Synthesis, spectroscopic characterization and Antibacterial screening of some new cefotaxime sodium derivatives

Sheela Joshi; Anju Shukla; Rahul Singh Jhala

doi:10.1016/j.arabjc.2014.12.031

View/Download PDF

Buy Reprints

PDF

Translate this page into:

Original article

12 (

); 2133-2140

doi:

10.1016/j.arabjc.2014.12.031

Synthesis, spectroscopic characterization and Antibacterial screening of some new cefotaxime sodium derivatives

Sheela Joshi^⁎, Anju Shukla, Rahul Singh Jhala

School of Chemical Sciences, Devi Ahilya University, Takshashila Campus, Khandwa Road, Indore 452001, Madhya Pradesh, India1

⁎Corresponding author. Tel.: +91 9826085169; fax: +91 0731 2365782. spjoshi11@rediffmail.com (Sheela Joshi),

Received: 2013-4-30, Accepted: 2014-12-31, Published: 2015-12-01

Disclaimer:
This article was originally published by Elsevier and was migrated to Scientific Scholar after the change of Publisher.

1 http://www.chemical.dauniv.ac.in

Abstract

Biologically active compounds with heteroaromatic ring system of cefotaxime sodium have been synthesized via aminomethylation reaction. The aminomethylation of cefotaxime sodium with various biologically potent sulphonamides/secondary amines was carried out and then characterized by elemental analysis and spectral studies – IR, ¹H NMR, ¹³C NMR, Powder X-ray diffraction and Scanning Electron Microscopy. The compounds were screened for their antibacterial activity against various pathogenic bacteria at varying concentrations. The antibacterial activity of cefotaxime sodium derivatives was compared with parent sulphonamides. The toxicity of synthesized cefotaxime sodium derivatives was ascertained by LD₅₀ test.

Keywords

Cefotaxime sodium

Sulphonamides

Powder XRD

Antibacterial activity

Show Related Articles from PubMed

1 Introduction

Since the introduction of the first antibiotic (penicillin, 1942) into medical practice, to date, there has been an ongoing “race” between scientists creating new drugs and pathogenic bacteria. This specific “arms race” causes that thousands of potentially active chemicals are synthesized in laboratories around the world every day.

Cefotaxime sodium drug belongs to the third generation of cephalosporin. It has potent broad spectrum of activity against important pathogens (Zhang et al., 2006) and (Raddatz et al., 1995). It is considered as one of the first choice antibiotics in the therapy of spontaneous bacterial peritonitis in cirrhosis (Gilbert and Kamath, 1995) and (Rimola et al., 1995). Cefotaxime is a semi-synthetic cephalosporin consisting of an acetyl side-chain on aminothiazolyl ring and an alpha-syn-methoxyimino group (Bucourt et al., 1980). The molecular formula is C₁₆H₁₆N₅NaO₇S₂ and molecular weight is 477. Its IUPAC name is sodium (6R, 7R)-7-[(Z)-2-(2-amino-4-thiazolyl)-2-(methoxyimino) acetamido]-3-acetyl-oxymethyl-3-cephem-4-carboxylate.

In addition to this the sulphonamide is well-known antimicrobial agents (Joshi et al., 2004; Wilkinson et al., 2007; Kamal et al., 2007; Joshi et al., 2012 ), anti-inflammatory (Bashir et al., 2011), antiproliferative (Hu et al., 2007), Carbonic anhydrase inhibitors (Garaj et al., 2004; Lehtonen et al., 2004; Puccetti et al., 2005; Zimmerman et al., 2004; Güzel et al., 2009 ) anti-tumour Crespo et al., 2010) and radiosensitizing agents (Ghorab et al., 2010).

The aminomethylation incorporated with sulphonamides is reported to be potent antibacterial agents and less toxic than parent sulphonamide (Joshi et al., 2007, 2010). Keeping in view the unique features of cefotaxime sodium and sulphonamide were condensed via aminomethylation reaction. A series of cefotaxime sodium derivatives were synthesized with different sulphonamides/secondary amines (Schemes 1 and 2). The synthesized compounds were characterized by elemental analysis and spectral studies – IR, ¹H NMR, ¹³C NMR, Powder XRD, SEM and screened for in vitro antibacterial activity gram-positive and gram-negative bacteria at arbitrarily chosen concentrations.

Synthesis of cefotaxime sodium methyl sulphonamide (3a–3f).

Synthesis of cefotaxime sodium methyl amines (3g–3j).

2 Experimental

All the melting points were determined in open capillary tubes and were uncorrected. Thin layer chromatography was used for monitoring the reaction and to check purity. IR spectra (KBr) were recorded as potassium bromide pellets on Schimadzu 820 IPC FTIR spectrometer and ¹H NMR spectra on Bruker DRX-300 FT NMR Spectrometer and chemical shifts were expressed as (ppm) values against tetramethylsilane (TMS) as internal reference. The XRD measurements were carried out on Bruker D8 Advance X-ray diffractometer using Cu Kα at a wavelength of 1.54 Å. SEM studies were performed with a Jeol JSM 5600 instrument having magnification range ×18 to ×300,000 and at an accelerating voltage of 0.5–30 kV. The chemical reagents used in the synthesis were purchased from E. Merck and Aldrich. All substituted sulphonamides were obtained as pure samples from reputed pharmaceutical establishment.

2.1

2.1 Chemistry

The reaction routes for synthesis of cefotaxime sodium derivatives were described as shown in Schemes 1 and 2.

2.1.1

2.1.1 Synthesis of cefotaxime sodium methyl sulphonamide (3a–3f) (Scheme 1)

To the ethanolic solution of 0.1 mol of cefotaxime sodium was added to 0.1 mol of sulphonamide slowly with constant stirring under rigorous ice cooling. The reaction mixture was cooled well and 2.5 ml of formaldehyde solution (37% v/v) was added slowly with constant stirring. The reaction mixture was then adjusted to the pH of 3.5 with hydrochloric acid. The reaction mixture was kept in efficient ice cooling for half an hour to avoid losses of formaldehyde and then refluxed on water bath. The reflux time was dependent upon the sulphonamide chosen. After refluxing, the refluxed mixture was cooled in refrigerator for 4 days, till crystallized product was obtained, which was recrystallized with dry distilled ethanol and DMF (1:1). Melting points were recorded and uncorrected. The purity of the compounds was ascertained by single spot during TLC where mobile phase was chloroform/methanol mixture (90:10) and stationary phase was silica gel-G (chromatographic grade).

