新型稠环嘧啶类HIV-1 NNRTIs的设计、合成、抗病毒活性与初步成药性研究

艾滋病(AIDS)是由人类免疫缺陷病毒(HIV)感染引起的全球重大流行性传染病。据世界卫生组织(WHO)的数据报道,截至2018年底,因感染艾滋病毒而死亡的人数达到了3200万,全球现存约有3790万艾滋病毒感染者,其中2018年新增感染人数约170万人,77万人死于艾滋病引起的并发症。逆转录酶(RT)在HIV-1的生命周期起着必不可少的作用,是开发抗HIV新药的重要靶点之一。非核苷类逆转录酶抑制剂(NNRTIs)以其抗病毒活性强、选择性高、作用机制明确以及可与其它药物协同作用等特点而备受关注,成为高效抗逆转录病毒治疗(HAART)的重要组分之一,其中以上市药物Etravirine和Rilpivirine为代表的二芳基嘧啶类化合物(DAPYs)表现格外耀眼。尽管NNRTIs在HIV-1患者的治疗上取得了巨大成功,但是由于其变构作用机制和较低的遗传屏障,在临床应用中迅速出现了耐药株。

Design and optimization of piperidinesubstituted thiophene[3,2-d]pyrimidine-based HIV-1 NNRTIs with improved drug resistance and pharmacokinetic profiles

HIV-1 reverse transcriptase(RT)has received great attention as an attractive therapeutic target for acquired immune deficiency syndrome(AIDS),but the inevitable drug resistance and side effects have always been major challenges faced by non-nucleoside reverse transcriptase inhibitors(NNRTIs).This work aimed to identify novel chemotypes of anti-HIV-1 agents with improved drugresistance profiles,reduced toxicity,and excellent druggability.A series of diarylpyrimidine(DAPY)derivatives were prepared via structural modifications of the leads K-5a2 and 25a.Among them,15a with dimethylphosphine oxide moiety showed the most prominent antiviral potency against all of the tested viral panel,being 1.6-fold(WT,EC_(50) Z 1.75 nmol/L),3.0-fold(L100I,EC_(50) Z 2.84 nmol/L),2.4-fold(K103N,EC_(50) Z 1.27 nmol/L),3.3-fold(Y181C,EC50 Z 5.38 nmol/L),2.9-fold(Y188L,EC_(50) Z 7.96 nmol/L),2.5-fold(E138K,EC_(50) Z 4.28 nmol/L),4.8-fold(F227L/V106A,EC_(50) Z 3.76 nmol/L)and 5.3-fold(RES056,EC_(50) Z 15.8 nmol/L)more effective than that of the marketed drug ETR.Molecular docking results illustrated the detailed interactions formed by compound 15a and WT,F227L/V106A,and RES056 RT.Moreover,15a-HCl carried outstanding pharmacokinetic(t1/2 Z 1.32 h,F Z 40.8%)and safety profiles(LD_(50)>2000 mg/kg),which demonstrated that 15a HCl is a potential anti-HIV-1 drug candidate.

Design,synthesis,and biological evaluation of benzo[4,5]thieno[2,3-d]pyrimidine derivatives as novel HIV-1 NNRTIs

Inspired by our previous studies to discover novel human immunodeficiency virus-1(HIV-1)nonnucleoside reverse transcriptase inhibitors(NNRTIs)by targeting the tolerant region II of the NNRTIs binding pocket(NNIBP),a series of novel benzo[4,5]thieno[2,3-d]pyrimidine derivatives were designed through structure-based drug design as novel potent HIV-1 NNRTIs.The results showed that compound16b was the most active inhibitor,exhibiting 50% effective concentration(EC50)values from 0.021μmol/L to 0.298μmol/L against wild-type(WT)and a panel of NNRTIs-resistant HIV-1 strains.Moreover,16b was demonstrated with a significantly low 50% cytotoxicity concentration(CC_(50))value(>200μmol/L)and high selectivity index(SI)values.In addition,16b yielded moderate reverse transcriptase(RT)enzyme inhibition with a 50% inhibition concentration(IC_(50))value of 0.183μmol/L,which demonstrated that it acted as HIV-1 NNRTIs.The binding mode of 16b with RT was also illustrated via molecular docking.Overall,this work provided a novel lead compound for developing potent HIV-1 NNRTIs.

Development of non-nucleoside reverse transcriptase inhibitors(NNRTIs):our past twenty years

Human immunodeficiency virus(HIV)is the primary infectious agent of acquired immunodeficiency syndrome(AIDS),and non-nucleoside reverse transcriptase inhibitors(NNRTIs)are the cornerstone of HIV treatment.In the last 20 years,our medicinal chemistry group has made great strides in developing several distinct novel NNRTIs,including 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine(HEPT),thio-dihydro-alkoxy-benzyl-oxopyrimidine(S-DABO),diaryltriazine(DATA),diarylpyrimidine(DAPY)analogues,and their hybrid derivatives.Application of integrated modern medicinal strategies,including structure-based drug design,fragment-based optimization,scaffold/fragment hopping,molecular/fragment hybridization,and bioisosterism,led to the development of several highly potent analogues for further evaluations.In this paper,we review the development of NNRTIs in the last two decades using the above optimization strategies,including their structure-activity relationships,molecular modeling,and their binding modes with HIV-1 reverse transcriptase(RT).Future directions and perspectives on the design and associated challenges are also discussed.

