麗豐樟芝發酵產品抑制B型肝炎病毒表面抗原和e抗原活性檢測及有效成分之測定

Inhibition of HBsAg and HBeAg Activities from Fermentation Products & Its Effective Compounds of Antrodia cinnamomea by NEW BELLUS ENTERPRISES

B型肝炎是國人常見的肝疾，會引起急性及慢性肝炎，慢性肝炎患者長期染病，極易轉為猛暴性肝炎、肝硬化及肝癌。B型肝炎目前雖然有疫苗，但是國內仍有約300萬病毒帶原者需要治療，目前臨床上使用之治療藥物，以肝安能（lamivudme；即3TC）有非常肯定的療效，可惜仍然有抗藥性的問題。本論文以體外細胞培養模式，利用酵素連結免疫吸附分析法(Enzyme-Linked Immunosorbent Assay)測定由麗豐實業股份有限公司提供的樟芝發酵產品抑制B型肝炎病毒表面抗原和e抗原活性及有效成分之分離及測定。結果顯示：麗豐實業股份有限公司經由發酵槽大量培養之樟芝菌絲體，以95%乙醇粗提得到四個粗萃物AN-L-EA、AN-L-W、AN-M-EA及AN-M-W，抗病毒試驗結果顯示：其中AN-M-EA對B型肝炎病毒具極顯著抑制作用。進一步取AN-M-EA分離得到ACEA M37、ACEA M41、ACEA M43、ACEA M53、ACEA M55、ACEA M65、ACEA M69、ACEA M74、ACEA M78、ACEA M84及一個純化合物(4-acetylantroquinonol B)，抗病毒測試結果顯示：ACEA M55、ACEA M65 及4-acetylantroquinonol B均具有非常顯著抑制B型肝炎野生型病毒株及肝安能抗藥性突變株之表面抗原及e抗原之活性。

Viral Hepatitis B is known to be a major cause of acute and chronic hepatitis. Chronic infection has been identified as a precursor for cirrhosis and hepatocellular carcinoma (HCC). Despite the availability of effective preventive vaccine for HBV in recent years, there are still 300 million existing chronic HBV carriers that urgently need therapy. Although lamivudine (also called 3TC), a cytosine nucleoside analogue, is very effective in therapeutic treatment of hepatitis B, HBV can mutate and develop resistance to the drug after long-term administration. In this paper, the in vitro cell culture model was used to investigate the inhibition of HBsAg and HBeAg activities from the fermentation products of Antrodia cinnamomea manufactured by NEW BELLUS ENTERPRISES as well as the isolation and detection of its effective compounds. The expression of HBsAg and HBeAg was detected by Enzyme-Linked Immunosorbent Assay system. Through 95% methanol extracts from a large fermentation culture of Antrodia cinnamomea (manufactured by NEW BELLUS ENTERPRISES), we obtained 4 crude extracts, AN-L-EA, AN-L-W, AN-M-EA, and AN-M-W. Among them, AN-M-EA shows very significant inhibition of HBsAg activities. By further extraction from AN-M-EA, we obtained ACEA M37, ACEA M41, ACEA M43, ACEA M53, ACEA M55, ACEA M65, ACEA M69, ACEA M74, ACEA M78, ACEA M84 and a pure compound (4-acetylantroquinonol B). Our experimental results from anti-viral test show that ACEA M55, ACEA M65 and 4-acetylantroquinonol B all depict significant inhibition of HBsAg and HBeAg activities for both wild-type and lamivudine-resistant mutations.