2.1.2

2.1.2 Synthesis of cefotaxime sodium methyl amines (3g–3j) (Scheme 2)

Secondary amines (0.01 mol) were added to an ethanolic solution (50 ml) of Cefotaxime sodium (0.01 mol) in a flat-bottom flask. One half of 0.015 mol of formaldehyde solution (37%) was added slowly with constant stirring. The reaction mixture was stirred at 70–75 °C on a magnetic stirrer for 5.5 and 8.5 h, except for diethanolamine (3h), depending upon the secondary amine taken. The remaining portion of formaldehyde solution was added in two instalments at an interval of 1 h, where first instalment was added immediately and second was added after one hour from start of experiment. The reaction mixture was kept overnight in the refrigerator. Next day excess of solvent was distilled off from the reaction mixture through vacuum pump which is under reduced pressure. Next day it was again kept for crystallization in the refrigerator. The product obtained was purified by recrystallization with dry distilled ethanol. Melting point was recorded and found uncorrected.

The compounds thus synthesized are presented in Schemes 1 and 2.

2.2

2.2 Spectral studies

2.2.1

2.2.1 IR and NMR studies

Compound 3a: cefotaxime sodium methyl sulpha methoxazole; C₂₇H₂₇N₈NaO₁₀S₃; yield 80%, m.p. 160–163 °C. Anal. Calcd C, 43.66; H, 3.66; N, 15.09 Found C, 43.62; H, 3.42; N, 15.03. IR (KBr) ν_max in cm⁻¹: 3442 ν_s N—H, 3398 ν_as N—H in SO₂NH, 2972 C—H Aliphatic, 2940 ν_as C—H in CH₂, 1779 ν_s C⚌O in beta lactam, 1657 ν_s C⚌O in amide, 1622 ν_s C⚌O in COONa, 1586 ν_s C⚌N, 1538 N—H bending, 1345 ν_s S⚌O, 1130 C—H in plane bending vibration of 1:4 disubstituted benzene, 1033 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.73 (s, 3H, CH₃—COO), 2.94 (s, 2H, CH₂—COO), 3.57 and 3.41 (ABq, 2H, CH₂ attached to thiazine ring), 5.05 (d, 1H, CH attached to thiazine ring), 5.59 (dd, 1H, CH attached to azetidine ring), 9.53 (d, 1H, NH—C⚌0 of cefotaxime), 3.84 (s, 3H, NO—CH₃), 6.74 (d, 1H, CH attached to thiazole ring), 7.84 (s, 1H, NH—CH₂—NH), 4.83 (d, 2H, N—CH₂—N′), 6.34 (s, 1H, CH₂—NH′), 9.03 (s, 1H, SO₂NH), 6.6–7.2 (m, ring proton of sulphonamide). ¹³C NMR (DMSO), δ ppm: 12.58 (CH₃ attached to oxazole), 25.78 (CH₂ attached to thiazine), 37.11 (CH₂—COO), 52.06 (CH₃—COO), 52.32 (—N—CH₂—N—), 57.72 (CH attached to thiazine), 59.64 (CH attached to azetidine), 62.98 (N—O—CH₃), 114.74 (CH attached to thiazole), 168.22 (COO—Na), 171.67(COO), 113.47, 128.88, 149.50, 130.24.

Compound 3b: cefotaxime sodium methyl Sulphacetamide; C₂₅H₂₆N₇NaO₁₀S₃; yield 72%, m.p. 105–107 °C. Anal. Calcd. C, 42.67; H, 3.72; N, 13.93; Found C, 42.61; H, 3.66; N, 13.90. IR (KBr) ν_max in cm⁻¹: 3440 ν_s N—H, 3392 ν_as N—H in SO₂NH, 2970 ν_s C—H Aliphatic, 2946 ν_as C—H in CH₂, 1774 ν_s C⚌O in beta lactam, 1655 ν_s C⚌O in amide, 1620 ν_s C⚌O in COONa, 1582 ν_s C⚌N, 1530 N—H bending, 1342 ν_s S⚌O, 1133 C—H in plane bending vibration of 1:4 disubstituted benzene, 1035 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.74 (s, 3H, CH₃—COO), 2.96 (s, 2H, CH₂—COO), 3.55 and 3.42 (ABq, 2H, CH₂ attached to thiazine ring), 5.04 (d, 1H, CH attached to thiazine ring), 5.57 (dd, 1H, CH attached to azetidine ring) 9.54 (d, 1H, NH—C⚌0 of cefotaxime), 3.82 (s, 3H, NO—CH₃), 6.71 (d, 1H, CH attached to thiazole ring), 7.80 (s, 1H, NH—CH₂—NH′), 4.82 (d, 2H, N—CH₂—N′), 6.30 (s, 1H, CH₂—NH′), 9.06 (s, 1H, SO₂NH), 6.6–7.07 (m, ring proton of sulphonamide). ¹³C NMR (DMSO), δ ppm: 22.43 (CO—CH₃) 25.70 (CH₂ attached to thiazine), 37.16 (CH₂—COO), 52.06 (CH₃—COO), 52.33 (—N—CH₂—N—), 57.77 (CH attached to thiazine), 59.67 (CH attached to azetidine), 62.97 (N—O—CH₃), 114.71 (CH attached to thiazole), 162.90 (CO—NH), 168.06 (CO—CH₃), 168.20 (COO—Na), 171.66 (COO), 113.47, 128.88, 149.50, 130.28.

