124 Nano Biomed. ng. , Vol Iss N ano B iomed Eng

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Nano Biomed Eng2019, 11(2): 124-137. doi: 10.5101/nbe.v11i2.p124-137.

Research Article

124 Nano Biomed. Eng., 2019, Vol. 11, Iss. 2

Synthesis and Antimicrobial Activity of Some New Barbituric Acid Derivatives Containing Thiazole Moiety from Sulfadiazine

Abstract

A new series of barbituric acid derivatives w prepared and characterized by Fourier-transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance (1H-NMR) and nuclear magnetic resonance (13C-NMR). The antimicrobial studies of synthesized compounds were screened for antibacterial activity against three different types of bacteria (Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli), and against two types of fungi (Candida albicans and Aspergillus flavus). In this research, a reaction of sulfadiazine with chloroacetylchloride gave 2-chloro-N-(4-(N-pyrimidin-2-ylsulfamoyl) phenyl) acetamide [A] which was reacted with thiourea, and with K2CO3 to give thiazole derivative [B]. The last compound (thiaole derivative) was reacted with chloroacetyl chloride in basic medium to form compound [R]. 5,5-Diethyl barbituric acid and 5-phenyl,5-ethyl barbituric acid were treated with formaldehyde to give 1,3-bis(hydroxyl methyl)barbituric acids derivatives [B1]and [B2]. Finally, the compound [R] reacted with 1,3-bis(hydroxyl methyl) barbituric acids derivatives via Williamson reaction to form new barbituric acid derivatives [RB1]and [RB2]. By the reaction of 5,5-diethyl barbituric acid with 5-phenyl,5-ethyl barbituric acid, new compounds of barbituric acids derivatives [RB3] and [RB4] were obtained. The biological applications of the synthesized compounds showed a greater effect in antimicrobial activities from the starting material (sulfadiazine).

Keywords: Sulfadiazine; Thiazole; Barbituric acid; Antimicrobial activity

Mahmood Mohi Fahad, Ezzat Hussein. Zimam , Majed Jary Mohamad

Department of Chemistry, Faculty of Science, University of Kufa, Iraq.

Corresponding author. E-mail: [email protected]

Received: Jun. 4, 2018; Accepted: Oct. 2, 2018; Published: Apr. 19, 2019.

Citation: Mahmood Mohi Fahad, Ezzat Hussein Zimam, and Majed Jary Mohamad, Synthesis and Antimicrobial Activity of Some New Barbituric Acid Derivatives Containing Thiazole Moiety from Sulfadiazine. Nano Biomed. Eng., 2019, 11(2): 124-137.DOI: 10.5101/nbe.v11i2.p124-137.

Introduction

Sulfadiazine is one of the antibiotic drugs that belong to the sulfonamide group; it has been utilized in veterinary and human therapy over 60 years [1]. Also, it is utilized for the treatment of urinary tract infections (UTIs) [2], and in clinic as a topical agent either alone or in combination with other compounds in the treatment of wound and burn infections [3]. Thiazoles are a group of heterocyclic compounds that possess a wide variety of biological activities and

their utility as medicaments is very much established [4]. The applications of thiazoles were found in drug development for the treatment of inflammation, bacterial, allergies, HIV infections hypertension, schizophrenia, hypnotics and in the treatment of pain [5]. Barbituric acid, or pyrimidine-2,4,6-(1H,3H,5H)-trione,H3BA, that contains five hetero atoms (three O and two N) is commonly known as barbiturates [6]. It was prepared by Adolf Von Baeyer in 1864 from a combination of the urea and malonic acid [7]. Barbituric acid has been extensively utilized in

125Nano Biomed. Eng., 2019, Vol. 11, Iss. 2


biological and medical studiws for many years, the best known of which is their sedative activity in the central nervous system [8]. They are a favour in medicinal chemistry as anticonvulsants, sedatives, hypnotics and anxiolytic agents [9].

Experimental

All reagents and solvents were purchased from commercial sources and used without purification. Melting points (m.p.) were recorded using Electro thermal melting point apparatus (UK). Fourier-transform infrared spectra were recorded using Shimadzu FTIR-8400S infrared spectrophotometer by KBr disc (Kufa University). Proton nuclear magnetic resonance (1H-NMR) and nuclear magnetic resonance (13C-NMR) were recorded by Bruker spectrometer, operating at (400MHZ for 1HNMR and 75 MHZ for 13C NMR ) with DMSO-d6 as a solvent (Abu Ali Center Lab, Mashhad, Iran). Thin layer chromatography (TLC) was performed on aluminum plates and coated with a layer of (0.25 mm) silica gel; compounds were detected by iodine vapor. Autoclave was used to sterilize agar media, supplied by Prestige Medical-England. Incubator was used to maintain different temperatures required for the growth of organism, supplied by Memert-Germany.

