Publication: Molnupiravirin olası hepatotoksisitesinin in vitro araştırılması
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Amaç: Viral enfeksiyonlarda; karaciğer hasarı, önemli komplikasyonlardan biri olduğundan tedavi ilacına bağlı olup olmadığının kapsamlı değerlendirilmesi önemlidir. Molnupiravir, COVID-19 tedavisinde ilk oral antiviral olarak, ülkemiz dahil birçok Sağlık otoritesinin tedavi şemasında yer almaktadır. Bazı klinik araştırmalarda mevcut hepatik advers olaylar sebebiyle toksisite şüphesinin yer aldığı görülmektedir. Bu yüksek lisans tez çalışmasının amacı, molnupiravirin hepatotoksisitesinin in vitro değerlendirilerek, ilaca bağlı karaciğer hasarına neden olma potansiyelinin aydınlatılmasıdır. Gereç ve Yöntem: İnsan karaciğer hücre dizisinde (HepG2) uygulanacak dozların belirlenmesi için MTT testi, reaktif oksijen türlerinin (ROS) seviyelerinin belirlenmesi için DCF testi, alanin aminotransferaz (ALT), aspartat aminotransferaz (AST), üre ve bilirubin biyokimyasal göstergelerinin ölçümü ve apoptotik/ nekrotik hücre ölümü değerlendirmesi için Anneksin testi yapılmıştır. Bulgular: HepG2 hücre dizisinde MTT testi sonucu molnupiravirin IC50 değeri 93,27 μg/ mL olarak Cmax değerinin 30 katı bulunmuştur. Çalışmada ilaç konsantrasyonu 10 μg/ mL ve 30 μg/ mL (Cmax değerinin sırasıyla 3 ve 10 katı) uygulanmıştır. DCF testi sonucu; molnupiravirin 10 μg/ mL ve 30 μg/ mL konsantrasyonlarında 24 ve 4 saatlik maruziyetlerde ROS seviyelerinde kontrole kıyasla azalma görülmüştür, istatistiksel olarak anlamlı bulunmamıştır (p>0.05). Bilirubin seviyeleri ve hem süpernatantta hem hücre ekstraktında ölçülen AST ve ALT değerleri her iki konsantrasyonda kontrole kıyasla artış göstermekle birlikte istatistiksel olarak anlamlı bulunmamıştır (p>0.05). 24 saatlik molnupiravir uygulamasının her iki konsantrasyonunda geç apoptotik ve nekrotik hücre popülasyonu kontrol grubuna oranla artmış ancak istatiksel olarak anlamlı bulunmamıştır (p>0.05). Sonuç: Molnupiravirin tek sefer 24 saat uygulanmasında karaciğer hasarı ile ilgili parametrelerdeki istatistiksel olmayan artış, karaciğer hasarı açısından potansiyelinin gözardı edilmemesi gerektiğini düşündürmektedir.
Objective: Since liver injury is one of the important complications in viral infections, it is important to comprehensively evaluate whether it is related to the treatment drug. Molnupiravir is included in the treatment scheme of many Health Authorities, including our country, as the first oral antiviral in the treatment of COVID-19. It is seen that there is a suspicion of toxicity due to existing hepatic adverse events in some clinical studies. The aim of this master's thesis study is to evaluate the hepatotoxicity of molnupiravir in vitro and to elucidate its potential to cause drug-induced liver injury. Materials and Methods: MTT test to determine the doses to be applied, DCF test to determine reactive oxygen species (ROS) levels, measurement of alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea and bilirubin biochemical indicators and Annexin test to evaluate apoptotic/ necrotic cell death were performed in human liver cell line (HepG2). Results: As a result of the MTT test in HepG2 cell line, the IC50 value of molnupiravir was found to be 93.27 μg/ mL, which is 30 times the Cmax value. In the study, drug concentrations of 10 μg/ mL and 30 μg/ mL (3 and 10 times the Cmax value, respectively) were applied. As a result of the DCF test; ROS levels decreased compared to control at 10 μg/ mL and 30 μg/ mL molnupiravir concentrations for 24 and 4 hours of exposure, but it was not found to be statistically significant (p>0.05). Bilirubin levels and AST and ALT values measured in both supernatant and cell extract increased compared to control at both concentrations, but it was not found to be statistically significant (p>0.05). Late apoptotic and necrotic cell population increased compared to control group at both concentrations of 24-hour molnupiravir administration, but it was not found to be statistically significant (p>0.05). Conclusion: The non-statistical increase in liver injury-related parameters following a single 24-hour administration of molnupiravir suggests that its potential for liver injury should not be ignored.
Objective: Since liver injury is one of the important complications in viral infections, it is important to comprehensively evaluate whether it is related to the treatment drug. Molnupiravir is included in the treatment scheme of many Health Authorities, including our country, as the first oral antiviral in the treatment of COVID-19. It is seen that there is a suspicion of toxicity due to existing hepatic adverse events in some clinical studies. The aim of this master's thesis study is to evaluate the hepatotoxicity of molnupiravir in vitro and to elucidate its potential to cause drug-induced liver injury. Materials and Methods: MTT test to determine the doses to be applied, DCF test to determine reactive oxygen species (ROS) levels, measurement of alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea and bilirubin biochemical indicators and Annexin test to evaluate apoptotic/ necrotic cell death were performed in human liver cell line (HepG2). Results: As a result of the MTT test in HepG2 cell line, the IC50 value of molnupiravir was found to be 93.27 μg/ mL, which is 30 times the Cmax value. In the study, drug concentrations of 10 μg/ mL and 30 μg/ mL (3 and 10 times the Cmax value, respectively) were applied. As a result of the DCF test; ROS levels decreased compared to control at 10 μg/ mL and 30 μg/ mL molnupiravir concentrations for 24 and 4 hours of exposure, but it was not found to be statistically significant (p>0.05). Bilirubin levels and AST and ALT values measured in both supernatant and cell extract increased compared to control at both concentrations, but it was not found to be statistically significant (p>0.05). Late apoptotic and necrotic cell population increased compared to control group at both concentrations of 24-hour molnupiravir administration, but it was not found to be statistically significant (p>0.05). Conclusion: The non-statistical increase in liver injury-related parameters following a single 24-hour administration of molnupiravir suggests that its potential for liver injury should not be ignored.