In vitro: The Antibacterial Activity of Some Heterocyclic compounds

Al-Mustansiriyah Journal of Science Vol. 26, No 2, 2015

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In vitro: The Antibacterial Activity of Some Heterocyclic compounds Kudair A.H. Al-Ameri1, Muna S. Al-Rawi2 and Doaa A. Qasim3 1 Department of Biology, College of Education for Pure Science/ Ibn Al-Haitham, University of Baghdad 2 Department of Chemistry, College of Education for Pure Science/ Ibn Al-Haitham, University of Baghdad 3 Center for Market Research and Consumer Protection, University of Baghdad

Articleinfo Received 7/12/2014 Accepted 14/4/2015

ABSTRACT In this present study ten heterocyclic compounds were prepared from Azetidine (Two compounds) and Oxazepine (Eight compounds) and tested their ability to inhibit bacteria growth. The prepared compounds were tested for their antibacterial activity against Bacillussubtilis, Staphylococcusaureus (Grampositive) Pseudomonasaeruginosa,Escherichiacoli, and Proteusmirabilis (Gram negative) by agar well diffusion method. Results showed that azetidine derivatives had higher growth inhibition zone diameter (GIZD) than oxazepine derivatives by comparing with antibiotics Lincomycin, Amoxicillin, Ampicillin and Cloxacillin. ‫الخالصة‬ ‫ وثمانية من مشتقات الـ‬Azetidine ‫درست الفعالية التثبيطية لعشرة مركبات حلقية غير متجانسة اثنان من مشتقات الـ‬ ‫ (موجبة لملون‬BacillussubtilisStaphylococcusaureus ‫ ضد الجراثيم المرضية وهــي‬Oxazepine ‫ أظهرت‬.)‫ (سالبة لملون كرام‬Proteusmirabilis‫و‬Escherichiacoli،Pseudomonasaeurginosa،)‫كرام‬ ‫يمتلكان فعالية مضادة للجراثيم أعلى من تلك التي للمركبات المحضرة‬Azetidine ‫النتائج أن المركبين المحضران من الـ‬ .Cloxacillin‫ و‬Ampicillin،Amoxicillin،Lincomycin‫ بالمقارنة مع المضادات الحيوية‬Oxazepine ‫من الـ‬ because of the use of β-lactam derivatives as antibacterial agents. A large number of 3-chloromonocyclic β-lactam possesses powerful, antimicrobial, anti-inflammatory, anticonvulsant and antitubercular activities[9,10,11]. The aim of this study is testing the antibacterial activity of the prepared compounds against P. aeruginosa, E. coli, P. mirabilis, B. subtilis and S. aureus.

INTRODUCTION The development of simple synthesis to widely used organic compounds ring readily available reagents isone of the main objectives of organic synthesis-nitrogen heterocyclic of especial interest because they constitute an important class of natural and non natural products, many of which exhibit useful biological activities, one-pot efficient synthesis of heterocyclic derivatives, many permit the development of novel therapies for the treatment of epilepsy, pain and other neurogenerative disorder [1]. Some Schiff bases bearing aryl groups [2] or heterocyclic residues possess excellent biological activities [3] which has attracted many researches attention in recent year. They have been reported to be used analgesic, anthelmintic, antitubercular, plant growth regulator, antiviral, antifungal and anticancer [4]. Oxazepine derivatives was introduced in 1965 for use in relief of the psychoneuroses characterized by anxiety and lension, oxazepam is non-homologous seven member ring contain two heteroatoms (Oxygen and nitrogen) [5].Oxazepine compounds have medical and biological importance and they have medical and pharmaceutical application. Among the wide chemical derivatives are heteropolymers which have activity and effectiveness against cancer [6], they also are effective against fungi and bacteria [7]. It was found that some oxazepine derivatives are considered a medical drug against the diseases [8]. Azetidinones which are part of antibiotics structure are known to exhibit interesting biological activities. Azetidinones or β-lactam chemistry is of great importance

MATERIAA AND METHODS Preparation of compounds All compounds used in the present study were prepared by [12] showed in Table (1). Concentration of compounds Concentration used 30 mg/ml (w/v) of each compound and prepared by using Dimethyl Sulfoxide (DMSO). Bacterial cultures Fivepathogensbacteria species used in this study as tested organisms. These are Escherichiacoli, Proteus mirabilis, Pseudomonas aeruginosa (Gram negative) and Bacillus subtilis,Staphylococcus aureus (Gram positive).These bacterial species were obtained from Center for Market Research and Consumer Protection, Baghdad University. Determination of antibacterial activity The agar well diffusion method was used to detect antibacterial activity for ten prepared compounds of derivatives oxazepine and azetidinones against five pathogenic bacteria species used in present study as described by Barefoot and Klaenhammer [13]. Young bacterial cultures suspension equivalent of 0.5 tube McFarland turbidity standards (108 cfu/ml) were spread on Muller-Hinton agar plates using sterile cotton swabs

