Bazı sübstitüe-2.4- dihidro-3H- 1,2,4- triazol -3-tiyon türevi bileşiklerinin in vivo metabolizması

Abstract

1. ÖZET 4,5-Disübstitüe-l,2,4-triazol-3-tiyonlar, antidepresan, antimikro- biyel, antitüberküler, analjezik, antiinflamatuar, diüretik ve hipoglisemik aktivite göstermektedir. Bu nedenle bu yapıların biyotransformasyonunu incelemeyi amaçladık. Bu araştırmada, 5-(4-nitrofenü)-4-fenetil-2,4-dihidro-3H-l,2,4- triazol-3-tiyon ve 5-(4-ni1rofenü)^-feml-2,4-dihidro-3H-l,2,4-triazol-3- tiyonun in vivo metabolik yollarmı araştırdık. Bu bileşiklerin, antibakteriyel ve antifungal etkileri daha önce incelenmiş ve aktif oldukları saptanmıştır. İlk aşamada, substratlann sodyum tuzlan, substratlar ve olası metabolitler sentezlenmiştir. Bu bileşiklerden, sodyum 5-(4-nitrofenil)- 4-fenetü-l,2,4-triazol-3-tiyolat ve sodyum 5-(4-nitrofenü)-4-fenil- 1,2,4- triazol-3-tiyolat, karşılık gelen tiyosernikarbazidlerin 2N sodyum hidroksid ile 4 saat ısıülmasıyla oluşturulmuştur. Bu maddeler, 5-(4- nitrofenil)-4-fenetn-2,4-dihidro-3H-l,2,4-triazol-3-tiyon (substrat) ve 5-(4-nitrofenil)-4-fenil-2,4-dihidro-3H-l,2,4-triazol-3-tiyonu (substrat) elde etmek için hidroklorik asid ile asidlendirürniştir. Substratlann sodyum tuzlan, sıçanlara enjeksiyon için ; substratlar ise in vivo çalışmada standart olarak kııUanılmıştır.Asetilasyon metabolitleri; 5-(4-asetilaminofenil)-4-fenetil-2,4- dihidro-3H-l,2?4-triazol-3-tiyon ve 5-(4-asetilaminofenil)-4-fenil-2,4- dihidro-3H-l,2,4-triazol-3-tiyon, karşılık gelen tiyosemikarbazidleriıı 2N sodyum hidroksid ile 1 saat ısıtılmasıyla sentezlenmiştir. Diğer metaboMer, laboratuarımızdan sağlanmıştır. Orijinal bileşikler, elemental analiz ve UV, ^-NMR spektroskopik metod- lar kuUamlarak aydınlatılmıştır. In vivo metabolizma çalışmasında, erkek Wistar sıçanlar (250+20 g) laulamlmıştır. Sodyum 5-(4-nitrofenil)-4-fenetil-l,2,4-triazol-3-tiyolat, 20 mg/ml isotonik serum fizyolojik içinde çözülmüştür ; Sodyum 5-(4- nitrofenil)-4-fenil-l,2,4-triazol-3-tiyolat 2 mg/ml isotonik serum fizyo lojik içinde çözülmüş ve 2 mi hacimde intraperitonal olarak enjekte edilmiştir. Kan (0.3±0.2 mi), eter anestezisi aranda, sıçanların oftabnik venlerinden steril kapiler tüp ile toplanmıştır. Santrifîijlendikten sonra, plazma ayınlmıştır. Bu bileşikler, İTK ve ters faz YBSK metodlan kullanılarak ayınlmıştır. Çalışmamızda, 5-(4-mtrofenü)-4-fenetil-2,4-dihidro-3H-l,2,4- triazol -3-tiyondan, asetil metabolit [ 5-(4-asetüaminofeml)-4-fenetil- 2,4-dihidro-3H-l,2,4-triazol-3-tiyon], amin N-dealkil metabolit [5-(4- ammo-fenü)-2,4-d^hidro-3H-l,2,4-triazol-3-tiyon] ve bilinmeyen bir metabolit bulunmuştur. 5-(4-Nitrofenil)-4-fenü-2,4-dihidro-3H-l,2,4-triazol-3-tiyondaa, amin metabolit [5-(4-aminofenil)-4-fenil-2,4-dihidro- 3H-l,2,4-triazol-3-tiyon], asetil metabolit [5-(4-asetilaminofenil)-4- fenil-2,4-dihidro-3H-l,2,4-triazol-3-tiyon] ve bilinmeyen bir metabolit bulunmuştur.

