOCD Obsessive–compulsive disorderOFT Open field testPD Panic disorderPTZ Pentylenetetrazolep.o. Per oralPTSD Post-traumatic stress disorderSARA Self Assessment of Resilience and AnxietySI Social interactionSAD Social phobia or Social anxiety disorderSP Specific phobias.c. SubcutaneouslyTDS Thrice daily


Review Article



*Correspondence: Tel.: +61 7 37357637; fax: +61 7 37355282. E-mail: [email protected] (I. E. Cock).DOI: 10.5530/pc.2011.1.3

semi-synthetic analogues of phytochemicals. It has been estimated that approximately 25% of all prescription drugs currently in use are of plant origin.[2,3] Furthermore, approximately 75% of new anticancer drugs marketed between 1981 and 2006 were derived from plant compounds.[3]

Traditionally, plant based medicines have been used as crude formulations such as infusions, tinctures and extracts, essential oils, powders, poultices and other herbal preparations. The current trend is to isolate and characterise the individual phytochemical components with the aim of producing an analogue of increased bioactivity/bioavailability. Such studies have given rise to many useful drugs such as quinine (from Cinchona spp.) and digoxin (from Digitalis spp.) as well as the anticancer drugs vincristine and vinblastine (from Vinca rosea). However, the bioactivities seen for crude extracts are often much enhanced, or even totally different to those seen for the individual components.[4,5] Crude plant extracts may contain hundreds, or even thousands of different chemical constituents that interact in complex ways. Often it is not known how an extract works, even when its therapeutic benefit is well established.

The study of crude extracts is itself fraught with difficulties. Plants grown under varied conditions will often produce different phytochemical profiles, or at least different quantities of the individual components.[6,7] Similarly, different cultivars within a

Problems of Reproducibility and Efficacy of Bioassays Using Crude Extracts, with reference to Aloe veraI. E. Cocka,b*aBiomolecular and Physical Sciences, Nathan Campus, Griffith University, 170 Kessels Road, Nathan, Queensland 4111, Australia. bEnvironmental Futures Centre, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Queensland 4111,Australia

IntRodUCtIon

Plants have a long history of usage as medicinal agents and were the main source of medicines prior to the advances of modern medicine. In many developing countries, herbal medicinal systems remain important in the treatment of many ailments. Ayuvedic medicine is still commonly practiced within India with an estimated 85% of Indians still using crude plant preparations for the treatment of a wide variety of diseases and ailments.[1] Traditional Chinese medicine (TCM) and African medicinal systems also account for a major portion of health care in these regions. Even in countries where allopathic/Western medicine is dominant, much is also owed to plant medicinal systems. Furthermore, many users are returning to herbal medicinal systems due to the perception that natural medicines are often safer than allopathic drugs, as well as seeking treatments to diseases for which modern medicine does not yet have solutions.

Many of the prescription drugs currently marketed for a wide variety of ailments were originally isolated from plants or are

ABSTRACT: Aloe vera has a long history of medicinal usage and its biological activities have been well documented in a variety of bioassays. However, isolated Aloe vera leaf components generally do not display the same bioactivities, or have lower efficacies than crude juice/extracts. It is likely that several components work in a synergistic manner in the crude mixture, resulting in increased bioactivities. Furthermore, different laboratories often report varying bioactivities using the same extraction procedure on plant material from the same species. Individual Aloe vera cultivars may have widely varying levels of the bioactive phytochemicals. Due to the structure and chemical nature of many of the Aloe vera phytochemicals, it is likely that many of its reported medicinal properties are due to anti-oxidant or pro-oxidant effects. The anti-oxidant/pro-oxidant activities of many of Aloe vera’s phytochemicals is dependent not only on their individual levels, but also on the ratios of various components, and on their individual redox states. Therefore, discrepancies between bioactivity studies are likely when using different crude mixtures. The potential differences between these crude mixtures need to be taken into account when analysing the reproducibility and efficacy of bioassays of crude extracts.

KEY WORDS: Aloe barbadensis Miller, Aloe vera, anti-oxidant, pro-oxidant, medicinal plant, crude extracts.

Review Article

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Cock: Problems of Reproducibility and Efficacy of Bioassays

species may also produce different levels of other bioactive components or other constituents which enhance/counteract their medicinal activities.[8] Therefore, the bioactivity of crude extracts may be reliant on the conditions in which the plant grows, the season, and the individual plant itself. Other contributing factors may even include induced chemical defences against predators or pathogens. The extraction procedure, treatment and handling of crude plant extracts may also affect the condition and therefore the bioactivity/efficacy of the phytochemical components.

Most plant extracts contain a complex mixture of terpenes, phenolic compounds and alkaloids, many of which can undergo oxidation/reduction processes.[6] The alteration of the redox state may change the behaviour of phytochemicals. Indeed, the maintenance of cellular redox state has been associated with the treatment and prevention of many diseases and ailments including atherosclerosis, inflammatory injury and cancer,[9,10] cardiovascular disease[11] and neurological degenerative disorders.[12] Redox control is also linked with diabetes/anti-diabetic bioactivities[13] and has been associated with the reduction of obesity.[14] Anti-oxidants can directly scavenge free radicals, protecting cells against oxidative stress related damage to proteins, lipids and nucleic acids.[15] The following discussion will examine some problems associated with reproducibility and efficacy of using crude extracts in bioassays, with reference to the well characterised medicinal plant, Aloe vera.

VARIABIlIty In BIoACtIVIty And EffICACy of CRUdE AloE VERA ExtRACts

Aloe barbadensis Miller (commonly known as Aloe vera) has a long history of usage as a food, cosmetic and as a medicinal agent. Amongst its noted therapeutic activities, Aloe vera has been reported to have anti-bacterial,[16,17] anti-fungal,[16] anti-viral,[18,19] immune-stimulatory,[20] anti-inflammatory[21,22] and

anti-diabetes[23] bioactivities. However, many studies examining the therapeutic potential of Aloe extracts report conflicting results, showing either a lack of therapeutic bioactivity for some Aloe species,[24] or even toxicity associated with some Aloe vera preparations.[25-27]

It is well known that plant age is an important determinant of Aloe vera bioactivities. With respect to anti-oxidant potential, the bioactivity has been shown to fluctuate within a given cultivar in relation to the age of the plant, with highest anti-oxidant levels reported for 3 year old plants.[28] This is complicated further as the relative levels of a plants anti-oxidant phytochemicals also fluctuates seasonally.[29] Furthermore, the phytochemical profiles of individual plants will change, dependent on a variety of other environmental and growth conditions.[6,7]

Plants may produce a wide variety of secondary metabolites which have no apparent role in primary plant growth or development processes. These molecules are often unique to plants from a single species and increase during times of high stress such as drought, fire and bacterial infection.[6] Therefore, whilst Aloe vera plant growth may be optimal during times of good growth conditions, it is likely that the level of useful bioactive phytochemicals will be elevated in conditions which stress the plant. Many of these secondary metabolites may exhibit anti-microbial, anti-oxidant, cytotoxic and other medicinally useful properties.[6]

A. bArbAdensis PhytoChEmIstRy

AnthraquinonesMany bioactive phytochemical components have been isolated from Aloe vera leaves and their bioactivities extensively examined. In particular, the anthraquinones, anthrones and chromones have been particularly well studied and have been shown to be effective at counteracting various disease states.[21,30] The anthraquinones aloe emodin (Figure 1a) and aloin (Figure 1b)

Figure 1: Chemical structures for the anthraquinones (a) aloe emodin and (b) aloin, the chromone (c) aloesin, (d) anthrone, (e) cinnamic acid and(f) β-sitosterol.

54


acid derivatives also have concentration dependent anti-oxidant/pro-oxidant activities. Cinnamic acid derivatives behave as anti-oxidants at lower concentrations, but convert to pro-oxidants at concentrations above 5 µM.[44]

In contrast, Yen et al. (2000) demonstrated that the chemical structure of anthrone (Figure 1d) predisposes it to function as an electron acceptor (electrophile), hence as a strong anti-oxidant, independent of its concentration within an extract.[31] It therefore remains possible that Aloe vera extracts with high concentrations of anthrone may maintain anti-oxidant potential, even under conditions which would otherwise predispose the extract to function as a pro-oxidant. For example, Aloe vera extracts containing high aloe emodin and low aloin concentrations (both of which favour pro-oxidant bioactivity) may still function as an anti-oxidant if high enough levels of anthrone are present to maintain the redox state of these anthraquinones. Conversely, low levels of anthrone may predispose an extract to display pro-oxidant activities. It is therefore likely that the redox character of an extract is not only dependent on the levels of the different phytochemicals present, but also on the ratios of several important components within the mixture.

Aloe vera leaves also contains a number of other medicinally important phytochemicals including β-sitosterol (Figure 1f) and β-sitosterol glucosides. These phytosterols have been shown to promote arterial endothelial cell proliferation.[45] They also promote the expression of proteins involved in angiogenesis and thus have potential applications in the management of chronic wounds. Recently, β-sitosterol has also been trialled for the treatment of breast cancer[46] and diabetes,[47] although the efficacy is still under investigation. It appears that these therapeutic bioactivities may be due, at least in part, to their redox state of the molecule. A recent study has indicated that β-sitosterol treatment results in glutathione reduction as well as maintaining the anti-oxidant enzymes superoxide dismutase and glutathione peroxidise in a reduced state.[48] This bioactivity in turn is related to the redox state of the sterol. Interactions between the various components within the crude extracts may also play a role in converting otherwise anti-oxidant molecules into pro-oxidants in the extract or vice versa.

Non-phenolic componentsOther phytochemical components of Aloe vera leaf extracts include acemannan (Figure 2), a long chain polymer of β (1→4) linked galactomannan saccharides.[49,50] Acemannan has been reported to accelerate wound healing,[51-54] activate macrophages[55,56] and have synergistic anti-viral activity in combination with azidothymidine and acyclovir.[19] It has been reported that acemannan also has anti-oxidant properties and that these properties may be responsible for its therapeutic activities.[57] Furthermore, the anti-oxidant potential of Aloe vera polysaccharides is dependent upon the concentration of the molecule and the degree of acetylation of the monomeric units. [58] High polysaccharide concentrations (>8 mg mL-1) were found

are thought to exert their reported therapeutic potentials via an anti-oxidant mechanism. For example, aloe emodin has high inhibitory free radical scavenging activity and has been shown to act as an anti-oxidant by inhibiting lipid peroxidation.[31]

Interestingly, aloe emodin and aloin have been shown to be capable of behaving as either an anti-oxidant or as a pro-oxidant, with their action being dependent upon their concentration.[32] Aloe emodin exerts anti-oxidant behaviour at lower concentrations, yet acts as a pro-oxidant at high concentrations. In contrast, aloin has an anti-oxidant effect at higher concentrations, yet a pro-oxidant effect at low concentrations. Thus, the variable effects reported for crude Aloe vera extracts in various studies may be due to differing level of aloe emodin and/or aloin present in the extract.

Other phenolic Aloe vera constituentsSimilar pro-oxidant effects have been reported for other anti-oxidant phytochemicals including flavonoids,[33] tannins[34] and curcumin.[35] Previous studies have shown that transition metal ions, such as copper or iron, can enhance the conversion of the anti-oxidant to the pro-oxidant state.[36,37] The pro-oxidant/anti-oxidant effect of plant extracts is due to a balance between the free radical scavenging activities and reducing power of their phytochemical components. This can be explained using the anti-oxidant vitamin ascorbic acid as an example. Although ascorbic acid has well characterised anti-oxidant bioactivities, it is also known to act as a pro-oxidant at high concentrations.[38] This is due to the greater reducing power of ascorbic acid compared to its free radical scavenging activity. In the presence of transition metal ions, ascorbic acid will function as a reducing agent, reducing the metal ions. In the process, it is converted to a pro-oxidant. Therefore, high dietary intake of ascorbic acid in individuals with high iron levels (e.g. premature infants) may result in unexpected negative health effects due to the induction of oxidative damage to susceptible biomolecules.[39-41]

The anti-oxidant activity of aloesin (Figure 1c) and other chromones has also been extensively described.[42,43] In contrast, a literature search did not reveal any studies examining the potential pro-oxidant activity of these compounds. One study reported several chromones to have higher reducing power than ascorbic acid.[42] The relatively high reducing power of ascorbic acid is believed to be responsible for its ability to function as a pro-oxidant.[38] It is therefore possible that aloesin and other Aloe vera chromones may have a similar anti-oxidant/pro-oxidant profile to ascorbic acid (i.e. anti-oxidant activity at lower concentrations and pro-oxidant activity at higher concentrations). However, it must be emphasised that this possibility is based on the reported higher reducing power of the chromones compared to their free radical scavenging activity[42] and has not been adequately tested.

Cinnamic acid (Figure 1e) and its derivatives are phenolic molecules which are present in many fruits, vegetables and whole grains, as well as in Aloe vera leaves. Studies indicate that cinnamic

55


problems with reproducibility when analysing crude extracts by bioassay due to differences in the levels of specific phytochemicals, their redox state, and their ratio to other components.

Anti-inflammatory ActivityInflammation is a complex response by the body to injury. It typically follows a variety of insults including burns, wounds, bites and stings etc. It is characterised by a wide variety of symptoms[59] including:

• Swelling. Injury may result in increased capillary permeability which allows leukocyte migration and fluid accumulation in the damaged tissue. This accumulation results in the swelling characteristic of inflammation.

• Redness and heat are caused by vasodilation, reducing blood pressure and increasing circulation.

• Pain is a complex reaction resulting from the release of short peptides and prostaglandins.

These inflammatory processes require the cellular release of several classes of molecules. Vasoactive substances (e.g. bradykinin, prostaglandins and vasoactive amines) are required to dilate blood vessels, opening junctions between cells to allow leukocytes to pass through capillaries. Any compound capable of blocking these vasoactive substances would potentially have a therapeutic effect on the symptoms of inflammation. β-sitosterol is the most abundant phytosterol in Aloe vera extracts. β-sitosterol stimulates smooth muscle cells to release of prostacyclin (PGI2).

[60] However, β-sitosterol treatment blocks the release of PGI2 and prostaglandin E2 (PGE2) from macrophages.[60] Thus, β-sitosterol treatment would be expected to affect vasodilation and, therefore, have a therapeutic effect on inflammation. The Aloe vera leaf chromone aloesin, and its derivatives, inhibit cyclooxygenase-2 and thromboxane A2 synthesis through their anti-oxidant activities. [61,62] Thus, Aloe vera chromones produce anti-inflammatory effects. In contrast, anthraquinones have been shown to stimulate PGE2 release[63] and would, therefore, be expected to promote pro-inflammatory activity.

to be necessary for Aloe vera polysaccharides to display anti-oxidant activity. The same study also showed that increased acetylation enhances the anti-oxidant activity of Aloe vera polysaccharides.[58] However, the polysaccharide components within Aloe vera leaves are not constant. Instead, the composition and concentration of the polysaccharides fluctuate with changes in the growing environment and conditions.

Aloe vera leaves also contain inorganic minerals in variable concentrations (e.g. calcium, magnesium, zinc, iron and copper). As previously discussed, the redox state of many Aloe vera phytochemicals is affected by the presence of metal ions, converting otherwise anti-oxidant components into pro-oxidants. Thus, Aloes growing in soil containing elevated levels of metallic ions would be expected to have higher concentrations of metal ions, and thus tend towards pro-oxidant rather than anti-oxidant bioactivities. Other molecules (such as vitamins, amino acids and proteins) may also have an effect on the redox state of the phytochemical components.

mEdICInAl EffECts of AloE VERA REqUIRIng mUltIPlE PhytoChEmICAls

The multitude of phytochemicals present in an Aloe vera crude extract not only affect each others redox state and ability to function as an anti-oxidant/pro-oxidant, but several phytochemicals may also be required for different aspects of the same therapeutic effect. Some of the medicinal properties associated with plant extracts require the concerted action of several bioactivities. The following discussion examines several therapeutic properties of Aloe vera extracts that require the synergistic action of several bioactivities, each of which may be reliant on multiple phytochemicals. This is by no means a complete examination of the therapeutic properties of Aloe vera extracts, but instead serves to illustrate the difficulties of assigning a therapeutic effect to a single component. Similarly, it further illustrates the

Figure 2: The structure of acemannan (a major polysaccharide component of Aloe vera leaves) consists of a polymer of β (1→4) linked galactomannan sugars.

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Aloe vera leaf extracts have been previously shown to display good anti-bacterial[16,74,75] and anti-fungal bioactivities.[16,76]

Early anti-bacterial studies of Aloe vera extracts have provided confounding and even contradictory results. Some of these studies indicate that the bioactive agent(s) are anthraquinones,[77,78] whilst other studies found Aloe vera anthraquinones to be inactive as anti-bacterial agents.[79] Numerous subsequent studies have demonstrated the anti-bacterial activity of isolated anthraquinones from Aloes[80,81] and various other plant species.[82-84] Whilst the mechanism of anti-bacterial activity is still subject to investigation, it has been suggested that aloe emodin and aloesin function by inducing bacterial membrane disruption.[80] This study also determined that the form of aloe emodin and aloesin tested also affects their anti-bacterial activity. It was demonstrated that anthraquinone loaded liposomes had strong anti-bacterial activity, whilst the purified free anthraquinones did not. It is, therefore, possible that some of the observed differences in the anti-bacterial activities of anthraquinones and Aloe vera extracts may be due to the form of anthraquinones that the bacteria were tested against. Whilst this study showed that anti-bacterial activity is dependent on the form of anthraquinone tested, the effect of concentration was not extensively examined. MIC values were determined by testing across a range of concentrations, although only relatively low concentrations were tested. It is possible that higher concentrations may have a very different effect, analogous to the concentration effects already described for anthraquinone anti-oxidant/pro-oxidant activity.

