Design,Synthesis And Anti-HBV Research Of2-Pyridone Derivatives

Hepatitis, especially the chronic hepatitis B caused by HBV is threatening the human health seriously. According to World Health Organization (WHO), about3.4-4.0million people were chronic infected with hepatitis B, and there are about100million people die from chronic hepatitis B related diseases every year. Appropriate in areas of high incidence of hepatitis B, there were over120million HBV carriers in China, and about30million among them were chronic hepatitis B patients. In China, the medical cost of treatment of chronic hepatitis B is up to50billion yuan per year, which has brought a heavy economic burden to the family and society. How to overcome hepatitis B has become a major public health problem in our country.Now the drugs used to treat chronic hepatitis B could be divided in to two categories: interferon and nucleoside analogues. The application of interferon is limited because of its expensive cost, limited for the crowd as well as the serious side effects. And for nucleoside, the main problem is the drug-resistant. Therefore, looking for new non-nucleoside anti-HBV drugs has become a hot spot inside and outside of China.Based on the information of pharmacophore model, we designed several2-pyrindone derivatives. Started with paeonol, through Claisen Condition reaction, closed loop reaction, open loop reaction and iodo-reaction, we got the intermediate3-iodo-7-methoxy-4H-chromen-4-one (5). Then the intermediate5was treated to yield the key intermediate (E)-methyl3-(7-methoxy-4-oxo-4H-chromen-3-yl)acrylate through Heck reaction. And we got the serial LK011-1～Lk011-5by treating the key intermediate through nitrification reaction, catalytic hydrogenation, acylation and open loop rearrangement reaction. The intermediate5was refluxed with AICl3to take off the7’methyl, and then we got the intermediate12through Heck reaction, and nitrification reaction. The intermediate12was treated through catalytic hydrogenation, cyclization condensation reaction and open loop rearrangement reaction to yield the serial LK011-6and LK011-8. LK011-10was synthesized through open loop rearrangement reaction from intermediate12. LK011-9was obtained through catalytic hydrogenation reaction from the serial LK011-10, and LK011-9was treated with1,1,1-triethoxythane to give the serial LK011-7. LK041-04、LK041-10、 LK04502、LK04505、LK04506、LK04508、LK04512、LK04522、LK011-8-1and LK011-8-7were reacted with hydroxylamine hydrochloride to yield serial LK011-11. All the target molecules were reported firstly and their structures were determined by1H NMR,13C NMR、LC-MS and element analysis.HBV DNA inhibitory efficacies of the target compounds were evaluated in HepG2.2.15cells transfected by HBV. MTT and probe PCR methods were used to detect the cytotoxicity and the replication of HBV DNA, respectively. Most of the compounds showed modest to good ant-HBV activity. Especially, serial LK011-6and LK011-8show the best anti-HBV DNA replication. The anti-HBV DNA activity of LK011-6-2, LK011-6-10, LKO11-6-14, LK011-8-1and LK011-8-5were in the range of3.0to10.0μg/ml. And the anti-HBV DNA activity of LK011-6-3, LK011-6-3b, LK011-6-4, LK011-6-6, LK011-6-6b, LK011-6-12, LK011-6-13, LK011-6-15and LK011-8-2were in the range of10.0μg/ml and20.0μg/ml.From the biological data, preliminary structure-activity relationships of2-pyridones derivatives of anti-HBV activity were summarized as below:(1) when introducing amide linkage on the hydrophobic group HY1, especially the electron-withdrawing groups, the anti-HBV activity decreased or even disappeared.(2) introducing oxazolyl on the hydrophobic group HY1was good to the anti-HBV activity.(3) the electron-donating groups which were connected at the1N atom were very important to the anti-HBV activity.In this dissertation, with the guidance of pharmacophore model of anti-HBV activity,106of new compounds have been designed and synthesized for the first time. Based on the results of anti-HBV assay, the affection of multi-site modification was investigated and some valuable structures were obtained. The anti-HBV mechanism will be the main task in the future.