Synthesis, Characterisation and Pharmacological Studies of Some New Heterocyclic Hybrid Chalcones and Its Derivative - Isoxazole as Anti-Infective Agents

INTRODUCTION

Various biological activity of heterocyclic compound are extensively distributed in human health. Nitrogen containing heterocyclic compound are important part in both medicinal and organic chemistry [1]. Large number of biological active compounds having common part of chalcones which containe α,β-unsaturated ketone moiety. Therefore, chalcone derivative from synthetic or nature origin exerted biological activity such as antimicrobial [2], antitumor [3], anticancer [4,5], radical scavenger [6], and inhibitor of topoisomeraseI [7]. Chalcone derivative attracts many researchers to development of an efficient synthetic protocol and pharmacological activity. Further chalcone are key intermediate in the synthesis of heterocyclic compound. In this paper, we have synthesised hybrid-chalcone and isoxazole and screen for biological proficiency. Five member heterocyclic compound isoxazole is containing nitrogen atom next to oxygen in ring system. Isoxazole forms several biological active agents and contains broad spectrum of biological activities [8].The substituted isoxazole derivatives possesses displayed antiviral[9], anticancer activities [10], anti-inflammatory, antibacterial [11], hypoglycaemic [12] and antifungal [13]. So studies of novel hybrid molecular targets for new antibacterial drugs are major field of research. Isoxazole gives various types of reaction like complexation, oxidation, reduction, carbanionic condensations, thermolysis, quaternization, protonation, photolysis, thermolysis, transformation of other heterocyclic ring system and reaction with electrophiles, grignard reagents and nuclephiles [14]. Several novel (substituted phenyl) Isoxazole derivatives are synthesized and exists herbicidal activities towards various weeds like Echinochloa, Crusgalli, SetariaViridis, Abutilon theoprastil [15-18]. Isoxazole derivative having property of inhibiting the porphyrinogen oxidase. The product have isoxazole rings have property to cure malaria, inflammatory and microbial activity [19, 20], some isoxazole compounds treat diseases like allergies, cholesterol level and viral [21-23]. Recently, the effort have been made with continuation of previous work to synthesis the biologically significant isoxazole derivatives, we demonstrate a

A 100 ml round bottomed flask, fitted with a reflux condenser was charged with a mixture of 2-chloroisonicotinohydrazide (1) (0.01 mol) and 5-acetylthiophene-2-carbaldehyde (2) (0.01 mol) in presence of acid catalyst in ethanol. Then the reaction mixture was heated under reflux temperature for 5-6 hours. After completion of the reaction as monitored by TLC, the reaction mixture was cooled, and poured onto water⋅ The precipitated solid was filtered off, washed with water, dried and recrystallized from ethanol gives N'-((5-acetylthiophen-2-yl) methylene)-2-chloronicotinohydrazide (3).

By applying classical Claisen-Schmidt condensation reaction, substituted aldehyde (4a-h) (0.01 mol) and N'-((5-acetylthiophen-2-yl)methylene)-2-chloronicotinohydrazide (0.01 mol) (3) dissolved in isopropylalcohol in a 100 ml conical flask. To make it alkaline solution of 40% KOH (5ml) was added in it. Then the reaction mixture was stirred for 24 hours on a magnetic stirrer at room temperature. The progress of reaction was monitored by TLC. After completion of the reaction, the reaction mixture was poured into crushed ice, neutralized with dilute hydrochloric acid and the mixture was agitated for 4 hours a yellow solid was obtained. Finally, the product was isolated yl)methylene)nicotinohydrazide (5a-h).