Synthesis of 2-chloro-N-(4-(N-pyrimidin-2-ylsulfamoyl)phenyl) acetamide[A] [10]

To a stirred mixture of sulfadiazine (0.01 mol) and triethylamine (1.67 mL) in dimethylformamide (DMF) as solvent, chloroacetyl chloride (0.01 mol) was added drop-wise by micropipette. After the addition was completed, the mixture was stirred for 3 h without heating. Finally, the solvent was evaporated, and the

precipitate was filtered, dried and washed with distilled water-ether to give compound [A]. The crystalline precipitates were re-crystallized with ethanol. Yield: Pale yellow, 87 %; m.p. = 220 – 222 °C; retardation factor (Rf) = 0.75; the TLC for the reaction was completed by using benzene : methanol = 4 : 1.

Synthesis of 4-(2-aminothiazol-4-ylamino)-N-(pyrimidin-2-yl)benzene sulfonamide [B] [11]

A mixture of [A] (0.01 mol), thiourea (0.01 mol) and anhydrous potassium carbonate (0.01 mol) in absolute ethanol (25 mL) was heated under reflux on water bath for 12 h. The excess of ethanol was removed by distillation and the residue was treated with 5% sodium carbonate solution to remove acid impurities. The yellow precipitate was filtered, washed with water several times and dried at 50 °C. Finally, the product was crystallized from ethanol. Yield: 78%; m.p.=178 – 180 °C; Rf = 0.58; benzene : methanol = 4 : 1 (Scheme 1).

2-chloro-N-(4-(4-(N-pyrimidin-2-ylsulfamoyl)phenylamino)thiazol-2-yl)acetamide[R] [12]

Preparation method of compound [R] was the same as preparation method of compound [A] in step 1: 0.01 mol of compound [B] and 0.01 mol of triethylamine in DMF; 0.01mol of chloroacetyl chloride was added drop-wise to the mixture. The reaction mixture was stirred for 4 h without heating. The product obtained after the evaporation of the solvent was filtered, washed with ether and re-crystallized with ethanol. Yield: Dark brown, 65%; m.p. = 211– 213 °C; Rf = 0.7; TLC for the reaction was completed by using benzene : methanol = 4 : 1.

Synthesis of 1 ,3-bis(hydroxyl methyl ) barbituric acids derivatives [B1]-[B2] [13]

5,5-diethyl barbituric acid and 5-phenyl,5-ethyl

Scheme 1 Preparation of thiazole derivative [B].

O

SHN

N

NHN

OS

Thiourea

N

NH2

K2CO3[B]

O

SHN

N

N

NH2ClCH2COCl

O

O

SHN

N

N

NH

O

Cl

O

TEA

[A][A0]



barbituric acid (0.01 mol) was dissolved in ethanol (20 mL) and distilled water (10 mL). To this solution, formaldehyde (0.02 mol) was added. The mixture was refluxed at 60 – 70 °C for 12 h. The solvent was evaporated; the solid precipitate was suspended in water (10 mL), and extracted with chloroform to give 5,5-diethyl-1,3-bis(hydroxylmethyl)pyrimidine-2,4,6(1H,3H,5H)-trione [B1] and 5-ethyl-5-phenyl-1,3-bis(hydroxylmethyl)-pyrimidine-2,4,6(1H,3H,5H) trione [B2]. Yield: White, 85% and 87%; m.p. = 93 – 95 and 163 - 165 °C; Rf = 0.7 and 0.65, respectively.

Synthesis of barbituric acids derivative [RB1]-[RB2] [14]

0.01 mol of compounds [R] in 15 mL DMF was

added drop wise to the stirred suspension of 0.01 mol [B1]-[B2] respectively and 0.01 mol NaOH in 15 mL DMF. The mixture was refluxed for 5 h. Then, the reaction mixture was diluted with water (40 mL), andthen extracted with ether (2×20 mL). The combined organic layers were washed successively with water (30 mL), filtered and evaporated to dryness to give compounds [RB1]-[RB2] (Table 1).