Compound 3c: cefotaxime sodium methyl Sulphacetamide sodium; C₂₅H₂₅N₇Na₂O₁₀S₃; yield 84%, m.p. 180–181 °C, Anal. Calcd. C, 41.38; H, 3.47; N, 13.51; Found C, 41.33; H, 3.45; N, 13.43. IR (KBr) ν_max in cm⁻¹: 2973 ν_s C—H Aliphatic, 2938 ν_as C—H in CH₂, 1775 ν_s C⚌O in beta lactam, 1659 ν_s C⚌O in amide, 1618 ν_s C⚌O in COONa, 1585 ν_s C⚌N, 1533 N—H bending, 1338 ν_s S⚌O, 1130 C—H in plane bending vibration of 1:4 disubstituted benzene, 1038 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.75 (s, 3H, CH₃—COO), 2.90 (s, 2H, CH₂—COO), 3.57 and 3.41 (ABq, 2H, CH₂ attached to thiazine ring), 5.03 (d, 1H, CH attached to thiazine ring), 5.54 (dd, 1H, CH attached to azetidine ring), 9.59 (d, 1H, NH—C⚌0 of cefotaxime), 3.81 (s, 3H, NO—CH₃) 6.74 (d, 1H, CH attached to thiazole ring), 7.82 (s, 1H, NH—CH₂—NH′), 4.82 (d, 2H, N—CH₂—N′), 6.31 (s, 1H, CH₂—NH′), 6.6–7.2 (m, ring proton of sulphonamide). ¹³C NMR (DMSO), δ ppm: 23.93 (CO—CH₃), 25.70 (CH₂ attached to thiazine), 37.11 (CH₂—COO), 52.01 (CH₃—COO), 52.33 (—N—CH₂—N—), 57.77 (CH attached to thiazine), 59.66 (CH attached to azetidine), 62.99 (N—O—CH₃), 114.70 (CH attached to thiazole), 167.50 (CO—CH₃), 168.20 (COO—Na), 171.69 (COO), 112.47, 123.88, 145.50, 132.20.

Compound 3d: cefotaxime sodium methyl silver sulphadiazine; C₂₇H₂₅AgN₉NaO₉S₃; yield 78%, m.p. 90 °C, Anal. Calcd. C, 38.31; H, 2.98; N, 14.89 Found C, 38.30; H, 2.95; N, 14.85. IR (KBr) ν_max in cm⁻¹ 2972 ν_s C—H Aliphatic, 2946 ν_as C—H in C—H in CH₂, 1770 ν_s C⚌O in beta lactam, 1657 ν_s C⚌O in amide, 1622 ν_s C⚌O in COONa, 1580 ν_s C⚌N, 1538 N—H bending, 1342 ν_s S⚌O, 1133 C—H in plane bending vibration of 1:4 disubstituted benzene, 1033 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.77 (s, 3H, CH₃—COO), 2.89 (s, 2H, CH₂—COO), 3.55 and 3.49 (ABq, 2H, CH₂ attached to thiazine ring), 5.10 (d, 1H, CH attached to thiazine ring), 5.50 (dd, 1H, CH attached to azetidine ring), 9.50 (d, 1H, NH—C⚌0 of cefotaxime), 3.82 (s, 3H, NO—CH₃), 6.78 (d, 1H, CH attached to thiazole ring), 7.88 (s, 1H, NH—CH₂—NH), 4.80 (d, 2H, N—CH₂—N′), 6.34 (s, 1H, CH₂—NH′), 6.6–7.2 (m, ring proton of sulphonamide). ¹³C NMR (DMSO), δ ppm: 25.88 (CH₂ attached to thiazine), 37.10 (CH₂—COO), 52.09 (CH₃—COO), 52.28 (—N—CH₂—N—), 57.71 (CH attached to thiazine), 59.66 (CH attached to azetidine), 62.93 (N—O—CH₃), 114.76 (CH attached to thiazole), 168.20 (COO—Na), 171.71 (COO), 113.44, 128.80, 149.51, 130.25, 156.43.

Compound 3e: cefotaxime sodium methyl Sulphadoxine; C₃₁H₃₀N₉NaO₁₁S₃; yield 85%, m.p. 222 °C. Anal. Calcd. C, 45.20; H, 3.67; N, 15.30 Found C, 45.12; H, 3.61; N, 15.24. IR (KBr) ν_max in cm⁻¹: 3454 ν_s N—H, 3381 ν_as N—H in SO₂NH, 2976 ν_s C—H in C—H Aliphatic, 2952 ν_as C—H in CH₂, 1772 ν_s C⚌O in beta lactam, 1653 ν_s C⚌O in amide, 1628 ν_s C⚌O in COONa, 1583 ν_s C⚌N,1536 N—H bending, 1346 ν_s S⚌O, 1136 C—H in plane bending vibration of 1:4 disubstituted benzene, 1035 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.73 (s, 3H, CH₃—COO), 2.95 (s, 2H, CH₂—COO), 3.57 and 3.42 (ABq, 2H, CH₂ attached to thiazine ring), 5.04 (d, 1H, CH attached to thiazine ring), 5.59 (dd, 1H, CH attached to azetidine ring) 9.53 (d, 1H, NH—C⚌0 of cefotaxime), 3.84 (s, 3H, NO—CH₃), 6.70 (d, 1H, CH attached to thiazole ring), 7.82 (s, 1H, NH—CH₂—NH), 4.79 (d, 2H, N—CH₂—N′), 6.35 (s, 1H, CH₂—NH′), 9.03 (s, 1H, SO₂NH), 6.6–7.2 (m, ring proton of sulphonamide). ¹³C NMR (DMSO), δ ppm: 25.78 (CH₂ attached to thiazine), 37.11 (CH₂—COO), 52.06 (CH₃—COO), 52.32 (—N—CH₂—N—), 57.72 (CH attached to thiazine), 59.64 (CH attached to azetidine), 62.98 (N—O—CH₃), 114.74 (CH attached to thiazole), 168.22 (COO—Na), 171.60 (COO), 110.63, 113.47, 128.88, 149.50, 157.70, 130.24, 192.97, 199.13.