A 100 ml round bottomed flask, fitted with a reflux condenser was charged with a mixture of an appropriate chalcone (5a-h) (0.01 mol) and hydroxyl amine hydrochloride (0.015 mol) in 40 ml ethanol was taken in a 250 ml round bottomed flask, fitted with a reflux condenser. To make the mixture basic catalytic amount of 40 % KOH (5 ml) solution was added. Then the reaction mixture was heated under reflux for 6-7 hours. The progress of the reaction was monitored by TLC. After completion of the reaction, the reaction mixture was then cooled and poured into crushed ice. Thus the solid mass separated out was collected by filtration, washed with water and recrystallized product (6a-h) from methanol. Reaction Scheme

Scheme. Methodical synthetic route for the target compounds (5a-h) and (6a-h)

SPECTRAL ANALYSIS DATA

Yield 86%; m.p. 1250C; Anal. Calcd. for C13H10ClN3O2S: C, 50.73; H, 3.28; N, 13.65%. (N-H assymetric streching), 1595 (aromatic C=C streching);1H NMR (400 MHz, CDCl3, δ ppm): 2.5 (s, 3H, -CH3), 7.0 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.2 to 8.7 (m, 5H, 3Ar-H and 2-CH of Thiophene moiety);13C NMR (400 MHz, CDCl3, δ ppm) :26.4 (CH3), 148.3(CH), 124.2 (CH), 137.9 (CH), 135.1 (C), 146.2 (C), 163.3 (CO),125.1 (CH), 153.5 (C), 131.3 (CH), 133.4 (CH), 141.6 (C), 190.7 (CO);LCMS (m/z): 308.4 (M+1).

Yield 82%; m.p. 1560C; Anal. Calcd. for C20H13Cl2N3O2S: C, 55.82; H, 3.05; N, 9.77%. Found: C, 55.96; H, 3.16; N, 9.98%; IR (KBr, vmax, cm-1): 1510 (C=N streching, pyridine ring moiety),1661 (C=O streching,amide ketone), 3364 (N-H assymetric streching), 1595 (aromatic C=C streching), 1508 (C=C streching, Chalcone), 3012 (C-H Aromatic ring streching);1H NMR (400 MHz, CDCl3, δ ppm): 7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.8 (m, 9H, 7Ar-H and 2-CH of Thiophene moiety), 7.9 (d, 1H, AR-CH=), 6.6 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) :121.8 (CH), 123.9(CH), 125.2 (CH), 131.7 (CH), 127.9 (CH), 126.5 (CH), 129.2 (CH),129.8 (CH), 138.0 (CH), 145.2 (CH), 148.2 (CH), 137.6 (CH), 146.2 (C), 145.4 (C), 133.3 (C),134.3 (C),154.8 (C), 145.4 (C), 163.4 (C), 180.5 (CO);LCMS (m/z): 431.1 (M+1).

Yield 75%; m.p. 1160C; Anal. Calcd. for C20H14ClN3O3S: C, 58.23; H, 3.43; N, 10.20%. Found: C, 58.61; H, 3.67; N, 10.43%; IR (KBr, vmax, cm-1): 1507 (C=N streching, pyridine ring moiety),1666 (C=O streching,amide ketone), 3361 (N-H assymetric streching), 3426 (phenolic –OH streching), 1592 (aromatic C=C streching), 1501 (C=C streching, Chalcone), 3017 (C-H Aromatic ring streching);1H NMR (400 MHz, CDCl3, δ ppm): 5.3 (s, 1H, -OH), 7.0 (s, 1H, -NH), 8.6 (s, 1H, =CH),7.1 to 8.9 (m, 9H, 7Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) : 121.6 (CH), 123.4(CH), 125.3 (CH), 131.8 (CH), 127.5 (CH), 126.6 (CH), 129.4 (CH), 129.6 (CH), 138.2 (CH), 145.4 (CH), 148.1 (CH), 137.3 (CH), 146.4 (C), 145.6 (C), 133.1 (C), 134.7 (C), 154.9 (C), 145.2 (C), 163.1 (C), 180.3 (CO);LCMS (m/z): 412.2 (M+1).