Other Aloe vera components have also been implicated in the antibacterial activity of leaf extracts. A recent study tested anthraquinone free leaf extracts and isolated components.[85] This study showed that cinnamic acid, coumaric acid, ascorbic acid and pyrocatechol purified from Aloe vera gel all display good anti-bacterial activity, especially towards Gram-positive bacteria. It was postulated that the phenolic anti-bacterial agents functioned by disrupting bacterial cell membranes, as well as by denaturing bacterial proteins. Furthermore, cinnamic acid is known to block bacterial glucose uptake and ATP production,[86] therefore, inhibiting bacterial growth. Coumaric acid has been shown to inhibit bacterial enzymatic activity.[87] A number of other phenolic components were also found to have low to moderate anti-bacterial activity.

In addition to direct inhibitory effects on bacteria, Aloe vera components may also function by selectively modulating the cells of the immune system (described in detail in section 4.4). Furthermore, acemannan also inhibits bacteria adhering to epithelial cells and establishing an infection.[88] It is likely that the anti-bacterial activity of Aloe vera extracts in vivo is due to the synergistic effects of multiple bioactive components, functioning through several mechanisms.

Anti-fungal activity has received less attention, although some studies have demonstrated the ability of Aloe vera extracts to

The peptidase bradykinase has been isolated from Aloe vera leaves and has been shown to break down the vasoactive peptide bradykinin.[64,65] As bradykinin treatment results in vasodilation, hydrolysing this protein would result in decreased vasodilation and, therefore, inhibit leukocyte passage and fluid leakage from the capillaries into the surrounding tissue. Aloe vera leaf bradykinase would, therefore, be expected to contribute to the therapeutic effects on the symptoms of inflammation.

Chemotactic factors, including several proteins and peptides, are required to increase cell motility, especially the motility of leukocytes during inflammation. Blocking these chemotactic factors, or blocking their effects, prevents inflammatory swelling. Several compounds in Aloe vera extracts have been shown to be capable of blocking chemotaxis. Anthraquinones suppress cytolytic T-lymphocytes in favour of suppressor cells.[66,67] Furthermore, anthraquinones decrease cytokine production and IL-2 mRNA expression in activated T lymphocytes,[68] thereby decreasing chemotaxis. More recent studies have demonstrated that the anthraquinone emodin decreases plasma levels of the cytokines IL-2 and TNF-α, whilst increasing IL-10 (which itself down-regulates IL-2 and TNF-α cytokine activity).[69] None of these studies, however, examined the relationship of the redox state of the anthraquinones with these effects. Furthermore, these studies have not rigorously examined the effects of a range of doses of these phytochemicals.

In contrast, Aloe vera polysaccharides (including acemannan) have a stimulatory effect on chemotaxis. Acemannan exposure stimulates cytokine production and activates lymphocytes.[70,71] Specifically, pure acemannan isolated from Aloe vera leaves has been shown to stimulate macrophages to release IL-1, IL-6, interferon, GM-CSF and TNF-α in vitro.[72] Similarly, Aloe vera lectins stimulate cytokine production. Aloctin A, the best characterised of the Aloe lectins, has been shown to stimulate the production of IL-2[73] and to enhance the production and activation of macrophages.[73] Therefore, Aloe vera extracts contain both chemotactic stimulatory and inhibitory compounds. The chemotactic effect of Aloe vera extracts would, therefore, be dependent on the levels and ratios of the factors affecting chemotaxis as well as their redox state.

Aloe vera extracts contain multiple active phytochemicals. It is likely that several of these may be required to address different aspects of the inflammatory process. Failure to consider this is likely to be responsible for past ambiguities about the efficacy of Aloe extracts in relation to its anti-inflammatory activity.

Antiseptic activityThe interruption of the external epidermal barrier by a wound, burn or other such event allows microbes to enter and infect the wound. The invasion of microorganisms may cause or intensify inflammation (described in section 4.1.) and may hinder wound healing (described in section 4.3) and/or cause disease.

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Other Aloe vera phenolic compounds have also been implicated in the wound healing effects of Aloe vera extracts. β -sitosterol and β-sitosterol glucosides promote endothelial cell proliferation and angiogenesis,[45] although their activity appears to be dependent on its redox state.[48] The reduced sterol has anti-oxidant activity and stimulates wound healing processes, whilst oxidised sterols are pro-oxidants and induce cell death. β -sitosterol and β-sitosterol glucosides, therefore, have potential applications in wound management in their reduced state. The Aloe vera chromone aloesin has also been reported to stimulate cellular proliferation. [51,61,99] It is possible that the proliferative effect of aloesin is due to its high anti-oxidant activity.[42,43] In contrast, cinnamic acid has been shown to down-regulate expression of cell proliferation and anti-apoptotic gene products, although the affects of both high and low concentrations were not examined. [100,101]

The redox environment affects cellular signal transduction, DNA and RNA synthesis, protein synthesis, enzyme activation, regulation of the cell cycle, ligand binding, DNA binding and nuclear translocation, and therefore ultimately cell proliferation/death.[102,103] Transcription factors are active in their reduced form and their translocation to the nucleus is also redox dependent. [104] A reducing environment favours cellular proliferation whilst an oxidising environment results in an increase in reactive oxygen species, initiating cell death.[105,106] Therefore, extract conditions favouring anti-oxidant activity (e.g. low aloe emodin, high aloin, low cinnamic acid, low ascorbic acid, low transition metal and high anthrone concentrations) would be expected to favour cellular proliferation whilst conditions favouring pro-oxidant activity (e.g. high aloe emodin, low aloin, high cinnamic acid, high ascorbic acid, high transition metal and low anthrone concentrations) would favour cell death.

The non-phenolic components, particularly acemannan, have also been shown to have a role in wound healing. For example, the stimulation of gingival fibroblast proliferation has been demonstrated when treating oral wounds with high doses of acemannan.[107] This stimulatory effect was found to be due to an induction in expression of the growth factors KGF-1, VEGF and an increase in collagen expression. This study only examined the effects of relatively high concentrations of acemannan, in the range that would correlate to anti-oxidant activity. As lower concentrations may correlate to pro-oxidant activities, it is possible that the induction of fibroblast proliferation may not be seen at these concentrations. Indeed, as lower concentrations of acemannan correspond to pro-oxidant effects, it is possible that at lower concentrations, cell death may be induced. The concentration dependent redox effect of acemannan may also contribute to the discrepancies seen between proliferative studies of Aloe vera extracts.

As well as requiring cellular growth and proliferation, wound healing also requires matrix remodelling. Aloe vera gel extracts have been shown to stimulate and speed up the production of hyaluronic acid and dermatan sulphate.[52] Activities of the enzymes

inhibit fungal growth.[16,76,89] Anthraquinones, especially aloe emodin and aloesin, were implicated in this anti-fungal activity,[76] however, the identity of anti-fungal components and their mechanisms of action have not been extensively examined. Similarly, the anti-viral activity of Aloe vera leaf extracts has been demonstrated,[18,90] although detailed purification, identification and mechanistic studies are required.

Wound HealingWhilst anti-inflammatory and anti-microbial bioactivities are complex processes requiring the synergistic action of several bioactivities, wound healing is more so. Wound healing, a relatively well studied therapeutic property of Aloe vera, is the result of several bioactivities including:

• Inflammation, which has summarised in section 4.1.• Antiseptic bioactivity, which has summarised in section 4.2.• Cell growth and proliferation• Matrix remodelling

The growth of endothelial, epithelial and fibroblast cells are critical in wound healing. As a first step in wound healing, a fibrin clot is formed as a temporary repair. This step is vital as it helps avoid microbial infection which may retard the healing process. The wound is subsequently invaded by a variety of cell types, some of which stimulate an inflammatory response, and others which are directly involved in the repair mechanism.[91] The effects of Aloe vera extract components on inflammation processes and chemotaxis have already been summarised in section 4.1. Wound repair itself occurs in three phases: the migration of epithelial cells and fibroblasts to the wound site, proliferation of cells and cellular maturation. It is likely that the wound healing effect of Aloe vera extracts involves the synergistic action of multiple components on several pathways.

Aloe vera anthraquinones reportedly possess contradictory effects on cell growth and proliferation. For instance, Aloe emodin has been shown to stimulate a 2.5 fold increase in rat hepatocyte DNA synthesis with a corresponding increase in cell growth. [92] Additionally, aloe emodin has been shown to protect hepatocytes from apoptosis.[69] In contrast, other studies have shown aloe emodin to induce apoptosis in pro-myeloleukemic HL-60 cells[93] and human lung squamous cell carcinoma,[94,95] and to inhibit human neuroectodermal tumour growth.[96] Some studies have postulated that the pro-apoptotic effect of aloe emodin is due to an induction of caspase 3 activity, together with a decrease in the levels of the anti-apoptotic protein Mcl-1. [93] Another study has implicated caspase 8 mediated cleavage in the apoptotic activity of emodin.[97] Studies into the pro-apoptotic mechanism of aloe emodin are ongoing. Similarly, anthrones have also been shown to induce cell death. In a recent study, an anthrone from the Ethiopian medicinal plant Kniphofia foliosa was shown to induce rapid death in mouse and human cancer cells via necrosis. [98]

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cells.[114] Interestingly, this study found the Aloe gel extract lacks this activity at either higher or lower concentrations, indicating a concentration dependence similar to that reported for the redox effects of Aloe vera components.[32] It is, therefore, possible that the variable immune-modulatory effects reported for Aloe vera extracts in different studies may be due to the concentrations, ratios and redox states of several important compounds in the tested extracts, with extract conditions favouring anti-oxidant bioactivity resulting in immune-stimulation. Conversely, conditions favouring pro-oxidant activity would be expected to result in immune-suppression, although this has not been extensively tested.

Anti-Diabetic activityDiabetes mellitus refers to a group of metabolic disorders that result in increased blood glucose concentrations, either because the pancreas does not produce enough functional insulin (type 1 diabetes), or because cells do not respond to the insulin which is produced (type 2 diabetes). The causes of diabetes mellitus include the auto-immune destruction of pancreatic cells,[115] viral infections,[116] genetic and environmental factors,[117] insulin or insulin receptor gene mutations[118] and altered pancreatic prostaglandin metabolism.[119] Diabetes has significant health effects, impacting on the quality of life and life expectancy of those suffering with it.

A number of studies have indicated the beneficial effects of Aloe vera extracts in diabetic patients.[23,120] Administration of Aloe vera extracts to streptozotocin-induced diabetic rats resulted in a decrease in blood glucose and a corresponding increase in liver glycogen.[120] The maintenance of glucose homeostasis by Aloe vera extracts in diabetic rats was shown to involve a number of mechanisms. Aloe vera extract treatment altered the activities of multiple enzymes: glycogen phosphorylase activity was decreased and glycogen synthetase increased, resulting in increased hepatic glycogen stores.[120] Hexokinase activity and mRNA levels were decreased in diabetic rats,[121] yet treatment with Aloe vera extract returned these parameters towards normal levels.[120] Similarly, increased lactate dehydrogenase, glucose-6-phosphatase and fructose-1, 6-bisphosphatase activities were seen in diabetic rats.[122] Aloe vera extract treatment significantly restored these enzyme activities.[120]

Glycosylation of blood proteins including haemoglobin, albumin and lipoproteins is also characteristic of diabetes mellitus.[123] Under the hyperglycaemic conditions of diabetes mellitus, blood glucose interacts with specific amino acids on the proteins surface, forming glycosylated protein products which may undergo a series of further chemical modifications resulting in the production of advanced glycation end products (AGE).[124] The binding of AGEs to their receptors results in altered cell signalling which in turn results in free radical production.[125] Indeed, diabetes mellitus has been shown experimentally to be associated with an increase in free radical formation and an associated decrease in anti-oxidant potential.[126,127] Studies have directly linked oxidative stress with

β-glucuronidase and N-acetyl glucosaminidase are increased during wound healing, resulting in increased carbohydrate turnover at the site of the wound. Other studies also demonstrated that wounded diabetic rats treated with Aloe vera gel show increased collagen formation[53] and cross linking.[54] It is evident that a synergistic action is required by several Aloe vera extract components on multiple wound healing associated bioactivities. The reported discrepancies between different studies may be due to differences in concentrations, ratios and redox states of these components.

ImmunomodulationManipulation of the immune system has therapeutic potential in the treatment of a variety of diseases. Aloe vera leaf extracts have been reported to have both good immuno-stimulatory[20] and immune-suppressive activities (as reviewed in Boudreay and Beland[108]); however, rigorous scientific examination of these effects is limited. Much of the studies into the immune-modulatory potential of Aloe vera extracts have focused on the immune-stimulatory effects, particularly of the polysaccharide components. Whilst numerous Aloe vera polysaccharide components have been shown to have immune-modulatory effects,[109-111] acemannan has been particularly well studied. The immune-modulatory effects of acemannan are thought to be due to activation of macrophage cells and antigen processing. The activated macrophages secrete cytokines including IL-1, IL-6, interferon, GM-CSF and TNF-α in vitro.[72] The release of these cytokines is itself associated with further pathology through the induction of inflammation. Acemannan also enhances macrophage sensitivity to IFN-γ, inducing apoptosis.[20] Neither acemannan nor IFN-γ was capable of inducing apoptosis alone. Instead, a synergistic effect is required and this effect appears to function through the inhibition of the expression of Bcl-2 proteins.[20]

Studies have also highlighted the immune-modulatory properties of the smaller phenolic components of Aloe vera leaves. Aloe emodin and other anthraquinone derivatives have been shown to have an immune-suppressive effect by blocking lymphocyte proliferation.[66,67] Emodin also reduced IL-1, IL-2 and IL-2 receptor expression.[66] It was suggested that emodin suppresses both macrophages and lymphocytes. Further studies have identified 37 other anthraquinones with the ability to block cytolytic T lymphocyte induction and the ability to prevent antibody production.[67] The effect of concentration and the ratio between anthraquinones were not tested in these studies.

It has been postulated that Aloe vera extracts may exert immune-modulatory effects through their functioning as anti-oxidants, inhibiting/stimulating the production of free radicals.[28] Treating streptozotocin induced diabetic[112] or gamma-irradiated rats[113] with Aloe vera leaf extracts reduces lipid peroxidation and the formation of hydroperoxides whilst increasing the levels of anti-oxidant enzymes (e.g. reduced glutathione, glutathione peroxidise, glutathione-S-transferase, catalase, superoxide dismutase) in the liver, lungs and kidney. Similarly, Aloe vera gel has been shown to inhibit ROS production in colorectal mucosa

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that average tumour weights increased in severe combined immune-deficient (SCID) mouse tumour xenografts from cells over expressing catalase or thioredoxin.[137] Tumours from both transfectants contained fewer apoptotic cells but mitotic cell numbers were similar. This suggested that anti-oxidant over expression resulted in increased tumour size due to a decrease in apoptosis.

The cell proliferation/apoptosis inducing abilities of Aloe vera extracts and isolated components have been described in Section 4.3. Briefly, ROS based tumour therapy may induce regression in apoptosis/oxidatant sensitive tumour cells. Thus, if Aloe vera components were present in concentrations and ratios consistent with pro-oxidant activity, the extract would induce apoptosis and, therefore, would have anti-cancer activity. If the levels of components were consistent with a reducing environment, anti-oxidant activity would result and the extract would not have anti-cancer activity. Conversely, should the protocol be repeated on a tumour with apoptotic resistant/oxidant resistant cells, the converse would apply and tumour progression would be likely.

ConClUsIons

The problems associated with reproducibility and efficacy of bioassays using Aloe vera juice and/or crude extracts illustrates some of the difficulties encountered in natural products research. Individual extract batches may vary widely with regards to individual phytochemical profiles, ratios between various components, and the redox state of these components. These variances may have profound effects on the reported bioactivities and are likely to account for the reported discrepancies between different studies bioassaying crude mixtures. Despite these difficulties, the use of crude extracts is often necessary as the individual components often do not show the same bioactivities, or have different efficacy to crude extracts. This is true for Aloe vera. Aloe vera juice, or Aloe vera crude extracts, often display higher efficacy than the purified components. It is likely that the biological activity of Aloe vera is a synergistic and perhaps additive action of the different classes of compounds found within the plant, rather than a single constituent or just a few compounds. Furthermore, these compounds are required in the correct levels/ratios/redox states for bioactivity to be observed.

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the impaired maintenance of glucose homeostasis and the enhanced lipid peroxidation seen in diabetes mellitus.[127] Furthermore, increased total anti-oxidant levels have been measured in the blood and saliva of diabetic patients, further supporting the proposed role of oxidative stress in diabetes mellitus.[128]

Oxidative stress induction has also been suggested to be the common link between the diverse medical complications (including cardiovascular disease, renal and neural degeneration, impaired vision and erectile dysfunction) seen in diabetes mellitus. [129,130] Therefore, treatment with anti-oxidants would be expected to counteract many of these complications. Aloe vera has a number of compounds (both phenolics and non-phenolic compounds) that can act as anti-oxidants (as described in section 3. - A. barbadensis phytochemistry). As many of these compounds can potentially behave as either anti-oxidant or pro-oxidant dependant on their concentration, redox state and ratio between compounds, it is not surprising that studies using Aloe vera crude extracts to treat diabetes mellitus have had mixed success.

Anti- Cancer activityThe growth and development of healthy cells depends on fine regulation of growth promoting and inhibiting pathways. Proto-oncogenes and tumour suppressor genes are responsible for encoding proteins that regulate cell division/cell cycle, as well as for the repair of damaged DNA and cell programmed death by apoptosis. Mutations within these genes have been implicated in the onset of cancer.[131] Such mutations result in cells that no longer require external signals to proliferate. Furthermore, these cells fail to recognise signals that restrict cell division, resulting in uncontrolled cell growth. In tumour genesis, multiple genes may be altered and transmitted to daughter cells, which subsequently escape normal growth restraints and form a tumour, which may be benign or malignant.