Compound 3f: cefotaxime sodium methyl sulphadiazine; C₂₇H₂₆N₉NaO₉S₃; yield 80%, m.p. 160–162 °C. Anal. Calcd. C, 43.84; H, 3.54; N, 17.04 Found C, 43.73; H, 3.51; N, 17.07. IR (KBr) ν_max in cm⁻¹: 3460 ν_s N—H, 3386 ν_as N—H in SO₂NH, 2979 ν_s C—H in C—H Aliphatic, 2945 ν_as C—H in CH₂, 1778 ν_s C⚌O in beta lactam,1654 ν_s C⚌O in amide, 1624 ν_s C⚌O in COONa, 1585 ν_s C⚌N, 1538 N—H bending, 1349 ν_s S⚌O, 1130 C—H in plane bending vibration of 1:4 disubstituted benzene, 1031 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.77 (s, 3H, CH₃—COO), 2.90 (s, 2H, CH₂—COO), 3.59 and 3.44 (ABq, 2H, CH₂ attached to thiazine ring), 5.10 (d, 1H, CH attached to thiazine ring), 5.60 (dd, 1H, CH attached to azetidine ring), 9.55 (d, 1H, NH—C⚌0 of cefotaxime), 3.87 (s, 3H, NO—CH₃), 6.79 (d, 1H, CH attached to thiazole ring), 7.81 (s, 1H, NH—CH₂—NH), 4.75 (d, 2H, N—CH₂—N′), 6.34 (s, 1H, CH₂—NH′), 9.07 (s, 1H, SO₂NH), 6.6–7.2 (m, ring proton of sulphonamide). ¹³C NMR (DMSO), δ ppm: 25.70 (CH₂ attached to thiazine), 37.09 (CH₂—COO), 52.05 (CH₃—COO), 52.33 (—N—CH₂—N), 57.77 (CH attached to thiazine), 59.65 (CH attached to azetidine), 62.90 (N—O—CH₃), 114.76 (CH attached to thiazole), 168.23 (COO—Na), 171.69 (COO), 113.47, 128.88, 149.50, 130.24.

Compound 3g: cefotaxime sodium methyl diethanol amine; C₂₇H₂₇N₆NaO₉S_2; yield 71%, m.p. 225–227 °C, Anal. Calcd. C, 42.42; H, 4.58; N, 14.13 Found C, 42.41; H, 4.51; N, 14.10. IR (KBr) ν_max in cm⁻¹: 3465 ν_s N—H, 3342 ν_s O—H, 2977 ν_s C—H in C—H Aliphatic, 2937 ν_as C—H in CH₂, 1765 ν_s C⚌O in beta lactam, 1650 ν_s C⚌O in amide, 1620 ν_s C⚌O in COONa, 1584 ν_s C⚌N,1531 N—H bending, 1032 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.73 (s, 3H, CH₃—COO), 2.94 (s, 2H, CH₂—COO), 3.56 and 3.38 (ABq, 2H, CH₂ attached to thiazine ring), 5.04 (d, 1H, CH attached to thiazine ring), 5.57 (dd, 1H, CH attached to azetidine ring) 9.53 (d, 1H, NH—C⚌0 of cefotaxime), 3.56 (t, CH₂—OH) 3.84 (s, 3H, NO—CH₃), 4.25 (s, CH₂—OH), 6.78 (d, 1H, CH attached to thiazole ring), 7.84 (s, 1H, NH—CH₂—N), 3.57 (d, 2H, N—CH₂—N′). ¹³C NMR (DMSO), δ ppm: 25.78 (CH₂ attached to thiazine), 37.11 (CH₂—COO), 52.06 (CH₃—COO), 52.32 (—N—CH₂—N—), 56.20 (CH₂—OH), 57.72 (CH attached to thiazine), 59.64 (CH attached to azetidine), 62.98 (N—O—CH₃), 114.74 (CH attached to thiazole), 168.22 (COO—Na), 171.67 (COO), 162.10, 164.96.

Compound 3h: cefotaxime sodium methyl morpholine; C₂₁H₂₅N₆NaO₈S₂; yield 79%, m.p. 215–218 °C, Anal. Calcd. C, 43.75; H, 4.37; N, 14.58 Found C, 43.72; H, 4.33; N, 14.55. IR (KBr) ν_max in cm⁻¹: 3462 ν_s N—H, 2975 ν_s C—H Aliphatic, 2933 ν_as C—H in CH₂, 1762 ν_s C⚌O in beta lactam, 1653 ν_s C⚌O in amide, 1618 ν_s C⚌O in COONa, 1582 ν_s C⚌N, 1528 N—H bending, 1038 ν_s N—O. ¹H NMR (DMSO) δ ppm: 2.40 (t, 2H, N—CH₂), 3.52 (t, 2H, O—CH₂₎, 3.75 (s, 3H, CH₃—COO), 2.94 (s, 2H, CH₂—COO), 3.57 and 3.41 (ABq, 2H, CH₂ attached to thiazine ring), 5.02 (d, 1H, CH attached to thiazine ring), 5.52 (dd, 1H, CH attached to azetidine ring), 9.51 (d, 1H, NH—C⚌0 of cefotaxime), 3.84 (s, 3H, NO—CH₃) 6.72 (d, 1H, CH attached to thiazole ring), 7.86 (s, 1H, NH—CH₂—N), 3.57 (d, 2H, N—CH₂—N′). ¹³C NMR (DMSO), δ ppm: 25.78 (CH₂ attached to thiazine), 37.11 (CH₂—COO), 52.06 (CH₃—COO), 51.63 (N—CH₂), 66.81 (O—CH₂) 52.32 (—N—CH₂—N—), 57.72 (CH attached to thiazine), 59.64 (CH attached to azetidine), 62.98 (N—O—CH), 114.74 (CH attached to thiazole), 168.22 (COO—Na), 171.67 (COO).

Compound 3i cefotaxime sodium methyl Piperazine; C₂₁H₂₆N₇NaO₇S₂; yield 82%, m.p. 235–236 °C, Anal. Calcd. C, 43.82; H, 4.55; N, 17.03 Found C, 43.80; H, 4.52; N, 17.05. IR (KBr) ν_max in cm⁻¹: 3463 N—H, 2972 ν_s C—H Aliphatic, 2930 ν_as C—H in CH₂, 1762 ν_s C⚌O in beta lactam, 1653 ν_s C⚌O in amide, 1626 ν_s C⚌O in COONa, 1583 ν_s C⚌N, 1525 N—H bending, 1038 ν_s N—O. ¹H NMR (DMSO) δ ppm: 2.18 (t, 2H, N—CH₂), 2.64 (t, 2H, NH—CH₂), 3.72 (s, 3H, CH₃—COO), 2.93 (s, 2H, CH₂—COO), 3.50 and 3.41 (ABq, 2H, CH₂ attached to thiazine ring), 5.12 (d, 1H, CH attached to thiazine ring), 5.58 (dd, 1H, CH attached to azetidine ring), 9.50 (d, 1H, NH—C⚌0 of cefotaxime), 3.87 (s, 3H, NO—CH₃), 6.73 (d, 1H, CH attached to thiazole ring), 7.85 (s, 1H, NH—CH₂—N), 3.54 (d, 1H, N—CH₂—N′). ¹³C NMR (DMSO), δ ppm: 25.76 (CH₂ attached to thiazine), 37.14 (CH₂—COO), 52.78 (N—CH₂), 45.90 (NH—CH₂), 52.02 (CH₃—COO), 52.37 (—N—CH₂—N—), 57.79 (CH attached to thiazine), 59.62 (CH attached to azetidine), 62.93 (N—O—CH₃), 114.75 (CH attached to thiazole), 168.26 (COO—Na), 171.61(COO).