Yield 75%; m.p. 1850C; Anal. Calcd. for C20H13ClN4O4S: C, 54.49; H, 2.97; N, 12.71%. (N-H assymetric streching), 1595 (aromatic C=C streching), 1511 (C=C streching, Chalcone), 3015 (C-H Aromatic ring streching), 1492 (asymetric streching nitro group);1H NMR (400 MHz, CDCl3, δ ppm):7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.9 (m, 9H, 7Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) : 121.5 (CH), 123.2(CH), 124.2 (CH), 125.2 (CH), 127.5 (CH), 129.8 (CH), 131.5 (CH),134.8 (CH), 137.4(CH), 138.1 (CH), 145.3 (CH), 148.3 (CH), 137.9 (C), 134.8 (C), 145.4 (C),146.2 (C),147.6 (C), 154.8 (C), 163.3 (CO), 180.5 (CO);LCMS (m/z): 441.5 (M+1). 2-Chloro-N'-((5-(3-(4- (diethylamino) phenyl) acryloyl) thiophen - 2 - yl) methylene) nicotinohydrazide (5d) Yield 78%; m.p. 126 0C; Anal. Calcd. for C24H23ClN4O2S: C, 61.73; H, 4.96; N, 12.00%. Found: C, 61.77; H, 5.13; N, 12.36%; IR (KBr, vmax, cm-1): 1501 (C=N streching, pyridine ring moiety),1658 (C=O streching,amide ketone), 3376 (N-H assymetric streching), 1591 (aromatic C=C streching), 1512 (C=C streching, Chalcone), 3007 (C-H Aromatic ring streching), 3050 (C-H streching aliphatic);1H NMR (400 MHz, CDCl3, δ ppm):1.2 (tt, 6H, -CH3), 3.4 (qq, 4H, -CH2), 7.2 (s, 1H, -NH), 8.6 (s, 1H, =CH),7.1 to 8.8 (m, 9H, 7Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) :13.1 (2CH3), 47.2 (2CH2), 121.1 (CH), 123.3(CH), 124.1 (CH), 125.3 (CH), 129.7 (CH), 111.8 (CH), 131.5 (CH),129.5 (CH), 137.5(CH), 138.3 (CH), 145.2 (CH), 148.3 (CH), 124.8 (C), 134.7 (C), 145.4 (C),146.1 (C),111.8 (C), 154.8 (C), 163.3 (CO), 180.5 (CO);LCMS (m/z): 467.3 (M+1).

Yield 78%; m.p. 1050C; Anal. Calcd. for C21H16ClN3O2S: C, 61.53; H, 3.93; N, 10.25%. Found: C, 61.69; H, 4.06; N, 10.50%; IR (KBr, vmax, cm-1): 1519 (C=N streching, pyridine ring moiety),1651 (C=O streching,amide ketone), 3369 (N-H assymetric streching), 1599(aromatic C=C streching), 1501 (C=C streching, Chalcone), 3016 (C-H Aromatic ring streching), 3072 (C-H streching aliphatic);1H NMR (400 MHz, CDCl3, δ ppm):2.3 (s, 3H, -CH3), 7.2 (s, 1H, -NH), 8.6 (s, 1H, =CH),7.1 to 8.8 (m, 9H, 7Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) :21.3 (CH3), 121.1 (CH), 123.3(CH), 124.2 (CH), 125.3 (CH), 128.4 (CH), 128.7 (CH), 131.5 (CH),128.6 (CH), 137.5 (CH), 138.3 (CH), 145.2 (CH), 148.3 (CH), 132.3 (C), 134.7 (C), 145.4 (C),146.1 (C),128.6 (C), 154.8

Yield 72%; m.p. 1490C; Anal. Calcd. for C19H12Cl2N4O2S: C, 52.91; H, 2.80; N, 12.99%. Found: C, 53.09; H, 3.02; N, 13.23%; IR (KBr, vmax, cm-1): 1502 (C=N streching, pyridine ring moiety),1680 (C=O streching,amide ketone), 3372 (N-H assymetric streching), 1601 (aromatic C=C streching), 1500 (C=C streching, Chalcone), 3019 (C-H Aromatic ring streching), 653 (C-Cl streching);1H NMR (400 MHz, CDCl3, δ ppm):7.2 (s, 1H, -NH), 8.6 (s, 1H, =CH),7.1 to 8.8 (m, 8H, 6Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) :121.1 (CH), 124.2 (CH), 125.3 (CH), 128.4 (CH), 117.8 (CH), 148.1 (CH),128.6 (CH), 128.7 (CH), 138.3 (CH), 145.2 (CH), 148.3 (CH), 132.3 (C), 144.3 (C), 145.4 (C),146.1 (C),128.6 (C), 148.4 (C), 163.3 (CO), 180.5 (CO);LCMS (m/z): 432.4 (M+1).

