Ayşen TEZCANER, Nesrin HASIRCI, Aysel KIZILTAY, Gülhan IŞIK

pH duyarlı hidrazon bağıyla mPEG-b-PCL kopolimerine konjuge edilen doksorubisin miktarının belirlenmesi

Amaç: Bu çalışmanın amacı, antikanser bir ilaç olan Doksorubisinin (DOX) sentezlenen metoksi poli(etilen glikol)-blok-polikaprolakton (mPEG-b-PCL) polimerine pH-duyarlı hidrazon bağları aracılığıyla bağlanabileceği etkin ortamı bulmak ve konjuge edilmiş ilaç miktarını belirlemektir. Yöntem: DOX konjugasyon işlemi iki farklı ortamda [dimetil sülfoksit (DMSO) ve methanol trifloroasetik asit (MeOH-TFA)] gerçekleştirilmiştir. Konjuge ilaç miktarı iki farklı yöntemle belirlenmiştir. Bir metot, ph duyarlılığını göz ardı ederek konjuge yapının kloroform:methanol (Ch:MeOH, 1:1 v/v) çözeltisi içerisinde çözünmesi sonrasında, diğer metot pH’ya duyarlı hidrazon bağlarının asidik ortamda [0.1 M hidroklorik asit (HCl), derişik HCl (12 M HCl) ve derişik sülfürik asit (18.3 M H2SO4) olarak üç farklı ortamda] kırılması sonrasında uygulanmıştır. Bulgular: En yüksek konjugasyon verimliliği MeOHTFA çözeltisi içerisinde konjugasyon sağlandığında ve polimer-ilaç konjugatları konsantre sülfürik asit ile muamele edildikten sonra elde edilmiştir. Sonuç: MeOH-TFA yönteminin DOX’un mPEG-b-PCL kopolimerine konjugasyonu için iyi bir yöntem olduğu ve H2SO4 (18.3 M) yönteminin hidrazon konjugasyonu yoluyla polimere bağlı DOX miktarının belirlenmesi için literatürde mevcut diğer yöntemlerden daha iyi olduğu bulunmuştur.

Determination of doxorubicin amount conjugated to mPEG-b- PCL copolymer via pH sensitive hydrazone bond

Objective: The aim of this study was to find the efficient medium to bind an anticancer drug, Doxorubicin (DOX), to a synthesized polymer, methoxy poly(ethylene glycol)-block-polycaprolactone (mPEG-b-PCL) via pH sensitive hydrazone bonds and to determine the amount of conjugated drug. Methods: DOX conjugation was carried out in two different media [dimethyl sulfoxide (DMSO) and methanol-trifluoroacetic acid (MeOH-TFA)]. The amount of conjugated drug was determined with two different methods. One method was applied the dissolution of the conjugate in chloroform: methanol (Ch: MeOH, 1: 1 v/v) solution without considering pH responsiveness, and the other method was after breaking pH sensitive hydrazone bonds in acidic medium [using three different media as 0.1 M hydrochloric acid (HCl), concentrated HCl (12 M HCl) and concentrated sulfuric acid (18.3 M H2SO4)]. Results: The highest conjugation efficiency was obtained when the conjugation was achieved in MeOHTFA solution, and for the polymer-drug conjugates after the treatment with concentrated sulfuric acid. Conclusion: It was concluded that, MeOH-TFA method was a good method for conjugation of DOX to mPEG-b-PCL copolymer, and H2SO4 (18.3 M) method was better than any other present in literature for determination of the amount of DOX linked to the polymer via hydrazone conjugation.

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