Compound 3j: cefotaxime sodium methyl diphenyl amine; C₂₉H₂₇N₆NaO₇S₂; yield 80%, m.p. 220 °C. Anal. Calcd. C, 52.88; H, 4.13; N, 12.76. Found C, 52.80; H, 4.10; N, 12.72. IR (KBr) ν_max in cm⁻¹: 3457 ν_s N—H, 2974 ν_s C—H Aliphatic, 2943 ν_as C—H in CH₂, 1768 C⚌O in beta lactam, 1658 ν_s C⚌O in amide, 1620 ν_s C⚌O in COONa, 1587 ν_s C⚌N, 1520 N—H bending, 1038 ν_s N—O. ¹H NMR (DMSO) δ ppm: 3.78 (s, 3H, CH₃—COO), 2.98 (s, 2H, CH₂—COO), 3.51 and 3.48 (ABq, 2H, CH₂ attached to thiazine ring), 5.20 (d, 1H, CH attached to thiazine ring), 5.53 (dd, 1H, CH attached to azetidine ring), 9.52 (d, 1H, NH—C⚌0 of cefotaxime), 3.80 (s, 3H, NO—CH₃) 6.79 (d, 1H, CH attached to thiazole ring), 7.86 (s, 1H, NH—CH₂—N), 3.57 (d, 2H, N—CH₂—N′). ¹³C NMR (DMSO), δ ppm: 25.68 (CH₂ attached to thiazine), 37.17 (CH₂—COO), 52.03 (CH₃—COO), 52.42 (—N—CH₂—N—), 57.78 (CH attached to thiazine), 59.66 (CH attached to azetidine), 62.99 (N—O—CH₃), 114.69 (CH attached to thiazole), 126.45, 128.96 and 146.08 (CH attached to phenyl), (168.24 (COO—Na), 171.59 (COO), 143.06, 164.10, 163.32, 162.12.

2.2.2

2.2.2 Powder X-ray diffraction studies

Powder X-ray diffraction patterns of three synthesized compounds namely cefotaxime sodium methyl silver sulphadiazine (3d), cefotaxime methyl morpholine (3h) and cefotaxime methyl Piperazine (3i) (Figs. 1–3 respectively) were reported.

Powder X-ray diffraction pattern of cefotaxime sodium methyl silver sulphadiazine (3d).

Powder X-ray diffraction pattern of cefotaxime sodium methyl morpholine (3h).

Powder X-ray diffraction pattern of cefotaxime sodium methyl Piperazine (3i).

Average particle size of the synthesized compounds was determined with the help of the Scherrer formula, in which particle size D is defined as: $0.9 λ / B \cos θ,$ where 0.9 = constant, λ = wavelength, B = angular width and θ = diffraction angle. Average particle size of the compounds determined was 53.33, 17.26 and 22.99 nm respectively.

2.2.3

2.2.3 Scanning Electron Microscopy (SEM) studies

Scanning Electron Microscopy Studies of two of the synthesized compound namely cefotaxime sodium methyl silver sulphadiazine (3d) and cefotaxime methyl Piperazine (3i) were carried out at magnification of ×1000–10 μm and ×600–2 μm. SEM uses a focused beam of high energy electrons to generate a variety of signals from the surface of signals revealing information about the sample, including external morphology, topography, chemical composition crystalline structure, and orientation of materials making up the sample. SEM was recorded different images of each compound at different magnifications, as described above.

Compound cefotaxime sodium methyl silver sulphadiazine (3d) (Figs. 4 and 5) has a different topography and morphology at two different magnifications. In images, the particles exhibit approximately round Shape with different sizes.

SEM image of cefotaxime sodium methyl silver sulphadiazine (magnification of ×1000–10 μm).

SEM image of cefotaxime sodium methyl silver sulphadiazine (magnification of ×600–2 μm).

Cefotaxime sodium methyl Piperazine (3i) is shown in Figs. 6 and 7 with different magnifications. This compound has also round particles in different sizes. SEM studies show microstructure of the compounds, which mainly include surface morphology.

SEM image cefotaxime sodium methyl Piperazine (magnification of ×1000–10 μm).

SEM image cefotaxime sodium methyl Piperazine (magnification of ×600–2 μm).

2.3

2.3 Antimicrobial activity and LD₅₀ test

The newly synthesized cefotaxime sodium derivatives (3a–3j) were screened for their antibacterial activity against pathogenic strains of Escherichia coli and Staphylococcus aureus at varying concentrations – 80 μg/ml, 160 μg/ml and 320 μg/ml using corresponding sulphonamide as their standards by cup plate method. Nutrient agar media were prepared for bacterial growth. The media were autoclaved at 15 lbs pressure (121.6.C) for 30 min. The culture of bacterium was mixed with autoclaved media and poured in plates and bored. The solution of cefotaxime sodium derivatives was poured in these cups in triplicate and incubated at 37 °C for 24 h. Antibacterial activity was ascertained by the zone of inhibition measured in mm as shown in Table 1. The similar procedure was followed for the parent sulphonamide.

Table 1 Antibacterial screening of synthesized cefotaxime sodium derivatives and sulphonamides against E. Coli and S. aureus (zone of inhibition in mm).