Yield 69%; m.p. 1190C; Anal. Calcd. for C18H12ClN5O2S: C, 54.34; H,3.04; N, 17.60%. Found: C, 54.55; H, 3.33; N, 17.88%; IR (KBr, vmax, cm-1): 1503 (C=N streching, pyridine ring moiety),1669 (C=O streching,amide ketone), 3348 (N-H assymetric streching), 1595 (aromatic C=C streching), 1521 (C=C streching, Chalcone), 3028 (C-H Aromatic ring streching);1H NMR (400 MHz, CDCl3, δ ppm):7.2 (s, 1H, -NH), 8.6 (s, 1H, =CH),7.1 to 9.4 (m, 8H, 6Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) :121.1 (CH), 140.5 (CH), 125.3 (CH), 128.4 (CH), 142.6 (CH), 147.3 (CH),128.6 (CH), 128.7 (CH), 145.2 (CH), 148.3 (CH), 132.3 (C), 153.1 (C), 145.4 (C),146.1 (C),128.6 (C), 148.4 (C), 163.3 (CO), 180.5 (CO);LCMS (m/z): 397.9 (M+1).

Yield 83%; m.p. 1370C; Anal. Calcd. for C22H18ClN3O4S: C, 57.96; H, 3.98; N, 9.22%. Found: C, 57.79; H, 4.21; N, 9.49%; IR (KBr, vmax, cm-1): 1515 (C=N streching, pyridine ring moiety),1658 (C=O streching,amide ketone), 3361 (N-H assymetric streching), 1595 (aromatic C=C streching),1716 (asymmetric C-O-C streching of ether linkage),1522 (C=C streching, Chalcone), 3001 (C-H Aromatic ring streching);1H NMR (400 MHz, CDCl3, δ ppm):3.7 (ss, 6H, -OCH3), 7.2 (s, 1H, -NH), 8.6 (s, 1H, =CH),7.1 to 8.8 (m, 9H, 7Ar-H and 2-CH of Thiophene moiety), 7.6 (d, 1H, AR-CH=), 6.7 (d, 1H, -CO-CH=);13C NMR (400 MHz, CDCl3, δ ppm) (CH), 118.2 (CH), 132.3 (C), 134.7 (C), 145.4 (C),106.5 (C),128.6 (C), 154.8 (C), 163.3 (CO), 180.5 (CO);LCMS (m/z): 456.2 (M+1). 2-Chloro-N'-((5-(5-(2-chlorophenyl)isoxazol-3-yl)thiophen-2-yl)methylene nicotine hydrazide (6a) Yield 76%; m.p. 1540C; Anal. Calcd. for C20H12Cl2N4O2S: C, 54.19; H, 2.73; N, 12.64%. Found: C, 54.47; H, 3.01; N, 12.87%; IR (KBr, vmax, cm-1): 1514 (C=N streching, pyridine ring moiety),1675 (C=O streching,amide ketone), 3368 (N-H assymetric streching), 1599 (aromatic C=C streching),3023 (C-H Aromatic ring streching), 659 (C-Cl streching);1H NMR (400 MHz, CDCl3, δ ppm):7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.9 (m, 10H, 7Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) :148.2 (CH), 124.2(CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH),144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 136.8 (C),128.2 (CH),127.4 (CH), 130.2 (CH), 129.2 (CH), 132.3 (C);LCMS (m/z): 442.8 (M-1). 2-Chloro-N'-((5-(5-(3-hydroxyphenyl) isoxazol-3-yl) thiophen-2-yl) methylene) nicotinohydrazide (6b) Yield 80%; m.p. 1690C; Anal. Calcd. for C20H13ClN4O3S: C, 56.54; H,3.08; N, 13.13%. Found: C, 56.74; H, 3.30; N, 13.34%; IR (KBr, vmax, cm-1): 1514 (C=N streching, pyridine ring moiety),1665 (C=O streching,amide ketone), 3372 (N-H assymetric streching), 1598(aromatic C=C streching),3020 (C-H Aromatic ring streching), 3435 (phenolic –OH streching);1H NMR (400 MHz, CDCl3, δ ppm):5.3 (s, 1H, -OH), 7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.9 (m, 10H, 7Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) : 148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 131.7 (C), 117.8 (CH), 130.7 (CH), 115.8 (CH), 157.6 (C), 111.4 (CH);LCMS (m/z): 424.4 (M+1).

