Research Articles

2019  |  Vol: 5(3)  |  Issue: 3 (May- June)  |  https://doi.org/10.31024/ajpp.2019.5.3.5
Synthesis, characterization and biological activity of some glycolylurea and β-lactam merged heterocyclic compounds

Bhadreshkumar R. Sudani

Department of Chemical Engineering,

Government Engineering College, Valsad, Gujarat, India-396001.

Address for Corresponding Author

Bhadreshkumar R. Sudani

Department of Chemical Engineering, Government Engineering College, Valsad, Gujarat, India-396001.


Abstract

Objective: The chemistry of β-lactams has established a prestigious role in organic and medicinal chemistry. The combination of these derivatives with other medicinally important class like glycolylurea might give some more applicable compounds. The objective of the present work was the synthesis and characterization of glycolylurea derivatives of some β-lactams. The synthesis involving formation of glycolylurea derivatives followed by the condensation was performed as per previously used methods. Materials and Methods: The synthesized compounds were characterized by various physico-chemical and analytical methods including FTIR and NMR spectroscopy. All the compounds were also investigated for their antimicrobial activity on gram negative and gram positive microbial cultures. Results and conclusions: It was found that from the investigated compounds one compound G5 mono-chloro derivative with one methyl group at ortho position was found active against gram nagetive microbes P. aeruginosa and G7 di-chloro derivative with one methyl group at ortho position was found active against gram positive microbes B. subtilis. Four out of eight compounds are moderately active against E. coli. Except G8 all the synthesized compounds are less or very less active against selected fungal stains.

Keywords: Glycolylurea, β-lactams, antimicrobial activity


Introduction

Glycolylurea is also known as Hydantoin and its derivatives are widely applicable heterocyclic compounds for many diseases like anticunvalsant (Singh et al., 2005; Mistry and Desai, 2012; Sudani and Desai, 2015), antiepilepsy (Anger et al., 2001; Rogawski and Loscher, 2004), antihypertensive (Menendez et al., 1992; Dylag et al., 2004) and many more. It is now well known that the β-lactam ring is part of the core structure of many antibiotic families and due that some of them are also called β-lactam antibiotics (Donowitz and Mandell, 1988). Aruna and Indra, (2018) showed the importance of the β-lactam compounds in various antibiotics including urinary tract infection (UTI). This class of compounds has played an important role in medicinal chemistry as broad spectrum antibiotic compound maker. Both of these classes had performed well in synthetic pharmacy.

There are many derivatives synthesized with these moieties in order to get better activities. In present work some derivatives of glycolylurea were synthesized as per the known route and than they were condensed with cyclization to form β-lactam merged compounds with the prediction of better antibacterial activities

Materials and Methods

All the chemicals and reagents were of analytical reagent (AR) grade, they were used without further purification. IR spectra were recorded on Bruker ALPHA FTIR spectrophotometer in KBr pellets. The H-NMR spectra were recorded on Bruker Avance II spectrometer in d-DMSO. Chemical shifts relative to TMS used as internal standard were obtained in d unit. The melting points were determined in open capillary tubes on SUNBIM apparatus and are uncorrected.

Synthesis of different compounds

Compounds h1 to h4 were synthesized with the previously used method.  These eight compounds were than treated with hydrazine hydrate at (1:1.2 mol) at 35-40 oC for 4 to 5 hours. After continuous stirring and cooling to room temperature compounds a1 to a4 were obtained. All these four intermediate products were allowed to react with two aromatic aldehydes. This condensation reaction was carried out at 70-80oC for 8 hours refluxing in acetonitrile. After checking the completion of reaction by TLC the each mixture was allowed to remove acetonitrile with vacuum distillation at room temperature and the products (az1 to az8) further used to react with chloroacetyl chloride (0.0016 mol) at 0-5 oC during addition in 30 minutes after 2-3 hours on adjustment of pH from 6.5 to 7.5 by diluted NaHCO3 all the products (G1 to G8) were filtered and washed with chilled water. After drying for 1 to 2 hours they were recrystallized in alcohols. After further purification all the samples were used to measure their melting points, samples were sent for analytical testing. Scheme 1 indicates the complete procedure of the reaction in figure 1.