Comp. No.	E. coli concentration in μg/ml				S. aureus concentration in μg/ml
Comp. No.	80	160	320	Avg.	80	160	320	Avg.
3a	5.6	7.6	8.8	7.3	16.4	20.0	25.3	20.5
3b	–	–	–	–	9.3	10.6	13.1	11.0
3c	–	–	–	–	15.3	18.0	21.0	18.1
3d	11.6	20.3	23.3	18.4	9.3	11.0	14.3	11.5
3e	13.3	14.0	17.6	14.9	7.3	10.6	12.3	10.0
3f	12.3	17.6	21.6	17.1	16.4	19.2	24.3	19.96
3g	–	–	9.0	3.0	–	–	8.3	2.7
3h	–	–	14.6	4.8	–	9.0	11.6	6.8
3i	6.8	8.0	10.3	8.3	–	11.0	16.6	9.2
3j	–	–	15.3	5.1	–	–	11.6	3.8
2a	12.0	16.6	20.0	16.2	15.6	18.2	21.3	18.3
2b	14.5	17.0	19.0	16.8	15.3	19.0	20.0	18.1
2c	14.0	16.3	20.0	16.7	10.6	16.0	19.3	15.3
2d	9.5	11.8	17.6	12.9	9.0	11.0	14.6	11.5
2e	9.6	10.3	16.0	11.7	7.3	8.3	11.0	8.8

∗Avg: average value of antibacterial activity (for 80, 160 and 320 μg).

The toxicity of synthesized cefotaxime sodium derivatives was ascertained by LD₅₀ test. The test was performed on white mice weighing 25 g. Doses were given orally as well as intraperitoneally and mice were kept under observation for 72 h for each trial. The cefotaxime sodium derivatives showed no adverse toxic effect even of an oral dose of 1400 mg/kg of the body weight of mice. However, when dose was administered intraperitoneally they proved to be lethal at the dose level of 750 mg/kg of the body weight of mice.

3 Results and discussion

The cefotaxime sodium derivatives synthesized by aminomethylation reaction were obtained in good yield (⩾85%). They were analysed for elemental analysis and results were found to be in full agreement with the calculated values. The anticipated structure was in agreement with the spectral data of IR and NMR. The purity of synthesized compounds was assured with aid of chromatographic technique. The stationary phase was silica gel-G. It was of chromatographic grade. The solvents used for mobile phase were methanol and chloroform. They were distilled before using. The spectral studies have shown characteristic band of methylene group incorporated between cefotaxime sodium and the amine component due to aminomethylation. This shows the presence of amino methyl linkage in the synthesized cefotaxime sodium derivatives. The NMR also confirms amino methyl linkage (—CH₂) between amine and active hydrogen. Some of the synthesized compounds were nanosize. The use of nanoparticle in medicine offers some exciting possibilities. Some techniques are only imagined, while others are at various stage of testing, or actually being used today. The cefotaxime sodium derivatives were screened for their biological significance. They were evaluated for antibacterial activity against pathogenic strains of E. Coli, S. aureus at varying concentrations – 80, 160 and 320 μg/ml. These pathogens were subcultured on specific media. The cefotaxime sodium derivatives and the standard compound (sulphonamide and secondary amines) were dissolved in DMF. The reported activities were mean of zone of inhibition in millimetre (in triplicate). All the reported compounds exhibit remarkable in vitro activity against these pathogens. Their activity was also compared with their parent sulphonamide.

Table 1 reflects that most of the compounds had shown remarkable activity only at 320 μg/ml. Antibacterial screening of cefotaxime sodium derivatives against E. coli shows interesting results. 3d was superior to other followed by 3f and 3e in inhibiting the growth of this pathogen. Comparison with parent sulphonamide shows that compound 3d and 3e were superior to the corresponding sulphonamide.

Cefotaxime sodium derivatives had shown significant activity against S. aureus. The compounds 3a, 3f, and 3c were significantly superior to other compounds in exhibiting antibacterial activity against S. aureus. Comparative study with sulphonamides indicates that compounds 3a, 3c and 3e are superior to the corresponding sulphonamides. Moreover, concentration 320 μg/ml was superior for inhibiting the growth of the bacterium.

Comparison of cefotaxime sodium derivatives with sulphonamides shows that, some cefotaxime sodium derivatives are having more antibacterial activity, but former is less toxic than latter as revealed by LD₅₀ test on white mice of weight 25 gm. The newly synthesized compounds seems to be really promising compounds for their antibacterial activity. In the light of those finding we will undertake further synthetic studies on the new compounds in the future.

4 Conclusion

This work shows that cefotaxime sodium derivatives are a potential source of compounds for inhibition of bacteria and could be used as efficient drugs with minimum side effects.

Acknowledgements

We dedicate our sincere thanks to SAIF Chandigarh and UGC-DAE CSR Indore for spectral studies. We also extend our sincere thanks to Govt. Holkar Science College Indore for providing facilities to conduct antibacterial studies.

