642 Indian Journal of Forensic Medicine & Toxicology, January-March 2020, Vol.DOI 14, No.Number: 1 10.37506/v14/i1/2020/ijfmt/192974 Study the Toxicity and Anticancer activity of Some New Amic Acid and Their Derivatives of Mefenamic acid Ali A. Sabah1, Muna S. Al-Rawi1, Jumbad H. Tomma1 1Department of Chemistry, College of Education for Pure Science Ibn Al- Haitham, University of Baghdad, Baghdad, Iraq Abstract A series of amic acid derivatives of mefenamic acid were synthesized with the aim of inhibitting topical gastrointestinal toxicity of mefenamic acid. The key intermediate amic acid (III) was prepared from the reaction of acid hydrazid of mefenamic acid(II) with phthalic anhydrides in dry actone. The new type of imide compound (IV) was dehydrated the amic acid(III) with acetic anhydrous and sodium acetate. The esterification of hydroxyl groups of amic acid (III) produce corresponding ester(V), which was condensed with hydrazine hydrate to give acid hydrazide (VI), then the later compound reacted with syringaldehyde in dry benzene to yield new schiff base (VII). The new derivatives containing heterocyclic unit (VIII)- (X), four and five, member ring were successfully formed such as azetidin-2-one, thiazolidin-4-one, and,imidazolidin-4-one. The structures of the synthesized compounds were confirmed using FTIR,1 HNMR, Mass and CHN-S. The antibacterial activities of some synthesized compounds were screened and showed a highest or low inhibition against Staph.aureus (G+), Bacillus subtilisa (G+), Klebsiella pneumoniae (G-), and E.coli (G-). Also, The cytotoxic effect of different concentrations of some the synthesized compounds was tested against MCF-7 cell line (human breast carcinoma cells) and positive results were obtained for some of them, which encouraged us to study the toxicity using living organisms (mice) to evaluate its acute toxicity and proved the resules of non-toxicity of the derivatives. Keywords: Mefenamic acid, imidazolidin-4-one, antibacterial activities, MCF-7 cell line, acute toxicity study. Introduction for different applications, including the presence of effective anti-cancer drugs, antimicrobial, inflammatory Mefenamic acid is one of the anthranilic acid and cytotoxic activities[2]. derivatives class of non-steroidal anti-inflammatory drugs (NSAID) ) is thought to be due to their interference Amic acid and its derivatives have been extremely with prostaglandin biosynthesis, and is used to treat used in drug industry, advanced materials and strong analgesic and antiinflammatory agents in the biologically active heterocycles. In fact, amic acid treatment of degeneration of joint cartilage, rheumatoid carrying suitable position of carboxylic group are prime [1] arthritis and musculosketal disease . precursors for synthesis of heterocyclic derivatives[3]. The general view in the literature shows that A large number of heterocyclic compounds have been mefenamic acid undergoes a number of reactions, this explored for developing pharmaceutically molecules. has been used in the preparation of many compounds Among this the derivatives: azetidin-2-one (β-lactam), containing heterocyclic moiety of therapeutic molecules thiazolidin-4-one, and imidazolidin-4-one, have been playing an important function in the pharmaceutical chemistry [4]. This study is aimed at synthesis new Corresponding Author: active molecules with fewer side effects that can be Muna S. Al-rawi, successfully used as drugs, the cytotoxic effect and non- Assistant Professor, toxicity proved the resules of this derivatives. E-mail: [email protected] Indian Journal of Forensic Medicine & Toxicology, January-March 2020, Vol. 14, No. 1 643 Experimental Shimadzo (Ir prestige-21) Fourier Transform Infrared Spectrometer. 1HNMR spectra were recorded on Materials and instruments: Bruker, Ultra Shield (300)MHz, Switzerland and using All solvents and chemicals were purchased from DMSO-d6, and CDCl3 with reference to TMS as internal Sigma- Aldrich, and GCC Chemicals company. reference. Elemental micro-analyses were carried out Melting points were recorded using electrothermal using an EuroEA Analyzer. Mass spectra of compounds melting point apparatus and are uncorrected. The were measured by Electron Impact (EI) 70eVmass using FTIR spectrums were registered using KBr discs on a MS Model: 5973 spectrometer. Synthetic Procedures The routes for synthesis new derivatives as depending on to the Scheme (1): HN O HN HN O Methanol O NH2NH2.H2O H2N HO H3CO HN H2SO4 (I) (II) Phthalic anhydride acetone O HN OH O HN O O anh.(CH CO) O H 3 2 N , N CH3COONa H N NH (IV) O (III) O H SO Methanol2 4 OCH3 O O HN H N N H O (V) Cl Ar NH NH H O C 2 2. 