Synthesis, characterization and biological activity of carbamide, semicarbazide, carbazide and ester derivatives of amino acids and nonsteroidal antiinflammatory drugs

Abstract

U okviru ovog doktorskog rada, koji je nastavak istraživanja o mogućnostima korištenja benzotriazola u sintetskoj kemiji na Zavodu za Farmaceutsku kemiju Farmaceutsko-biokemijskog fakulteta u Zagrebu, sintetizirano je nekoliko serija spojeva. Polazni spoj u sintezi svih prekursora bio je klorid 1- benzotriazolkarboksilne kiseline (BtcCl) koji daje vrlo reaktivne derivate. BtcCl vrlo lako reagira s nukleofilima dajući cijeli niz reaktivnih spojeva iz kojih je do sada pripravljen velik broj organskih spojeva. Derivati aminokiselina ureidoamidi 4a-o pripravljeni su reakcijom klorida N-(1-benzotriazolkarbonil)aminokiselina 3 i odgovarajućih aminoalkohola, a N- i O- supstituirane hidroksiuree 6a-l iz amida N-(1- benzotriazolkarbonil)aminokiselina i odgovarajućih hidroksilamina. Derivati nesteroidnih protuupalnih lijekova (NSAID, ibuprofena, fenoprofena i ketoprofena) sintetizirani su iz NSAID hidrazida 11 koji su dobiveni aminolizom NSAID benzotriazolida 10 i hidrazina. Hidrazidi 11 su u reakciji s 1-karbamoilbenzotriazolima (aktivne uree, 7) dali 1-acilsemikarbazidne derivate NSAID 13a-v, a u reakciji s 1-(1-benzotriazolkarbonil)-4- benziloksisemikarbazidom (9) 1-acil-5-benziloksikarbamoilkarbazide 14a-c. Reakcije u talini uz TEA bitno su skratile vrijeme reakcije. 4-Hidroksisemikarbazidni derivati NSAID 13w-y i 14d-f pripravljeni su katalitičkim hidrogeniranjem O-benzilnih derivata 13t-v i 14a-c uz Pd/C. Iz benzotriazolida ketoprofena (10d) i odgovarajućih ureidoamida (4b,i,j,m) pripravljeni su esteri s dvije molekule ketoprofena kovalentno povezane ureidoamidima u jednu molekulu („dvojni lijekovi“). Svi sintetizirani spojevi karakterizirani su uobičajenim analitičkim postupcima i spektroskopskim metodama te biološki ispitani in vitro na citostatsko, antivirusno, antimikrobno i antioksidativno djelovanje. Nadalje, ispitana je inhibicija lipooksigenaze i lipidne perkosidacije. Najbolje antitumorsko djelovanje pokazao je spoj 6l, antivirusno 13a,g,m, a antiooksidativno spoj 9. Najjači inhibitor lipooksigenaze bio je spoj 13s, a lipidne peroksidacije 13c,l,t. Uočeno je ponavljanje benzhidrilnog supstituenta u spojevima s istaknutom aktivnosti, što je dobar pokazatelj za daljnje usmjeravanje istraživanja.The research described in the thesis is a continuation of the previous work on the use of benzotriazole in the synthetic chemistry which has been carring out at Department of Medicinal Chemistry, Faculty of Pharmacy and Biochemistry in Zagreb for many years. The thesis includes synthesis of several types of compounds. The initial compound for the synthesis of all the intermediates was benzotriazole carboxylic acid chloride (BtcCl), which upon the reaction with nucleophiles gives reactive products, useful in the preparation of various organic compounds. Amino acid derivatives ureidoamides 4a-o were syntehsized by aminolysis of N-(1- benzotriazolecarbonyl)amino acid chlorides 3 with corresponding aminoalcohols and N- i O- substituted hydroxyureas 6a-l were synthesizes from N-(1-benzotriazolecarbonyl)amino acid amides 5 and hydroxylamines. Nonsteroidal antiinflammatory drugs (NSAID, ibuprofen, fenoprofen and ketoprofen) derivatives were prepared from NSAID hydrazides 11 which were obtained by aminolysis of NSAID benzotriazolides 10 and hydrazine hydrate. The reaction of hydrazides 11 and 1-benzotriazole carboxylic acid amides 7 gave 1-acylsemicarbazides 13a-v, while with 1-(1-benzotriazolecarbonyl)-4-benzyloxysemicarbazide (9) gave 1-acyl-5- benzyloxycarbamoyl-carbazides 14a-c. Reactions in melted state in the presence of TEA significantly reduced the reaction time. 4-Hydroxysemicarbazide compounds 13w-y and 14d-f were prepared by catalytic hydrogenation of O-benzylated compounds 13t-v i 14a-c with Pd/C. Ketoprofen benzotriazolide (10d) reacted with several ureidoamides (4b,i,j,m) and gave esters with two molecules of ketoprofen combined covalently in one („twin drugs“). The compounds were evaluated for their biological activity in vitro: antitumor, antimicrobial, antiviral and antioxidant activity, inhibition of lipoxygenase and linoleic acid lipid peroxidation. The best antitumor activity was exerted by 6l, antiviral by 13a,g,m, antioxidant by compound 9. The strongest inhibitor of lipoxygenase was 13s. Compounds 13c,l,t showed the highest inhibition of linoleic acid lipid peroxidation. It is worth to note that compounds with the highest biological activity contained benzhydryl substituent