Figure 1. Scheme-1 for the reaction procedure to synthesize G1 to G8

 

 

Characterization

Thin Layer Chromatography was used to detect the reaction completion at every stage of each reaction. Suitable mobile phases were prepared including polar non polar solvents for the better upliftment and separation of the reaction mass. All the prepared samples were recrystallized from alcohol and dried samples were sent for the elemental analysis where percentages of carbon, hydrogen and nitrogen were determined. Results were compared and found resemble with calculated data as shown in table-1. In order to know the combination and groups of atoms in the synthesized compounds IR spectra of each compound was recorded on FTIR spectrophotometer with the help of KBr pellets. The frequencies were recorded in cm—1 and compared with the corresponding standard data. To know the positions of different types of hydrogen atoms in the carbon chain and particular groups H1 NMR was also recorded on Bruker Avance II spectrometer in d-DMSO as solvent for each of the prepared compound where TMS was used as internal standard.   

Antimicrobial activity

The newly synthesized compounds, as shown in table 1 were tested for their antimicrobial activity against the following microorganisms: two gram positive stains B. subtilis and S. aureus, two gram negative stains P. aeruginosa and E. coli., three fungal stains viz. P. piricola, A. niger and F. oxysporum. The preliminary screening of the investigated compounds was performed using Broth dilution method. The minimum inhibitory concentrations were recorded accordingly. Secondary screening was carried out as earlier but with lower concentration in microgram/mL. The highest dilution showing at least 99 % inhibition zone is taken as MIC. The test mixture was containing 108 organisms per mL. The result of this is much affected by the size of the inoculum. The results of the testing are given in the table 2. 

Results and discussion

Physicochemical analytical data given in the table indicates R2 and R3 as per the reaction scheme shown in figure 1. Yield is calculated for the final step of the reaction.

Table 1. Physicochemical analytical data

Compounds

R2

R3

M.F. /M.W. g/mol

M.P.

oC*

Yield

%

Elemental analysis

Cal & Found (%)

C

H

N

G1

-CH3

-2-CH3

C17H19ClN4O4

378.11

67

42

53.90, 53.90

5.06, 5.07

14.79, 14.80

G2

-CH3

-4-CH3

78

40

53.90, 53.91

5.06, 5.06

14.79, 14.79

G3

-C2H5

-2-CH3

C18H21ClN4O4

392.84

77

39

55.03, 55.04

5.39, 5.38

14.26, 14.28

G4

-C2H5

-4-CH3

55

43

55.03, 55.03

5.39, 5.37

14.26, 14.27

G5

-C6H5

-2-CH3

C22H21ClN4O4

440.88

67

28

59.93, 59.92

4.80, 4.81

12.71, 12.69

G6

-C6H5

-4-CH3

82

47

59.93, 59.93

4.80, 4.80

12.71, 12.70

G7

-4-Cl-C6H5

-2-CH3

C22H20Cl2N4O4

474.09

66

45

55.59, 55.58

4.24, 4.24

11.79, 11.80

G8

-4-Cl-C6H5

-4-CH3

79

29

55.59, 55.60

4.24, 4.23

11.79, 11.78

*Melting points are uncorrected

Spectral analysis of different compounds

G1: 3-(2-((3-chloro-2-oxo-4-(o-tolyl)azetidin-1-yl) amino)acetyl)-5,5-dimethylimi- dazolidine-2,4- dione: IR (KBr, cm–1): 3256(NH), 1748, 1765 (β lactam)(C=O), 1535(C=C Ar), 1428, 1408(-CH3); 1Н NMR (400 МHz, DMSO-d6): δ ppm = 1.47(s, 6H), 2.33(s, 3H), 3.53(d, 2H) 5.08(d, 1H), 5.48(d, 1H), 6.80-7.40(m, 4H Ar), 9.38(s, br, 1H), 10.08(t, 1H).

G2: 3-(2-((3-chloro-2-oxo-4- (p-tolyl)azetidin-1-yl)amino)acetyl) -5,5-dimethyl imidazolidine -2,4-dione: IR (KBr, cm–1): 3260(NH), 1751, 1771 (β lactam)(C=O), 1538(C=C Ar), 1431, 1412(-CH3); 1Н NMR (400 МHz, DMSO-d6): δ ppm = 1.48(s, 6H), 2.35(s, 3H), 3.55(d, 2H) 5.11(d, 1H), 5.51(d, 1H), 7.19-7.20(m, 4H Ar), 9.87(s, br, 1H), 10.21(t, 1H).