2 N O NH2 HN HN O O O HN O HN H H N N N N ClCH COCl H H 2 O (VI) O (VIII) Ar ArCHO Ar CH S GAA NH N HN HN HSCH2COOH O O HN O H O O HN N H N N H N O (VII) H O (IX) glycine Ar NH Et3N N HN O O O HN H N N H Ar = 4- OH,3,5-OCH3-C6H2 O (X) 644 Indian Journal of Forensic Medicine & Toxicology, January-March 2020, Vol. 14, No. 1 Scheme (1): The synthetic route for target Calcd. for C23H19N3O3: C, 71.68; H, 4.93; N, 10.90; derivatives(I)-(X) Found: C, 71.98; H, 5.11; N, 11.03. Preparation of methyl 2-(2,3-dimethylanilino) Synthesis of methyl 2-(2-(2-((2,3-dimethylphenyl) benzoate (I) amino) benzoyl) hydrazine -1-carbonyl) benzoate (V): This compound (I)was prepared following the procedure described by Lit. [5] yield 80%; m.p(96-98)0C. A mixture of compound (III) (4.03 g, 0.01mol), absolute methanol 30mL and H2SO4 5.4 mL was refluxed Preparation of 2-(2,3-dimethylphenylamino) 6 hrs. After cooling washed the mixture with NaHCO3 benzohydrazide (II) solution, The resulting of a white crystals solid was This compound(II) was prepared following the filtered off, washed with water, dried and recrystallized procedure described by Lit. [5]. yield 78%,; m.p (118- by ethanol to give compound (V), yield 71%; mp ( o -1 120)ºC. 137-139) C; FTIR (ν ,cm ): 3330-3309 (NH), 2987 (C-H arom.), 2939-2922(C-H aliph.), 1732, 1685, 1657 Synthesis of 2-(2-(2-((2,3-dimethylphenyl)amino) (C=O), and 1255 (C-O); 1H NMR (δ ppm): 2.08 (s, 3H, benzoyl)hydrazine-1-carbonyl ) benzoic acid(III) CH3), 2.27 (s, 3H, CH3),3.86(s,3H,OCH3), 6.66-8.09 (m,11H,Ar-H), 9.66,and 9.18 for (s,1H,NH); Elemental In a flask with magnetic stirrer, a solution of analysis for this compound : Calcd. for C24H23N3O4: C, (2.55g, 0.01 mol) of acid hydrazide (II) in 20 mL of 69.06; H, 5.51; N, 10.07; Found: C, 69.45; H, 5.80; N, acetone was dripped into a solution of (1.48g, 0.01 10.30. mol) phthalic anhydride in 20 mL of acetone for 30 min at room temperature. Stirring was continued for 5 hrs. Synthesis of N’-(2-((2,3-dimethylphenyl) amino) The brown solid precipitate was filtered, washed, dried benzoyl) phthalohydrazide (VI): and recrystallized from ethanol to give a new amic acid [III], yield 74% , m.p (167-169)o C; FTIR (ν ,cm-1): A mixture of ester compound (V) (4.17gm,0.01mol), 3392-2571(OH), 3340-3309 (NH), 3012 (C-H arom.), was dissolved in absolute ethanol 3mL. hydrazine 2974-2912(C-H aliph.), 1712, 1653 (C=O), and 1255 hydrate(0.5gm,0.01 mol) was added slowly and the 1 mixture was refluxed for 6hrs. The mixture was cooled (C-O); H NMR (δ ppm): 2.09 (s, 3H, CH3), 2.28 (s, and the pale yellow solid was filtered, then used ethanol 3H, CH3), 6.66-8.08 (m,11H,Ar-H), 9.45, 7.91,and 7.58 for (s,1H,NH), 12.06(s, 1H, OH, broad); mass spectra to recrystallized, yield 70%, m.p ( 158-160)°C, FTIR (ν -1 (relative intensity%): m/z=403(5), 223 (75), 178(70), ,cm ): 3305-3145(NH2,NH), 3001 (C-H arom.), 2941- 1 75(100), 104(68), 51(25):Elemental analysis: Calcd. 2822(C-H aliph.), 1660,1639(C═O amide); H NMR (δ ppm): 2.11 (s, 3H, CH3), 2.28 (s, 3H, CH3), 6.56- for C23H21N3O4: C, 68.48; H, 5.21; N, 10.42; Found: C, 68.18; H, 5.41; N, 10.73. 8.05 (m,11H,Ar-H),11.13, 9.18,and 8.13(s,1H,NH); Elemental analysis: Calcd. for C23H23N5O3: C, 66.18; H, Synthesis of 2-((2,3-dimethylphenyl)amino) -N- 5.51; N, 16.78; Found: C, 66.70; H, 5.78; N, 17.01. (1,3-dioxoisoindolin-2-yl) benzamide (IV) Synthesis of (Z)-N’1-(2-((2,3-dimethylphenyl) A mixture of 5 mL acetic anhydride and (0.82g , amino) benzoyl) - N’2-(4-hydroxy - 3,5- 0.01mol) anhydrous sodium acetate was allowed to dimethoxybenzylidene) phthalohydrazide (VII) dropping with stirring to a solution of amic acid(III) (4.03 g, 0.01 mol) in 10 mL ethanol for1hrs. The A mixture of new compound (VI)(4.17g, 0.01 mol) , resulting mixture was refluxed over water path for Syringaldehyde (1.82g, 0.01mol) with EtOH 15 mL and 3hr. The reaction poured into ice water, the pale beige four drops of GAA glacial acetic acid was refluxed for precipitated was collected filtered and washed with 6hrs, cooled after that filtered the product and crystallized ice water and recrystallization from ethanol, yield from ethanol,dark yellow powder, yield 70% , m.p. (150- -1 70% , mp. (188-190)oC; FTIR (ν ,cm-1): 3344 (NH), 152)°C, FTIR (ν ,cm ): 3548(OH), 3469-3344(NH), 2978 (C-H arom.), 2885-2735(C-H aliph.), 1747, 1645 3016 (C-H arom.), 2968-2935(C-H aliph.), 1651(C═O 1 (C=O), and 1238 (C-O); 1H NMR (δ ppm): 2.09 (s, amide), 1626(C=N); HNMR(δ,ppm,CDCl3): 2.10 (s, 3H, CH3), 2.29 (s, 3H, CH3), 2.51(s, 1H,OH) , 3.84 (s,6H, 3H, CH3), 2.27 (s, 3H, CH3), 6.66-8.06 (m,11H,Ar- H),9.57(s,1H,NH), 8.14(s,1H,NH); Elemental analysis: OCH3),6.68-8.58 (m,13H,Ar-H),7.32(s,1H,CH-N) Indian Journal of Forensic Medicine & Toxicology, January-March 2020, Vol.