Yield 81%; m.p.1760C; Anal. Calcd. for C20H12ClN5O4S: C, 52.93; H, 2.66; N, 15.43%. Found: C, 53.16; H, 2.87; N, 15.61%;IR (KBr, vmax, cm-1): 1514 (C=N streching, pyridine ring moiety),1670 (C=O streching,amide ketone), 3367 (N-H assymetric streching), 1598 (aromatic C=C streching),3020 (C-H Aromatic ring streching), Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) : 148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 131.3 (C),131.5 (CH), 130.0 (CH), 123.8 (CH), 148.5 (C), 122.6 (CH);LCMS (m/z): 454.7 (M+1).

Yield 79%; m.p. 1320C; Anal. Calcd. for C24H22ClN5O2S: C, 60.06; H,4.62; N, 14.59%. Found: C, 60.23; H, 4.83; N, 14.72%;IR (KBr, vmax, cm-1): 1506 (C=N streching, pyridine ring moiety),1662 (C=O streching,amide ketone), 3352 (N-H assymetric streching), 1591 (aromatic C=C streching),3010 (C-H Aromatic ring streching), 3064 (C-H streching aliphatic);1H NMR (400 MHz, CDCl3, δ ppm): 1.2 (t, 6H, -CH3), 3.4 (q, 4H, -CH2),7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.7 (m, 10H, 7Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) :12.8 (2CH3), 47.3 (2CH2), 148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 136.0 (C),125.7 (CH), 112.8 (CH), 149.6 (C), 112.8 (CH), 125.6 (CH);LCMS (m/z): 479.1 (M-1).

Yield 74%; m.p. 1070C; Anal. Calcd. for C21H15ClN4O2S: C, 59.64; H, 3.58; N, 13.25%. Found: C, 59.88; H, 3.79; N, 13.50%;IR (KBr, vmax, cm-1): 1503 (C=N streching, pyridine ring moiety),1676 (C=O streching,amide ketone), 3359 (N-H assymetric streching), 1597 (aromatic C=C streching),3009 (C-H Aromatic ring streching), 3071 (C-H streching aliphatic);1H NMR (400 MHz, CDCl3, δ ppm): 2.2 (s, 3H, -CH3),7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.7 (m, 10H, 7Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) :21.4 (CH3), 148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 123.6 (C),124.8 (CH), 129.3 (CH), 131.7 (C), 129.6 (CH), 124.7 (CH);LCMS (m/z): 423.2 (M+1).

Yield 60%; m.p. 1590C; Anal. Calcd. for C19H11Cl2N5O2S, C, 51.36; H, 2.50; N, 15.76%. Found: C, 51.64; H, 2.73; N, 15.87%;IR (KBr, vmax, cm-1): 1519 (C=N streching, pyridine ring (C-Cl streching);1H NMR (400 MHz, CDCl3, δ ppm):7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.7 (m, 9H, 6Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) : 148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 143.4 (C),120.4 (CH), 148.5 (CH), 148.2 (CH), 130.1 (CH);LCMS (m/z): 445.1 (M+1).