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Al- Mustansiriyah J. Sci., Vol. 26, No 2, 2015

Table 1: The ten prepared heterocyclic compounds. Comp. Nomenclature Molecular No. Formula Azetidine Compounds 3-chloro-1-(4C18H18N2OCl dimethylaminophenyl)-41 (4-methylphenyl) azetidine-2-one 3-chloro-1-(4C16H13N2OCl hydroxyphenyl)-4-(42 methylphenyl) azetidine-2one Oxazepine Derivatives Benzene1,2,4,5-{2(4dimethylaminophenyl) -11 C26H20N2O6 tolyl -2,3-dihydro [1,3 ] oxazepine -4,7 -diones} Benzene 1,2,4,5- { 2- (4hydroxyphenyl ) -1- tolyl 2 2 ,3 - dihydro [1,3]C24H15NO7 oxazepine - 4 , 7dinones} 2-(4-dimethyl aminophenyl) - 1 – totyl - 2, 3 3dihydrobenz- [1,2e] C28H24N2O3 [1,3] - oxazepine-4,7diones 2 - (4-hydroxyphenyl)- 1tolyl-2,3-dihydrobenz-[1, 4 C26H19NO4 2e] [1,3]-oxazepine -4,7diones 2(4dimethylaminophenyl) -15 tolyl-2,3- dihydro naptha C24H22N2O3 [2,3e] [1,3]- oxazepine 4,7- diones 2-(4-hydroxyphenyl)-1tolyl-2,3-dihydro naptha6 C22H17NO4 [2,3e] [1,3]- oxazepine-4,7diones 2-(4dimethylaminophenyl)-17 C20H20N2O3 tolyl-2,3-dihydro-[1,3] oxazepine-4,7-diones 2-(4-hydroxyphenyl) -18 tolyl-2,3-dihydro-[1,3] C18H15NO4 oxazepine-4,7-diones

. Wells of 6mm diameter were cut in solidified agar and filled with 50 μl 0f each concentration. The Dimethyl Sulfoxide also used as control. The plates were incubated aerobically at 37°C for 24 hours, then inhibition zones diameter (mm) around wells were measured by rule. All testes were applied as triplicate. Antibiotic sensitivity test Antibiotics susceptibility of P. aeruginosa,E. coli, P. mirabilis, B. subtilis and S. aureus was determined also by the agar well diffusion method (13) which mentioned previously. Antibiotics solutions were prepared by using DMSO. These antibiotics with their respective concentrations are Lincomycin (10μg/50μl), Amoxicillin (25μg/50μl), Ampicillin (25μg/50μl), and Cloxacillin (5μg/50μl). RESULTS AND DISCUSSION Small ring heterocycles containing nitrogen and sulfur have been under investigation along time because of their important medicinal properties. Among the wide range of heterocycles explored to develop pharmaceutically important molecules, Azetidine played an important role in medicinal chemistry. Table (2) and Fig. (1, 2, 3) showed results of antibacterial activity of azetidine compounds and oxazepine derivatives at 30mg/ml against E. coli, P. mirabilis, P. aeruginosa, B. subtilis and S. aureus. Azetidinecompounds had the strongest antibacterial activity. While the oxazepine derivatives had lowest activity by comparing with azetidine compounds (Table 2, Fig. 1, 2, 3). The strongest compound from the oxazepine derivatives was compound No.1 against S. aureus while compounds No. 6 and No. 2 had the highest growth inhibition activity against B. subtilis and P. aeuroginosa. Compounds No. 1 and No. 2 had the best growth inhibition activity against E. coliP.mirabilis resectively (Table 2, Fig. 1, 2, 3). The results of sensitivity test for five bacteria species to antibiotics Lincomycin, Amoxicillin, Ampicillin and Cloxacillin showed in Table (3).

