Preparation, Identification and Evaluation of Biological Activity of some new β-Lactam compounds derived from Schiff bases

 

Nadia Sadiq Majeed , Fatima Naeem Abdul-Hussein

Department of Chemistry, College of Education for Girls, University of Kufa, Iraq.

*Corresponding Author E-mail: nadia.albobaid@uokufa.edu.iq

 

ABSTRACT:

A series of four- membered rings has been synthesized from Dapsone drug. In the first step examination Dapsone has been taken as initial material and treated with 4-aminoacetophenone to prepare new Azo compound. then Azo compound reacts with various aromatic amines in absolute ethanol to prepare Schiff bases derivatives. In the second step these derivatives react with Chloroacetylchloride in the presence of tri ethyl amine(Et3N) to prepare new four- membered heterocyclic compounds . All these derivatives were tested against different bacteria(Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Bacillus anthracis). The high efficacy results of these derivatives proved significantly greater than the efficacy of the same Dapsone drug.These compounds were identified and confirmed by FT- IR, 1HNMRand 13C-NMR.

 

KEYWORDS: Dapsone, Schiff bases, β-Lactam, Anti-bacterial activity.

 

 


INTRODUCTION: 

Dapsone or diaminodiphenyl sulfone (DDS), a sulfones class antibiotic drug1 reduces swelling (inflammation) and stops the bacteria’s development. Generally, it is administered with rifampicin and clofazimine in the treatment of leprosy, elimination half-life 20–30 hours2. Dapsone is widely employed as effective antibiotic for prophylaxis agent pneumocystis and an opportunistic disease in (HIV).

 

Schiff bases are condensation products of primary (aromatic) amines with aldehydes or ketones carrying the azomethine (imino) moiety (-CR=N-). They are considered versatile pharmacophores for various pharmacological activities where the azomethine group has been demonstrated to be critical to the bioactivity. For example, Schiff bases, whether of natural or non-natural origin, have exhibited promising antibacterial, antitubercular, antifungal, antiparasitic, antiviral, antioxidant, anticancer3,4.

 

 

Infection. It has been approved as antibiotic by food and drug administration since 1963.Its molecular formula is C12H12N2O2S, molar mass 248.30 g.mol−1and melting point (175–176°C). Dapsone is widely employed as effective antibiotic for prophylaxis agent pneumocystis and an opportunistic disease in HIV infection. It has been approved as antibiotic by food and drug administration, since 19635.

 

β-lactam (2-Azetidinone) are four-membered cyclic amide that which derived from 3-amino-propanoic acid6. It is named so because the (N) atom is connected to the β-carbon atom close to the carbonyl7. The β-lactam ring is portion of the core structure of several antibiotic families, the major ones being the cephalosporins, penicillins ,carbapenems, and mono bactams, which are also called (β-lactam antibiotics). Almost all of these antibiotics work by inhibition bacterial cell wall biosynthesis8. The first synthetic β-lactam ring was prepared by Hermann Staudinger in(1907) by reaction of the Schiff base ofbenzaldehyde and aniline with diphenylketene in a [2+2] cyclo addition9,10. A new research has suggested that β-lactams can subject ring-opening polymerization to form the amide bonds, to become “nylon-3 polymers”. The basis of these polymers are similar to peptides, which offer them biofunctionality. nylon-3 polymers can either mimic host defense peptides or act as signals to stimulate 3T3 stem cell function11.

 

Experimental:

All chemicals were of highest purity and supplied by Merck and Fluka-company. Measurements melting points were recorded by using electro thermal 9300,” melting point engineering LTD , U.K”. Thin layer chromatography (TLC)was performed on silica gel for and spots were visualized by Iodine vapors.” FTIR” spectra, Fourier transform infrared shimadzu (8400), H1-NMR and C13-NMR-spectra in (ppm) unit were operating in DMSO -d6 as solvent using (BrukerUltra Shield 400 MHz Switzerland)- (Iran).

 

Synthesis ofAzo Compound( F1)12:

1,1'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))diethanone

Compound F1 was prepared by dissolving (3g, 0.012mol) of Dapsone in a mixture consisting of 5ml of concentrated HCl acid and 20ml of cooled distilled water, the mixture in Can ice bath at a temperature of 0-5, then a solution was added to it. It consists of (0.828g, 0.012mol) of sodium nitrite NaNO2 dissolved in 10ml of cold distilled water, drop by drop with continuous stirring and making sure that the temperature does not rise above (5)C. leave the formed solution for a period of 20 min to settle to complete The dialysis process, after which the formed diazonium solution was gradually added with continuous stirring to a solution consisting of (1.62g, 0.12mol) from 4-aminoacetophenone dissolved in 20ml ethanol and 10ml sodium hydroxide solution 10% NaOH and it was noticed that the solution was colored orange at pH = 3, the precipitate was filtered and washed several times after the sedimentation process was completed with distilled water, then dried and recrystallized using absolute ethanol.