Synthesis of barbituric acids derivative [RB3]-[RB4] [15]

To a solution of 5,5-diethylbarbituric acid and 5-ethyl-5-phenyl barbituric acid (0.01 mol), respectively, 15 mL of dry benzene and 0.01 mol compound [R] and 0.01 mol potassium carbonate

Table 1 Some physical properties of barbituric acids derivativeComposition General formula Color Yield (%) M.P. (°C) Rf

RB1 C26H32N8O8S2 Pale yellow 76 170-172 0.62

RB2 C30H32N8O8S2 Pale yellow 69 280-282 0.71

RB3 C24H27N8O6S2 Light brown 72 237-239 0.65

RB4 C28H27N8O6S2 Light brown 64 163-165 0.62

Scheme 2 Preparation of barbituric acid derivatives [RB1]-[RB4].

O

SHN

N

NHN

O

CH2-OHHO-H2C

SN[R]

HN

N

O

N

O

NaOH, D

MFOR'

HN

O

NH

OOR'

HN

SN

HN

HN

SN

HN

N

O

N

OOR'

CH2-OH

O

SHN

N

N

O

[RB1]-[RB2][RB3]-[RB4] N

O

NH

OOR'

R'=Ethyl, Phenyl

O

O

SNH

N

N

O

Cl

O

O O

K2 CO

3 , Dry benzene



was added. The mixture was refluxed for 4-5 h with stirring. The residue obtained was treated with ice-cold water (5 mL), filtered, dried in oven and recrystallized (Table 1, Scheme 2).

Test of biological activity

Bacterial suspension was prepared from used bacteria and compared with McFarland tube 1.5 × 108 cell/mL which refers to the sensitivity prepared and the number of bacteria cultivated. Bacterial suspension was spread on Muller Hinton Agar homogeneously (0.1 mL) to cover the whole medium. Afterwards, three holes were made in the paten dish by the cork piercing to diameter of 6 mm at the concentration used. Diluted solutions of 0,03 and 0.06 mg/mL were prepared for each compound at pH 7. Then, the concentrated solutions prepared from chemical compounds were put in holes to reveal their effectiveness of biological activity. The paten dish was incubated at temperature 37 °C for 24 h. Finally, the diameter of inhibition zone for each disc was measured by the ruler to determine the effectiveness of each compound and compare it with the standard limits of sensitivity of the same species of bacteria against antibiotics [16].

To examine the antifungal activity, 0,03 and 0.06 mg/mL of synthesized compounds were dissolved in 5 mL dimethyl sulfoxide as solvent, and evaluated in vitro employing the filter paper disc method against Candida albicans and Aspergillus flavus by measuring inhibition zone in millimetre [17].

Results and Discussion

In this research, a series of new barbituric acid derivatives [RB1]-[RB4] containing thiazole moiety were synthesized from sulfa drug. Sulfadiazine is the starting material for this research, it was converted to 2-chloro-N-(4-(N-pyrimidin-2-ylsulfamoyl)phenyl)acetamide [A] by the reaction with chloroacetyl chloride and triethylamine in DMF as solvent. The synthesized compound [A] was characterized by sodium fusion test and the result was positive. The chlorine element was identified using saturated solution of AgNO3 reagent; white precipitate was observed, which was due to the presence of chlorine in compound [A]. FTIR spectrum showed the disappearance of one absorption band at 3425 cm-1 which was due to the stretching vibrations of -NH2 group of sulfadiazine, the remaining of the band at 3381 cm-1 which was due to the N-H group of amide, the absorption band at

3080 cm-1 which was attributed to the -CH2 group, the absorption band at 1670 cm-1 due to the stretching band of C=O amide group, and the absorption band at 665 cm-1 due to the stretching band of C-Cl group. All these changes in absorption bands are good evidence to the formation of compound [A].

The compound 4-(2-aminothiazol-4-ylamino)-N-(pyrimidin-2-yl) benzene sulfonamide [B] was prepared by the reaction of [A] with thiourea and anhydrous potassium carbonate (Scheme 1). The structure of 4-(2-aminothiazol-4-ylamino)-N-(pyrimidin-2-yl) benzene sulfonamide [B ]was confirmed by FTIR, 1H-NMR and 13C-NMR.

The FTIR spectrum of [B] showed absorption of the band at 3394-3358 cm-1 due to the NH2 group and absence of the band at 665 cm-1 due to C–Cl for compound [A]. The appearance of stretching vibration band at 3275 cm-1 was due to the -NH group. FTIR showed absorption peaks at 1543 cm-1 (C=C thiazole), 1583 cm-1 (C=C aromatic), 1622 cm-1 (C=N thiazole) and 1662 cm-1(C=N pyrimidine).

