Screening of new non-nucleoside reverse transcriptase inhibitors of HIV-1 based on traditional Chinese medicines database

HIV- 1 RT is an important target for the treatment of AIDS. There are two major classes of antiviral agents that inhibit HIV- 1 RT have been identified, nucleoside RT inhibitors （NRTIs） and non-nucleoside RT inhibitors （NNRTIs）. In this report, a noval class of non-nucleoside compound with potential RT inhibitory activity were found from the traditional Chinese medicines database （TCMD） using a combination of virtual screening, docking, molecular dynamic simulations, where results were ranked by scoring function of the docking tool. The result indicates that M4753 （a compound derived from TCMD） has not only the lowest bonding energy but also the best match in geometric conformation with the forthcoming NNRTIs. Accordingly M4753 might possibly become a promising lead compound of NNRTIs for AIDS therapy.

Synthesis and anti-HIV-1 activity evaluation of N-1-alkyl-5-halogeno-6-alkylamino uracils as novel non-nucleoside HIV-1 reverse transcriptase inhibitors

N-1-alkyl-5-halogeno-6-alkylamino uracils,which are novel 1-[(2-hydroxyethoxy) methyl]-6-(phenylthio) thymine (HEPT) analogues,were synthesized as the selective and potent non-nucleoside human immunodeficiency virus(HIV)-1 reverse transcriptase inhibitors.Some of the compounds showed potent inhibitory activity against HIV-1 reverse transcriptase.For instance,compounds 1d,1m and 1n exhibited potent anti-HTV-1 activity with the IC_(50) values of 13.3,11.7 and 3.15μM,respectively, which are comparable to that of nevirapine(IC_(50) 8.38μM).

Synthesis and biological evaluation of novel 1-aryl-5-iodo-6-benzyluracils as potent HIV-1 non-nucleoside reverse transcriptase inhibitors

We have synthesized the novel compounds 1a-1i,which are a series of hybrid analogues to 6-benzyl-1-（benzyloxymethyl）- 5-iodouracil,a compound showing strong activity against HIV-1.We also evaluated the activity of these compounds as the inhibitors of HIV-1 reverse transcriptase（HIV-1 RT）,and they have demonstrated moderate activity.

Liver injury in HIV-1-infected patients receiving non-nucleosides reverse transcriptase inhibitors-based antiretroviral therapy

Background Liver injury is one of the most important adverse effects of antiretroviral therapy, leading to therapy changing or discontinuation. Data on liver injury in human immunodeficiency virus-1-infected patients receiving antiretroviral therapy are limited in China. The purpose of this study was to investigate the features of liver injury in human immunodeficiency virus type 1-infected patients receiving non-nucleosides reverse transcriptase inhibitors-based antiretroviral therapy in China.Methods Seventy-five patients on antiretroviral therapy containing non-nucleosides reverse transcriptase inhibitors were retrospectively studied. The patients were divided into 2 groups： group 1 （with liver injury, n=45） and group 2（without liver injury, n=30）. The features of liver injury were analyzed. The sex, age, baseline CD4 counts, hepatitis B virus （HBV） and/or hepatitis C virus （HCV） co-infection, hepatotoxic drug use and nevirapine or efavirenz use were compared between two groups.Results Forty-five patients （60.0%）, 31 （68.9%） males and 14 （31.1%） females, aged 12 to 52 years （averaged （3g±9）years）, experienced at least one episode of liver injury. Forty （53.3%） patients were co-infected with HBV and/or HCV, 42 （56%） patients had concomitant use of antituberculosis drugs or cotrimoxazole, 46 （61.3%） and 29 （38.7%） patients received regimen containing nevirapine and efavirenz, respectively. Grade 1 liver injuries were observed in 26 （57.8%）patients, grade 2 in 16 （35.6%）, grade 3 in 2 （4.0%） and grade 4 in 1 （2.2%）. Three （6.7%） patients discontinued highly active antiretroviral therapy （HAART） due to liver injury. In group 1, there were 29 （64.4%） patients co-infected with HBV and/or HCV, 32 （71.1%） patients received regimen containing nevirapine, and 30 （66.7%） patients had concomitant use of anti-tuberculosis drugs or cotrimoxazole, respectively, significantly higher than those in group 2 （11 （36.7%）, 14 （46.7%）and 12 （40%）, respectively; P=0.018, 0.033, 0.023, respectively）. The sex, age, baseline CD4 counts and disease stage were not factors associated with liver injury.Conclusions Liver injury associated with HAART containing non-nucleosides reverse transcriptase inhibitors was mild to moderate and those who were co-infected with HBV and/or HCV, had concomitant use of antituberculosis drugs or cotrimoxazole and received a regimen containing nevirapine were prone to liver injury while receiving HAART.