References

Bashir R., Ovais S., Yaseen S., Hamid H., Alam M.S., Samim M., Javed K., . Synthesis of some new 1, 3, 5-trisubstituted pyrazolines bearing benzene sulfonamide as anticancer and anti-inflammatory agents. Bioorg. Med. Chem. Lett.. 2011;21(14):4301-4305.
[Google Scholar]
Bucourt R., Bormann D., Heymes R., Perronnet M., . Chemistry of cefotaxime. J. Antimicrobiol. Chemother.. 1980;6(suppl A):63-67.
[Google Scholar]
Crespo R., de Bravo M.G., Colinas P.A., Bravo R.D., . In vitro antitumor activity of N-glycosyl sulfonamides. Bioorg. Med. Chem. Lett.. 2010;20(22):6469-6471.
[Google Scholar]
Garaj V., Puccetti L., Fasolis G., Winum J.-Y., Mon-tero J.-L., Scozzafava A., Vullo D., Innocentia A., Su-purana C.T., . Carbonic anhydrase inhibitors: synthesis and in-hibition of cytosolic/tumor-associated carbonic anhy-drase isozymes I, Ii, and Ix with sulfonamides incorporat-ing 1,2,4-triazine moieties. Bioorg. Med. Chem. Lett.. 2004;14:5427-5433.
[Google Scholar]
Ghorab M.M., Ragab F.A., Heiba H.I., Youssef H.A., El-Gazzar M.G., . Synthesis of novel pyrrole and pyrrolo [2,3-d] pyrimidine derivatives bearing sulfonamide moiety for evaluation as anticancer and radiosensitizing agents. Bioorg. Med. Chem. Lett.. 2010;20(21):6316-6320.
[Google Scholar]
Gilbert J.A., Kamath P.S., . Spontaneous bacterial peritonitis: an update. In Mayo Clin. Proc.. 1995;70(4):365-370.
[Google Scholar]
Güzel Ö., Innocenti A., Scozzafava A., Salman A., Supuran C.T., . Carbonic anhydrase inhibitors. Phenacetyl-, pyridylacetyl-and thienylacetyl-substituted aromatic sulfonamides act as potent and selective isoform VII inhibitors. Bioorg. Med. Chem. Lett.. 2009;19(12):3170-3173.
[Google Scholar]
Hu L., Li Z.R., Wang Y.M., Wu Y., Jiang J.D., Boykin D.W., . Novel pyridinyl and pyrimidinylcarbazole sulfonamides as antiproliferative agents. Bioorg. Med. Chem. Lett.. 2007;17(5):1193-1196.
[Google Scholar]
Joshi S., Khosla N., Tiwari P., . In vitro study of some medicinally important Mannich bases derived from antitubercular agent. Bioorg. Med. Chem.. 2004;12(3):571-576.
[Google Scholar]
Joshi S., Bilgaiyan P., Pathak A., . Synthesis and in-vitro study of some medicinally important mannich bases derived from 2-amino-9 [{(1, 3 dihydroxy propane-2yl) oxy} methyl] 6–9 dihydro-3h-purin-6-one. J. Chil. Chem. Soc.. 2012;57(3):1277-1282.
[Google Scholar]
Joshi S., Manikpuri A.D., Tiwari P., . Synthesis and biological study of medicinally important Mannich bases derived from 4-(dimethylamino)-1, 4, 4a, 5, 5a, 6, 11, 12a-octahydro-3, 6, 10, 12, 12a pentahydroxy naphthacene carboxamide. Bioorg. Med. Chem. Lett.. 2007;17(3):645-648.
[Google Scholar]
Joshi S., Bilgaiyan P., Manikpuri A.D., Pathak A., Vyas K., . Synthesis, characterization and Antibacterial screening of aminomethylated derivatives of 7-azaspiro [4.5] decane-6 8-dione. Der Pharma Chem.. 2010;2(3)
[Google Scholar]
Kamal A., Khan M.N.A., Reddy K.S., Rohini K., Sastry G.N., Sateesh B., Sridhar B., . Synthesis, structure analysis, and antibacterial activity of some novel 10-substituted 2-(4-piperidyl/phenyl)-5, 5-dioxo [1, 2, 4] triazolo [1,5b] [1, 2,4] benzothiadiazine derivatives. Bioorg. Med. Chem. Lett.. 2007;17(19):5400-5405.
[Google Scholar]
Lehtonen J.M., Parkkila S., Vullo D., Casini A., Scozzafava A., Supuran C.T., . Carbonic anhydrase inhibitors. Inhibition of cytosolic isozyme XIII with aromatic and heterocyclic sulfonamides: a novel target for the drug design. Bioorg. Med. Chem. Lett.. 2004;14(14):3757-3762.
[Google Scholar]
Puccetti L., Fasolis G., Vullo D., Chohan Z.H., Scozzafava A., Supuran C.T., . Carbonic anhydrase inhibitors. Inhibition of cytosolic/tumor-associated carbonic anhydrase isozymes I, II, IX, and XII with Schiff’s bases incorporating chromone and aromatic sulfonamide moieties, and their zinc complexes. Bioorg. Med. Chem. Lett.. 2005;15(12):3096-3101.
[Google Scholar]
Raddatz J.K., Ostergaard B.E., Rotschafer J.C., . Therapeutic options for cefotaxime in the management of bacterial infections. Diagn. Microbiol. Infect. Dis.. 1995;22(1):77-83.
[Google Scholar]
Rimola A., Salmerón J.M., Clemente G., Rodrigo L., Obrador A., Miranda M.L., Guarner C., Planas R., Solá R., Vargas V., Casafont F., Marco F., Navasa M., Bañares R., Arroyo V., Rodés J., . Two different dosages of cefotaxime in the treatment of spontaneous bacterial peritonitis in cirrhosis: results of a prospective, randomized, multicenter study. Hepatology. 1995;21:674-679.
[Google Scholar]
Wilkinson B.L., Bornaghi L.F., Wright A.D., Houston T.A., Poulsen S.A., . Anti-mycobacterial activity of a bis-sulfonamide. Bioorg. Med. Chem. Lett.. 2007;17(5):1355-1357.
[Google Scholar]
Zhang H., Wang J., Chen Y., Nie Q., . Solubiliy of sodium cefotaxime in aqueous 2-propanol mixtures. J. Chem. Eng. Data. 2006;51(6):2239-2241.
[Google Scholar]
Zimmerman S., Innocenti A., Casini A., Ferry J.G., Scozzafava A., Supuran C.T., . Carbonic anhydrase inhibitors. Inhibition of the prokariotic beta and gamma-class enzymes from Archaea with sulfonamides. Bioorg. Med. Chem. Lett.. 2004;14(24):6001-6006.
[Google Scholar]

Introduction

Experimental

Chemistry
Spectral studies
Antimicrobial activity and LD50 test

Results and discussion

Conclusion

Fulltext Views
598

PDF downloads
205

View/Download PDF
Download Citations

BibTeX
RIS

Show Sections

[1] Bashir R., Ovais S., Yaseen S., Hamid H., Alam M.S., Samim M., Javed K., . Synthesis of some new 1, 3, 5-trisubstituted pyrazolines bearing benzene sulfonamide as anticancer and anti-inflammatory agents. Bioorg. Med. Chem. Lett.. 2011;21(14):4301-4305.
[Google Scholar]

[2] Bucourt R., Bormann D., Heymes R., Perronnet M., . Chemistry of cefotaxime. J. Antimicrobiol. Chemother.. 1980;6(suppl A):63-67.
[Google Scholar]

[3] Crespo R., de Bravo M.G., Colinas P.A., Bravo R.D., . In vitro antitumor activity of N-glycosyl sulfonamides. Bioorg. Med. Chem. Lett.. 2010;20(22):6469-6471.
[Google Scholar]

