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PROTEIN AND PEPTIDE DRUG DELIVERY SYSTEM

PRESENTED BY

BRAJESH KUMAR

M. PHARM (1ST YEAR)

PHARMACEUTICS

BBAU LUCKNOW

,

2 Contents

I. Protein & Peptides

II. Structure of protein

III. Classification of protein

IV. Drug delivery system

V. Stability testing

VI. Conclusion

VII. References

3 1. Protein & Peptides

PROTEINS: Proteins are the large organic compounds made of amino acids arranged in a linear chain and joined together by peptide bonds.

Protein > 50 amino acids PEPTIDES: These are short polymers formed from the linking,

in a defined order of amino acids.

peptide < 50 amino acids

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Why protein and peptide are used;

The protein and peptides are very important in biological cells.

Lack of proteins and peptides causes diseases like Diabetes mellitus.

Diabetes mellitus is caused due to the lack of protein called INSULIN.

Now a days R-DNA technology and hybridoma techniques also used in protein and peptide based pharmaceuticals.

5 Continue…ADVANTAGES

Erythropoietin used for production of RBC.

Tissue plasminogen activator is used for Heart attack, Stroke.

Oxytocin maintain labor pain.

Bradykinin increases the peripheral circulation.

Somatostatin decrease bleeding in gastric ulcer.

Gonadotropin induce ovulation.

Insulin maintain blood sugar level.

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FUNCTIONS

Transport and storage of small molecules.

Coordinated motion via muscle contraction.

Mechanical support from fibrous protein.

Generation and transmission of nerve impulses.

Enzymatic catalysis.

Immune protection through antibodies.

Control of growth and differentiation via hormones

7 2. Structure of protein

The structure of protein divided in to four types:

1. Primary structure : The amino acid sequence.

2. Secondary structure: Regularly repeating local structures stabilized by hydrogen bond.

3. Tertiary structure : Three dimensional structure of polypeptide.

4. Quaternary structure: The structure formed by several protein molecules (polypeptide chains).

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9 3. Classification of protein

Depending on the number of amino acids they are classified as follows:

Polypeptides protein

Oligopeptides protein

Fibrous proteins

Globular proteins

Oligo meric proteins

10 4. Delivery of proteins

D R U G D E L IV E R Y C L A S S IF IC A T IO N

P u lm o n a ry P a re nte ra l T ra n sd e rm al

Im p la n ts O c u lar N a sal

M isce lla ne o us O ra l

R o u te o f A dm in is tra tion

P E G y la tion P ro -d ru g P o ly m er d e p ot

D ru g M o d ific a tion

D ru g D e liv e ry

11 Continue…Parenteral routes of administration;

Parenteral route is most efficient way for systemic delivery of proteins and peptides.

This is the best choice to achieve therapeutic activity

Mainly 3 routes of administration Intravascular Intramuscular Subcutaneous

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Advantages Route of delivery for 95% of proteins

Allows rapid and complete absorption .

Avoids first pass metabolism

Disadvantages

Problems with overdosing, necrosis

Local tissue reactions/hypersensitivity

Everyone hates getting a needle

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INTRAVENEOUS ROUTE: Excessively metabolized and tissue drug bound at the site of

IM can be administered by this route such as Insulin , Interferon etc.

DISADVANTAGES:

Causes pain, tissue necrosis and thrombocytopenia.

ADVANTAGES: Antibiotics can be administered.

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INTRAMUSCULAR ROUTE:

Gamma globulins given by this route are proved to have long-term protection from hepatic infection. some drugs given by this route include long acting insulin, GH.

DISADVANTAGES:

Not used for all proteins and peptide drugs because of metabolism of drugs at the site of injection.

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SUBCUTANEOUS ROUTE;

Controlled release is obtained from implantable polymeric devices.

These are prepared from crossed linked polymers which are biocompatible and biodegradable e.g. Poly lactic acid.

Release of Insulin, bovine serum albumin, LH was prolonged by this route.

16 4. PARENTERAL DRUG DELIVERY SYSTEM

Polymer based drug delivery system. Liposome based drug delivery system. Hydro gel based drug delivery system. Emulsion based drug delivery system.

PUMPS: Implantable infusion pumps Mechanical pumps

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18Parenteral routes of administration

Parenteral route is most efficient way for systemic delivery of proteins and peptides.

This is the best choice to achieve therapeutic activity. Mainly 3 routes of administration

INTRAVASCULAR INTRAMUSCULAR SUBCUTANEOUS

19• Route of delivery for 95% of proteins• Allows rapid and complete absorption

.• Avoids first pass metabolism

Advantages

• Problems with overdosing, necrosis• Local tissue

reactions/hypersensitivity• Everyone hates getting a needle

Disadvantages

20 INTRAVENEOUS ROUTE Excessively metabolized and tissue drug bound at the site

of IM can be administered by this route.