The induction of oxidative stress has been linked with several types of cancer.[132,133] Chromosome instability is also a common feature of many of the cancers that have been linked with oxidative stress, suggesting that increased oxidative stress may contribute to development of genetic instability. Oxidative stress leading to genetic instability may result in the emergence of new tumour phenotypes. In such populations, a decrease in apoptosis but an increase in tumour growth and subsequent tumour progression is observable.

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Research Article




*Correspondence: [email protected]; +233 204620000DOI: 10.5530/pc.2011.1.4

Cassane-type diterpenoids from the genus CaesalpiniaR. A. Dickson1*, T. C. Fleischer1, P. J. Houghton2

1Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical, KNUST Kumasi, Ghana. 2Pharmacognosy Research Laboratories, Pharmaceutical Sciences Research Division, King’s College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK

INTRODUCTION

Caesalpinia is the name of a genus of enormous size and of ancient origin. It is named after the Italian naturalist, Andreas Caesalpino, of Arezzo (1519-1603). He was also a botanical collector, systematist and philosopher, chief physician to Pope Clement VIII, and a professor of medicine and botany in Pisa and Rome.[1] Caesalpinia consists of about 200 species, consisting of shrubs, tall climbers, small and tall trees, mostly armed with spines and curves, hooked, sharp thorns and rarely unarmed. Their leaves are bipinnate, lacy, and attractive, while the leaflets are few to many, opposite, rarely alternate, small or large, herbaceous or leathery. The flowers are yellow, red, or variegated, showy, handsome, medium to large and are multiflowered. Finally, the pods are variable, often prickly, flat, straight or beaked.[2,3]

THE CAESALPINACEAE FAMILY

The leguminous trees fall under the sub-families Caesalpinaceae, Fabiaceae and Mimosaceae. The trees of Caesalpinaceae are by far the most scenic, exhibiting many-coloured splendour. The leguminosae family is currently divided into three subfamilies and 36 tribes. Subfamily Caesalpinioideae comprises of four tribes and 2,250 species, subfamily Mimosoideae four tribes and 3,270 species, and subfamily Papilionoideae 28 tribes and 13,800 species.[2,3] Polhill and Vidal,[4] divided the Caesalpinieae

into 8 informal generic groups: the Gleditsia group (2 genera), the Acrocarpus group (monogeneric), the Sclerolobium group (3 genera), the Peltophorum group (13 genera), the Caesalpinia group (16 genera), the Poeppigia and Pterogyne groups (both monogeneric) and the Dimorphandria group (10 genera). These authors stated that the tribe is ‘a remarkable mixture of relics and complexes of relatively recent speciation, providing many pitfalls for formal systematics and biogeographical interpretations. [4] Since 1980, several studies have cast new light on intergeneric relationships within the Caesalpinieae, necessitating the restructuring of some of the nine informal generic groups.[5,6] Without doubt, the genus with the greatest taxonomic and nomenclatural complexity within the Caesalpinieae is the genus Caesalpinia, which in its broadest sense comprises about 140 species and contains 25 generic names in synonymy.[4]

GEOGRAPHICAL DISTRIBUTION OF THE GENUS CAESALPINIA

Members of Caesalpinia are widely distributed throughout the tropics and subtropics, primarily in America and Asia, and extending to Australia, Polynesia Madagascar and Africa[7,8] (Table 1). In Africa it is widespread in the western and southern areas with C. benthamiana being the most common. About 25 species are found in the Caribbean, 10 in Cuba and the Bahamas, 2-3 species in Mexico and a few extending to Central America.[4] C. pulcherrima is found in the Philippines and the Caribbean where it is an ornamental plant. It is red with yellow margins, and this variety is the national flower of Barbados, known as ‘Pride of Barbados’.[9]

ABSTRACT: Medicinal plants belonging to the Caesalpinia (Ceasalpiniaceae) genus are widely distributed in most tropical countries and have been frequently employed in folkloric medicine worldwide in the treatment of various ailments including skin diseases, malaria, cancer, infections, erectile dysfunction, pain and wounds. Interest in this genus has increased considerably over the years and the biological properties of different phytoconstituents, such as the cassane-type diterpenoid isolates, have been studied. Over the past 60 years, a number of cassane-type diterpenoids have been isolated from species of this genus and some of them possess interesting biological activities. Recently, three novel cassane-type diterpenoids, benthaminin 1, 2 and 3, which demonstrate antimicrobial and antioxidant properties, have been isolated from Caesalpinia benthamiana growing in Ghana. This review seeks among other things to collate all these isolated compounds, recognising their diversity and commenting on their relevance as bioactive compounds.

KEY WORDS: Cassane-type diterpenoids, Caesalpinia, Ceasalpiniaceae, Biological activity.

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Dickson, et. al.: Review on C. major

Caesalpinia is also found in Colombia, Ecuador, Peru, Paraguay and Argentina and is very popular in Brazil. Indeed, the name ‘Brazil’, had its origin in the Portuguese word ‘Bresil’ or ‘brazil’ which means bright red and resembling glowing coals, and was used to describe the colour of Caesalpinia wood abundant in this area.[10] Nine species are widespread in Asia with about two confined to China.[11] C. major is widely distributed in Southeast Asia.[12] The genus Caesalpinia is also popular in Thailand and Indonesia[13] and C. minax, is found in China.[14]

Apart from their medicinal importance, Caesalpinia species may also serve as garden ornamentals and hedge plants. The beauty of C. pulcherrima, whose showy red flowers are borne in long spikes, is reflected in its common names: pride of Barbados, paradise flower, Spanish carnation, peacock’s crest, flower tree, and others. This semi-drought-resistant species flowers in favourable habitats when only 8 months old.[10]

ETHNOPHARMACOLOGICAL USES

The roots of C. benthamiana are considered to be an effective dysentery remedy in Ghana.[15] The powdered roots are mixed with shea butter or palm kennel oil to treat skin diseases and wounds.[15] An infusion of the dried root is consumed or used in bathing for general malaise in Senegal.[16]

In the Philippines and the Caribbean, decoctions of the leaves, bark and roots of C. pulcherrima are used traditionally to treat liver disorders, ulcers of the mouth and throat. It also reduces fevers, acts as an abortificient, and alleviates fungal infections. The fruit is also used to check bleeding and prevent diarrhoea and dysentery.[17] The flowers have also been used to combat oxidative stress by eliminating free radicals from the system.[18]

Within southeast Asia, C. major has traditionally been implemented as a tonic, anthelmintic, and for rheumatism and back-ache.[19] In Thailand, the seeds of this plant are used as an expectorant and antitussive agent.[19] C. minax finds use in Chinese folk medicine in the treatment of common colds, fever and dysentery.[20] Also in folk medicine in Kagoshima in Japan, C. decapetala is used in the treatment of neuralgia. A decoction from the pods of C spinosa is used in eye washes in the Callera district of the Czech

Republic.[21] In addition, plant part decoctions are employed in folk medicine to treat intermittent fever and as an abortifacient, emmenagogue, and as a general tonic.

Bonducin, an amorphous, white bitter glycoside, is abundant in the seed cotyledons of C. bonduc Roxb., C. bonducella Flem., and C. crista L.[22,23] It is sometimes referred to as “poor man’s. quinine” because it is used as a substitute for quinine in the treatment of intermittent fever. The seeds of C. bonducella are grey, round, smooth and stony. The buoyancy of the seed accounts, in part, for this species being widely dispersed tropically by ocean currents. They are used as talismans and beads, and also by children as marbles. They yield oils for cosmetics and use in medical preparations. It is a shrubby tree of Argentina and Chile, exuding a golden yellow gum that contains approximately 80% arabin. It is completely soluble in water and is an acceptable substitute for gum Arabic.[24] C. echinata Lam. is the national tree of Brazil. The name ‘Brazil’, had its origin in the Portuguese words ‘Bresil’ or ‘brazil’ which means bright red, resembling glowing coals and were used to describe the colour of caesalpinia wood abundant in this area.

BIOLOGICAL ASPECTS OF THE GENUS CAESALPINIA

The genus Caesalpinia (Ceasalpiniaceae) has been associated with a number of biological activities. Plants in this group have been employed globally in folkloric medicine in the treatment of numerous diseases. Various investigations have been carried out on plants belonging to the genus Caesalpinia in order to validate the folkloric uses of these plants to determine its antimicrobial, antimalarial and anticancer properties, among others.

Antimicrobial ActivityExtracts and compounds from the seeds have been screened against pathogenic organisms that include viruses, bacteria and fungi. Cassane furanoditerpenes, designated as caesalmin C-G, were evaluated for their effects on the proliferation of the Para 3 virus. The tetracyclic furanoditerpenoid isolates showed significant activity against the Para 3 virus, with IC50 values ranging between 7.8 and 14.8 µg/mL. However, caesalmin G, which is the only furanoditerpenoid lactone, is highly toxic, with a therapeutic index (TI) value of 3.0.[25] It is noteworthy that the

Table 1: Distribution of Caesalpinia species

Selected species Geographical location References

C. benthamiana (Baill.)Herend. & Zarucchi

W. Africa Irvine (1961), Burkill (1994)

C. brevifolia Baill. France Naudin (1894)C. coriaria (Jacq.) Willd. Philippines Banados & Fernandez (1954)C. crista L. Hawaii, USA Allen & Allen (1936b)C. decapetala (Roth) Alst. Zimbabwe

S. AfricaCorby (1974) Grobbelaar & Clarke (1974)

C. japonica S. & Z. Japan Asai (1944)C. percherrima (L.) Sw. Philippines Banados & Fernandez (1954)C. sappan L. Hawaii, USA Allen & Allen (1936b)C. spinosa (Mol.) Ktze. S. Africa Grobbelaar & Clarke (1974)

65


abdominal cramps.[29] In the Philippines, decoctions of the leaves, bark and roots are used to manage liver bleeding and prevent diarrhoea and dysentery, whilst the flowers are utilized to combat oxidative stress.[30] These plants are used in the treatment of common cold, fever and dysentery in China.[31] In South Sulawesi of Indonesia, the seed kernel of the plant has been traditionally used as an anthelminthic and antimalarial.[32] The seeds of plants belonging to this genus are used as expectorant and antitussive agents in the herbal medicine practice of Thailand.[33] Their usefulness in the treatment of rheumatism and back-ache and as a tonic has also been reported in Indonesia.[34,35] Antiviral and anticancer activities from these plants have also been reported. [36,37] In Caribbean folk medicine, plants of this genus have been employed extensively.[38] Medicinal plants belonging to this group have been used in traditional medicine in the management of diseases in African countries including Senegal, Nigeria, Sudan and Liberia.[39] Ghana is not an exception in this regard as plants belonging to the genus are extensively employed in folkloric practice in the treatment of various ailments such as skin diseases and wounds, gonorrhoea, sleeping sickness and constipation.[40] The taxonomy of the family Ceasalpiniaceae has been the subject of much debate. It was previously referred to as Fabaceae, and prior to this was known as Leguminosae.

Antimalarial ActivityForty four cassane-and norcassane-type diterpenes isolated from Caesalpinia crista of Myammar and Indonesia were evaluated for their antimalarial activity against the malaria parasite Plasmodium falciparum (FCR – 3/A2 clone in vitro). Most of the tested diterpenes displayed antimalarial activity, and norcaesalpinin E showed the most potent activity with an IC50 value of 0.090µM, a greater potency than the clinically used drug chloroquine (IC50, 0.29µM).[41]

Ten new cassane diterpenes including caesalpinins H-P and norcaesalpinin F were tested for their inhibitory activities on the growth of Plasmodium falciparum (FCR – 3/A2 in vitro. All displayed activity in a dose dependent manner. Among the newly isolated compounds, caesalpinin K and norcaesalpinin F showed the most potent inhibitory activity with an IC50 value of 120 and 140nM respectively, which is lower than the value reported for the well-characterizedantimalarial drug, chloroquine (IC50, 282 – 291nM).[42] Three new cassane furanoditerpenoids (1-3) exhibiting antimalarial activity against the multidrug-resistant K1 strain of Plasmodium falciparum have been isolated from kernels of Caesalpinia bonduc.

Anticancer ActivityCaesaldekarin J possesses inhibitory activity against glutathione S-transferase, an enzyme that has been implicated in resistances during treatment of cancer and parasitic infections, and can be isolated from the ethanolic extract of Caesalpinia bonduc bark.[43] Two new cassane butenolides, caesalpinolide A (1) and B (2), epimeric at the hemiketal position, were isolated from the marine

therapeutic index (TI) value of caesalmin C is almost the same as that of ribavirin (an inhibitor of DNA and RNA viruses), which serves as a positive control in the bioassay. It can be concluded that the anti-Para3 virus activity of tetracyclic furanoditerpenoids is better than that of the furanditerpenoid lactone.[25] Since the major components of the seed of C. minax possess such potent activity, it may be feasible to develop a new antiviral agent from this medicinal plant.

Furthermore, macrocaesalmin, a cassane furanoditerpenoid lactone from the seeds of C. minax, was evaluated for antiviral activities against RSV, Para-3 and influenza Type A viruses according to an established protocol which showed inhibitory activity against the RSV (IC50 = 24.2µg/mL, TC 50 = 138.3µg/ mL and SI = 5.7) in cell culture, and the corresponding values for the positive control (ribarivin) were 3.4, 60.6 and 17.8µg/mL, respectively. The antiviral activity of the compound was less than the positive control; however, the selectivity index for natural products was considered significant (SI > 4). However, it is inactive against the para-3 virus (IC 50 = 51.9µg/mL, TC 50 = 137.5µg/mL and SI = 2.6) with the corresponding values for the positive control being 2.7 µg/mL, 62.5µg/mL and 23.1µg/ mL, respectively. Similarly, macrocaesalmin was inactive against the influenza Type A virus. Respiratory viral infections have long been recognized as important contributors to morbidity and mortality in young children and older adults, and the search for natural products as antiviral agents against respiratory viruses has attracted considerable attention in recent years. The isolation of macrocaesalmin and the evaluation of its efficacy on three major respiratory pathogens thus provide useful clues in the search for antiviral drugs against RSV infection.[26]

Four new cassane-type furanoditerpenoids possessed antimicrobial activities against several bacteria including S. aureus, E. coli, P. aeruginosa and B. subtilis and fungi (C. albicans and T. mentagrophytes have been isolated from the air-dried leaves of C. pulcherrima. A cassane-type diterpene ester, pulcherralpin, isolated from the stems of this plant has potential fertility regulating and antitumor activities.[27]

Two antitubercular cassane furanoditerpenoids, namely 6 β-benzoyl-7 β-hydroxyvouacapen-5 α-ol and 6 β cinnamoyl-7β-hydroxyvouacapen-5 α-ol, have been isolated from the root of Caesalpinia pulcherrima. It was observed that 6β-cinnamoyl-7β-hydroxyvouacapen-5 α-ol possessed stronger antitubercular activity demonstrating a minimum inhibitory concentration (MIC) of 6.25 µg/mL, while the benzoyl analogue 6 β-benzoyl-7 β-hydroxyvouacapen-5 α-ol was less active (MIC 25 µg/mL). Both compounds exhibited moderate cytotoxic activity towards KB (human oral carcinonoid cancer), BC (human breast cancer) and NCl-H187 (small cell lung cancer) cell lines.[28]

According to Mexican folklore, plants belonging to the genus Caesalpinia have found use in the treatment of kidney ache, cystitis, urethritis, prostate inflammation, fever, tooth-ache and

66


plant, two cassane diterpenes neocaesalpin A (18) and B (19) were isolated.[55] From the roots of the same plant in the following year, caesaldekarin F (20) and G (21), were isolated.[56] In the same year, and again from the roots of this plant, seven cassane diterpenoids that included caesaldekarin A (15), H, I, J, K and L (22-26) and demethylcaesaldekarin C (27) were isolated.[57] Additional diterpenoid lactone compounds, caesalmins A, B, C, D, E, F and G (28-34) were isolated from the seeds of C. minax. [58] Four novel diterpenoid compounds (35-38), possessing both antibacterial and antifungal activities, have been isolated from the leaves of C. pulcherrima.[59]

A novel diterpenoid named macrocaesalmin (39), together with caesalmin B, D and H possessing antiviral and anticancer activities were isolated from the seeds of C. minax.[31,60] This was followed in the following year by the isolation of cassane diterpenoid compounds (+)-vouacapenic and (+)- vouacapenate (2, 7) from Vouacapoua americana belonging to the Leguminosae.[61]

Novel norcassane-type diterpenes norcaesalpinin A, B and C (44-48), have also been obtained from the seed kernels of C. crista. [61] In the following year, five new cassane-type diterpenes, caesalpinins MA-ME 1-5 (49-53) and three new norcassane-type diterpenes, norcaesalpinins MA-MC (54-56) together with known cassane-type diterpenes ( 29, 30, 32 and 51) were isolated from C. crista.[62] Nine new cassane-type diterpenes taepeenin A-I

creeper Caesalpinia bonduc. They exhibited inhibitory effects on MCF-7 breast cancer cell lines, with IC50 values of 12.8 and 6.1 (μM), respectively, and also inhibited endometrial and cervical cancer cell lines.[44] Similarly, Phanginin I, a cassane-type diterpenoid isolated from the seeds of Caesalpinia sappan exhibited cytotoxic effects against KB cell line with IC50 value of 4.4μg/ ml. [45]

Basic Molecular Skeleton of Cassane-type DiterpenoidsPlants belonging to the genus Caesalpinia have proven to be a rich source of cassane-type diterpenoids.[46,47-51] These cassane diterpenoids are characterized by a molecular skeleton constructed from the fusion of three cyclohexane rings A, B and C and a furan ring (1) .Ring C may sometimes be aromatic as in compounds 24, 38, 59-62. The existence of an exocyclic methylene group at position 14 is a characteristic of some of these cassane-type diterpenoid compounds (see 62, 76, 81, 83, 88). Generally, these diterpenoids give a red colour with Ehrlich reagent, suggesting the presence of a furan ring in their molecular structure. However, not all cassane-type diterpenes have this furan ring (e.g 5, 6) and therefore will not respond to this test.