1H-NMR spectrum (DMSO-d6, δ ppm) of [B]: 7.25 (s, 2H, NH2 ), 5.33 (s, 1H, C-5 thiazole), 7.46-7.71 (m, 4H, Ar-H), 11.48 (s, 1H, NH-SO2), and 10.46 (s, 1H, NH).

13C-NMR spectrum (DMSO-d6, δ ppm) of [B]: 160.5 (C-NH2), 109.7 (C-5 thiazole), 139.6 (C-1 thiazole), 112.3-133.4 (C-aromatic ring), 164.7 (N=C–N pyrimidine), and 157.7(C–N pyrimidine).

Compound [R] was synthesized by treatment of 4-(2-aminothiazol-4-ylamino)-N-(pyrimidin-2-yl) benzene sulfonamide [B] with chloro acetyl chloride using triethylamine as a base triethylamine in DMF.

FTIR spectrum (KBr, cm-1) of [R] showed appearance of absorption bands at 3404 cm-1 of stretching vibration of rhe NH group, and absorption bands at 2924 and 657 cm-1 of stretching vibrations of -CH2 and C-Cl groups, respectively. Absorption band at 1674 cm-1 was due to the stretching vibration of C=O amide group. FTIR showed absorption peaks at 1543 cm-1 for C=C thiazole, and at 1593 cm1 for C=C aromatic.

1H-NMR spectrum (DMSO-d6, δ ppm) of [R]: 8.52 (s, 1H, NH amide), 4.37 (s, 2H,CH2–Cl), 5.33 (s, 1H, C-5 thiazole), 7.39-7.73 (m, 4H, Ar-H), 12.47 (s,1H, NH-SO2), and 11.36 (s, 1H, NH).

13C-NMR spectrum (DMSO-d6, δ ppm) of [R]:



164.0 (C=O amide), 43.6 (CH2-Cl), 161.5 (C-NH2), 107.1 (C-5 thiazole), 140.3 (C-1 thiazole), 111.5-133.4 (C aromatic ring), 164.6 (N=C–N pyrimidine), and 157.6 (C–N pyrimidine).

1,3-bis(hydroxyl methyl) barbituric acid derivatives [B1]-[B2] were prepared from the reaction of 5,5-diethyl barbituric acid and 5-phenyl,5-ethyl barbituric acid with formaldehyde. According to the FTIR spectrum for [B1] , H-bonding of the OH)band was a broad peak at 3452 cm-1. The absorption band at 1674 cm-1 was due to the -C=O amide group, and the absorption bands at 1714 and 1766 cm-1 was due to the stretching vibration for -C=O barbituric acid. And for compound [B2], the H-bonding of OH band was at 3464 cm-1; the C=O group of amide was shown at 1635 cm-1. Also, absorption band was at 1724 cm-1 for -C=O barbituric acid.

Williamson etherification of 2-chloro-N-(4-(4-(N-pyrimidin-2 ylsulfamoyl)phenylamino)thiazol-2-yl)acetamide [R] with [B1]-[B2] in DMF as solvent and the presence of NaOH offered [RB1]-[RB2] in very good yields, while the compounds [RB3]-[RB4] were prepared from the reaction of intermediates [R] with 5,5-diethylbarbituric acid and 5-ethyl-5-phenyl barbituric acid.

[RB1]: 2-((5,5-diethyl-3-(hydroxymethyl)-2,4,6-trioxotetrahydropyrimidin-1(2H)yl)methoxy)-N-(4-(4-(N-pyrimidin-2-ylsulfamoyl)phenylamino)thiazol-2-yl)acetamide.

FTIR spectrum (KBr, cm-1) of [RB1]: 1668 (υ(C=O) amide), 3412 (υ(O-H), 1712, 1689 (υ(2(C=O)) barbituric), 1595 (υ(C=C) aromatic), 1548 (υ(C=C) thiazole), 2968 and 2929 (υ(C–H) aromatic and (CH2-O)) , 2831(υ(C–H) al iphat ic) , 1363 and 1134(υ(SO2)), 1255 and 1232 (υ(C–O) and (C–O–C)), 1051 (υ(C–O) and (C–O-H)).

1H-NMR spectrum (DMSO-d6, δ ppm) of [RB1]: 1.67 (q, 4H, CH2 barbituric acid), 0.64 (t, 6H, CH3

barbituric acid), 5.45 (s, 2H, O-CH2-N), 5.31 (s, 2H, N-CH2), 4.24 (s, 2H, COCH2), 8.49 (s,1H, NH amide), 7.32-7.70 (m, 4H, Ar-H), 5.68 (s, 1H, C-5 thiazole), 10.93 (s, 1H, NH), and 11.36 (s, 1H, NH pyrimidine).