Synthesis and biological evaluation of novel 2-arylalkylthio-5-iodo-6-benzyl S-DABOs as potent non-nucleoside HIV-1 reverse transcriptase inhibitors

A series of novel dihydro-alkylthio-benzyl-oxopyrimidines （S-DABOs） 7a-f have been designed and synthesized with an efficient method. Biological evaluation of their HIV-1 reverse transcriptase inhibitory activities was performed using Nevirapine （NVP） as a reference compound. Among the series, compound 7d shows the highest reverse transcriptase inhibitory activity, which is better than Nevirapine.

Synthesis and anti-HIV-1 activity evaluation of N-l-alkyl-5-halogeno-6- alkylamino uracils as novel non-nucleoside HIV-I reverse transcriptase inhibitors

N-l-alkyl-5-halogeno-6-alkylamino uracils, which are novel l-[（2-hydroxyethoxy） methyl]-6-（phenylthio） thymine （HEPT） analogues, were synthesized as the selective and potent non-nucleoside human immunodeficiency virus （HIV）-I reverse transcriptase inhibitors. Some of the compounds showed potent inhibitory activity against HIV-1 reverse transcriptase. For instance, compounds ld, lm and In exhibited potent anti-HIV-1 activity with the ICso values of 13.3, 11.7 and 3.15 μM, respectively, which are comparable to that of nevirapine （IC50 8.38 μM）.

比较以蛋白酶抑制剂或非核苷类反转录酶抑制剂为主方案治疗HIV/HCV合并感染患者的安全性

目的探讨以PIs或NNRTIs为主方案治疗HIV／HCV合并感染的患者比较其安全性。为临床治疗药物的选择提供依据。方法2007～2008年分别在上海、广州等传染病医院的门诊，选择确诊的HIV／HCV合并感染，CD4’T淋巴细胞≤350／mm。的100例患者，随机分为两组（PIs组和NNRTIs组），各50例患者接受1年的治疗，临床观察肝肾功能、代谢等指标。结果治疗前、后两组进行了比较，NNRTIs组天冬酸转氨酶（AST）、丙氨酸转氨酶（ALT）、总胆红素（TBIL）、甘胆酸（CG）上升（P〈0．01），血糖、肌苷上升（P〈0．05）。Pls组总胆固醇、载脂蛋白Al、载脂蛋白B、低密度脂蛋白上升（P〈0．01）。结论NNR-TIs比PIs对肝脏、血糖、肌苷的不良反应大。PIs比NNRTIs对血脂的不良反应大。

以蛋白酶抑制剂或非核苷类逆转录酶抑制剂为主的方案治疗HIV/HCV合并感染患者的临床疗效比较

目的 比较以蛋白酶抑制剂（PIs）或非核甘类逆转录酶抑制剂（NNRTIs）为主的方案治疗HIV/HCV合并感染患者的临床疗效.方法 将CD4＋T淋巴细胞≤350/mm3的100例HIV/HCV合并感染患者随机分为2组,每组各50例,分别接受以PIs或NNRTIs为主的方案治疗,观察CD4＋ T淋巴细胞计数和HIV RNA载量等指标的变化.结果 治疗后两组患者的HIV RNA均下降（P＜0.01）,外周血CD4＋T淋巴细胞计数均有不同程度的升高（P＜0.01）,两组比较差异无显著性（P＞0.05）.结论 以PIs或NNRTIs为主的治疗方案治疗HIV/HCV合并感染者均有效.

基于逆转录酶结构和耐药突变的新型非核苷类逆转录酶抑制剂研究进展

为了应对艾滋病治疗过程中严重的耐药性问题,开发新型抗耐药性非核苷类逆转录酶抑制剂(NNRTI)势在必行。本文通过分析2015~2018年文献报道的新型抗耐药性NNRTI的代表性实例,探讨了抗耐药性NNRTI与逆转录酶的结合模式,并讨论了药物化学策略在发现和优化新型抗HIV-1药物中的作用。本文指出基于逆转录酶晶体结构信息的药物化学策略是快速发现具有新结构的NNRTI的有效途径,前期构效关系研究能为后续的药物开发提供一定的支持。