Cassane-type diterpenoids from the genus CaesalpiniaThe isolation of the cassane-type diterpenoids may have begun in the mid 1950’s from other sources other than the genus Caesalpinia. However, Jiang et al (2001), isolated pulcherrimin A (3) and ε-caesalpin (4) from Caesalpinia pulcherrima.[52] In 1992, the roots of Caesalpinia decapetala yielded caesaljapin (9), a cassane diterpenoid.[53] Caesaldekarin A (15), C (16), D (16i) and E (16ii) were also isolated from the roots of Caesalpinia major.[54]

Caesalpinin 1 (17), a cassane furanoditerpene, has been isolated from Caesalpinia bonducella roots.[48] From the seeds of the same

O

1

2 9

3

45

6

7

810

11 1213

14

15

16

17

Figure 1: Carbon skeleton of cassane-type diterpenoids

O

H

HO

H

OHHOOC O O

O O

CH3

Figure 2: Pulcherrimin A

O

OH

AcOOAc

OHH

H

Figure 3: e-caesalpin

O

H

H

HOOC

MeOOC

H

Figure 4: Caesaljapin

67


O

OHOAc

H

Figure 5: Caesaldekarin A

O

OHMeOOC

H

Figure 6: Caesaldekarin C

O

O

OAc

OH

OOAc

Figure 7: Caesalpinin 1

O

AcO

OAc

OH

H

O

HO

OHH

Figure 8: Neocaesalpin A

H

HO

O

H

OH

OAc

AcO

O

H

Figure 9: Neocaesalpin B

O

OH

H

H

MeOOC

Figure 10: Caesaldekarin F

OH

OH

H

O

H

MeOOC

Figure 11: Caesaldekarin G

H

O

OH

H

AcO CH2

Figure 12: Caesaldekarin H

68


H

O

OH

H

CH2OH

OH

Figure 13: Caesaldekarin I

O

OHMeOOC

Figure 14: Caesaldekarin J

H

O

OH

OH

MeOOC

Figure 15: Caesaldekarin A

H

H

OH

OH

HOH

HOCH2

Figure 16: Caesaldekarin B

H

O

OH

H

HOOC

Figure 17: Demethylcaesaldekarin C

O

OH

OAc

H

OH

OH

H

H

Figure 18: Caesalmin A

OH

H

OAc

H

H

O

OH

H

Figure 19: Caesalmin B

O

H

H

OAcOH

OAc

OAc

Figure 20: Caesalmin C

OAc

OAcOH

OAc

O

H

H

OH

Figure 21: Caesalmin D

OAc

OAcOH

OAc

O

H

H OH

Figure 22: Caesalmin E

69


O

H

H

OAcOH

OAc

OAc

OMe

Figure 23: Caesalmin F

OH

OH

H

OAc

O

H

H

H

Figure 24: Caesaldekarin G

O

H

OHO

H

O

Figure 25: Isovouacapenol A

H

OOH

H

O

OH

O

Figure 26: Isovouacapenol B

H

OOH

H

O

OH

H

O

Figure 27: Isovouacapenol C

OOH

O

O

Figure 28: Isovouacapenol D

O

O

OO

OAc

O

H

H

H

H

Figure 29: Macrocaesalmin

O

H

H

OH OH

H

OAc

Figure 30: Caesalmin H

70


O

H

OH

OAc

H

H

O

OH

Figure 31: Bonducellpin D

O

O

OH

OAc

AcO

H

H

Figure 32: Norcaesalpinin A

OAc

O

O

OH

Figure 33: Norcaesalpinin B

O

OH

OH

OAcH

Figure 34: Norcaesalpinin C

AcO

O

OH

H

AcO

CH3

H

Figure 35: Caesalpinin MA

H

O

OH

H

AcO

HCOOCH3

Figure 36: Caesalpinin MB

OH

O

AcO

AcO

Me

Figure 37: Caesalpinin MC

AcO

OAcOH

O

AcOMe

Figure 38: Caesalpinin MD 4

O

H

H

OAc

O

H

AcO

Figure 39: Caesalpinin ME

OH

OOH

AcOH

Figure 40: Norcaesalpinin MA

71


AcO

OH O

OH

Figure 41: Norcaesalpinin MB

O

OAc

H

OH

H O

OAc

AcO

Figure 42: Norcaesalpinin MC

O

HMeOOC

Figure 43: Taepeenin A

O

HHOOC

Figure 44: Taepeenin B

O

HOHMeOOC

Figure 45: Taepeenin C

O

HMeOOC COOMe

Figure 46: Taepeenin D

OHH

H

H

Figure 49: Taepeenin G

O

H

H

H

CHOMeOOC

Figure 50: Taepeenin H

O

HCHOMeOOC

Figure 47: Taepeenin E

OO

HCOOMe

Figure 48: Taepeenin F

72


O

H

H

H

CH2OHMeOOC

Figure 51: Taepeenin I

O

OH

HMeOOC

H

Figure 52: Nortaepeenin A

O

OH

HOHMeOOC

H

Figure 53: Nortaepeenin B

O

OH

OAc

AcO

Figure 54: Caesaldekarin e

OAc

O

OH

AcO

AcO

Figure 55: 2-Acetoxycaesaldekarin e

OAc

O

OH

AcO

HO

Figure 56: 2-Acetoxy-3-deacetocaesaldekarin e

OAc

O

OHOAc

Figure 57: 6-Acetoxy-3-deacetoxycaesaldekarin e

O

H

OH

H

OAc

Figure 58: 14 (17)-Dehydrocaesalmin F

OAc

O

OH

OH

H

OH

COOMe

Figure 59: Bonducellpin B

O

H

OH

H

OAc

H

H

H

OH

COOMe

Figure 60: Bonducellpin C

73


O

H

OH

H

OAc

H

H

OAc

COOMe

Figure 61: 7-Acetotoxybonducellpin C

O

O

H

OH

H

OAc

OAc

Figure 62: 1-Deacetoxy-1-oxocaesalmin C

O

O

H

OHOH

H

OAc

Figure 63: S-Caesalpin

O

O

H

OH

H

OAc

OAc

Figure 64: 1-Deacetylcaesalmin C

O

H

OH

H

AcO

OAc

OAc

OAc

O

Figure 65: Caesalpinin C

O

O

O

H

OAc

OAc

H

OH

Figure 66: Caesalpinin D

O

OH

H

HOAc

H

AcO

AcO

Figure 67: Caesalpinin E

O

O

OH

O

OAc

OAc

Figure 68: Caesalpinin F

O

O

O

OAc

OH

OH

H

H

Figure 69: Caesaldekarin H

O

O

O

O

OAc

H

H

OH

Figure 70: Caesaldekarin I

74


O

O

OH OAc

OAc

COOMeH

H

Figure 71: Caesalpinin J

O

OH

H

HOAc

H

OH

Figure 72: Caesalpinin K

O

OH

H

H

OAc

OAc

H

COOMe

OH

Figure 73: Caesalpinin M

O

OH

H

H CHOOAc

OH

H

Figure 74: Caesalpinin N

O

O

O

OHOH

OAc

Figure 75: Caesalpinin O

O

OH

AcO

OAc

H

H

Figure 76: Caesalpinin P

O

H

OH

H

OAc

OAc

H

COOMe

Figure 77: Caesalpinin MF

O

H

OH

H

OAc

OAc

OAc

H

COOMe

Figure 78: Caesalpinin MG

O

H

OH

H

OAc

OH

COOH

OAc

H

Figure 79: Caesalpinin MH

O

H

OH

H

OH

H

Figure 80: Caesalpinin MI

75


O

H

H

H

MeOOC

Figure 89: Benthaminin 3

O

OH

H

H

OAc

OAc

Figure 81: Caesalpinin MJ

OAc

O

H

OH

H

O

OAc

Figure 82: Caesalpinin MO

O

O

O

OH

H

H

OAc

OAc

Figure 83: Norcaesalpinin MD

O

H

OH

H

O

OAcO

AcO

Figure 84: Norcaesalpin D

Figure 85: Norcaesalpin E

O

H

OH

H

O

O

OH

O

H

OH

H

O

O

OH

OAc

Figure 86: Norcaesalpin F

MeOOC

O

H


MeOOC

O

H

H

H


(57-65) and two new norcassane-type diterpenes nortaepeenin A-B (66-67) were also isolated from the stems and roots of C. crista.[63] From the seed kernels of the same plant, known cassane and norcassane-type diterpenes including compounds 29, 30, 32, 34, 46-49, 54, 68-78 and new cassane-type diterpenes, namely caesalpinins C-K (65-68), M-P (73-76), caesalpinins MF-MJ (77-81), MO (82) and norcaesalpins MD (83), D-F (84-86) possessing antimalarial activity, have been isolated. Norcaesalpinin E (85), displayed the most potent antimalarial activity.[62]

76


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CONCLUSION

Cassane-type diterpenoids continue to be isolated from medicinal plants. Three novel cassane-type diterpenoids- benthaminin 1 (87), 2 (88) and 3 (89) possessing antimicrobial and antioxidant properties have been isolated from Caesalpinia benthamiana.[51] Similarly, two novel cassane-type diterpenoids designated magnicaesalpin and neocaesalpin O together with three known ones named caesalmin D and E and neocaesalpin L have been isolated from the seeds of Caesalpinia magnifoliolata.[52] A number of these cassane-type furanoditerpenoids have been found to manifest various biological activities including antibacterial, antifungal,[29] anti-inflammatory, anti-analgesic,[52,53] antiviral and anticancer,[31] antimalarial[54] and antituberculosis activities.[36] Thus, these cassane diterpenoids are of interest due to their structural diversity and their broad spectrum of biological activities. Further studies should be performed to indicate which of these isolated bioactive chemical constituents may serve as lead compounds in the synthesis of biomolecules to tackle the numerous global health challenges due to the emerging and ongoing drug resistance associated with long term use of conventional medicines used in the treatment and management of infectious diseases.

ACKNOWLEDGEMENTS

R. A. Dickson is grateful to the Commonwealth Scholarship Commission, UK and the Kwame Nkrumah University of Science and Technology, Ghana for sponsorship.

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Research Article



*Correspondence: +913222-282220/282657; Fax: +913222-282221;Email: [email protected], [email protected]: 10.5530/pc.2011.1.5

Azadirachtolide: An anti-diabetic and hypolipidemic effects from Azadirachta indica leavesDinesh kumar B1, Analava Mitra2*, Manjunatha M2

1Department of Pharmaceutics, PSG College of Pharmacy, Coimbatore-641 004, India. 2School of Medical Science and Technology, Indian Institute of Technology, Kharagpur -721302, West Bengal, India

IntRoductIon

Diabetes is a group of metabolic diseases characterized by hyperglycemia resulting from defects in insulin secretion or insulin action, or both.[1] Broad research on diabetes has resulted in the development of a number of oral hypoglycemic agents including biguanides, sulphonylureas and thiozolidinediones which are available commercially for the management of diabetes. However, these drugs also produce nondesirable side effects.[2] Hence, there is a need to develop alternative anti-diabetes medicines. The herbal medicines are widely used for the treatment of disease because of their effectiveness, safety, affordability and acceptability.[3] Medicinal plants including their phyto-compounds have been used in the Indian traditional systems of medicine for treatment of diabetic populace all around the world with less known scientific basis of their functioning.[4-7] Hence, phyto-products from medicinal plants need to be investigated by scientific methods for their anti-diabetic activity. Various medicinal effects have been reported for anti-inflammatory, anti-arthritic, antipyretic,

antifungal, anti-bacterial, diuretic, immunomodulatory and anti-tumor properties. Phyto-compounds such as azadirachtins, nimocinol, isomeldenin, 2, 3′-dehydrosalanol gedunin, nimbin, nimolicinol from Azadirachta indica have been reported in the leaves.[8]

Tetranortriterpenoids has been reported for anticancer, antiviral, anti-allergic and anti-inflammatory activities.[9-12] There is no report on azadirachtolide (tetranortriterpenoid from Azadirachta indica leaves) for antidiabetic and hypolipidemic activities. Therefore, the effect of azadirachtolide (tetranortriterpenoid from Azadirachta indica leaves) on blood glucose and serum lipid profiles on streptozotocin-induced diabetic rats was investigated. Further, in vitro alpha amylase and alpha glucosidase an inhibitory effect of azadirachtolide was evaluated.

MAteRIAls And Methods

Chemicals and reagentsStreptozotocin, starch azure, porcine pancreatic amylase, alpha glucosidase from yeast Saccharomyces cerevisiae, para-nitrophenyl gluco-pyanoside and Tris-HCl buffer were procured from Sigma Chemicals, USA. Dimethyl sulfoxide, acetic acid, calcium chloride, ethanol, chloroform, petroleum ether, potassium bromide,

ABSTRACT: Introduction: Azadirachta indica (Meliaceae) leaves are used traditionally in the Indian Ayurvedic medicinal system to treat diabetes. The aim of the present study is to investigate the effect of azadirachtolide (tetranortriterpenoid from Azadirachta indica leaves) on blood glucose and serum lipid profiles on streptozotocin-induced diabetic rats. Methods: Streptozotocin-induced diabetic rats were used for the study. Azadirachtolide (at a dose 50 and 100 mg/kg) was administrated intra-peritoneally in diabetic rats once a week for 30 days. Biochemical parameters notably fasting blood sugar, total cholesterol, triglycerides, low-density lipoprotein, very low-density lipoprotein and high-density lipoprotein were determined. The in vitro alpha amylase and alpha glucosidase inhibitory effects of azadirachtolide were measured and IC50 values were determined. Results: Azadirachtolide exhibited significant (P < 0.05) anti-diabetic as well as hypolipidemic effects by lowering FBS, TC, TG, LDL, and VLDL levels; but also with elevation of HDL level in diabetic rats. Azadirachtolide showed appreciable alpha amylase (IC50 value of 55.80 ± 1.7 µg/ml) and alpha glucosidase inhibitory effects (IC50 value of 47.85 ± 1.4 µg/ ml) compared with acarbose (IC50 value of 83.33 ± 1.8 µg/ml). Conclusion: The present study indicated that azadirachtolide possesses anti-hyperglycemic and anti-lipidemic effects. Thus, results suggested azadirachtolide has a beneficial effect in the management of diabetes associated with abnormal lipid profile and related cardiovascular complications.

KEYWORDS: Azadirachta indica, Azadirachtolide, Anti-diabetic, Hypolipidemic

Research Article

79

Kumar, et. al.: Azadirachtolide: An anti-diabetic and hypolipidemic effects from Azadirachta indica leaves

twin-trough glass chamber previously saturated with mobile phase vapor for 20 min. After developing the plate, it was dried at 105ºC for 15 min and then it was scanned using Scanner 3 (CAMAG, Switzerland) at 254nm using WinCATS 4 software. IR spectrum was recorded using a Thermo Nicolet Nexus 870 FT-IR Spectrophotometer using potassium bromide pellets. Mass spectrum was recorded on Electro-Spray Ionization Mass Spectroscopy (Waters, UK). NMR spectra were recorded in CDCl3 in a Bruker 400 MHZ spectrometer using Topspin software.

In vitro alpha amylase inhibitory assayThe assay was carried out following the standard protocol with slight modifications.[13] Starch azure (2 mg) was suspended in a tube containing 0.2ml of 0.5 M Tris-Hcl buffer (pH 6.9) containing 0.01 M calcium chloride (substrate). The tube was boiled for 5 min and then pre-incubated at 37º C for 5 min. Azadirachtolide was dissolved in 0.1% of dimethyl sulfoxide in order to obtain concentrations of 10, 20, 40, 60, 80 and 100 µg/ml. Then 0.2 ml of azadirachtolide of a particular concentration was put in the tube containing the substrate solution. 0.1 ml of porcine pancreatic amylase in Tris-Hcl buffer (2units/ml) was added to the tube containing the azadirachtolide and substrate solution. The reaction was carried out at 37 ºC for 10 min. The reaction was stopped by adding 0.5 ml of 50% acetic acid in each tube. The reaction mixture was then centrifuged (Eppendorf -5804 R) at 3000 rpm for 5 min at 4ºC. The absorbance of resulting supernatant was measured at 595 nm (Perkin Elmer Lambda 25 UV-VIS). The concentration of the azadirachtolide required to inhibit 50% of alpha amylase activity under the conditions was defined as the IC50 value. The experiments were repeated thrice with the same protocol.

The alpha amylase inhibitory activity was calculated as follows:

Alpha amylase inhibitory activity = (Ac+) – (Ac–) – (As–Ab)

(Ac+) – (Ac–) × 100

Where, Ac+, Ac–, As, Ab are defined as the absorbance of 100% enzyme activity (solvent with enzyme alone), 0% enzyme activity (solvent without enzyme), a test sample (with enzyme) and a blank (a test sample without enzyme) respectively.

In vitro alpha glucosidase inhibitory assayThe assay was performed using a standard protocol.[14] Alpha glucosidase (2U/ml) was premixed with 20 µl of azadirachtolide at various concentrations (10, 20, 40, 60, 80 and 100 µg/ml) and incubated for 5 min at 37ºC. 1mM para-nitrophenyl gluco-pyanoside (20 µl) in 50mM of phosphate buffer (pH 6.8) was added to initiate the reaction. The mixture was further incubated at 37ºC for 20 min. The reaction was terminated by addition of 50 µl of 1 M sodium carbonate and the final volume was made up to 150 µl. Alpha glucosidase activity was determined spectrophotometrically at 405nm on a Biorad microplate reader

deuterated chloroform, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, Whatmann filter paper and sodium carbonate were purchased from Merck, India. Thin layer chromatography plates were obtained from Merck (silica gel 60 F254 grade, Germany). Diagnostic kits and reagents for fasting blood sugar, total cholesterol, triglyceride, high density lipoprotein, low density lipoprotein and very low density lipoprotein were obtained from Merck, India. Acarbose was gifted by Zota Pharmaceutical Pvt. Ltd., Chennai. Glibenclamide (Aventis Pharma- Mumbai) was procured from local medical market.