 

Synthesis of Schiff’s bases(F2,F3,F4,F5,F6)13,14:

2,2'-(1E,1'E)-(1,1'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(ethan-1-yl-1-ylidene))bis(azan-1-yl-1-ylidene)bis(4-nitrophenol) [F2]

1,1'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(ethan-1-yl-1-ylidene))dipyridin-2-amine [F3]

1,1'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(ethan-1-yl-1-ylidene))bis(2-nitroaniline) [F4]

(1,1'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(ethan-1-yl-1-ylidene))bis(4-methoxy-2-nitroaniline) [F5]

1,1'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(ethan-1-yl-1-ylidene))bis(4-bromoaniline) [F6]

 

0.02mol from (2-amino-4-nitrophenol, 2-aminopyridine, 4-methoxy-2-nitroaniline, 4-bromoaniline)respectively which was mixed with 0.01mol from Azo compound F1 in 30ml absolute ethanol. After that 2-3 drops of glacial acetic acid. The whole mixture were stirred under refluxed for 20-24h. After reflux the products were obtained, filtered, driedand recrystallized using absolute ethanol are given in Table1.

 

Preparation of azetidinone from Schiff base( F2B,F3A,F4E,F5C,F6D) 15,16:

4,4'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(3-chloro-1-(2-hydroxy-5-nitrophenyl)-4-methylazetidin-2-one) [F2B]

4,4'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(3-chloro-4-methyl-1-(pyridin-2-yl)azetidin-2-one) [F3A]

4,4'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(3-chloro-4-methyl-1-(2-nitrophenyl)azetidin-2-one) [F4E]

4,4'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(3-chloro-1-(4-methoxy-2-nitrophenyl)-4-methylazetidin-2-one) [F5C]

4,4'-(3,3'-(4,4'-sulfonylbis(4,1-phenylene)bis(diazene-2,1-diyl))bis(4-amino-3,1-phenylene))bis(1-(4-bromophenyl)-3-chloro-4-methylazetidin-2-one) [F6D]

 

To a mixture of compounds (F2, F3, F4, F5, F6) (0.2g, 0.001mol) in dry1,4- dioxane (20 ml), triethylamine (1 ml, 0.002mol), was added chloroacetyl chloride (1ml, 0.002mol) drop-wise at 5-100C. The reaction mixture was then poured into crushed ice. The solid separated was dried and recrystallized from ethanol and water Thereaction was monitored by TLC (ethanol:benzene) (2:3). The physicochemical data for synthesized azetidinone compounds are given in Table 1.

 

Biological activity assay17-19:

Antibacterial activity of the synthesized compounds [F1-F6D] have been carried out against four typesof bacteria like, (staphylococcusaureus, Bacillus anthracis) gram positive and (E. coli, Klebsiella pneumoniae) gram negative using nutrient agar medium viawell diffusion method. All derivatives were suspendedin aqueous solutions in dissimilar concentrations ranged from 0.001g and 0.1g in 10ml of DMSO, the results are voiced on MIC (minimal inhibitory concentration), and exhibited high biological activity of all these synthesized compounds of these microorganisms more than the effectiveness of (Dapsonedrug). The biological investigation data is given in Table 2.

 

RESULTS AND DISCUSSION:

Schiff bases and their Azetidinone derivatives were synthesized from Azo compound (F1) which prepared from Dapsone drug. Thin layer chromatography was performed on pre-coated silica gel G, glass plates using benzene : ethanol (3:2) solvent systems to ascertain the purity of these compounds. The compounds gave single spots. The structure of synthesized compounds was confirmed by FT-IR, 1H-NMR, 13C-NMRspectroscopy, The assignment of FT-IR for compound (F1) appeared absorption band (õ=cm-1 ) at 1691 for C=O (ketone) and exhibited singlet signal at 1.91ppm for protons of (CH3) and singlet signal at 6.40ppm for protons of ( NH2) in 1H-NMR spectra as well as exhibited singlet signal at 27ppm for carbon of (CH3) and singlet signal at 195ppm for carbon C=O ketone in 13C-NMR spectra.these bands were disappeared in the spectra of compounds (F2, F3, F4, F5, F6) and appeared new bands at 1624-1635 for C=N ( imine)20,21 and 1515-1595 for C=C aromatic in FT-IR spectra ,the structures of compounds were identified by the 1H-NMR spectra and showed exhibited singlet signal at (1.80-1.99) ppm for the hydrogen of ( CH3) and singlet signal at (6.17-6.78)ppm related to free amine group NH2 while exhibited singlet signal at 11.02 ppm for proton of hydroxyl group in compound F2 and appearedsinglet signal at 3.71 ppm for protons of methoxy group in compound F5 but in 13-C-NMR exhibited singlet signal at (24-28) ppm for carbon of methyl groups and singlet signal at (156-158) ppm for carbon of imine groups (C=N).Scheme (1)

