Combining Oxymatrine or Matrine with Lamivudine Increased Its Antireplication Effect against the Hepatitis B Virus In Vitro

Evid Based Complement Alternat Med. 2013;2013:186573.

http://dx.doi.org/10.1155/2013/186573

Zhi-Jie Ma,1, 2 Qi Li,1 Jia-Bo Wang,1 Yan-Ling Zhao,1 Yan-Wei Zhong,3 Yun-Feng Bai,4 Rui-Lin Wang,4 Jian-Yu Li,4 Hui-Yin Yang,4 Ling-Na Zeng,1 Shi-Biao Pu,1 Fei-Fei Liu,1 Da-Ke Xiao,5 Xin-Hua Xia,6 and Xiao-He Xiao4

1 China Military Institute of Chinese Medicine, 302 Military Hospital, Beijing 100039, China

2 College of Traditional Chinese Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichnan 610075, China

3 Pediatric Liver Disease Therapy and Research Center, 302 Military Hospital, Beijing 100039, China

4 Integrative Medical Center, 302 Military Hospital, No. 100 4th Ring Road, Beijing 100039, China

5 College of Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu 211198, China

6 College of Pharmacy, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China

Abstract

Some recent clinical reports have shown that the combination of oxymatrine, a phyto-derived drug, with lamivudine (3TC) could improve its curative effect against hepatitis B virus (HBV) infection. However, the experimental data in support of this combination strategy are lacking. In this study, we investigated the anti-HBV activity of the combination of 3TC and either oxymatrine or matrine on HepG2 2.2.15 in vitro. The activities of the combination and the solo compound, each in different concentrations, were compared on the 3rd, 6th, and 9th experimental days. The cytotoxicity results showed that the nontoxic concentrations of both oxymatrine and matrine to HepG2 2.2.15 cells were 800 μg/mL. We found that the single use of oxymatrine below 100 μg/ml, matrine below 200 μg/ml, and 3TC below 30 μg/ml showed weak inhibitory effects on the secretion of hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg), and HBV-DNA in culture media; the combination of 3TC (30 μg/ml) with oxymatrine (100 μg/ml) or matrine (100 μg/ml) showed significant inhibitory effects that were higher than or equivalent to the single use of 3TC at 100 μg/ml. The results provide a new impetus to develop novel, multicomponent anti-HBV drugs through the combination of natural products with nucleoside analogs to enhance their activity.

 

Supplement:

Given the on-going threat of hepatitis B virus (HBV) infection and continuing emergence of new drug-resistant HBV virus strains, novel strategies for preventing and treating HBV-infected disease are urgently needed. Combinational therapies of different drugs are usually adopted in clinic to conquer HBV. Interestingly, some traditional Chinese medicines (TCMs) play synergetic effect to antireplication agent, although those TCMs are usually considered no antiviral activity.

In this study, Wang et al. presented an attractive example regarding oxymatrine or matrine increased lamivudine’s antireplication effect against HBV in HepG2 2.2.15 cell line.

As shown in the bottom figure, The solo use of either matrine below 400 μg/ml (1.61 μmol/ml) or 3TC at 30 μg/ml (0.13 μmol/ml) revealed weak inhibitory effects on the secretion of HBeAg into culture media after 9 days of treatment. However, the combinations of 3TC (30 μg/ml) with matrine at 100, 200 or 400 μg/ml all showed significant inhibitory effects (63%, 68% and 75%, respectively). Furthermore, the effects of such combinations were all superior to the effects of 3TC in solo use at 100 μg/ml.

Jia-bo Wang-1

Similarly, the oxymatrine also showed synergetic effect to 3TC though the solo use of oxymatrine is not effective. The results are shown in the below figure.

Jia-bo Wang-2

And there is an interesting phenomenon that the dose-effect relationships of the combinations between 3TC and matrine or oxymatrine were not significant. That is, the synergistic effect between 3TC and matrine or oxymatrine is nearly steady over a relatively wide range of doses, at least from 1:1 to 1:4. The author therefore considered that the wide range of synergistic dose is very important for the clinical application of combinatorial therapy. As we known, most of combinatorial therapies are only effective in vitro but ineffective in vivo. Different drugs have differential pharmacokinetic characteristics that are time-dependent and tissue-specific. Although we can combine two synergistic drugs at the correct proportion in one capsule, the two drugs will not achieve the expected proportion in the blood, tissue, cell or target. Their proportion will certainly vary over time (shown in the bottom figure). If the range of the synergistic proportion is narrow, it could be anticipated that the combination will not achieve expected in vivo synergistic effects; a phenomenon which has been confirmed in vitro.

Jia-bo Wang-3The importance of this study is two-fold. First, it provides experimental evidence for the first time in support of the superiority of the combination of 3TC with matrine or oxymatrine over 3TC alone. Since the cheaper price of combination of matrine or oxymatrine with 3TC than the new launched antiviral agents (e.g. entecavir), the study indicates a new avenue using combinatorial therapy in treating HBV infection in undeveloped regions of low income.

Second, the study shows how advantageous a wide range of synergistic proportions is for combinatorial therapy due to pharmacokinetic concerns. The results provide new inspiration with concerning pharmacokinetics in screening novel and patient-effective combinatorial therapies in the future.

Acknowledgements: The authors acknowledge Professor Juan Wang for her useful advices to their study. This work was funded by the National Natural Science Foundation of China (Nos. 30973947, 81274026, and 81173571), the National Industry Program of China (Nos. 200807020 and 201207002), and by the National Key Technology R&D Program (Nos. 2012BAI29B02 and 2012ZX10005010-002-002).

Contact:
Jia-bo Wang, Ph.D.,
Assistant Professor
China Military Institute of Chinese Materia Medica
302 Military Hospital of People’s Republic of China
100#, 4th West Ring Rd., Beijing, 100039

pharm_sci@126.com (J.b. Wang)
http://www.302hospital.com

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