[4] Garaj V., Puccetti L., Fasolis G., Winum J.-Y., Mon-tero J.-L., Scozzafava A., Vullo D., Innocentia A., Su-purana C.T., . Carbonic anhydrase inhibitors: synthesis and in-hibition of cytosolic/tumor-associated carbonic anhy-drase isozymes I, Ii, and Ix with sulfonamides incorporat-ing 1,2,4-triazine moieties. Bioorg. Med. Chem. Lett.. 2004;14:5427-5433.
[Google Scholar]

[5] Ghorab M.M., Ragab F.A., Heiba H.I., Youssef H.A., El-Gazzar M.G., . Synthesis of novel pyrrole and pyrrolo [2,3-d] pyrimidine derivatives bearing sulfonamide moiety for evaluation as anticancer and radiosensitizing agents. Bioorg. Med. Chem. Lett.. 2010;20(21):6316-6320.
[Google Scholar]

[6] Gilbert J.A., Kamath P.S., . Spontaneous bacterial peritonitis: an update. In Mayo Clin. Proc.. 1995;70(4):365-370.
[Google Scholar]

[7] Güzel Ö., Innocenti A., Scozzafava A., Salman A., Supuran C.T., . Carbonic anhydrase inhibitors. Phenacetyl-, pyridylacetyl-and thienylacetyl-substituted aromatic sulfonamides act as potent and selective isoform VII inhibitors. Bioorg. Med. Chem. Lett.. 2009;19(12):3170-3173.
[Google Scholar]

[8] Hu L., Li Z.R., Wang Y.M., Wu Y., Jiang J.D., Boykin D.W., . Novel pyridinyl and pyrimidinylcarbazole sulfonamides as antiproliferative agents. Bioorg. Med. Chem. Lett.. 2007;17(5):1193-1196.
[Google Scholar]

[9] Joshi S., Khosla N., Tiwari P., . In vitro study of some medicinally important Mannich bases derived from antitubercular agent. Bioorg. Med. Chem.. 2004;12(3):571-576.
[Google Scholar]

[10] Joshi S., Bilgaiyan P., Pathak A., . Synthesis and in-vitro study of some medicinally important mannich bases derived from 2-amino-9 [{(1, 3 dihydroxy propane-2yl) oxy} methyl] 6–9 dihydro-3h-purin-6-one. J. Chil. Chem. Soc.. 2012;57(3):1277-1282.
[Google Scholar]

[11] Joshi S., Manikpuri A.D., Tiwari P., . Synthesis and biological study of medicinally important Mannich bases derived from 4-(dimethylamino)-1, 4, 4a, 5, 5a, 6, 11, 12a-octahydro-3, 6, 10, 12, 12a pentahydroxy naphthacene carboxamide. Bioorg. Med. Chem. Lett.. 2007;17(3):645-648.
[Google Scholar]

[12] Joshi S., Bilgaiyan P., Manikpuri A.D., Pathak A., Vyas K., . Synthesis, characterization and Antibacterial screening of aminomethylated derivatives of 7-azaspiro [4.5] decane-6 8-dione. Der Pharma Chem.. 2010;2(3)
[Google Scholar]

[13] Kamal A., Khan M.N.A., Reddy K.S., Rohini K., Sastry G.N., Sateesh B., Sridhar B., . Synthesis, structure analysis, and antibacterial activity of some novel 10-substituted 2-(4-piperidyl/phenyl)-5, 5-dioxo [1, 2, 4] triazolo [1,5b] [1, 2,4] benzothiadiazine derivatives. Bioorg. Med. Chem. Lett.. 2007;17(19):5400-5405.
[Google Scholar]

[14] Lehtonen J.M., Parkkila S., Vullo D., Casini A., Scozzafava A., Supuran C.T., . Carbonic anhydrase inhibitors. Inhibition of cytosolic isozyme XIII with aromatic and heterocyclic sulfonamides: a novel target for the drug design. Bioorg. Med. Chem. Lett.. 2004;14(14):3757-3762.
[Google Scholar]

[15] Puccetti L., Fasolis G., Vullo D., Chohan Z.H., Scozzafava A., Supuran C.T., . Carbonic anhydrase inhibitors. Inhibition of cytosolic/tumor-associated carbonic anhydrase isozymes I, II, IX, and XII with Schiff’s bases incorporating chromone and aromatic sulfonamide moieties, and their zinc complexes. Bioorg. Med. Chem. Lett.. 2005;15(12):3096-3101.
[Google Scholar]

[16] Raddatz J.K., Ostergaard B.E., Rotschafer J.C., . Therapeutic options for cefotaxime in the management of bacterial infections. Diagn. Microbiol. Infect. Dis.. 1995;22(1):77-83.
[Google Scholar]

[17] Rimola A., Salmerón J.M., Clemente G., Rodrigo L., Obrador A., Miranda M.L., Guarner C., Planas R., Solá R., Vargas V., Casafont F., Marco F., Navasa M., Bañares R., Arroyo V., Rodés J., . Two different dosages of cefotaxime in the treatment of spontaneous bacterial peritonitis in cirrhosis: results of a prospective, randomized, multicenter study. Hepatology. 1995;21:674-679.
[Google Scholar]

[18] Wilkinson B.L., Bornaghi L.F., Wright A.D., Houston T.A., Poulsen S.A., . Anti-mycobacterial activity of a bis-sulfonamide. Bioorg. Med. Chem. Lett.. 2007;17(5):1355-1357.
[Google Scholar]

[19] Zhang H., Wang J., Chen Y., Nie Q., . Solubiliy of sodium cefotaxime in aqueous 2-propanol mixtures. J. Chem. Eng. Data. 2006;51(6):2239-2241.
[Google Scholar]

[20] Zimmerman S., Innocenti A., Casini A., Ferry J.G., Scozzafava A., Supuran C.T., . Carbonic anhydrase inhibitors. Inhibition of the prokariotic beta and gamma-class enzymes from Archaea with sulfonamides. Bioorg. Med. Chem. Lett.. 2004;14(24):6001-6006.
[Google Scholar]

Synthesis, spectroscopic characterization and Antibacterial screening of some new cefotaxime sodium derivatives