Plant materialsAzadirachta indica leaves (Rutaceae) were collected from the locality of IIT Kharagpur campus, West Bengal, India in the month of September and October 2007. The leaves were inspected to be healthy and botanically identified and authenticated by Dr. M. Senthilkumar, Plant Biotechnologist, Prathyusha Institute of Technology and Management, Chennai. The herbarium Azadirachta indica leaves was deposited in the Prathyusha Institute of Technology and Management (PITAM) against voucher no. PITAM/ CH/00015/ 2007. Azadirachta indica leaves after collection were dried at room temperature (27-30ºC) for 25-30 days. After complete drying (inspection), the dried materials were ground into fine powder using a domestic electric grinder (Product: GX 21, Bajaj appliances, Mumbai, India) and used for extraction.

Extraction and isolationDried plant powder of Azadirachta indica leaves (500 g) was extracted with ethanol (1 L) at room temperature. Then extract was filtered (Whatmann filter paper, 110mm, Cat. no 1001 110). The filtrate was evaporated by rotary evaporation (Buchi Rotavapor R-210) to get a dark greenish solid residue. These greenish solid residues (15 g) was successively extracted with petroleum ether (3.5 g) and chloroform (5.2 g) and subjected for in vitro alpha amylase inhibitory activity. The chloroform fraction showed appreciable alpha amylase inhibitory compared to petroleum ether fraction. The chloroform fraction was subsequently subjected to column chromatography using gradient elution using acetone and chloroform as solvents (10% acetone in chloroform for 15 mins, 20% acetone in chloroform for 15 mins and 30% acetone in chloroform for 15 mins). The fractions obtained with 20% acetone in chloroform afforded compound-I (10 mg). These fractions were subjected to preparative TLC with mobile phase hexane: ethyl acetate (8.5:1.5) for isolation of compound-I. Compound-I was identified as azadirachtolide by comparing its FTIR, ESI-MS and NMR with previously published literature (Ragasa et al., 1997).

General experimental procedureHPTLC (CAMAG, Switzerland) analyzes was performed using silica gel 60 F254 TLC plate. All collected fractions were spotted (10 µl) on a silica gel 60 F254 (Merck, Darmstadt, Germany) TLC plate. The plate was air dried and then developed using the solvent system hexane: ethyl acetate (8.5:1.5) in a CAMAG-

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Statistical analysisAll values were expressed mean ± standard deviation. Statistical analysis of in vivo results were performed by one-way analysis of variance (ANOVA) followed by Student’s t-test. P < 0.05 was considered statistically significant. In vitro inhibitory assay statistical difference and linear regression analysis were performed using Graphpad prism 5 statistical software.

Results

Azadirachtolide (10 mg) was isolated from 500 g of dried leaves of Azadirachta indica (Figure 1). HPTLC analyzes indicated that F2 contained azadirachtolide and the retention factor (Rf) values of azadirachtolide was found to be 0.31 (Figure 2). The F2 fractions were subjected to preparative TLC with the solvent system hexane: ethyl acetate (8.5:1.5) to get the compound-1 (azadirachtolide).

FTIR (KBr disc) is shown in Figure 3: peak at 3444 cm-1 indicated presence of OH group, peak at 2925 cm-1, 2854 cm-1 was due to presence of C-H, peak at 1370 cm-1 showed C-H bending, peak at 1736 cm-1 indicated presence of ester carbonyl group, peak at 1666 cm-1 showed presence of C-O group and peak at 1458 cm-1 indicated presence of CH-CH bending (Figure 3).

ESI-mass spectroscopy showed the presence of a molecular weight peak of azadirachtolide at 593. ESI-MS (m/z, % intensity): m/z 593 [M-H]-. Proton NMR (CDCl3 solvent) showed senecioyloxy subtitutent δ 1.88 (3H), δ 2.20 (3H), δ 5.70 (1H). an acetate δ 1.97 (3H). Four additional methyl singlet δ 0.8, δ 1.25, δ 1.28, δ 1.30, two olefinic hydrogen δ 5.57, δ 5.71, methylene hydrogen bonded to oxygenated carbons δ 4.15 (1H), δ 3.81 (1H), δ 3.68 (1H), δ 3.59 (1H) and methine hydrogen bonded to oxygenated carbons δ 4.12 (1H), δ 4.15 (1H), δ 5.30 (1H), δ 5.47 (1H).

Azadirachtolide showed appreciable alpha amylase (IC50 value of 55.80 ± 1.7µg/ml) and alpha glucosidase inhibitory effects (IC50 value of 47.85 ± 1.4µg/ml) as compared with acarbose (IC50 value of 83.33 ± 1.8µg/ml) (Figure 4). The body weight was slightly increased in normal control rats compared to initial body weight whereas streptozotocin-induced diabetic rats showed loss of body weight (172.6 ± 2.05 g) after 30 days as compared with initially weight of diabetic rats (186.6 ± 1.24 g). However, body weight of diabetic rats was restored by treating with

by measuring the quantity of para-nitrophenol released from pNPG. The assay was performed in triplicate. The concentration of azadirachtolide required to inhibit 50% of alpha glucosidase activity under the conditions was defined as the IC50 value. The experiments were repeated thrice with same protocol.

Animal studiesAdult male Wistar Rats (weighing 150-200 g) were used for this investigation. The animals were acclimatized to the laboratory conditions for a period of 2 weeks prior to the experiment. They were maintained at an ambient temperature (25 ± 2 ºC) and relative humidity (40-60%), with 12/12 h of light/dark cycle. The animals were maintained on balance diet and water ad libitum. Institutional Animal Ethical Committee (IAEC) approved the study and all the experiments were carried out by following the guidelines of CPCSEA, India.

Induction of diabetes and blood sample collectionA freshly prepared solution of streptozotocin (45mg/kg) in 0.1M citrate buffer pH 4.5 was injected intra-peritoneally in overnight fasted rats. After 3 days, blood was collected from the tail vein of overnight fasting rats under the supervision of a veterinary surgeon using aseptic conditions. The FBS level of blood was checked regularly up to the stable hyperglycemia stage, usually one week after streptozotocin injection. Animals with marked hyperglycemia (FBS 250 mg/dl) were selected for the study.[15]

Experimental designGroup I - Normal controlGroup II - Diabetic controlGroup III - Diabetic +50 mg/kg (i.p.) azadirachtolideGroup IV – Diabetic +100 mg/kg (i.p.) azadirachtolideGroup V - Diabetic + 0.5 mg/kg (i.p.) glibenclamide

The experiment was carried on five groups (I, II, III, IV and V) of six rats each. Group-I served as normal control. Group-II served as diabetic control. Group III-diabetic + 50 mg/kg (i.p.) of azadirachtolide. Group IV-diabetic + 100 mg/kg (i.p.) of azadirachtolide. Group V-diabetic + 0.5 mg/kg (i.p.) of glibenclamide and served as positive control. The azadirachtolide was suspended in 0.3% w/v sodium carboxy methyl cellulose (Sodium CMC) as a vehicle and injected intra-peritoneally into rats once a week for one month with a dose of 50 mg/kg and 100 mg/kg body weight. The blood samples were collected from each rat by retro-orbital vein-puncture. Biochemical parameters were estimated at the beginning and after 30 days of experiment.

Biochemical parametersBiochemical parameters notably fasting blood sugar (FBS), total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL) levels and high-density lipoprotein (HDL) level in blood serum were measured spectrophotometrically (Semi-Autoanalyzer, Microlab 300, Merck) as per the manufacturers instructions using diagnostic kits and reagents obtained from Merck, India. Figure 1: Structure of azadirachtolide.

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rats treated with azadirachtolide (at a dose of 50 mg/kg and 100 mg/ kg, i.p.) once a week for 30 days on being compared with diabetic rats exhibited significant (P < 0.05) reduction in fasting blood sugar levels (204.0 ± 2.94 and 198.3 ± 2.86 mg/dl respectively). The standard glibenclamide (0.5 mg/kg, i.p.) also showed anti-diabetic activity with reduction of fasting blood sugar level (215.0 ± 2.18 mg/dl) on 30 days as compared to the diabetic control. There was a significant (P < 0.05) reduction in triglycerides, total cholesterol, low density lipoprotein and very low density lipoprotein levels of diabetic rats treated with azadirachtolide (50 and 100 mg/ kg, i.p.) on being compared with diabetic control. Also, there was a significant (P < 0.05) elevation of HDL level in azadirachtolide (50 and 100 mg/kg, i.p.) treated diabetic rats.

glibenclamide (0.5 mg/kg) and azadirachtolide (at a dose of 50 mg/kg and 100 mg/kg, i.p.) for 30 days (Table 1).

Streptozotocin treatment resulted in elevation of fasting blood glucose, triglycerides, total cholesterol, low density lipoproteins, very low density lipoproteins and a reduction in high density-lipoprotein levels as compared to the normal control rats (Table 2).

Intra-peritoneal administration of azadirachtolide (at a dose of 50 mg/kg and 100 mg/kg, once a week for 30 days) exhibited significant (P < 0.05) reduction in fasting blood sugar levels (204.0 ± 2.94 and 198.3 ± 2.86 mg/dl in diabetic rats. Diabetic

Figure 2: HPTLC peaks of collected column fractions CE-Crude extract (Pink peak), F1-10% acetone in chloroform (Violet peak), F2-20% acetone in chloroform (Green peak), F3-30% acetone in chloroform (Orange peak).

Figure 3: FTIR Spectrum of azadirachtolide.

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role in occurrence of premature and severe atherosclerosis, which affects patients with diabetes.[20]

In the present study, an increase in blood sugar levels in diabetic rats was observed after the induction of diabetes by streptozotocin. This was prevented by treating diabetic rats with azadirachtolide (at a dose 50 and 100 mg/kg, i.p.) once a week for 30 days. The standard drug glibenclamide has been used to treat diabetes, which stimulate insulin secretion from pancreatic beta cells, it may be suggested that the mechanism of action of azadirachtolide is similar to glibenclamide. The azadirachtolide (at a dose 50 and 100 mg/ kg, i.p.) treated diabetic rats showed a significant reduction in both fasting blood sugar levels and some lipid parameters (TC, TG, LDL, and VLDL). Some biological active compounds such as mimbidin, sodium nimbidate, nimbin, nimbolide, gedunin, azadirachtin, mahmoodin, gallic acid, catechin, margoone, isomargolone, cyclic trisulphide, cyclic tetrasulphide and polysacharides were isolated from leaves and seeds of Azadirachta indica.[21] Aqueous extract of neem leaf significantly reduced the blood glucose level of male albino rats of Wistar strains.[22] The combined ethanolic extracts of Azadirachta indica and Vernonia amygdalina leaf extracts showed anti-hyperglycemic effect on alloxan induced albino wistar rats.[23] The weight loss in diabetic rats may be associated with lipid lowering activity of azadirachtolide or due to its influence on various lipid regulation systems. Treatment with azadirachtolide (at a dose 50 mg/ kg and 100 mg/ kg body weight) in diabetic rats may have potential role to prevent formation of atherosclerosis and coronary heart disease. The present in vivo study showed that intra-peritoneal

dIscussIon

The present study was designed to explore the effect of azadirachtolide (tetranortriterpenoid from Azadirachta indica leaves) on blood glucose and serum lipid profiles on streptozotocin-induced diabetic rats. A comparison of the FTIR, ESI-MS, NMR spectra of isolated fraction showed significant similarity with previously reported azadirachtolide data.[16] Intra-peritoneal administration of 50mg/kg and 100 mg/kg of azadirachtolide once a week for 30 days showed anti-diabetic and hypolipidemic effects in diabetic rats. Lipid abnormalities associated with atherosclerosis is the major cause of cardiovascular disease in diabetes. High level of TC and LDL are major coronary risk factors.[17] Further, several studies suggested that TG itself is interdentally related to coronary heart disease.[18,19] The abnormalities in lipid metabolism lead to elevation in the levels of serum lipid and lipoprotein that in turn play an important

Figure 4: Alpha amylase and alpha glucosidase inhibitory effects of azadirachtolide.

Table 1: Body weights of streptozotocin-induced diabetic rats after treatment with azadirachtolide.

Group Initial body weight

Final body weight

Normal control 191.0 ± 0.81 200.0 ± 0.63Diabetic control 186.6 ± 1.24 172.6 ± 2.0550 mg/kg of azadirachtolide 183.0 ± 1.60 178.3 ± 1.69*100 mg/kg of azadirachtolide 182.0 ± 1.63 177.6 ± 1.69*0.5 mg/kg of glibenclamide 180.3 ± 0.47 176.3 ± 1.24*

*(P <0.05) compared with treated diabetic groups Vs Diabetic control. n = 6/group. Values are expressed as mean ± S.D

83


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8. Atawodi SE, Atawodi JC. Azadirachta indica (neem): a plant of multiple biological and pharmacological activities. Phytochem Rev. 2009; 8:601-620.

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15. Gupta RK, Kesari AN, Murthy PS, Chandra R, Tandon V, Watal G. Hypoglycemic and antidiabetic effect of ethanolic extract of leaves of Annona squamosa L. in experimental animals. J Ethnopharmacol. 2005; 99:75-81.

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administration of azadirachtolide (at a dose 50 and 100 mg/kg) exhibited anti-diabetic and hypolipidemic effects in streptozotocin-induced diabetic rats.

One of the therapeutic approaches for type 2 diabetes is to reduce the post-prandial hyperglycemia. Alpha amylase and alpha glucosidase are the enzymes involved in the metabolism of carbohydrates. Alpha amylase degrades complex dietary carbohydrates to oligosaccharides and disaccharides, which are ultimately converted into monosaccharide. Liberated glucose is then absorbed by the gut and results in postprandial hyperglycemia. Inhibition of alpha amylase and alpha glucosidase limits postprandial glucose levels by delaying the process of carbohydrate hydrolysis and absorption.[24] The plant based alpha amylase and alpha glucosidase inhibitor offers a prospective therapeutic approach for the management of post-prandial hyperglycemia. [25] In this study, azadirachtolide showed appreciable alpha amylase and alpha glucosidase inhibitory effects compared with acarbose.

conclusIon

The present study indicated that azadirachtolide (at a dose 50mg/ kg and 100 mg/kg body weight) exhibited anti-diabetic and hypolipidemic effects in streptozotocin-induced diabetic rats. Therefore, azadirachtolide could be used as anti-diabetic agent in the management of diabetes associated with abnormalities of lipid profiles.

AcknowledgMents

Authors would like to acknowledge Prof. P.K. Dutta, Head, School of Medical Science and Technology, IIT Kharagpur and for his valuable support in the research work. The authors would like to acknowledge the Central Research Facility (CRF) of Indian Institute of Technology, Kharagpur for providing the facility of FTIR, ESI-MS and NMR.

Table 2: Effect of azadirachtolide on biochemical parameters in normal and diabetic rats.

BBP (mg/dl) Days Group 1 Group 2 Group 3 Group 4 Group 5

FBS 0 75.6 ± 1.60 271.0 ± 2.9 278.3 ± 1.24 280.0 ± 3.77 286.0 ± 1.4130 76.6 ± 1.69 294.3 ± 3.29# 204.0 ± 2.94* 198.3 ± 2.86* 215.0 ± 2.18*

TC 0 93.6 ± 1.75 200.3 ± 1.36 201.5 ± 1.47 201.7 ± 1.83 200.8 ± 1.9230 94.6 ± 1.24 224.3 ± 3.09# 127.3 ± 2.05* 125.3 ± 1.24* 126.6 ± 1.24*

TG 0 68.5 ± 1.47 182.3 ± 2.05 176.3 ± 1.24 170.0 ± 1.63 159.6 ± 1.2430 69.0 ± 2.94 200.0 ± 1.63# 133.3 ± 2.05* 121.6 ± 1.69* 115.6 ± 1.69*

HDL 0 31.6 ± 1.69 25.3 ± 2.05 22.5 ± 1.08 21.7 ± 0.98 21.1 ± 1.0430 33.6 ± 1.24 21.1 ± 0.62# 30.3 ± 1.69* 31.6 ± 1.24* 32.0 ± 1.63*

LDL 0 51.7 ± 1.28 110.0 ± 1.63 114.6 ± 2.05 113.0 ± 2.16 112.6 ± 2.8630 54.1 ± 1.30 117.8 ± 2.24# 81.3 ± 0.94* 77.3 ± 1.69* 83.5 ± 1.22*

VLDL 0 18.3 ± 1.30 40.7 ± 1.25 37.0 ± 1.50 33.5 ± 1.08 30.2 ± 1.2030 19.6 ± 1.69 48.6 ± 0.69# 25.9 ± 1.96* 21.6 ± 1.24* 20.8 ± 0.47*

Values are expressed as mean ± S.D. n = 6/group. Group -1 (rats treated with 0.3% w/v sodium carboxy methyl cellulose (i.p) - served as normal control), Group-2 (rats treated with (45mg/kg) of streptozotocin (i.p) - served as diabetic control), Group 3 - (diabetic+50 mg/kg (i.p.) of azadirachtolide), Group 4- (Diabetic+100 mg/kg (i.p.) of azadirachtolide), Group 5 - (diabetic+0.5 mg/kg (i.p.) of glibenclamide (positive control). BBP-Biochemical parameters, FBS-fasting blood sugar, TC-total cholesterol, TG-triglycerides, LDL-low-density lipoprotein, VLDL-very low-density lipoprotein, HDL-high-density lipoprotein. #P < 0.05, Group 1 vs. Group 2. *P < 0.05, treated diabetic groups vs. diabetic control group.

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22. Bajaj S, Srinivasan BP. Investigations into the anti-diabetic activity of Azadirachta indica. Indian J Pharmacol. 1999; 31:138-141.