2. SUMMARY THE IN VIVO METABOLISM OF SOME SUBSTITUTED-2,4-DIfflDRO-3H-l,2,4-TRIAZOLE-3-THIONE DERIVATIVES It has been reported that 4,5-disubstituted-l,2,4-triazoline-3- thiones exhibit antidepressant, antimicrobial, antitubercular, analgesic, antiinflammatory, diuretic and hipoglycemic activities. Therefore, we aimed to investigate of the biotransformation of these structures. In this research, we examined the in vivo metabolic pathways of 5-(4-nitrophenyl)^-phenemyl-2,4-dmydro-3H-l,2,4-1riazole-3-thione and 5-(4-nitrophenyl)-4-phenyl-2,4-dihydro-3H-l,2,4-triazole-3-thione. These compounds had been previously investigated for their antibacterial and antifungal activities and they were found to be active. In the first step, the sodium salts of substrates, substrates and their potential metabolites were synthesized. From these compounds, sodium 5-(4-mtrophenyl)^-phenemyl-l,2,4-triazole-3-thiolate and sodium 5-(4- nitrophenyl)-4-phenyl-l,2,4-triazole-3-thiolate were formed by heating appropriate tmosemicarbazides with 2N sodium hydroxide for 4 hours. These compounds were acidified with hydrochloric acid to obtain the substrates, 5-(4-nitrophenyl)^-phene1hyl-2,4-dihydro-3H-l,2,4-triazole-3-thione and 5-(4-nitrophenyl)-4-phenyl-2,4-dihydro-3H-l,2,4-triazole-3- thione. The sodium salt of substrates were used to inject rats and substrates were used as standarts for the in vivo study. The acetylated metabolites, 5-(4-acetylaminophenyl)-4-phen- emyl-2,4-dihydro-3H-l,2,4-triazole-3-thione and 5-(4-acetylamino- phenyl)^-phenyl-2,4-dihydro-3H-l,2,4-triazole-3-thione were synthe sized by hearing appropriate thiosemicarbazides with 2N sodium hydroxide for 1 hour. Other metabolites were supplied in our laboratory. The original compounds were elucidated using elemental analysis and UV, ^-NMR spectroscopic methods. In the in vivo metabolism study, male Wistar Rats (250 ±20 g) were used. Sodium 5-(4-nitrophenyl)-4-phenethyl-l,2,4-triazole-3- thiolate was dissolved 20 mg/ml in isotonic saline ; Sodium 5-(4- nitrophenyl)-4-phenyl-l,2,4-triazole-3-thiolate was dissolved 2 mg/ml in isotonic saline and these were injected intraperitoneally in 2ml volume. Blood (0.3±0.2 ml) was collected from ophthalmic vens of the rats by sterile capillary tube under ether anesthesia. After centrifugation, the plasma was separated. These compounds were seperated using TLC and reverse phase HPLC methods.From 5-(4-nitrophenyl)-4-phenethyl-2,4-dihydro-3H-l,2,4-triazo- le-3-thione, the acetylated metabolite [5-(4-acetylaminophenyl)-4- phenemyl-2,4-dihydro-3H-l,2,4-triazole-3-tmone], the N-dealkylated amine metabohte [5-(4-ammophenyl)-2,4-dihydro-3H-l,2,4-triazole-3- thione] and an unknown metabohte were found. From 5- (4-nitrophenyl)- 4-phenyl-2,4-dihydro-3H-l,2,4-triazole-3-thione, the amine metabohte [5-4-ammophenyl)^-phenyl-2,4-dihydro-3H-l,2,4-triazole-3-tiiione], the acetylated metabohte [5-(4-acetylaminophenyl)-4-phenyl-2,4- dihydro-3H-l,2,4-triazole-3-thione] and an unknown metabohte were also found.