Highly active antiretroviral therapy dysregulates proliferation and differentiation of human pre-adipocytes

AIM To investigate the mechanism(s) by which potential effects of multi-drug highly-active antiretroviral therapy contributes to lipodystrophy syndrome. METHODS Preadipocytes from healthy donors were assessed for proliferation and differentiation in the presence of nucleoside reverse transcriptase inhibitors(NRTIs), nonnucleoside reverse transcriptase inhibitors(NNRTIs), and protease inhibitors(PIs) individually and in combination. Effects on proliferation were assessed with a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay and effects on differentiation were assessed from glycerol-3-phosphate dehydrogenase(GPDH) activity and quantitation of Oil Red O staining for intracellular lipid. Data were analyzed with a randomized block ANOVA with post-hoc Fisher's Least Significant Difference test. RESULTS Preadipocyte proliferation was inhibited by a combination of NNRTI + NRTI(14% at 48 h, P < 0.001) and PI + NRTI(19% at 48 h, P < 0.001) with additional suppression when ritonavir(RTV) was added(26% at 48 h). The drug combination of atazanavir(ATV) + RTV + emtricitabine(FTC) + tenofovir(TDF) had the greatest inhibitory effect on proliferation at 48 h. Preadipocyte differentiation was most significantly reduced by the efavirenz + FTC + TDF assessed either by GPDH activity(64%) or lipid accumulation(39%), P < 0.001. Combining NRTIs with a PI(ATV + FTC + TDF) significantly suppressed differentiation(GPDH activity reduced 29%, lipid accumulation reduced by 19%, P < 0.01). This effect was slightly greater when a boosting amount of RTV was added(ATV + FTC + TDF + RTV, P < 0.001). CONCLUSION Although combination antiretroviral therapy is clinically more efficacious than single drug regimens, it also has a much greater inhibitory effect on preadipocyte proliferation and differentiation.

非核苷逆转录酶抑制剂新进展

治疗艾滋病的逆转录酶抑制剂主要包括核苷逆转录酶抑制剂(NRTIs,如齐多夫定)和非核苷逆转录酶抑制剂(NNRTIs,如奈韦拉平)。目前至少有30多类非核苷类化合物被发现具有选择性抑制HIV-1逆转录酶的作用,其中已经获得美国FDA批准上市的包括奈韦拉平、地拉夫定和埃法韦伦,还有一些品种正在进行临床试验,其中效果较好的包括MKC-442、trovirdine、loviride等。“第一代”NNRTIs的缺点是容易使病毒产生变异,出现耐药性。“第二代”NNRTIs则活性谱广,不容易产生耐药的变异病毒。属于“第二代”NNRTIs的化合物除了埃法韦伦外,还包括埃法韦伦的衍生物DPC083,咪唑衍生物capravirine,二芳基嘧啶类化合物etravirine、rilpivirine。此外,还有一些喹喔啉、吡嗪酮类、烯基二芳基甲烷等也表现出很好的抗HIV活性。相信在不久的将来,将有更多更有效的非核苷逆转录酶抑制剂为人类的健康带来福音。

Characterization of Genotypic Mutations and Antiretroviral Resistance among Viremic HIV-Infected Patients in a High HIV Prevalence Area: Treatment Challenge and Transmission Risk

There have been few reports evaluating the prevalence of genotypic mutations and antiretroviral resistance among chronic HIV-infected Veterans within the United States. This retrospective cross-sectional study characterizes the rates and changes in HIV genotypic mutations and antiretroviral resistance among viremic patients from 2001 to 2006 at the VA Medical Center located in Washington, DC. The District of Columbia is the metropolitan area with the highest HIV prevalence within the United States. De-identified, linked HIV RNA, genotypic reverse transcriptase (RT) and protease (Pr) mutations and antiretroviral resistance results were assessed for changes during the 6-year period. Aggregated clinic and antiretroviral utilization, and HIV acquisition risk data were evaluated for patients in care during this time. Among 990 viremic samples, the rate of any detected RT or Pr mutation fell from 100% in 2001 to 95% in 2006. This was primarily attributable to the 15% - 20% decrease seen for RT gene mutations against nucleoside/nucleotide class and non-nucleoside class during this period. Resistance to didanosine, stavudine, zidovudine, nevirapine and efavirenz decreased, and tenofovir resistance increased. Despite stable rates of Pr gene mutations, atazanavir resistance increased by 22% from 2003 to 2006. Some but not all changes in genotypic mutations and resistance patterns reflected our patients’ antiretroviral drug utilization. As sexual contacts (77%) and injection drug use (22%) were the leading acquisition risks disclosed by our HIV-infected patients, the high prevalence and changing patterns of HIV genotypic mutations and drug resistance among these patients have had pivotal impacts not only on HIV treatment but potential transmission into our community.