23. Ebong PE, Atangwho IJ, Eyong EU, Egbung GE. The anti-diabetic efficacy of combined extracts from two continental plants: Azadirachta indica (A.Juss) (Neem) and Vernonia amygdalina (Del.) (African bitter leaf). Am J Biochem Biotechnol. 2008; 4: 239-244.

24. Bell DS. Type 2 diabetes mellitus: What is the optimal treatment regimen? Am J Med. 2004; 116:23-29.

25. McCue P, Vattem D, Shetty K. Inhibitory effect of clonal oregano extracts against porcine pancreatic amylase in vitro. Asia Pac J Clin Nutr. 2004; 13:401-408.

18. Bainton D, Miller NE, Botton CH, Yarnell JWG, Suretman PM, Baker IA. Plasma triglycerides and high density lipoprotein cholesterol as predictors of ischemic heart disease in British man. Br Heart J. 1992; 68:60-66.

19. EI-harzmi MA, Warsy AS. Evaluation of serum cholesterol and triglyceride level in 1-6- year -old Saudi children. J Trop Pediatr. 2001; 47:181-185.

20. Ravi K, Rajasekaran S, Subramanian S. Anti-hyperlipidemia effect of Eugenia jambolana seed kernel on streptozotocin-induced diabetes in rats. Food Chem Toxicol. 2005; 43:1433-1439.

21. Biswas K, Chattopadhyay I, Banarjee RK, Bandyopadyay U. Biological activities and medicinal properties of neem (Azadirachta indica). Curr sci. 2002; 82:1136-1345.


Research Letter



#Correspondence: +233-3220-60359; +233-244597464;Email: [email protected]: 10.5530/pc.2011.1.6

Antimicrobial and anti-inflammatory activities of the leaves of Clerodendrum splendens leavesFleischer, TC1#, Mensah, AY2, Oppong, AB2, Mensah, MLK1, Dickson, RA2, Annan, K2

1Department of Herbal Medicine, Faculty of Pharmacy and Pharmaceutical Sciences, Kwame Nkrumah University of Science and Technology (KNUST), Kumasi, Ghana. 2Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical Sciences, Kwame Nkrumah University of Science and Technology (KNUST), Kumasi, Ghana

INTRODUCTION

Clerodendrum splendens G. Don (Family: Verbenaceae) also known as the Flaming Glory - bower is a woody or semi-woody evergreen vine which grows in the tropical and subtropical regions of the world. In ethnomedicine, the plant is used to treat wounds and burns,[1] haemorrhoids, diarrhoea and dysentery.[2] The leaves have been found to contain reducing sugars, glycosides, unsaturated sterols, triterpenoids and flavonoids.[3] Recently the plant has been reported to show wound healing, antioxidant and antimicrobial properties.[4] Various species of Clerodendrum, including C. trichotomum, C. indicum and C. serratum which are used traditionally in the management of inflammatory conditions, have been shown to possess potent anti-inflammatory activities.[5] We have investigated the antimicrobial and anti-inflammatory activities of the leaves of C. splendens and in this report provide further support for its ethnomedicinal uses.

MATERIALS AND METHODS

Plant materialThe leaves of C. splendens were collected from Asokore Mampong in Kumasi in May, 2008. The plant material was authenticated

by Mr. Ntim-Gyakari, the curator of the Herbarium of the Forestry Commission in Kumasi and a voucher specimen (KNUST/HM1/2010/L033) has been deposited at the herbarium of the Faculty of Pharmacy and Pharmaceutical Sciences, Kwame Nkrumah University of Science and Technology (KNUST) Kumasi, Ghana.

Extraction of plant materialThe leaves were air dried for four days and ground into a coarse powder. The powder (0.5 kg) was serially extracted using petroleum ether, ethyl acetate and 70% ethanol. The various extracts were evaporated under reduced pressure using a rotary evaporator until a viscous extract of each was produced. The petroleum ether extract gave a yield of 9.19 %w/w, that of ethyl acetate extract was 9.59 %w/w and 13.45 %w/w for the ethanolic extract. Phytochemical screening of the powdered leaves using methods described by Sofowora[6] and Harborne[7] showed the presence of flavonoids, tannins and alkaloids.

Test organismsThe microorganisms used in this study were obtained from the stocks of the Department of Pharmaceutics, Faculty of Pharmacy and Pharmaceutical Sciences, KNUST, Kumasi. They included; Staphylococcus aureus (NCTC 10788), Bacillus subtilis (ATCC 6633), Pseudomonas aeruginosa (NCTC 10662), Eschericia coli (ATCC 25922) and Candida albicans (ATCC 102321). A 24 hour broth culture of the organisms was used. The media used was Nutrient agar (MERCK) for the bacteria and Sabouraud agar (MERCK) for Candida.

ABSTRACT: Clerodendrum splendens is a West African climbing shrub used in traditional medicine for wounds and infectious conditions. The petroleum ether, ethyl acetate, and 70% ethanolic extracts of the leaves obtained by successive Soxhlet extraction, inhibited the growth of Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Eschericia coli and Candida albican. The ethyl acetate extract was the most active. Again, all the extracts dose-dependently inhibited carrageenan-induced foot paw oedema in 7-day old chicks. Again, the ethyl acetate extract showed the greatest inhibition. The results of this study provide scientific evidence for the ethnomedicinal use of the leaves of C. splendens.

KEY WORDS: Clerodendrum splendens, Antimicrobial, Anti-inflammatory activity.

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Fleischer, et. al.: Antimicrobial and anti-inflammatory activities of the leaves of Clerodendrum splendens leaves

intervals over the next 6 hours post carrageenan injection. The right footpads of the chicks were injected intraplantar with carrageenan (10 µl of a 1% solution in saline). The change in foot thickness for the various groups was recorded hourly for six hours by means of a digital caliper. The oedema component of inflammation was quantified by measuring the foot thickness before carrageenan injection and at the various time points.

Statistical analysis of dataThe extracts were tested against test organisms in triplicates and the results were presented as the mean ± the standard error of means (SEM). Raw scores for the right foot thickness were individually normalized as percentage of change from their values at time 0 and then averaged for each treatment group. The time-course curves for foot thickness were subjected to two-way (treatment × time) repeated measures analysis of variance with Bonferroni’s post hoc t test. Total foot thickness for each treatment was calculated in arbitrary units as the area under the curve (AUC) and to determine the percentage inhibition for each treatment, the following equation was used.

% Inhibition of oedema = AUC control – AUC treatment

AUC control × 100

RESULTS AND DISCUSSION

Undeniably, plants have played very important roles in the lives of humans for centuries. C. splendens enjoys traditional use as anti-inflammatory and antimicrobial agents. This study was conducted on the leaves of C. splendens to validate these folkloric uses.

All the extracts showed some level of antimicrobial activity against Staph. aureus, B. subtilis, P. aeruginosa, E. coli and C. albicans in vitro, with the ethyl acetate extract exhibiting the highest activity (Table 1). The zones of inhibition ranged from 4.00 ± 0.5 4 mm to 9.0 ± 0.16 mm. The activity of the petroleum ether and ethanolic extracts ranged between 3.50 ± 0.50 mm to 4.67 ± 0.33 mm and 3.33 ± 0.47 mm to 4.3 ± 0.47 mm respectively. The least susceptible organism to the extracts was P. aeruginosa. Staph. aureus which generally causes infections that are very difficult to combat due to their multi drug resistance[9,10] was found to be susceptible to all extracts. Generally, the activities of the extracts were weak compared to the activities of the standard antibiotics used in the study.

The anti-inflammatory activity of the leaves of C. splendens was established using the carrageenan-induced oedema in chicks, a common experimental animal model used to evaluate NSAIDs. [11] It is believed to act in a biphasic manner. The initial phase of inflammation (0-2 h) has been attributed to the release of histamine and kinins, followed by a late phase (2.5-6 h) mainly sustained by release of prostaglandins.[12] The second phase is sensitive to most clinically effective anti-inflammatory drugs.[13] In this study, the time course curves revealed a dose-dependent effect of the extracts on oedema (Figure 1). Furthermore, when

Materials used in Anti-inflammatory StudiesDay old post-hatched Cockerels (Gallus gallus; strain Shaver 579) were obtained from Akropong Farms, a commercial breeder, in Kumasi. The chicks were housed in standard environmental conditions at the Department of Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, KNUST. The standard drugs used for the positive control were diclofenac sodium and dexamethasone. Carrageenan sodium (Sigma - Aldrich Inc., St Louis, MO, USA) was used to induce oedema in the chicks.

Preparation of extractsExtracts of C. splendens (10 mg/ml) were prepared in Dimethyl sulphoxide (DMSO) for the antimicrobial assay. Ciprofloxacin and Ketoconazole were used as the positive controls at a concentration of 0.5 mg/ml each.

Agar well diffusion bioassayThe inocula were prepared by inoculating the test organisms in nutrient broth and incubating them for 24 hours at 37°C for the bacteria, while for Candida albicans in Sabouraud’s dextrose broth was incubated for 48 hours. One milliliter of the diluted cultures was inoculated into a sterile molten nutrient agar at 45ºC and poured into a sterile petri dish. Similarly, 1 ml of the diluted fungal suspension was poured into sterile Sabouraud’s dextrose agar plates. These were swirled gently and allowed to solidify. Wells were bored into the solidified inoculated nutrient agar plates using cork borer number 6. The wells were filled with equal volume of 0.1 ml of each extract. One hour was allowed for the extract to diffuse into the agar after which the plates were incubated overnight at 37ºC and 25ºC for fungi and bacteria respectively. At the end of the incubation period, the diameter of inhibition zone(s) were measured with a ruler and recorded. The extracts and standard antibiotics were tested in triplicate and mean zones of inhibition were calculated for each extract and the standard antibiotics.

Anti inflammatory AssayThe anti-inflammatory properties of the extracts were evaluated using the carrageenan-induced foot oedema in 7-day old chicks as described by Roach and Sufka[8] with some modifications. The experiment was performed to evaluate the prophylactic effects of the petroleum ether, ethyl acetate and 70% ethanolic extracts on the oedema component of inflammation. Dexamethasone, a steroidal anti-inflammatory drug and diclofenac, a non-steroidal anti-inflammatory drug (NSAID) were used as positive controls. In this method, chicks were randomly selected, grouped (5 per group) and fasted for 24 hours before the experiment. Water was available ad libitum. The test samples were prepared by dissolving the fluid extracts in 2% tragacanth in distilled water. Doses of 30, 100 and 300 mg/kg were prepared and given orally (p.o) 1h before the carrageenan challenge and for the diclofenac (10, 30 and 100 mg/kg) and dexamethasone (0.1, 1.0 and 3 mg/ kg) were given intraperitoneally (i.p) 30 minutes before the carrageenan challenge. The foot thickness of each chick was measured before carrageenan injection (baseline measurement) and then at hourly

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Table 1: Antimicrobial Activities of C. splendens extracts

Extracts Mean Zones of Inhibition (mm)

E.coli B. subtilis Staph aureus P. aeruginosa C. albicans

Pet ether 4.2 ± 0.17 4.3 ± 0.67 4.2 ± 0.17 3.5 ± 0.50 4.7 ± 0.33Ethyl acetate 9.0 ± 0.17 7.0 ± 0.17 5.2 ± 0.44 4.0 ± 0.50 6.3 ± 1.1770% ethanol 3.7 ± 0.33 4.0 ± 0.56 4.3 ± 0.33 3.3 ± 0.33 3.7 ± 0.33Ciprofloxacina 20 ± 0.67 24 ± 0.50 17.5 ± 0.71 23.5 ± 0.43 –Ketoconazoleb – – – – 18 ± 0.302% DMSO 0 0 0 0 0

–; no assay performed, the data are shown as mean ± Standard Error of the Mean (SEM), a,b; positive controls

pet-ether

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ethanol

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Figure1: Time course effects of Petroleum Ether, Ethyl acetate and Ethanol Extracts (10-300 mg kg-1 p.o), in the prophylactic protocol on carrageenan induced foot oedema in the chick and their respective total oedema responses for 6 h [defined as the area under the time course curve (AUC)]. Each point on the column represents the Mean ± S.E.M. (n = 5). ***P < 0.001, **P < 0.01, *P < 0.05.

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inhibiting chemical mediators such as prostaglandins. The ethyl acetate extract exhibited the highest inhibitory effect in a dose-dependent manner at all doses with a maximal effect of 66.1 ± 3.67% at 300 mg/kg body weight. The extent of inhibition of the foot oedema by the extracts was less than the standard anti-inflammatory drugs, diclofenac and dexamethasone. Phytochemical screening revealed the presence of tannins, alkaloids, flavonoids, glycosides and sterols in the leaves. Some of these metabolites have been reported to possess antimicrobial activity.[10] Our results agree with that observed by Gbedema et al.[4] and lend further support to the use of the leaves of C. splendens for the treatment of wounds and microbial infections in traditional medicine. The results again provide support for the ethnomedicinal use of C. splendens in the treatment of inflammatory diseases.

total oedema over the period of the experiment was represented arbitrarily as AUC of the time course curves, all the extracts significantly reduced total oedema with a maximal inhibitory effect of 47.29 ± 8.65%, 66.09 ± 13.13% and 45.19 ± 5.09% respectively at 300 mg/kg (Table 2). Diclofenac (10-100 mg/ kg, i.p) also showed significant effect on the time course curve and total oedema with maximal inhibitory effect of 79.56 ± 18.24% at 100 mg/kg as seen in Figure 2. Similarly, treatment with dexamethasone, a steroidal anti-inflammatory agent, (0.3-3 mg/ kg, i.p) exhibited a significant effect on the time course curve of carrageenan-induced oedema (Figure. 2) with a maximal inhibitory effect of 78.69 ± 3.91% at 3 mg/kg. Thus, all the extracts inhibited oedema from the second hour (Figure. 1). The extracts may therefore be acting in the late phase of the inflammation by

diclo auc

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Figure 2: Time course effects of Diclofenac (10-100 mg kg-1, i.p) and Dexamethasone (0.3-3.0 mg kg-1 i.p) in the prophylactic protocol on carrageenan induced foot oedema in the chick and the total oedema response for 6 h). Each point and column represents the mean ± S.E.M. (n = 5) ***P < 0.001, **P < 0.01, *P < 0.05

Table 2: Inhibitory effects of Petroleum ether, ethyl acetate and 70% ethanolic extract on carrageenan-induced oedema on 7-day old chicks.

Extract 300 mg/kg 100 mg/kg 30 mg/kg

Pet ether 47.29 ± 8.65% % 46.43 ± 2.98 24.41 ± 3.97%Ethyl acetate 66.1 ± 3.67% 50.57 ± 0.67% 44.65 ± 4.77%70% Ethanol 45.19 ± 5.09% 19 ± 5.34% 11.11 ± 9.77%Diclofenac (100 mg/kg) 79.56 ± 18.24%Dexamethasone (3 mg/kg) 78.69 ± 3.91%

Values are mean ± S.E.M (n = 5), P < 0.001

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Revision of Ethnobotanical and Floristic Studies in Ghana. Accra: Science and Technology Press, CSIR, 2000:587

2. Burkhill HM. Useful Plants of West Africa. Kew: Royal Botanic Gardens, 1985:130.

3. Shehata AH, Yousif MF, Soliman GA. Egypt J. Biomed. Sci. 2001; 7:145.

4. Gbedema, S. Y., Kisseih, E., Adu, F., Annan, K. and Woode, E. Pharmacognosy Research 2010; 2:63.

5. Jung-Ho C, Wan-Kyun W, and Hong-Jin K. Arch. Pharmaceut. Res. 2003; 27:189.

6. Sofowora A. Medicinal plants and Traditional medicine in Africa. Ibadan: Spectrum Books Ltd, 1993

7. Harborne JB. Phytochemical methods: A guide to modern techniques of plant analysis. London: Chapman and Hall, 1973.

8. Roach JT, Sufka KJ. Brain Res. 2003; 994:215

9. Afolayan AJ, Aliero AA. Afr. J. Biotechnol. 2006; 5:369.

10. Cowan MM. Clin. Microbiol. Rev.1999; 12:564.

11. Di Rosa M, Willoughby DA. J. Pharm. Pharmacol. 1971; 23:297.

12. Di Rosa M. J. Pharm. Pharmacol. 1972; 24:89.

13. Vinegar R, Schreiber W, Hugo R. J. Pharmacol Exp. Ther. 1969; 66:96.

CONCLUSION

The present study demonstrates a weak antimicrobial activity compared to standard antibiotics and a good anti-inflammatory activity in chicks. Of the various extracts tested, the medium polar EtOAc extract showed the highest activity. The results support the wound healing activities of earlier reports, and provide the rationale for the ethnomedicinal use of the leaves of C. splendens in the management of inflammatory disorders. Flavonoids, tannins and alkaloids which were found present in the leaves of the plant may be responsible for these antimicrobial and anti-inflammatory activities.

REFERENCES1. Mshana, N. R., Abbiw, D. K., Addae- Mensah, I., Adjanouhoum, E., Ahyi, M. R. A.,

Odunlami, H., et al. Traditional Medicine and Pharmacopoeia; Contribution to the


Research Letter



*Correspondence:Dr. G. Venkateswara Rao, Principal Scientist,CavinKare Research Centre, Chennai - 600 032.E-mail: [email protected]: 10.5530/pc.2011.1.7

activity at 1, 5 and 25% level. At all the three concentrations, the essential oil showed more significant activity than 1% permethrin based product.[7] Previous reports on this plant occurring in different regions yielded, furanoditerpenoids,[8] terpenoids,[9-10] steroids[11] and aromatic esters.[1] However, no information was available on the preparation of an appropriate selective extract or fraction of the plant and its efficacy directed towards promoting hair growth or retarding hair fall or isolation of hair growth active compounds based on bioassay. In continuation of our interest on the isolation of biologically active molecules from medicinal plants for personal care applications,[12-21] we have undertaken the chemical examination of the rhizomes of H. spicatum. The present study describes the isolation of two known compounds, pentadecane (1) and an aromatic ester, ethyl p-methoxycinnmate (2) and hair growth studies of crude hexane extract, fractions and active compound.