[RB2]: 2-((5-ethyl-3-(hydroxymethyl)-2,4,6-trioxo-5-phenyltetrahydropyrimidin-1(2H)-yl)methoxy)-N-(4-(4-(N-pyrimidin-2-ylsulfamoyl)phenylamino)thiazol-2-yl) acetamide.

FTIR spectrum (KBr, cm-1) of [RB2]: 1668 (υ(C=O) amide), 3323 (υ(O-H)), 1710 and 1691 (υ(2(C=O))

barbituric), 1629 (υ(C=N) thiazole), 1589 (υ(C=C) aromatic), 1548 (υ(C=C) thiazole), 3059, 2964 and 2927 (υ(C–H) aromatic and (CH2-O)), 3190 (υ(C–H) and (HC=N) pyrimidine), 2854 (υ(C–H) aliphatic), 1363 and 1134 (υ(SO2)), 1263 and 1238 (υ(C–O) and (C–O–C)), and 1097 (υ(C–O) amd (C–O-H)).



[RB3]: 2-(5,5-diethyl-2,4,6-trioxotetrahydrop- yr imidin-1(2H)-yl ) -N-(4-(4-(N-pyr imidin-2-ylsulfamoyl)phenylamino)thiazol-2-yl)acetamide,

FTIR spectrum (KBr, cm-1) of [RB3]: 1678 (υ(C=O) amide), 3404 (υ(N-H)), 1762 and 1712 (υ(2(C=O)) barbituric), 1629 (υ(C=C) thiazole), 1585 (υ(C=C)

aromatic), 1537 (υ(C=C) thiazole), 3080, 2974 and 2933 (υ(C–H) aromatic and (CH2-O)), 3170 (υ(C–H) and (HC=N) pyrimidine), 2862 (υ(C–H) aliphatic), 1375 and 1153 (υ(SO2)), 1240 (υ(C–O) and (C–O–C)), and 1091 (υ(C–O) and (C–O-H)).


barbituric acid), 4.98 (s, 2H, O-CH2-N), 4.70 (s, 2H, N-CH2), 4.29 (s, 2H, COCH2), 8.48 (s,1H, NH

Amide),7.01-7.95 (m, 4H, Ar-H), 5.52 (s, 1H, C-5 thiazole), 10.71 (s, 1H, NH), and 11.55 (s, 1H, NH

pyrimidine).

[ R B 4 ] : 2 - ( 5 - e t h y l - 2 , 4 , 6 - t r i o x o - 5 -phenyltetrahydropyrimidin-1(2H)-yl)-N-(4-(4-(N pyrimidin-2-ylsulfamoyl)phenylamino)thiazol-2-yl)acetamide

FTIR spectrum (KBr, cm-1) of [RB4]: 1670 (υ(C=O) amide), 3406 (υ(N-H)), 1710, Overlap (υ(2(C=O)) barbituric), 1625 (υ(C=C) thiazole), 1591 (υ(C=C)

aromatic), 1543 (υ(C=C) thiazole), 3062, 2962 and 2927 (υ(C–H) aromatic and (CH2-O)), 3099 (υ(C–H) and (HC=N) pyrimidine), 2854 (υ(C–H) aliphatic), 1355 and 1141 (υ(SO2)), 1253 and 1228 (υ(C–O) and (C–O–C)), and 1087 (υ(C–O) and (C–O-H)).





Antimicrobial activity

Ant ibac te r i a l ac t iv i ty o f t he syn thes i zed compounds was tested against three types of bacteria, Staphylococcus aureus as a gram-positive bacterium, Escherichia coli and Pseudomonas aeruginosa as gram-negative bacteria by using Muller Hinton Agar medium. The prepared compounds [B], [R], [RB1], [RB2], [RB3] and [RB4] were measured for zone of

inhibition around each disc. Solvent control (DMSO) did not inhibit bacteria or fungi. The test results are presented in Table 2 and Fig. 1, which showed that [B] and [R] exhibited high antibacterial active against S. aureous, P. aeruginosa and E. coli at 0.03 and 0.06 mg/mL. Compound [RB1] exhibited high antibacterial active against P. aeruginosa, and it was moderately active against E. coli whereas inactive against S.