Yield 69%; m.p. 1380C; Anal. Calcd. for C18H11ClN6O2S: C, 52.62; H,2.70; N, 20.46%. Found: C, 52.81; H, 2.91; N, 20.72%;IR (KBr, vmax, cm-1): 1505 (C=N streching, pyridine ring moiety),1666 (C=O streching,amide ketone), 3354 (N-H assymetric streching), 1599 (aromatic C=C streching),1711 (asymmetric C-O-C streching of ether linkage),1515 (C=C streching, Chalcone), 3011 (C-H Aromatic ring streching);1H NMR (400 MHz, CDCl3, δ ppm):7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.7 (m, 9H, 6Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) : 148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 152.4 (C),144.8 (CH), 142.3 (CH), 144.6 (CH);LCMS (m/z): 411.3 (M+1).

Yield 69%; m.p. 1460C; Anal. Calcd. for C22H17ClN4O4S: C, 56.35; H, 3.65; N, 11.95%. Found: C, 56.58; H, 3.75; N, 12.21%;IR (KBr, vmax, cm-1): 1504 (C=N streching, pyridine ring moiety),1669 (C=O streching,amide ketone), 3368 (N-H assymetric streching), 1595 (aromatic C=C streching),1709 (asymmetric C-O-C streching of ether linkage),1513 (C=C streching, Chalcone), 3016 (C-H Aromatic ring streching);1H NMR (400 MHz, CDCl3, δ ppm): 3.8 (s, 6H, -OCH3),7.1 (s, 1H, -NH), 8.7 (s, 1H, =CH),7.1 to 8.7 (m, 10H, 7Ar-H and 2-CH of Thiophene moiety, 1-CH isoxazole moiety);13C NMR (400 MHz, CDCl3, δ ppm) :55.5 (OCH3), 56.2 (OCH3)148.2 (CH), 124.2 (CH), 138.2 (CH), 135.1 (C), 146.2 (C), 163.2 (CO), 125.3 (CH), 144.1 (C), 140.0 (C), 127.5 (CH), 128.3 (C), 150.1 (CH), 98.2 (CH), 169.2 (C), 106.9 (C),127.4 (CH), 107.3 (CH), 161.4 (C), 98.7 (CH), 158.4 (C);LCMS (m/z): 469.5 (M+1).

All these new heterocyclic derivatives (5a-h) and (6a-h) were characterised by means of spectroscopic techniques FT-IR, 1H NMR,13C NMR and LCMS. FTIR spectrum of compound 5a, C=O and CH = CH functionality of chalcone moiety were observed at 1661 and 1508 cm-1 further the 1H NMR spectrum of same compound exerted a doublet at δ 76.6 and 7.9 ppm for the -CO-CH= and Ar-CH= of α, β proton. the, The most deshielded signal appeared in 13C NMR spectrum of compound 5a at δ 170.2, 148.2 and 137.6 ppm was mainly due to the carbonyl carbon and CH = CH functionality of the chalcone moiety. Isoxazole was confirmed by screened the compound 6a for FTIR, a strong absorption band was observe at 1603 cm-1which corresponds to the stretching vibration of the C-N functionality of of isoxazoloe ring. 1H NMR of isoxazole derivative 6a to 6h showed the CH proton of isoxazole ring at between 5.5 to 6.9 ppm as well as carbon NMR also depicted the C=N functionality of isoxazole ring at 157.6 δ ppm. Additionally, mass spectrum of all the compounds showed molecular ion peak M+ corresponding to their exact mass which is in agreement with its proposed structure. The obtained elemental analysis values are in good agreement with theoretical data.

All the synthesised compounds were screened for their antibacterial and antitubercular activity using standard broth micro dilution method according to National Committee for Clinical Laboratory Standards (NCCLS) [24] against Staphylococcus aureus MTCC 96 andStreptococcus pyogenes MTCC 442, Escherichia coli MTCC 443, Pseudomonas aeruginosa MTCC 441, Candida albicans MTCC 227, Aspergillusniger MTCC 282 and Aspergillusclavatus MTCC 1323. The minimal inhibitory concentration (MIC) of all the synthesised compounds are summarised in Table 1. The same all compounds screened for in vitro antitubercular activity by using Lowenstein-Jensen medium (conventional method) against Mycobacterial tuberculosis H37Rv strain [25]. The compunds who has more than 80 % zone of inhibition in primary scrring are selected for MIC and data obtained are summerized in Table 2.