MATERIALS AND METHODS

GeneralMelting points reported are uncorrected. UV spectra were recorded on Shimadzu UV spectrophotometer. IR spectra were recorded on a Shimadzu IR prestige 21. GC spectra were recorded in Shimadzu GC-17A capillary GC. 1H and 13C NMR spectra were recorded on a Bruker AMX 400 in CDCl3 with TMS an internal standard and the chemical shifts being represented in parts per million (ppm, d values). GC-MS mass spectrum on a Jeol SX 102/DA 6000 mass spectrometer. Column chromatography was performed on silica gel (100-200 mesh, Acme synthetic chemicals, Mumbai, India). Fractions and purity of the compounds were monitored by analytical thin layer chromatography (TLC) and the spots were visualized by exposure to iodine vapour or 5% sulphuric acid in methanol followed by heating the plate at

Chemical Examination and Hair Growth studies on the Rhizomes of Hedychium spicatum Buch.-hamG. Venkateswara Rao*, T. Mukhopadhyay, M. S. L. Madhavi, S. Lavakumar

M/s. CavinKare Research Centre, 12, Ekkattuthangal, Chennai-600 032, India

INTRODUCTION

Hedychium spicatum (Zingiberaceae), also known as spiked Ginger Lily is employed in the preparation of Abir, a fragrant coloured powder used during the Holi festival. The rhizomes possess strong aromatic odour and bitter camphoraceous smell. The rhizomes of the plant have been used in the preparation of cosmetic powders used for promoting hair growth. The rhizomes are also considered to have insect-repelling properties and are used for preservating clothes. The rhizomes are stomachic, carminative, stimulant and tonic, and are used in dyspepsia in the form of powder or decoction.[1] The rhizomes are much used in veterinary medicine.[2] The prior literature on Hedychium spicatum reveals that the cosmetic composition containing this plant extract regulates the firmness, tone or structure of skin or regulate wrinkles.[3] The compositions containing extract of Hedychium spicatum are useful for treating Tinea infections by topical application.[4] The ethanolic extract of rhizomes of H. spicatum possessed anti-inflammatory and analgesic activity. The anti-inflammatory activity was found in the hexane fraction and the compound hedychienone was found responsible for such activity and the analgesic activity was found in benzene fraction.[5] The cinnamic acid ester, obtained from the extracts of H. spicatum and Alpinia galanga and the same has been patented for natural sunscreen property.[6] The essential oil extracted from the rhizomes of H. spicatum was evaluated for in-vitro pediculicidal

ABSTRACT: The hexane extract of the rhizomes of H. spicatum yielded two known compounds, pentadecane, and ethyl p-methoxycinnamate. The structures of these compounds were established by spectroscopic data (UV, IR, GC, 1H and 13C NMR, Mass) and comparison with an authentic compounds. The crude extract, fractions and one of the isolated compounds showed hair growth property.

KEYWORDS: Hedychium spicatum, rhizomes, pentadecane, hair growth activity.

Research Letter

91

Rao, et. al.: Chemical Examination and Hair Growth studies on the Rhizomes of Hedychium spicatum Buch.-ham

food and water ad libitum. The floor mat husk in each cage was removed and laid afresh on daily basis.

Hair growth activity in vivoThe hair on the dorsal portion of the body of each animal was depilated using a standard, commercially available depilatory cream. After removal of the hair, the skin was cleaned with distilled water and wiped with surgical spirit. Four centimeter square area in the depilated dorsal skin was marked with permanent ink marker. The animals which showed any skin irritant response to the depilatory were removed from the experiment and new animal was replaced.

The rats were divided into 3 groups of 6 animals each. Group 1 animals were served as negative control without any treatment. The negative control comprised of the vehicle for application (only) without having any active extract/fraction/compound. Group 2 animals were applied 50 micro liters of commercial 2% Minoxidil solution in the pre defined area. The group 3 animals were applied samples (extract/fractions/compounds) prepared in liquid paraffin at 2%. The quantity of the solution used for the experiment was 50 micro liters per 4 cm sq area per animal. The application of the Minoxidil and the test samples were continued for 30 days. The observations such as hair growth initiation time in days and hair growth completion time in days were recorded for all the animals on daily basis. The hair growth initiation time was defined as the presence of new hair in the treated site of 4 cm sq area. The hair growth completion time was defined as complete filling of hair in the treated site of 4 cm sq area in each animal which become indistinguishable from the adjacent untreated portion of the body. The average of hair growth initiation time and hair growth completion time was calculated for each group along with control animals. The untreated control for hair growth initiation time (HGIT) is 10 days and hair growth completion time (HGCT) is 30 days. The percentage reduction in hair growth completion time (% Reduction in HGCT) for the treatment is calculated by the formula given below. The results of hair growth activity are shown in [Table 1].

Calculation = HGCT in untreated control – HGCT in test sample

HGCT in untreated control × 100

110ºC for 5 min. The TLC was performed on pre-coated silica gel plates (aluminium sheets 20X20 cm, silica gel 60 F254 plates of Merck KGaA, Germany). All solvents and reagents used were of analytical grade obtained from Merck. Pentadecane was obtained from M/s. Sigma aldrich, USA.

Plant materialThe rhizomes of Hedychium spicatum were obtained from bazaar in December, 2007 and was authenticated by Dr. P. Santhan, botanist, M/s. Durva Herbal Centre, Chennai. A voucher specimen was deposited in M/s. CavinKare Research Centre, Chennai.

Extraction and isolationThe air dried and finely powdered rhizomes (2.2 kg) were extracted with hexane through soxhlet apparatus for 8 hrs. The dilute extract was filtered and evaporated to dryness in vacuo using a rotary evaporator at 40oC to get crude hexane extract (33g). The crude hexane extract was submitted for hair growth studies and found to shown good hair growth.

Part of the crude hexane extract (30g) was subjected to column chromatography eluted with hexane, hexane: chloroform (1:1, 1:3) and chloroform to get corresponding fractions 4.3g (Fr. I), 15.5g (Fr. II) and 8.8g (Fr. III), respectively. All three fractions were submitted for hair growth studies along with crude hexane extract. Out of three, fraction I showed good hair growth promotion activity. Part of the fraction I, 1.0g was subjected to normal silica gel chromatography followed by repeated silver nitrate impregnated column chromatography with hexane: chloroform (95:5) yielded colorless compound 1 (130mg). Compound 2 (1.6 g) obtained from fraction II as colorless solid which was further crystallized in hexane to afford colorless crystalline compound.

Compound 1: Colorless oil; 1H NMR (400 MHz, CDCl3): d 0.86 (6H, s), 1.26 (26H, s). 13C NMR (100 MHz, CDCl3): d 14.3 (C-1,15), 22.9 (C-2,14), 29.3 (C-3,13), 29.6 (C- 4 to12).

Compound 2: Colorless crystals; mp = 49-50oC; IR (KBr): 2931, 1711, 1605, 1512, 1250, 1150, 830 cm–1; 1H NMR (400 MHz, CDCl3): d 7.62 (1H, d, J = 16.0 Hz), 7.45 (1H, d, J = 8.8Hz), 6.88 (1H, d, J = 8.8Hz), 6.29 (1H, d, J = 16.0Hz), 4.25 (2H, q, J = 7.1Hz), 3.82 (3H, s), 1.32 (3H, t, J = 7.1Hz) ; 13C NMR (100 MHz, CDCl3): d167.5, 161.1, 144.2, 129.8, 129.4, 127.3, 115.8, 114.4, 114.4, 60.5, 55.4, 14.3.

Hair growth promotion activityThe hair growth promotion activity was studied by using in vivo animal model[15],[22].

Animals: Female Wistar rats weighing 120-150 g, from Dr. MGR Janaki College, Chennai were used for hair growth study. Based on the guidelines of the ethical committee of the college, the animals were maintained in a clean cage and were provided with

Table 1: Comparison of in-vivo hair growth promotion activity

Extract/Fraction/

Compound

Hair growth initiation Time (HGIT in days)

Hair growth completion Time (HGCT in days)

% Reduction

in time

Hexane extract 8 20 33Fraction 1 8 20 33Pentadecane 7 21 30Minoxidil 6 16 47Untreated control

10 30 0

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The results of hair growth promotion (Table 1) showed that crude hexane extract was required less time than pure compound, pentadecane. It is worth mentioning that many crude extracts or active fractions are showing better activity than individual compounds.

CONCLUSION

To our best knowledge, the present study is the first report of the isolation of active compound from Hedychium spicatum for hair growth studies.

ACKNOWLEDGEMENT

We thank Mr. C.K. Ranganathan, CMD and of CavinKare Pvt. Ltd., Chennai for his interest, constant encouragement and providing necessary facilities. We are also thankful to Dr. K. S. Rao for isolating the compounds.

REFERENCES1. The Wealth of India, CSIR, New Delhi, 2001, vol. 5, 11,

2. Tayal JN and Dutt S. Proc Nat Acad Sci India 1940; 10A:47-51.

3. Martin KM and Saliou C. Compositions containing H. spicatum and use thereof. PCT Int Appl.WO 2002; 02056859.

4. Chuahan VS, Satyan KS and Kadam KP. Herbal compositions for Tinea infections. US patent 2009:7635493.

5. Srimal RC, Sharma SC and Tandon JS. Anti-inflammatory and other pharmacological effects of Hedychium spicatum (Buch-Hem). Ind J Pharmacol 1984; 16:143-147

6. Mitra SK, Babu UV and Ranganna MV. Natural sunscreen compositions and processes for producing the same. US Patent 2007; 7311896.

7. Varsha J, Anagha K and Kadam VJ. In-vitro pediculidical activity of H. spicatum essential oil. Fitoterapia 2007; 78:470-473.

8. Sharma SC, Tandon JS and Dhar MM. 7-Hydroxyhedychenone, a furanoditerpene from H. spicatum. Phytochem 1976; 15:827-828.

9. Joshi S, Chanotiya CS, Agrwal G, Prakash O, Pant AK and Mathela CS. Terpenoid compositions and antioxidant and antimicrobial properties of the rhizomes essential oils of different Hedychium species. Chemistry and Biodiversity 2008; 5:299-309.

10. Botini AT, Garfagnoli DJ, Delgado LS, Dev V, Duong ST, Kelley CG, Keyer R, Raffel R, Joshi P and Mathela CS. Sesquiterpene alcohols form H. spicatum var. acuminatum. J Nat Prod 1987; 50:732-734.

11. Shekhar CS, Shukla YN and Tandaon JS. Alkaloid and terpenoids of Ancitrocladus heyneanus, Sagittaria sagitifolia, Lyonia Formosa and H. spicatum. Phytochem 1975; 15:578-579.

12. Rao GV, Annamalai T, Mukhopadhyay T and Madhavi MSL. Chemical constituents and melanin promotion activity of stems of Cissus quadrasngularis Linn. Res J Chem Sci 2011; 1, 24-28.

RESULTS AND DISCUSSIONS

The initial screening of the hexane extract of the rhizomes of H. spicatum showed positive response in hair growth promotion activity. The bioassay guided purification of the hexane fractions of the rhizomes of H. spicatum repeated chromatography with a silica gel and re-crystallization with solvents furnished pentadecane and ethyl p-methoxycinnamate. The structure of the compounds were elucidated on the basis of UV, IR, GC, 1H and 13C NMR and Mass spectral data and comparison with an authentic samples.

The hair growth promotion activity of pentadecane showed good reduction in hair growth time, where as minoxidil, a positive control showed an excellent activity in the standard method but it had other side effects[23]. Even though the plant is being used in the preparation of hair oils, so far no reports on the compounds responsible for hair growth promotion activity.

The compound 1 was readily recognized as hydrocarbon by its preliminary spectral data. Its molecular formula was established as C15H32 by GC-MS, M+ 212. Its IR and UV spectra showed no characteristic peaks. Its proton spectrum showed only two peaks: methyl at d 0.86 (s) and methylene at d 1.26 (s). Its carbon spectrum showed the presence of four signals. It showed methyl and methylene carbons. Its mass spectrum showed m/z value 212. Based on the spectral data the compound has been identified as pentadecane.[24] To confirm further the compound, pentadecane has been purchased from M/s. Sigma-Aldrich, USA and analyzed by GC along with compound 1. The retention time of both the compounds were exactly matching with each other. Thus, the compound 1 has been established as pentadecane.

The compound 2 was identified as colorless crystals from hexane: chloroform, mp:49-50oC. It was readily recognized as aromatic acid ester based on its preliminary spectral data. Its molecular formula was established as C12H14O3 by GC-MS, M+ 206. The IR spectrum showed the presence of an ester peak at 1711cm-1 in the molecule. Its proton spectrum showed the presence of four aromatic protons at d 7.45 (2H, d, J = 8.8Hz) and 6.88 (2H, d, J = 8.8Hz), one aromatic methoxyl group at d 3.82 (3H, s), two double bond protons each showed as doublet at d 7.62 and 6.29 (J = 16.0Hz), one oxymethylene group at d 4.25 (2H, q, J = 7.1Hz), one methyl at d 1.32 (3H, t, J = 7.1Hz). Based on the aromatic proton integration, the molecule has 1,4 di-substitution patteren. The two olefinic protons showed large coupling constant indicates that these two protons are in trans position. The carbon spectrum showed total of 12 carbons including ester carbonyl at d 167.5. Out of twelve, eight double bond carbons at d 161.1, 144.2, 129.8, 129.4, 127.3, 115.8, 114.4, 114.4, of which six aromatic and two olefinic carbons. It also showed one methoxy carbon at d 55.4, one oxy methylene carbon at d 60.5 and one methyl carbon at d 14.3. By revealing the literature, the spectral data of the compound 2 is exactly matching with those of previously reported values. So, the compound 2 has been identified as ethyl p-methoxycinnamate.[24-25]

O

MeO

O

CH3

1

2

Figure 1: Compounds from H. spicatum

93


20. Rao GV, Annamlai T and Mukhopadhyay T. Nardal, a new sesquiterpene aldehyde from the plant, Nardostachys jatamansi DC. Ind J Chem 2008; 47B: 163-165.

21. Rao GV. Chemical constituents and biological studies of Chloroxylon swietenia DC: A review. Indian Drugs 2008; 45:5-15.

22. Adirajan N, Ravikumar T, Shanmugasundaram N and Mary B. In vivo and in vitro evaluation of hair growth potential of Hibiscus rosa-sinensis Linn. J. Ethnopharmacol 2003; 88:235-238.

23. Semalty M, Semalty A, Joshi GP and Rawat MS. Development and in vivo studies of Herbal hair oil for hair growth promotion. Indian Drugs 2010; 47:28-32.

24. Yu J, Yu D, Sun L, Zhang S, Zheng C and Chen Y. The chemical constituents of diterpenoids from Kaempferia marginata Carey. J Chinese Pharm Bull 2010; 10:61-64.

25. Benjamin L, Arno D, Maria THF, Andreas J and Ramon RT. A practical, efficient and atom economic alternative to the Wittig and Horner-Wadsword-emmons reactions for the synthesis of (E)- α,β-unsaturated esters from aldehydes. Tetrahedron 2006; 62:476-482.

13. Rao GV, Annamalai T and Mukhopadhyay T. Chemical examination and biological studies on the bark of Crataeva nurvala Buch.-Ham. Pharmcog J 2011; 3:1-4.

14. Rao GV, Mukhopadhyay T, Annamalai T, Radhakrishnan N and Sahoo MR. Chemical examination and biological studies of Origanum vulgare Linn.. Phar. Res 2011; 000

15. Rao GV, Annamalai T and Mukhopadhyay T. Phytochemical investigation and hair growth studies on the rhizomes of Nardostachys jatamansi DC. Pharm. Mag 2011; 7:142-146

16. Rao GV, Mukhopadhyay T and Radhakrishnan N. Artoindonesianin F, a potent tyrosinase inhibitor from the roots of Artocarpus heterophyllus Lam. Ind J Chem 2010; 49B:1264-1266.

17. Rao GV, Rao KS, Annamalai T, Radhakrishnan and Mukhopadhyay T. Chemical Constituents and Mushroom Tyrosinase Inhibition Activity of Chloroxylon swietenia Leaves. Turk J Chem 2009; 33:521-526.

18. 18. Rao GV, Rao KS, Annamalai T and Mukhopadhyay T. A new coumarin derivative from the leaves of Chloroxylon swietenia DC. Ind J Chem 2009; 48B: 1041-1044.

19. Rao GV. Chemical constituents of Adiantum genus: A review. Indian Drugs 2008; 45:837-858.


World Wide Web



*Correspondence: Tel.: +61 7 37357637; fax: +61 7 37355282E-mail: [email protected], [email protected]: 10.5530/pc.2011.1.8

Inside Pharmacognosy – A BlogI.E. Cocka,b*

Editor in Chief, Pharmacognosy Communications

aBiomolecular and Physical Sciences, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Brisbane, Queensland 4111, Australia. bEnvironmental Futures Centre, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Brisbane, Queensland 4111, Australia.

Pharmacognosy Network Worldwide (www.phcog.net) has established a blog for researchers interested in Pharmacognosy and medicinal plant research. The Inside Pharmacognosy – A Blog website can be accessed at http://www.pharmacognosy.in/. The blog combines reviews of new publications related to this expanding field with profiles of international departments and institutes that are engaged in Pharmacognosy research. A recent visit to the website showed 14 reviews of Pharmacognosy related books and texts, 4 profiles of Pharmacognosy research departments and details of an independent pharmacognosy consulting service, for the month of May 2011 alone. The blog began in October 2010 and has been steadily growing since, to reach its current size.

Archives of previous posts are also readily available for access by readers of the blog.