Table 2: Growth inhibition zone diameter(mm)of ten heterocyclic compounds against five pathogenic bacteria species Heterocyclic Compounds Mean of Growth Inhibition Zone Diameter (mm) Pseudomon Escherichia Proteus Bacillus Staphylococcus as coli mirabilis subtilis aureus aeruginosa Azetidinone comp. No.1 42 35 34 45 33 Azetidinone comp. No.2

35

33

40

40

23

Oxazepine derivative No.1

18

20

16

16

23


18

18

17

18

22


8

8

8

8

8


8

8

8

8

8


13

15

8

12

23


18

19

13

18

26


13

18

14

8

17


12

11

15

16

21

Dimethyl Sulfoxide (Control)

0.0

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Heterocyclic compounds are considered one of important types of organic compound due to their implication in drugs and industrial studies [14]. Mechanisms of action of antibacterial drugs include inhibition of cell wall synthesis, inhibition of protein synthesis, inhibition of nucleic acid synthesis, inhibition of metabolic path way, interference with cell membrane integrity [15,16]. The activity of β-lactam moiety was greatly influenced by substituent or fused rings [17]. The probable cause of potentiality of azetidinone compounds isdue to greater lipophilicity of chloro group which in turn is responsible for greater penetrability of the compound in the cell [18]. These interesting biological activities attached our attention to the chemistry of nitrogen heterocycles, some oxazepine derivatives acts as inhibitors of enzyme action. The antibacterial activity of new heterocyclic derivatives containing oxazepine ring with the hope of discovering new serving as antibacterial [19]. The antibacterial revealed that nature of substituent on phenyl ring viz, methyl,methoxy, hydroxyl, dimethyl amino, chloro, and hydroxyl group at the para position of the aryl moieties are determined for nature and extent of the antibacterial activity of the compound, which might have influences on their inhibiting mechanism of action [20].

Figure 1: Growth inhibition zone diameter(mm) of two Azetidinone andeight Oxazepine derivative heterocyclic compounds against three Gram negativepathogenic bacteria species at 30 mg/ml concentration.

Figure 2:Growth inhibition zone diameter(mm) of two Azetidinone andeight Oxazepine derivative heterocyclic compounds against two Gram positive pathogenic bacteria species at 30 mg/ml concentration.

Table 3:Growth inhibition zone diameter (mm) of four antibiotics against five pathogenic bacteria species. Concentration Mean of Growth Inhibition zone Diameter (mm) (μg/50μl) Antibiotic Pseudomonas Escherichia coli Proteus Bacillus aeruginosa mirabilis subtilis *Lincomycin 10 8

Staphylococcus aureus 17

Amoxicillin

25

11

8

8

8

23

Ampicillin

25

8

8

8

8

13

*Cloxacillin

5

-

-

-

8

8

Dimethyl Sulfoxide (Control)

100 %

0.0

*Used for gram positive bacteria only.

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Fig 3:Growth inhibition zone diameter (mm) of tenheterocyclic compounds against five pathogenic bacteria species.

CONCLUSION According to the results, biological activity data showed that azetidine derivatives had higher antibacterial activity than oxazepine derivatives. REFERENCES [1] Katrizky, A. R.; Vakulenko, A. V.; Gedu, R. A. and Roger, J. W. Thepharmacological basis of therapeutic. Arkivoci, 85: 1153-1158. (2007). [2] Katrizky, A. R.; Vakulenko, A. V.; Gedu, R. A. and Roger, J. W. The new derivative of 3-[(2morpholine-4-yl) ethyl]-4-substituted -1,2,4triazolline-5-thiole. Arkivoci, 8555-8560 (Cited from reference 1).(2007). [3] Ye, X.; Chen, Z.; Zhang, A. Synthesis and antiinflammatory activity of chalcone and related mannch bases medical chemistry. Molecules, 12: 1202-1210. (2007). [4] Khalaf, A. J. Synthesis andantibacterialactivity of metronidazole and imidiazole derivatives. Molecules, 14: 2431-2446.(2009). [5] Wilson, C. O.and Givold, O. Text book of organic medicinal and pharmaceutical chemistry. 5th ed., Ptiman Medical Publishing. J. B. Lippin Cott Company, USA. (1966). [6] Matsuzaki, H.; Takuchi, I.; Hamad, Y. and Hatano, K. Studieson the 1,4-oxazepine ring formation reaction using the molecular orbital method. Chem. Pharm. Bull., 48:755-756.(2000). [7] Al-Joubory, A, M. D. Synthesis and comparative study of some heterocyclic compounds containing five and seven-member rings byconventional and microwave method (MAOS) and evaluate thebiological activity of some of them. Tikrit J. Pure Sci., 1: 103-105. (2010). [8] Hüe, B.; Palomba, B.; Giacardy-Paty M.; Bottaï, T.; Alric, R. and Petit, P. Concurrent high-performance liquid chromatographic measurement of loxapine

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