Table 2 Antibacterial activity of some synthesized compounds

Sample

Inhibition zone diameter (mm/mg sample)

S. aureus P. aeruginosa E. coli0.03 0.06 0.03 0.06 0.03 0.06

[B] 20 28 20 25 20 20[R] 28 30 25 28 22 28

[RB1] -- -- 20 20 11 12

[RB2] -- 20 11 15 15 15

[RB3] 20 28 -- -- 17 20

[RB4] 18 20 -- 20 -- 20

AntibioticsGentamicin 31 28 22

Penicillin 32 20 23Control : DMSO 0 0 0

Fig. 1 Antibacterial activity of some synthesized compounds at (a) 0.03 mg/mL and (b) 0.06 mg/mL.

S. aureusP. aeruginosaE. coliDMSO

30

25

20

15

10

5

0 [B] [R]

(a)

[RB1] [RB2] [RB3] [RB4]

[B] [R] [RB1] [RB2] [RB3] [RB4]

DMSO Sulfadiazine

S. aureusP. aeruginosaE. coliDMSO

35

30

25

20

15

10

5

0

(b)

DMSO Sulfadiazine



aureous for both concentrations. [RB2] exhibited moderate antibacterial activity against P. aeruginosa and E. coli, while it exhibited high antibacterial activity against S. aureous at 0.06 mg/mL and inactive at 0.03 mg/mL. Compound [RB3] exhibited high antibacterial activity against S. aureous, and E. coli, while it exhibited inactivity against P. aeruginos.

Finally, compound [RB4] exhibited high antibacterial

activity against S. aureous, high antibacterial activity against P. aeruginosa and E. coli at 0.06 mg/mL, and inactivity against S. aureous at 0.06 mg/mL.

Antifungal activity of the synthesized compounds was tested against two types of fungi, Candida albicans and Aspergillus flavus by using potato dextrose agar (PDA) medium. The test results are presented in Table 3 and Fig. 2. Compounds [B], [R], [RB1], [RB3]

Table 3 Antifungal activity of some synthesized compounds

SampleInhibition zone diameter (mm/mg sample)

Candida alicans Aspergillus flavus0.03 0.06 0.03 0.06

[B] 22 25 -- --[R] 30 30 -- 15

[RB1] 20 28 15 --[RB2] -- -- 15 15[RB3] 23 30 25 15[RB4] 25 28 20 --

Antifungal

Fluconazole 22 16Voriconazole 28 --

Nystatin -- 13Control : DMSO 0 0

Fig. 2 Antifungal activity of some synthesized compounds at (a) 0.03 mg/mL and (b) 0.06 mg/mL.

Candida alicansAspergillus flavus

Candida alicansAspergillus flavus

35

30

25

20

15

10

5

0

(a)

DMSO Sulfadiazine

35

30

25

20

15

10

5

0

(b)

DMSO Sulfadiazine

[B] [R] [RB1] [RB2] [RB3] [RB4]

[B] [R] [RB1] [RB2] [RB3] [RB4]



and [RB4] exhibited high antifungal activity against Candida albicans, while compound RB3 was inactive against the same fungus at 0.03 and 0.06 mg/mL.

To Aspergillus flavus, compound [B] was inactive at 0.03 and 0.06 mg/mL, while compound [R] exhibited moderate antifungal activity at 0.06 mg/mL and inactivity at 0.03 mg/mL. Compound [RB1] was moderately active at 0.03 mg/mL and inactive at 0.06 mg/mL. [RB2] was moderately active at 0.03 and 0.06

mg/mL. Compound [RB3] was highly active at 0.03 mg/mL, while it was moderately active at 0.06 mg/mL. Compound [RB4] was highly active at 0.03 mg/mL whereas inactive at 0.06 mg/mL.

FTIR spectra for some compounds are presented in Fig. 3-6; 1H-NMR and 13C-NMR spectra for some compounds are presented in Fig. 7-12; and antimicrobial activities for some compounds are shown in Fig. 13 and 14.

Fig. 3 FTIR spectrum of compound [A].

3381

.21

3105

.39

3080

.32

3039

.81

2839

.52

2872

.01

2812

.21

2738

.92

1670

.35

1624

.06

1587

.42

1484

.83

1543

.05

1440

.83 14

08.0

4

1332

.81

1159

.22

1257

.59

1093

.64

1022

.27

945.

12

840.

9689

4.97

773.

4671

9.45

665.

4464

0.37

603.

7257

0.93

520.

78

449.

4143

2.05

800.

461467

.83

105

90

75

60

45

30

T (%

)

3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 8001 (cm)

600 400

Fig. 4 FTIR spectrum of compound [B].