S. a.: Staphylococcus aureus, S. p.: Streptococcus pyogenes, E. c.: Escherichia coli, P. a.: Pseudomonas aeruginosa, C. a.: Candida albicans, A. n.: Aspergillusniger, A. c.: Aspergillusclavatus.Ampi: Ampicillin, Chlo.: Chloramphenicol, Cipr.: Ciprofloxacin, Gris.: Greseofulvin, Nyst.: Nystatin. ‗-‗: not tested. Table 2. In vitro antitubercular activity (% inhibition) and MIC of the synthesized compounds (5a-h) and (6a-h) The antibacterial screening of compounds hybrid chalcone (5a-h), compound 5f and 5g showed an outstanding inhibitory effect i.e. MIC = 62.5 and 50 µg/ml against Staphylococcus aureusas compared ampicillin (MIC = 250 µg/ml) and moderate to chloramphenicol and ciprofloxacin (MIC = 50 µg/ml) while in isoxazole series compounds, compounds 6a, 6f and 6g exhibited good activity against same Staphylococcus aureusas compared ampicillin (MIC = 250 µg/ml). In the case of inhibiting Streptococcus pyogenes, compound 5a, 5b, 5c, 5e, 5f, 5h, 6e, 6g and 6h were found to be negative bacteria, compound 5g, 6g and 6h (MIC = 62.5 µg/ml) showed excellent activity against Escherichia coli as compared to ampicillin while remaining compounds poor activity against Escherichia coli upon comparison with the standard drug ampicillin and lowest to chloramphenicol (MIC = 50 µg/ml) and ciprofloxacin (MIC = 25 µg/ml). Against Pseudomonas aeruginosa, Compound 5a, 5d, 5e, 5f, 5g, 5h, 6e, 6f, 6g and 6h found to possesses better to ampicillin (MIC = 100 µg/ml) and modest to chloramphenicol (MIC = 50 µg/ml) and ciprofloxacin (MIC = 25 µg/ml).The remaining compounds showed moderate to good activity to inhibit the growth of bacterial pathogens and were found less effective than the employed standard drugs.

In vitro antifungal activity data summarized in table 1 shows that, it is found that compounds 5a, 5b,5f, 5g,6f and 6g displayed good antifungal activity against Candida albicans as compared to griseofulvin (MIC = 500 µg/ml) and nystatin (MIC = 100 µg/ml). Compound 5a, 5b, 5f, 5g, 6a, 6f and 6g showed better to griseofulvin (MIC = 100 µg/ml) and nystatin (MIC = 100 µg/ml) against Aspergillus niger. While compound 6g was found to be active against the fungal pathogen Aspergillus clavatus.

The antitubercular activities summarized in table 2 `indicate that the compounds 5a, 5f, 5g, 5h, 6a, 6b, 6f and 6h observed more than 80% inhibition in preliminary test and that compounds are selected for screening for MIC and showed good zone of

CONCLUSION

In this study, some new hybrid chalcone-imines clubbed compounds and their derivatives isozxazoles have been synthesized and act as a powerful template for making a potent antimicrobial and antitubercular agents as anti-infective agents. In addition to that each compound had been shown an excellent antimicrobial and antitubercular activity. Among the various analogues, compounds 5a, 5f, 5g, 6a, 6b and 6f had electron donating groups such as Cl,-OH present in the part of the molecular structure and offered as excellent antiinfective agents. Therefore, there is a need for further study of the above mentioned compounds for the development of the novel a typical antiinfective agents.

The authors are grateful thankful to, RSIC Punjab University for FTIR, 1H NMR, and 13C NMR spectral analysis as well as elemental analysis and Microcare Laboratory, Surat, for antimicrobial activity and antitubercular activity.

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Department of Chemistry, Veer Bahadur Singh Purvanchal University, Jaunpur