EX: Insulin, Interferon.

DISADVANTAGES: Causes pain, tissue necrosis and thrombopenia.

ADVANTAGES: Antibiotics can be administered.

21INTRAMUSCULAR ROUTE Gamma globulins given by this route are proved to have long-

term protection from hepatic infection.

some drugs given by this route include long acting insulin, GH.

DISADVANTAGES: Not used for all proteins and peptide drugs because of

metabolism of drugs at the site of injection.

22 Subcutaneous route Controlled release is obtained from implantable polymeric

devices.

These are prepared from crossed linked polymers which are biocompatible and biodegradable.

EX: polylactic acid

Release of Insulin, bovine serum albumin, LH was prolonged by this route.

23 Parenteral drug delivery system

Polymer based drug delivery system. Liposome based drug delivery system. Hydro gel based drug delivery system. Emulsion based drug delivery system.

PUMPS: Implantable infusion pumps Mechanical pumps

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25 Polymer based drug delivery system

polymers are used as carriers in this drug Pug delivery system.

CHARACTERS OF POLYMERS It should be biodegradable. It should be bio compatible. And non-toxic.

Two types of polymers are used widely natural polymers synthetic polymers

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Natural polymers: Collagen , hemoglobin and gelatin. Synthetic polymers: mainly poly esters like PLA and PGA are

used widely. Diffusion of drug out of the polymer

Drug Release by Polymer Degradation

o o oo o oo o oo

o o oo o

27Liposome based drug delivery

Spherical vesicles with a phospholipid bilayer Liposome's are microscopic vesicles composed of one or more

aqueous compartments.

Liposome’s in Proteins delivery : Example: Lecithin used in controlled drug release.

Liposome’s in peptide drug delivery: Bleomycin : A peptide with anti tumor activity, reduces normal

tissue toxicity. Negatively charged liposome's produces a prolonged

hypoglycemic effect in diabetic drugs, which are injected by subcutaneous injection.

28ADVANTAGES OF LIPOSOME DRUG DELIVERY Soluble in both organic and aqueous media. Liposome’s are important for targeting drugs directly to the

liver, and brain. Lipsosomes easily crosses blood brain barrier.

EXAMPLE: Dopamine converted to L-Dopa. Used as a vehicles for vaccines.

DISADVANTAGES Less stable , easily susceptible to oxidation. Hence liposome’s are replaced by noisome an alternate for

liposome’s.

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Hydrogel based dds

Hydrogels are three dimensional networks of hydrophilic polymers that are insoluble- Hydro gels are polymers which have the ability to swell in

water . Biodegradable hydro gels are used, due to its biocompatibility .

Examples: Hydroxymethylacrylate, used to minimize mechanical irritation to surrounding tissue.

30EMULSION BASED DELIVERY Emulsions can be used for parenteral drug delivery of proteins

and peptides used to prolong the release of drug. e.g. subcutaneous administration of muramyl dipeptide in a

w/o emulsion. It is used to potentiate immune system.

CELLULAR CARRIERS Protein and peptides can be incorporated in erythrocytes to

achieve the prolong release or targeting. Resealed erythrocytes as delivery system for c-reactive protein,

and mainly used to target liver and spleen.

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PUMPS

Types of pumps:

1. IMPLANTABLE PUMPS Drug is implanted subcutaneously, and delivered by I.V

infusion. Pumps are filled with drug through a septum with a needle. Pumps deliver drugs to central vein for 7-14 days a constant rate.

2. MECHANICAL PUMPS Easily manipulated to deliver protein and peptide drugs.

Example: insulin has been successfully delivered by portable syringe.

32NON PARENTERAL ROUTES OF ADMINISTRATION

Parenteral route is not properly achievable, hence other routes are preferred. Oral route. Rectal route. Nasal route. Pulmonary route. Buccal route. Transdermal route. Ocular route

33 ORAL ROUTE Encapsulated peptides or proteins in amino acids with

microsphere of approximately 10 micron in diameter , used for oral delivery.

Example: Insulin and heparin. Orally administered insulin produces hypoglycemic effect .

DISADVANTAGES:

Acid catalyzed degradation in stomach. Proteolysis in GIT.

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Transdermal route of administration

This is topical medication. Drug is absorbed through the skin.

EX: Insulin, vasopressin

ADVANTAGES: Controlled administration of drug is possible. Improved patient compliance. Drugs with short half lives can be administered.

DISADVANTAGES: High intra and inter patient variability. Low permeation because of high molecular weight. Hydrophilicity and lipophilicity of stratum corneum.