Evolution of blood lipids and risk factors of dyslipidemia among people living with human immunodeficiency virus who had received first-line antiretroviral regimens for 3 years in Shenzhen

Background:Lipid abnormalities are prevalent among people living with human immunodeficiency virus(HIV)(PLWH)and contribute to increasing risk of cardiovascular events.This study aims to investigate the incidence of dyslipidemia and its risk factors in PLWH after receiving different first-line free antiretroviral regimens.Methods:PLWH who sought care at the Third People’s Hospital of Shenzhen from January 2014 to December 2018 were included,and the baseline characteristics and clinical data during the follow-up were collected,including total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C).The risk factors of dyslipidemia after antiretroviral therapy were analyzed with the generalized estimating equation model.Results:Among the 7623 PLWH included,the mean levels of TC,HDL-C and LDL-C were 4.23±0.85 mmol/L,1.27±0.29 mmol/L and 2.54±0.65 mmol/L,respectively,and the median TG was 1.17(IQR:0.85-1.68)mmol/L.Compared with that in PLWH receiving tenofovir disoproxil fumarate(TDF)+lamivudine(3TC)+ritonavir-boosted lopinavir(LPV/r),zidovudine(AZT)+3TC+efavirenz(EFV),and AZT+3TC+LPV/r,the incidence of dyslipidemia was lower in PLWH receiving TDF+3TC+EFV.In multivariate analysis,we found that the risks of elevations of TG,TC,and LDL-C were higher with TDF+3TC+LPV/r(TG:odds ratio[OR]=2.82,95%confidence interval[CI]:2.55-3.11,P<0.001;TC:OR=1.24,95%CI:1.14-1.35,P<0.001;LDL:OR=1.06,95%CI:1.00-1.12,P=0.041),AZT+3TC+EFV(TG:OR=1.41,95%CI:1.28-1.55,P<0.001;TC:OR=1.43,95%CI:1.31-1.56,P<0.001;LDL:OR=1.18,95%CI:1.12-1.25,P<0.001),and AZT+3TC+LPV/r(TG:OR=3.08,95%CI:2.65-3.59,P<0.001;TC:OR=2.40,95%CI:1.96-2.94,P<0.001;LDL:OR=1.52,95%CI:1.37-1.69,P<0.001)than with TDF+3TC+EFV,while treatment with TDF+3TC+LPV/r was less likely to restore HDL-C levels compared with TDF+3TC+EFV(OR=0.95,95%CI:0.92-0.97,P<0.001).In addition to antiretroviral regimens,antiretroviral therapy duration,older age,overweight,obesity and other traditional factors were also important risk factors for dyslipidemia.Conclusion:The incidence of dyslipidemia varies with different antiretroviral regimens,with TDF+3TC+EFV having lower risk for dyslipidemia than the other first-line free antiretroviral regimens in China.

Route improvement of 3-substituted-4-(2-methylcyclohexyloxy)-6-phenethylpyridinone

trans-3-Isopropyl-4-（2-methylcyclohexyloxy）-6-phenethylpyridin-2（1H）-one, as reverse transcriptase （NNRTIs）, exhibited significant potent activity not only against wild-type HIV-1 strains but also on mutant strains. For furthering study this compound, the original synthetic route should be shorten to improve the total yield. In this report, we designed an efficient synthetic strategy to obtain the target compound with higher yield.

Steered molecular dynamics simulations of protein-ligand interactions

Studies of protein-ligand interactions are helpful to elucidating the mechanisms of ligands, providing clues for rational drug design. The currently developed steered molecular dy- namics (SMD) is a complementary approach to experimental techniques in investigating the biochemical processes occurring at microsecond or second time scale, thus SMD may provide dynamical and kinetic processes of ligand-receptor binding and unbinding, which cannot be ac- cessed by the experimental methods. In this article, the methodology of SMD is described, and the applications of SMD simulations for obtaining dynamic insights into protein-ligand interactions are illustrated through two of our own examples. One is associated with the simulations of bind- ing and unbinding processes between huperzine A and acetylcholinesterase, and the other is concerned with the unbinding process of α-APAfrom HIV-1 reverse transcriptase.