Design,synthesis And Activity Evaluation Of Benzamide HIV Latency-reversing Agents

Based on the "Shock & Kill" theory in therapeutic regimen of human immunodeficiency virus(HIV)treatment,this paper first studied the design,synthesis and activity evaluation of benzamide HIV latency-reversing agents(effective activation);then this paper explored the activation and drug-like property of 4-substituted-1,5-diarylamine compounds(effective elimination),aiming in activating the latent virus reservoir and the elimination of that.1.Correlational research on benzamide virus latency-reversing agentsSince 1981,HIV was first discovered in the United States,in period of more than 30 years,has spread to all continents,and infected tens of millions of people,causing social burden and economic losses.Highly active antiretroviral therapy(HAART)is the most clinically effective anti-HIV / AIDS treatment program,and HIV patients can reduce plasma viral load to undetectable levels after HAART treatment.But after stopping HAART treatment,the viral load will rebound quickly,leading to lifelong medication of AIDS patients.The rebound of HIV is mainly due to the virus latency in the cell's virus reservoir,and the latent reservoir can not be cleared by HAART.The existence of HIV latent reservoirs is the root cause of the difficulty of eradicating HIV.Based on the "shock & kill" theory,we first designed and synthesized a series of anti-latent compounds to activate latent HIV.North Carolina University Guoxiong Li research group in their study found a special anti-latency reversing agents: benzamides HDAC inhibitor Lead,it can activate latent HIV at a low concentration without toxic side effects,compared to those current anti-latency drug entered the clinical study,SAHA may be more effective.Therefore,we designed and synthesized a series of new derivatives to improve their anti-latency activity so as to achieve a good activation effect.As an exploratory research,we based on the reasonable design ideas,using computer aided design,bioisostere principle,skeleton transition and activity data guidance and a combination of a variety of methods,through continuous optimization of synthesis methods and modification of the reaction conditions,the different sites of the Lead which found in previous studies,anti-latent lead compounds,benzamide type HDAC inhibitors,had been modified.It included the changes in the ring size of groups bound to zinc ions(ZBG),a variety of changes to the diaryl structure fragment that interacts with the target pocket,and the modification of CAP and Linker which could identified protein targets.A total of 48 new compounds were synthesized and their structures were determined by MS and 1H-NMR.Modification of functional groups bound to zinc ions(ZBG)Considering the difference in the size of the ring formed by the chelating of ZBG with zinc ions in the inhibitor molecule,it is envisaged that if the size of the chelating ring is changed,it may be possible to change the intensity of binding between ZBG,thereby affecting the activity of the inhibitor.Therefore,we designed and synthesized three series of target compounds I,II and III,which can form a seven-membered,six-membered,five-membered chelate ring with zinc ions in the HDAC catalytically active central region,respectively,and evaluated these new compounds anti-latent activity,thus understanding the relationship between ZBG and its resistance to latent activity.Structural Modification of Diaryl Structural Fragments(Diaryl)We have designed and synthesized a series of compounds that replace B rings with other groups to increase the complementarity of small molecule inhibitors with the target and the shape of the molecular surface,including the introduction of: four-membered saturated heterocyclic ring,five-membered saturated heterocyclic ring,six-membered aromatic ring and six-membered aromatic ring with a substituent.the modification for CAP and LinkerThe CAP of the Lead was very small,and we only introduced a smaller group that the combination of the smaller CAP portion of the molecule and the target binding region was maintained,and for Linker,we had been modified by the replacement of the biological isotype.At the same time,we evaluated the latent activity of the total of 48 compounds of three categories which synthesized at the cellular and enzyme levels,respectively.The latent activity of the five compounds(5a,5b,29 a,45a,45b)was comparable to that of the positive control SAHA,in which compound 5a had the best latent activity and remained less toxic which was better than the positive control SAHA.It had the meaning of further study.In conclusion,the research of this subject has some reference significance for the future research on the anti-latency research of these compounds,and also lay a foundation for the study of its anti-latent mechanism.At present,we are studying the anti-latency mechanism of synthetic compound 5a in order to find a new type of benzamide latent activator with high efficiency and low toxicity,so as to finally eliminate the HIV latent library.2.Correlation research of 4-substituted-1,5-diarylamine compounds inhibitorsBased on the theory of "Shock & Kill",after the activation of virus reservoir the focus of research has been changed to the elimination of virus reservoir.The experimental data show that we have found with benzamide compounds with good activation activity for virus reservoir,for anti HIV therapy,"shock" steps have been completed,the focus of treatment transfer to "kill" steps,that is the design and synthesis and drug-likeness evaluation of virus inhibitorsCurrent clinical anti HIV drugs mainly fall into four categories: reverse transcriptase inhibitors,protease inhibitors,integrase inhibitors,and entry inhibitors.We have also found a series of HIV non-nucleoside reverse transcriptase inhibitors(HIV-NNRTIs)with highly activation,namely,4-substituted-1,5-diarylamine compounds.The experimental data show that the anti HIV activity of that is better than that of clinical drugs,so it is necessary for further development.In this paper,by continuous optimization of mass spectrometry and chromatographic separation conditions,we established a complete set of LC-MS/MS quantitative analysis,it is of strong specificity,high sensitivity,low detection limit,fast analysis speed.It can test drug metabolism of the 4-substituted-1,5-diarylamine compounds,also laid the foundation for us to carry out the late research of drug metabolism in vivo.At the same time,in order to enrich the drug-metabolism properties of 4-substituted-1,5-diarylamine compounds type HIV-NNRTIs,we selected human liver microsomes,human liver S9 and human plasma to preliminary study in vitro on the stability of the 23 generation of 4-substituted-1,5-diarylamine compounds with good activation,completing the evaluation of drug metabolism of the compounds in vitro,and obtained the elimination half-life of those in the human liver microsomes and human liver S9,plasma,thereby providing us some information about drug-metabolism nature of these compound.Moreover,it provides guidance for the optimization of our precursor,we can continue to promote the research on stability of drug metabolism in vivo.At present,the stage research progresses has been collated and published.