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Number of approaches are available for effective protein and peptide drug delivery. They are IONTOPHORESIS PHONOPHORESIS PENETRATION ENHANCERS PRODRUG Iontophoresis: Used for local and systemic delivery of proteins and

peptides. In this an electric current is used to drive the molecules across the skin surface.

Example: Transport of insulin using iontophoresis. Phonophoresis: The absorption is enhanced by thermal effect of

ultrasonic waves and subsequent alteration of physical structure of skin surface.

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Pulmonary route of administration

Lungs are attractive site for systemic delivery of proteins and peptides because of their enormous surface area(70 sq.m)

Alveoli and lungs are the absorption sites. Drugs are absorbed through lungs by simple diffusion, carrier

mediated transport

37ADVANTAGES: Decrease in dose requirement. Fast absorption Increased patient compliance

DISADVANTAGES: Inflammation may be observed in lungs. Degree of bioavailability was less due to hydrolytic enzymes

present in lungs

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Rectal route of administration

Rectum is highly vascularised body cavity. Rectal mucosa is devoid of villi. Drugs are in form of suppositories, gel, dry powders.

EX: Insulin, calcitonin

ADVANTAGES: Reduced proteolytic degradation. Improved systemic bioavailability with co-administration of

absorption enhancers.

EX: surfactants Large dose can be administered.

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OCULAR ROUTE In this route enkephalins, thyrotrophin releasing

hormones ,luteinizing hormones ,glucagon and insulin are administered

BUCCAL ROUTE Mucoadhesive dosage forms can be used for buccal route. Adsorption enhancers like salicylates or a surfactant is used for

protein and peptide delivery through buccal route.

Example: Oxytocin , vasopressin , insulin, are reported to be absorbed

through buccal mucosa . And adhesive gel, patches , tablets are used.

Insulin is absorbed through buccal mucosa in the presence of sodium glycolate.

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The drugs are absorbed through oral mucosa mainly through the non-keratinized regions.

ADVANTAGES: It can be attached or removed without any discomfort and pain. Well acceptability by patients. Drugs are absorbed rapidly.

DISADVANTAGES: Administration time is limited. Drug loss by accidental swallowing.

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Nasal route of administration

The nasal route has been employed for producing local action on the mucosa which is more permeable compared to oral mucosa.

Nasal absorption is through passive diffusion.

EX: Insulin, human growth hormone.

ADVANTAGES: Rapid onset of action First pass metabolism can be avoided Better drug absorption

DISADVANTAGES: Long-term usage causes toxicity. Size of proteins and peptide drugs reduces systemic

bioavailability.

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PROTEIN FORMULATIONS

1

• Protein sequence modification (site directed mutagenisis)

• PEGylation

2• Proteinylation• Microsphere encapsulation

3• Formulating with permeabilizers

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PEGYLATION PEG is a non-toxic, hydrophilic, FDA approved, uncharged

polymer. Increases in vivo half life. Decreases immunogenicity. Increases protease resistance. Increases stability.

CH

-CH

-CH

-CH

-CH

-CH

-CH

-CH

-CH

-CH

| | | | | | | | | |O

H O

H O

H O

H O

H O

H O

H O

H O

H O

H

+

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PROTEINYLATION

Attachment of additional or secondary (non-immunogenic) proteins for in

vivo protection.

Cross-linking with Serum Albumin.

Increases in vivo half life.

Cross-linking or connecting by protein engineering with antibody

fragments.

+

Protein drug

scfc (antibody)

45 FORMULATION WITH PERMEABILIZERS

Salicylates (aspirin)

Fatty acids

Metal chelators (EDTA)

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Stability testing

The capability of a particular formulation in a specific container/closure system to remain within its physical,chemical,microbiological,toxicological and protective specifications.

Evaluates the effect of environmental factors on the quality of the a drug substance or a formulated product which is utilized for prediction of its shelf life, determine proper storage conditions.

General

Selection of Batches

Container Closure System

Specification

Testing Frequency

Storage Conditions

Stability Commitment

Evaluation

Statements/Labelling

On-going Stability Studies  

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Conclusion

Protein and peptide based pharmaceuticals are rapidly becoming a very important class of therapeutic agents and are likely to replace many existing organic based pharmaceuticals in the very near future.

Peptide and protein drugs will be produced on a large scale by biotechnology processes and will become commercially available for therapeutic use.

Their need in the clinical & therapeutic regions has intensified the investigation for their convenient & effective delivery through noninvasive system.

48References

Controlled drug delivery concepts and advances by S.P vyas & Roop k.khar Understanding the Fundamentals of Peptides and Proteins

By GARY HU (Bio processing journal -trends in development in bio process technology)

PEPTIDES AND PROTEINS IN PHARMACEUTICALS    RATNAPARKHI M.P.,* CHAUDHARI S.P., PANDYA V.A. International Journal of Current Pharmaceutical Research

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