G3: 3-(2-((3-chloro-2-oxo-4-(o-tolyl)azetidin-1-yl)amino ) acetyl)-5-ethyl-5- methylimi dazolidine-2,4-dione: IR (KBr, cm–1): 3272(NH), 1745, 1776 (β lactam)(C=O), 1548(C=C Ar), 1432, 1417, 1402 (-CH2, -CH3);  1Н NMR (400 МHz, DMSO-d6): δ ppm = 1.01(t, 3H), 1.65(q, 2H), 1.81(s, 3H), 2.55(s, 3H), 3.58(d, 2H) 5.15(d, 1H), 5.58(d, 1H), 6.79-7.38(m, 4H Ar), 10.02(s, br, 1H), 10.51(t, 1H).

G4: 3-(2-((3-chloro-2-oxo-4-(p-tolyl) azetidin-1-yl)amino )acetyl) -5-ethyl-5-methyl imidazolidine-2,4-dione: IR (KBr, cm–1): 3253(NH), 1741, 1767 (β lactam)(C=O), 1551(C=C Ar), 1422, 1416, 1396 (-CH2, -CH3);  1Н NMR (400 МHz, DMSO-d6): δ ppm = 1.13(t, 3H), 1.71(q, 2H), 1.93(s, 3H), 2.65(s, 3H), 3.65(d, 2H) 5.19(d, 1H), 5.60(d, 1H), 7.23-24(m, 4H Ar), 10.12(s, br, 1H), 10.65(t, 1H).

From the above spectral data of compound G1 and G2 it is cleared that the similarity of the structure is also proven by the spectral data in both the graphs. In NMR spectra of compound G2 there is a sharp peak (m) at 7.19-7.20 ppm indicates the presence of methyl group in aromatic ring at para position. While in the graph of compound G1 it is broad at 6.80-7.40 ppm indicates the presence of methyl group in aromatic ring at ortho position. More over same similarity is also found in the comparison of NMR data of G3 and G4 compounds. In the comparison of G1 with G3 and G2 with G4, presence of one extra quartet peak in H-NMR for –CH2- group is also observed. Similar results are also observed for G5, G6, G7 and G8 compounds in order to establish the structure.

Antimicrobial activity

All the newly synthesized compounds were screened for their biological potential. The results obtained with minimal inhibition concentrations in μg/mL are given in table 2.

Table 2. Antimicrobial activity of different compounds

Compounds and Standards

Minimal Inhibitory Concentration in μg/mL

Bacterial Culture

Fungal Culture

P. aeruginosa

E. coli

B. subtilis

S. aureus

P. piricola

A. niger

F. oxysporum

G1

150

200

300

300

200

250

250

G2

150

250

250

500

500

500

500

G3

100

250

500

150

250

500

1000

G4

250

125

250

500

250

1000

125

G5

62.5

125

125

175

250

350

400

G6

400

250

175

250

200

150

1000

G7

125

125

62.5

150

250

125

1125

G8

125

175

250

200

125

250

100

Ciprofloxacin

125

125

250

250

ND

ND

ND

Chloramphenicol

50

50

50

50

ND

ND

ND

Nystatin

ND

ND

ND

ND

100

100

100

The results shows that G5 has some good activity against P. aeruginosa and G7 has activeness against E.coli while G8 has good to moderate activity against P. piricola and F. oxysporum. Bothe the compounds G5 and G7 contains one methyl group on phenyl ring attached to the lactam ring. Compound G3 is also showing some moderate activity against P. aeruginosa which also contain one ortho methyl group at same position. Compound G7 also moderate active against P. aeruginosa and E. coli. This activity might be due to the presence of one chloro group at R2 position. Similar activity for similar group is also found in the case of compound G8 against P. aeruginosa. Moreover it, the compound G8 is found moderately active against only fungal stain F. oxysporum. All other compounds are found less or very less active against all three fungal stains. It clearly indicates that this series of compounds is more sensitive against microbial stain than that of fungal stains.

Conclusions

From the experimental data it is proven that the synthesis of these compounds can done with condensation and cyclization reaction with selected aldehydes. The process adopted can give well to moderate yields. Some of the compounds have biological activity against selected stains. This series is more antimicrobial active than that of antifungal activity.

Acknowledgements

Authors want to express their sincere appreciation to all the staff members of Chemical Engineering Department of Government Engineering College, Valsad for their helps and moral supports in this work.

Conflicts of interest: Not declared.

References

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