97.5

90.0

82.5

75.0

67.5

60.0

52.5

45.0

T (%

)

3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 8001 (cm)

600 400

3394

.72

3358

.07

3275

.13

3182

.55

3105

.39

3078

.39

3037

.89 29

39.5

228

72.0

128

10.2

827

36.9

9 2663

.69

1681

.93

1622

.13

1583

.56

1411

.89

1438

.90

1494

.83

1543

.05

1323

.17

1261

.45

1091

.71 94

3.19

1004

.91

837.

1179

8.53

729.

0968

0.87

638.

44

530.

42 480.

2844

9.41

432.

05

549.

7157

2.86

1153

.43

1662

.64



Fig. 7 1H-NMR spectrum of compound [B].

Fig. 5 FTIR spectrum of compound [R].

97.5

90.0

82.5

75.0

67.5

60.0

52.5

T (%

)

3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 8001 (cm)

600

3404

.36

2924

.09

2854

.65

1674

.21

1627

.92

1593

.20

1543

.05

1516

.05

1433

.11

1402

.25

1328

.95

1255

.66

1145

.72

1083

.99

1024

.20

991.

4195

8.62

937.

40 902.

6986

7.97

837.

1181

5.89

798.

8077

9.24

759.

9574

0.67

723.

3170

5.95

657.

7363

6.51

619.

1557

2.86

549.

7153

6.21

516.

9249

7.63

470.

63

1568

.13

100

95

90

85

80

75

70

65

T (%

)

3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 8001 (cm)

600

3412

.08 29

68.4

529

29.8

728

31.5

0

1712

.79

1689

.64

1668

.43

1595

.13

1548

.84

1421

.54

1363

.67

1313

.52

1255

.66

1232

.51

1134

.14

1101

.35 10

51.2

010

18.4

1 985.

6294

7.05

916.

1987

9.54

812.

0377

5.38 71

7.52

680.

8764

4.22

623.

0158

6.36

549.

7151

8.85

486.

0645

3.27

1529

.55

Fig. 6 FTIR spectrum of compound [RB1].

4500

4000

3500

3000

2500

2000

1500

1000

500

0

4.0 4.5 5.0 6.0 7.0 8.0 8.5 9.0 9.5 10.0 11.0 11.5 12.0 13.0 14.013.512.510.57.56.55.5δ

1.72

4.92

1.23

12.6

7

3.34

8.93

10.3

83.

14

8.18

3.89

4.00

5.33

5.36

5.39

5.40

5.42

6.31

6.33

6.34

6.35

6.37

6.37

6.38

6.39

6.40

6.40

6.41

6.42

6.42

6.43

6.45

6.45

6.46

6.48

6.49

6.51

7.13

7.15

7.25

7.44

7.44

7.45

7.45

7.46

7.47

7.48

7.49

7.50

7.50

7.52

7.52

7.53

7.54

7.54

7.55

7.55

7.56

7.56

7.57

7.59

7.61

7.66

7.69

7.69

7.69

7.70

7.70

7.71

7.71

7.72

7.72

7.73

7.73

7.73

8.07

8.07

8.08

8.09

8.09

8.10

8.11

8.13

8.14

8.15

8.28

8.30

8.51

8.52

9.85

10.2

610

.46

11.4

8



6500000

6000000

5500000

5000000

4500000

4000000

3500000

3000000

2500000

2000000

1500000

1000000

500000

0

−500000

δ−2 −1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

0.62

1.93

10.1

31.

881.

12

3.34

0.86

7.87

0.93

4.18

2.65

2.16

5.32

2.25

2.50

DM

SO2.

50 D

MSO

2.50

DM

SO2.

51 D

MSO

2.94

3.06

3.24

3.26

3.27

3.30

3.34

3.40

HD

O3.

433.

503.

883.

894.

375.

335.

375.

395.

405.

426.

336.

346.

356.

376.

376.

396.

406.

426.

436.

456.

486.

506.

517.

137.

397.

407.

447.

467.

487.

507.

527.

537.

547.

557.

577.

597.

617.

667.

697.

697.

707.

717.

717.

727.

738.

088.

098.

108.

118.

468.

5211

.36

12.4

7

23002200210020001900180017001600150014001300120011001000900800700600500400300200100

0−100−200

δ−2 −1 0 1 2 3 4 5 6 7 8 9 10 11 12

22.8

15.

882.

695.

201.