The blog also serves to notify readers of the publication of new issues of journals under the Pharmacognosy Network umbrella. The names, scope and contacts of other related journals within the pharmacognosy/natural product/plant science fields are also provided in the blog. This is a valuable resource for researchers deciding for which journal their research is best suited. The blog is well set out and easy to use. The reader can either read the latest posts or search for relevant articles under the categories of books, databases, departments worldwide, journals, organisations/associations and resources. Whether you are involved in phytochemical studies, bioactivity investigations or ethnobotanical research, Inside Pharmacognosy – A Blog is worth visiting, bookmarking and/or signing up for the blog newsletter (this is a free notification service). I encourage all researchers in pharmacognosy and related fields to read this site and submit relevant articles. I reviewed this site on 3rd June 2011.


Medicinal Plant Images



DOI: 10.5530/pc.2011.1.9

Eucalyptus ficifolia and Xanthorrhoea johnsonii

Figure 1: Eucalypts are the most iconic Australian medicinal plants and are possibly the most useful commercially for their medicinal properties (including antimicrobial, insect repellent, pesticidal, anticough and decongestant bioactivities). Eucalyptus is a diverse genus of trees in the family Myrtaceae. Of the more than 700 species that comprise this genus, most are endemic to Australia. A smaller number are also native to New Guinea, Indonesia and the Philippines. Pictured is the red flowering species Eucalyptus ficifolia (also known as Corymbia ficifolia). Photograph taken in Brisbane Australia by Dr Ian Cock.

Figure 2: The genus Xanthorrhoea (Australian grasstrees) is a small genus of slow growing and very long living plants endemic to Australia. The leaves of Xanthorrhoea johnsonii (pictured) have recently been shown to have an anaesthetic effect (Cock and Kalt, 2010) similar to the effects previously described for tubocurarine, dimethyltubocurarine and alcuronium (collectively known as curare, a South American arrow poison) from Chondrodendron tomentosum. Photograph taken in Toohey Forest, Brisbane Australia by Dr Ian Cock.

Cock IE, Kalt FR, Toxicity evaluation of Xanthorrhoea johnsonii leaf methanolic extract using the Aretemia franciscana bioassay. Phcog Mag 2010; 6, 23: 166-171.


Department Profile



*Correspondence: Tel.: +61 7 37357637; fax: +61 7 37355282.E-mail: [email protected], [email protected]: 10.5530/pc.2011.1.10

Sciences (BPS) is one of four schools that comprise SEET (the others being Engineering, Environment and Information and Communication Technology). BPS offers degree programs and postgraduate studies in diverse fields including the physical sciences, biomolecular and biomedical sciences, medical science, forensic sciences and aviation. Both traditional and emerging science disciplines are taught. The facilities include modern research facilities, with access to most modern technologies.

BPS researchers undertake a diverse range of research with projects including but not limited to:

• Medicinal agents discovery from Australian and international plants and fungi.

• Mechanistic studies into the toxicity of Australian native plants.• Cancer drug discovery and cancer therapies.• Molecular modelling for drug discovery and design.• Novel antimicrobial agents, antimicrobial mechanisms and

antimicrobial therapies.• Antimalarial drugs and antimalarial therapies.• Ataxia telangiectasia.• Neurodegenerative disorders, treating Parkinson’s and

Alzheimer’s Diseases.• Stem cell research and stem cell therapies.• Cytotoxic natural products from marine invertebrates.• Ecosystem restoration.• Drug design with novel target proteins to fight parasitic diseases.• The molecular basis of symbiosis in insects.

Biomolecular and Physical Sciences, Griffith University, AustraliaI.E. Cocka,b*

Editor in Chief, Pharmacognosy Communications

aBiomolecular and Physical Sciences, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Brisbane, Queensland 4111, Australia. bEnvironmental Futures Centre, Nathan Campus, Griffith University, 170 Kessels Rd, Nathan, Brisbane, Queensland 4111, Australia

Upcoming issues of Pharmacognosy Communications will be feature departmental profiles from the authors and readers of Pharmacognosy Communications. To begin, I have included a profile of my own department, Biomolecular and Physical Sciences, at Griffith University, Australia. We welcome departmental profile contributions from all regions of the world where pharmacognosy research and studies occur.

Griffith University consists of five main campuses in the Brisbane and Gold Coast region of southeastern Queensland, Australia. The university has diverse and unique settings, with two campuses sitting in a bushland/nature conservation area (Nathan and Mt Gravatt campuses), one campus in a rural setting bordered by farmland and a golf course (Logan campus), one campus in an urbanised coastal region (Gold Coast campus) and another campus in the central business district (CBD) of Brisbane (Southbank campus).

The University currently has approximately 40,000 students and 4,000 full time equivalent staff. Science, Environment, Engineering and Technology (SEET) is one of four main academic groups/faculties that comprise the university. SEET is further divided into individual schools. The School of Biomolecular and Physical

Department Profile

97

Cock: Biomolecular and Physical Sciences, Griffith University, Australia

• Regulation of cell surface sialylation by targeting the CMP-sialic acid transporter: towards the development of anti-metastatic agents.

• The role of Semaphorins in the immune system, neuronal development and cancer development.

• The use of natural product scaffolds in the generation of novel chemical libraries.

• Transcriptional control of gametocytogenesis.• Wolbachia’s role in nematodes.• Natural compounds from traditional Chinese medicine (TCM).

This is by no means a complete listing of the research projects undertaken in BPS at Griffith University. For a more comprehensive and up to date listing, see the Griffith University web site.[1] New projects will be listed on this site as they become available.

• The replicative mechanisms of thermophilic bacteria.• Investigation of metal based small molecule drug targets.• Molecular probes for pancreatic cancer.• Novel antimicrobial agents from bacteriophage proteins that

interfere with DNA replication.• Novel therapeutics for Human African trypanosomiasis.• Protein engineering of variants of the Green Fluorescent

Protein (GFP).• Regulation of apoptosis (programmed cell death).

Figure 1: The diversity of Griffith University’s campuses: (a) Nathan campus, (b) and (c) the unique bushland setting of Nathan campus, (d) Mt Gravatt campus, (e) Mt Gravatt campus surrounded by Toohey Forest, (f) Logan campus, (g) the rural setting of farmland adjoining Logan campus, (h) Southbank campus, (i) the river side setting of Southbank campus, (j) Gold coast campus and (k) the coastal setting surrounding Gold Coast campus.

Figure 2: Research postgraduate students in a Biomolecular and Physical Sciences research laboratory at Nathan campus.

Figure 3: Dr Derek Kennedy of Biomolecular and Physical Sciences instructing a student in a laboratory on Gold Coast campus, Griffith University.

98

Cock: Biomolecular and Physical Sciences, Griffith University, Australia

in Australia and 30th in the Asia-Pacific region for research outputs. [3] The university is currently experiencing rapid growth and whilst it already outperforms many larger universities, the university’s administration is predicting further improvement in Griffith University’s ranking in future years.

RefeRenceS1. http://www.griffith.edu.au/science-aviation/school-biomolecular-physical-

sciences/research/research-projects

2. http://www3.griffith.edu.au/03/ertiki/tiki-read_article.php?articleId=28602

3. http://www3.griffith.edu.au/03/ertiki/tiki-read_article.php?articleId=29462

Griffith University in general and BPS specifically provides an outstanding research environment for its staff. Recently, Excellence in Research for Australia (ERA) ranked the university in the top eight research universities in Australia.[2] Forty five research disciplines within the university were regarded as performing above world standard with some research fields (including the physical sciences) awarded the highest possible ranking for outstanding research. Indeed, 93% of the university’s researchers have been assessed by ERA as being world standard or better. Furthermore, recent Nature rankings (based on the number of primary research articles published in the Nature family of journals in a one year period) ranked Griffith University seventh


Upcoming Events



DOI: 10.5530/pc.2011.1.11

August 11-14, 2011 8th Brazilian Symposium of Pharmacognosy and 11th International Symposium of the Brazilian Society of Pharmacognosy, Brazil.

http://www.unb.br/fs/farmacognosia/

August 14-18, 2011 7th European Conference on Marine Natural Products, Strömstad, Sweden. http://fkogserver.fkog.uu.se/7ecmnp/

August 17-19, 2011 International Symposium on Medicinal and Aromtaic Plants, Flores Petén, Guatemala. http://www.imaps2001-peten.org/

September 4-9, 2011 59th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Producy research, (GA2011), Antalya, Turkey.

http://www.ga2011.org/

September 11, 2011 42nd International Symposium on Essential Oils (IESO 2011), Antalya, Turkey. http://www.iseo2011.org

September 27-30, 2011 PSE Conference Bari: Phytochemicals in Nutrition and Health, Giovinazzo (Bari), Italy. http://www.phytochemicalsociety.org/bari

October 17-20, 2011 5th InternationaL Conference on Polyphenols and Health (ICPH), Barcelona, Spain. http://www.icph2011barcelona.com/

November 3-6, 2011 BIT’s 9th annual Congress of International Drug Discovery Science and Technology (IDDST), Shenzen, China. http://www.iddst.com/iddst2011-06-01

December 10-15, 2011 Phytochemical Society of North America, 50th annual meeting, Fairmont Orchid, Kohala Coast Hawaii. http://psna.uhhconferencecenter.com/


Instructions for Authors



DOI: 10.5530/pc.2011.1.12

Pharmacognosy Communications (www.phcogcommn.org) is a new journal published by Pharmacognosy Network Worldwide (www.phcog.net). It is a peer reviewed journal aiming to publish high quality original research articles, methods, techniques and evaluation reports, critical reviews, short communications, commentaries and editorials of all aspects of Pharmacognosy and medicinal plant research. The journal is aimed at a broad readership, publishing articles on all aspects of pharmacognosy, and related fields. The journal aims to increase understanding of pharmacognosy as well as to direct and foster further research through the dissemination of scientific information by the publication of quality manuscripts. The submission of original contributions in all areas of pharmacognosy are welcomed.

The journal aims to report the latest outstanding developments in the field of pharmacognosy and natural products and drug design covering but not limited to the following topics:


isolates• Natural product discovery and evaluation• Mechanistic studies• Method and technique development and evaluation• Isolation, identification and structural elucidation of natural


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This section should relate the results section to current understanding of the scientific problems being investigated in the field. Description of relevant references to other work/s in the field should be included here. This section also allows the author to discuss the significance of the results - i.e. does the data support the hypotheses you set out to test? This section should end with new answers/questions that arise as a result of the author’s work.

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Materials and/or subjects utilized in the study, as well as the procedures undertaken to complete the work should be included in this section. The methods should be described in sufficient detail such that they could be repeated by a competent researcher. The sources of all major instruments and reagents used (kits, drugs, etc) must be given with parentheses. Illustrations and/or tables may be helpful in describing complex equipment or elaborate procedures. Statistical tools used to analyze the data should be mentioned. All procedures involving experimental animals or human subjects must accompany a statement on ethical approval from appropriate ethics committee.

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new lead anti-malarial compounds, several research groups screen plant extracts to detect secondary metabolites with relevant biological activities that could serve as templates for the development of new drugs. Flavonoids have been isolated and characterized from many medicinal plants used in malaria endemic areas.[1-2] However, controversial data have been obtained regarding their antiplasmodial activity, probably because of their structural diversity.[3,5,6] More recently, several flavonoids have been isolated from Artemisia afra[7] and Artemisia indica[8] two plants related to Artemisia annua, the famous traditional Chinese medicinal plant from which artemisinin is isolated.

rEFErEncE stylE

Journal References1. Standard journal articleSingle/Multiple Authors:List the first six authors followed by et al. (Note: NLM now lists all authors.)Halpern SD, Ubel PA, Caplan AL. Solid-organ transplantation in HIV-infected patients. N Engl J Med. 2002 Jul 25; 347(4):284-7.

As an option, if a journal carries continuous pagination throughout a volume (as many medical journals do) the month and issue number may be omitted.

Halpern SD, Ubel PA, Caplan AL. Solid-organ transplantation in HIV-infected patients. N Engl J Med. 2002; 347:284-7.

More than six authors:Rose ME, Huerbin MB, Melick J, Marion DW, Palmer AM, Schiding JK, et al. Regulation of interstitial excitatory amino acid concentrations after cortical contusion injury. Brain Res. 2002; 935(1-2):40-6.

Organization as author:Diabetes Prevention Program Research Group. Hypertension, insulin, and proinsulin in participants with impaired glucose tolerance. Hypertension. 2002; 40(5):679-86.

Both personal authors and an organization as author:Vallancien G, Emberton M, Harving N, van Moorselaar RJ; Alf-One Study Group. Sexual dysfunction in 1,274 European men suffering from lower urinary tract symptoms. J Urol. 2003; 169(6):2257-61.

2. Journal article on the InternetSaraswathy A, Shakila R, Sunilkumar KN; Phcog.Net. HPTLC Fingerprint Profile Of Some Cinnamomum Species — Pharmacognosy Journal [Phcog J]. Pharmacognosy Journal. 2010 April; 2(8):211-215. Available from: http://phcogj.com/content/hptlc-fingerprint-profile-some-cinnamomum-species.

Hussain A, Mohammed S, Rizvi A, Wahab S; Phcog.Net. Pharmacognostical Standardization of Stem Bark of Adenanthera

figures are accepted provided they are of high quality. Please note that graphs produced by many statistical packages are rarely adequate. In particular, letter quality on axes and captions are often poor. Such figures should be exported into an accepted graphics package and lettering rendered using a text function. Authors should note that .dot, .bmp, and .pat fills should be avoided. Do not use postscript fill patterns. When filling illustrations, use fills such as lines, tints or solids. Line width minimum is 0.25 pt (0.09 mm). Also avoid the use of bitmap scans to render text and detail. Text should be saved as text at a minimum text size of 6 pt (2.1 mm). Submit line art as Corel Draw, Adobe Illustrator, or EPS files. These must be at a minimum resolution of 800 DPI at publication size. High resolution may be necessary where fine line detail is present. For graphs, Excel graphs are also acceptable. Note that vertical axes must all be at the same scale especially when the paper compares them. Otherwise they should be produced as separate figures. Avoid 3D plots when presenting 2D data. All tables and figures must be placed in appropriate places in the manuscript and when this is not possible, appropriate place must be indicated in the manuscript. Please note, good quality figures must be submitted as separate files as outlined above, apart from presenting a copy of the same at appropriate places in the manuscript.

Figures, tables or other materials copied verbatim or adopted from previously published materials, the author must have written permission from the the copyright holder of that material (publisher and/or authors) for reproduction in your article. A copy of the permission release must be submitted with the manuscript. It is the author’s responsibility to obtain permission.

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Figure captions/legends should be single spaced and typed in the journal format. Explanations should be brief and authors should keep in mind that captions/legends will be placed below figures. Tables are to be incorporated at the end of Manuscript.

AcKnowlEdgEMEnts

Those who have helped the authors carry out the study and/or prepare the manuscript but have not made significant intellectual contribution to deserve authorship must be acknowledged. Mention all applicable grants and other funding that supported the work.

rEFErEncEs

In-text citationCorrect/Acceptable FormatNatural products have proven to be a great source of new biologically active compounds. Thus, in an effort to discover

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8. Conference proceedingsHarnden P, Joffe JK, Jones WG, Editors. Germ cell tumours V. Proceedings of the 5th Germ Cell Tumour Conference; 2001 Sep 13-15; Leeds, UK. New York: Springer; 2002.

9. ThesisSenol FS. Pharmacognosic research on some Salvia species growing in Turkey. M.Sc. Thesis, Institute of Health Sciences, Gazi University, Ankara, Turkey, 2009.

10. WebsitesWebsite informationCancer-Pain.org [homepage on the Internet]. New York: Association of Cancer Online Resources, Inc.; c2000-01 [updated 2002 May 16; cited 2002 Jul 9]. Available from: http://www.cancer-pain.org/.

Manuscript SubmissionManuscripts may be submitted electronically through the online submission at the journals web site (http://phcogcommn.org/home). Alternately, manuscripts may be submitted by email - [email protected]. All submissions are peer reviewed by the editorial board and a select group of reviewers. Please make sure that all guidelines are followed carefully. All the accepted articles will be queued for publication and will appear in the futures issues based on the priorities set by the editorial board.

contActs

Editor-in-ChiefDr. Ian [email protected]

[email protected]

pavonina L. Pharmacognosy Journal. 2010 April; 2(8):240-246. Available from: http://phcogj.com/content/pharmacognostical-standardization-stem-bark-ad....

Abood S. Quality improvement initiative in nursing homes: the ANA acts in an advisory role. American Journal of Nursery [serial on the Internet]. 2002 June [cited 2002 Aug 12]; 102(6): [about 3 p.]. Available from:http://www.nursingworld.org/AJN/2002/june/Wawatch.htm

3. Book author(s)Murray PR, Rosenthal KS, Kobayashi GS, Pfaller MA. Medical microbiology. 4th ed. St. Louis: Mosby; 2002.

4. Editor(s), compiler(s) as authorGilstrap LC 3rd, Cunningham FG, VanDorsten JP, Editors. Operative obstetrics. 2nd Ed. New York: McGraw-Hill; 2002.

5. Author(s) and editor(s)Breedlove GK, Schorfheide AM. Adolescent pregnancy. 2nd Ed. Wieczorek RR, Editor. White Plains (NY): March of Dimes Education Services; 2001.

6. Organization(s) as authorRoyal Adelaide Hospital; University of Adelaide, Department of Clinical Nursing. Compendium of nursing research and practice development, 1999-2000. Adelaide (Australia): Adelaide University; 2001.

7. Chapter in a bookMeltzer PS, Kallioniemi A, Trent JM. Chromosome alterations in human solid tumors. In: Vogelstein B, Kinzler KW, Editors. The genetic basis of human cancer. New York: McGraw-Hill, p. 93-113; 2002.

Edited and Published by Dr. Mueen Ahmed KK on behalf of Pharmacognosy Network Worldwide [Phcog.Net], Bangalore 560 41

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