 

 

Scheme (1): Synthesis of Schiff’s bases from Azo compound drived from Dapsone drug

 

Schiff bases (F2- F6) that reacted with chloro acetyl chloride in the presence of tri ethyl amine in dry dioxane by a [2+2] cyclo addition reaction Scheme 2 to give corresponding β-lactam derivatives (F2B, F3A, F4E, F5C, F6D)22. appearance of the absorption bands of carbonyl lactam ring at (1687-1695) cm-1, these stretching frequencies in biscyclic lactams indicates that the carbonyl group in β-lactam behaves.thesecompounds were identified by the 1H-NMR spectra and showed exhibited singlet signal at (4.22- 4.55)ppm for the hydrogen oflactam ring( Cl—CH). . The spectra 13C-NMR of carbonyl Lactam Showed the typical resonance at δ (183-187) ppm related to carbon of amide C=O in β-lactams the values outside this range are possible if strong electron donating or electron withdrawing groups are present on the adjacent carbon atoms as well as appeared new peaks

 

 

 

at 46-48 ppm for carbon of C-Cl in (2-Azetidinone)rings.

 

 

Scheme(3): mechanisim of [2+2] cyclo addition

 


 

Table (1): physical properties of synthesized derivatives

S. No.

M.F

M.WT

M.P

Rf

color

%yield

Solvent

gm.\mol

 

F1

C 28H24N6SO4

440.59

172-174

 

Light brown

84

Ethanol

F2

C40H32N10SO8

812.8

126-128

0.66

Dark brown

75

Ethanol

F3

C38H32N10SO2

692.79

136-138

0.75

Orange

73

Ethanol

F4

C40H32N10SO6

780.8

139-141

0.61

Dark red

67

Ethanol

F5

C42H36N10SO8

840.86

122-124

0.67

Light red

77

Ethanol

F6

C40H32N8SO2Br

768.7

lukewarm

0.88

Light browm

82

Ethanol

F2B

C44H34N10SO10Cl2

965.77

151-153

0.8

Black

80

1,4-dioxane

F3A

C42H34N10SO4Cl2

845.75

134-136

0.6

Light brown

75

1,4-dioxane

F4E

C44H34N10O8SCl2

901.7

lukewarm

0.87

Red

70

1,4-dioxane

F5C

C46H38N10SO10Cl2

993.82

146-148

0.54

Light orange

81

1,4-dioxane

F6D

C44H34N8SO4Cl2Br

921.66

130-132

0.69

Brown

79

1,4-dioxane

 

Table (1):Biological activity of the prepared compounds

No.

Staph.

C1

C2

E.Coli

C1

C2

Bacil

C1

C2

Klebse.

C1

C2

mm

mm

mm

mm

mm

mm

mm

mm

10-Feb

10-Mar

10-Feb

10-Mar

10-Feb

10-Mar

10-Feb

10-Mar

F1

++

22

14

++

20

14

- +

18

0

+++

22

20

F3

++

26

14

+++

24

14

++

24

14

+++

28

20

F6

++

22

20

+++

22

20

++

22

20

++

22

18

F2B

++

20

17

+++

22

16

++

17

15

++

22

20

F3A

- +

16

0

- +

18

0

++

24

0

++

18

17

F4E

+++

22

20

+++

22

20

++

18

17

+++

28

22

F5C

+++

28

14

+++

22

17

++

22

14

+++

30

22

F6D

++

17

15

+++

17

16

+

17

16

++

22

20

 


 

CONCLUSIONS:

The synthesis of the designed compounds has been successfully achieved. Characterization& identification of the target compounds were confirmed by determination of the physical properties, FT-IR spectroscopy , 1H-NMR , 13C-NMR spectra . The synthesized compounds were thereafter evaluated for antibacterial activity against some strains of gram positive and gram negative bacteria. Most of the synthesized compounds show very promising antibacterial activity against some of the bacteria strains while some do not show any activity against some of the bacteria strains .

 

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Received on 04.04.2022          Modified on 26.06.2022

Accepted on 23.08.2022        © RJPT All right reserved

Research J. Pharm. and Tech 2023; 16(2):593-596.

DOI: 10.52711/0974-360X.2023.00101