8420

.73

1.52

1.94

1.80

2.12

1.53

1.25

1.01

5.06

1.88

2.35

9.30

3.64

6.72

0.73

0.62

0.64

0.66

0.74

0.79

0.80

0.82

0.84

0.85

1.08

1.09

1.23

1.42

1.64

1.66

1.68

1.69

1.88

2.03

2.08

2.26

2.27

2.49

DM

SO2.

50 D

MSO

2.50

DM

SO2.

51 D

MSO

2.51

DM

SO2.

91

3.36

HD

O

4.24

10.9

311

.26

5.31

5.45

5.68

6.33

6.34

6.35

6.36

6.37

6.42

6.44

7.30

7.32

7.39

7.41

7.43

7.45

7.52

7.54

7.58

7.60

7.62

7.65

7.68

7.70

8.06

8.07

8.08

8.33

8.49

Fig. 8 1H-NMR spectrum of compound [R].

Fig. 9 1H-NMR spectrum of compound [RB1].



Fig. 10 1H-NMR spectrum of compound [RB3].

Fig. 11 13C-NMR spectrum of compound [B].

δ−2 −1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

1.85

0.06

0.09

2800

2600

2400

2200

2000

1800

1600

1400

1200

1000

800

600

400

200

0

−200

0.72

0.74

0.76

0.85

1.23

1.47

1.78

1.80

1.82

1.84

1.98

2.27

2.49

DM

SO2.

50 D

MSO

2.50

DM

SO2.

51 D

MSO

2.51

DM

SO3.

35 H

DO

3.36

4.29

4.70

4.98

5.52

7.01

7.21

8.48

9.86

10.7

1

11.5

5

7.74

7.76

7.93

7.95

5500

5000

4500

4000

3500

3000

2500

2000

1500

1000

500

0

−500

δ0 20 40 60 80 100 120 140 160 180 200 220 240

36.4

939

.23

DM

SO39

.44

DM

SO39

.65

DM

SO39

.86

DM

SO40

.07

DM

SO40

.11

DM

SO40

.28

DM

SO40

.34

DM

SO40

.49

DM

SO45

.75

47.3

148

.06

63.5

9

109.

7811

0.24

110.

6211

0.91

112.

3511

6.42

118.

3511

8.55

128.

0112

8.15

128.

6612

8.72

132.

8313

3.41

139.

6314

0.35

141.

5814

1.82

149.

8415

0.43

157.

6615

7.71

160.

5416

3.05

164.

7016

7.33

168.

3016

9.77

172.

94



Fig. 12 13C-NMR spectrum of compound [R].

Fig. 13 Antibacterial activity of compounds [R], [RB1] and [RB2].

4500

4000

3500

3000

2500

2000

1500

1000

500

0

−500

δ30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190

36.9

239

.23

DM

SO39

.44

DM

SO39

.65

DM

SO39

.86

DM

SO40

.07

DM

SO40

.11

DM

SO40

.28

DM

SO40

.34

DM

SO40

.49

DM

SO45

.92

43.6

3

107.

1110

7.92

111.

5311

8.42

118.

72

128.

0112

8.15

128.

6612

8.72

133.

43

149.

8415

0.43

157.

6216

1.54

164.

0116

4.61

168.

3316

9.78

171.

94

183.

3218

4.12

140.

3314

1.57

141.

77

E. Coli E. Coli E. Coli

P. aeruginosaP. aeruginosa

R

R'

RB1'

RB2'

RB1'

RB2'

RB1'

RB1

RB2

C

R'

R' C

S. aurcus

S. aurcusS. aurcus

C

C

R

R RB1P. aeruginosa

RB2'

RB1

RB2

RB2C

C

C

CC



Conclusions

This research involved the synthesis of some new barbituric acid derivatives and the study of antimicrobial activity of the prepared compounds. FTIR, 1H-NMR and 13C-NMR techniques confirmed formation of these derivatives. Antifungal activities of the synthesized compounds were observed against two types of fungi, Candida albicans and Aspergillus flavus using PDA medium. Some of the synthesized compounds were shown to be highly active at 0.03 mg/mL whereas moderately active at 0.06 mg/mL.

AcknowledgementsGreat thanks to biologist Nadheema Abed Abbas,

Department of Ecology, Faculty of Science, University of Kufa for his help in the biological test.

Conflict of InterestsThe authors declare that no competing interest

exists.

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R'

RB1

RB1'RB2'

RB2'RB1'

RB1

CC

RB2

C

A. flavusA. flavusA. flavus

R

RB2

R'

C

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Copyright© Mahmood Mohi Fahad, Ezzat Hussein Zimam, and Majed Jary Mohamad. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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