Exploring the hydrophobic channel of NNIBP leads to the discovery of novel piperidinesubstituted thiophene[3,2-d]pyrimidine derivatives as potent HIV-1 NNRTIs

In this report,a series of novel piperidine-substituted thiophene[3,2-d]pyrimidine derivatives were designed to explore the hydrophobic channel of the non-nucleoside reverse transcriptase inhibitors binding pocket(NNIBP)by incorporating an aromatic moiety to the left wing of the lead K-5 a2.The newly synthesized compounds were evaluated for anti-HIV potency in MT-4 cells and inhibitory activity to HIV-1 reverse transcriptase(RT).Most of the synthesized compounds exhibited broad-spectrum activity toward wild-type and a wide range of HIV-1 strains carrying single non-nucleoside reverse transcriptase inhibitors(NNRTI)-resistant mutations.Especially,compound 26 exhibited the most potent activity against wild-type and a panel of single mutations(L1001,K103 N,Y181 C,Y188 L and E138 K)with an EC50 ranging from 6.02 to 23.9 nmol/L,which were comparable to those of etravirine(ETR).Moreover,the RT inhibition activity,preliminary structure-activity relationship and molecular docking were also investigated.Furthermore,26 exhibited favorable pharmacokinetics(PK)profiles and with a bioavailability of 33.8%.Taken together,the results could provide valuable insights for further optimization and compound 26 holds great promise as a potential drug candidate for the treatment of HIV-1 infection.

Boronic acid-containing diarylpyrimidine derivatives as novel HIV-1NNRTIs: Design, synthesis and biological evaluation

Drug resistance remains to be a serious problem with type Ⅰ human immunodeficiency virus(HIV-1) nonnucleoside reverse transcriptase inhibitors(NNRTIs). A series of novel boronic acid-containing diarylpyrimidine(DAPY) derivatives were designed via bioisosterism and scaffold-hopping strategies,taking advantage of the ability of a boronic acid group to form multiple hydrogen bonds. The target compounds were synthesized and evaluated for their anti-HIV activities and cytotoxicity in MT-4 cells.Compound 10 j yielded the most potent activity and turned out to be a single-digit nanomolar inhibitor towards the HIV-1 ⅢB [wild-type(WT) strain], L100 I and K103 N strains, with 50% effective concentration(EC_(50)) values of 7.19–9.85 nmol/L. Moreover, 10 j inhibited the double-mutant strain RES056 with an EC_(50) value of 77.9 nmol/L, which was 3.3-more potent than that of EFV(EC_(50)= 260 nmol/L) and comparable to that of ETR(EC_(50)= 32.2 nmol/L). 10j acted like classical NNRTIs with high affinity for WT HIV-1 reverse transcriptase(RT) with 50% inhibition concentration(IC_(50)) value of 0.1837 μmol/L. Furthermore,molecular dynamics simulation indicated that 10 j was proposed as a promising molecule for fighting against HIV-1 infection through inhibiting RT activity. Overall, the results demonstrated that 10 j could serve as a lead molecule for further modification to address virus-drug resistance.

Pyridin-2(1H)-ones as HIV-1 NNRTIs:a combinatorial optimization strategy

With rapid spread of HIV(human immunodeficiency virus) on a global scale and increasingly severe drug-resistance of it,it is urgently necessary to develop novel effective anti-HIV drugs.Non-nucleoside reverse transcriptase inhibitor(NNRTIs)is one of the most significant antiretroviral drugs for fighting against HIV infection due to their various structures,unique mode of action,good efficacy and low toxicity.Pyridinone derivatives,a type of NNRTIs,have been reported to achieve remarkable development in the past few decades.In this review,we summarized current drug design and medicinal chemistry efforts toward the development of next-generation pyridinones as HIV-1 NNRTIs.

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。

不同方案治疗丙型肝炎病毒和人类免疫缺陷病毒合并感染患者对血细胞影响的比较

目的探讨以蛋白酶抑制剂(PIs)或非核苷类反转录酶抑制剂(NNRTIs)为主方案治疗丙型肝炎病毒(HCV)/人类免疫缺陷病毒(HIV)合并感染的患者,比较其对血细胞的影响。方法采用随机、开放、对照方法收集初次就诊的273例HCV/HIV合并感染患者为研究对象。分别用PIs(PIs组)或NNRTIs(NNRTIs组)为主的方案治疗。比较治疗前、后血红蛋白(Hb)、白细胞(WBC)、红血胞(RBC)、血小板(PLT)等指标。结果治疗后PIs组和NNRTIs组Hb、WBC、PLT、RBC均有不同程度的上升,但是NNRTIs组比PIs组上升明显,Hb分别为137.0±24.5g/L和133.0±14.7g/L,WBC分别为(5.810±1.981)×109/L和(4.440±1.244)×109/L,PLT分别为(206.0±66.7)×109/L和(135.0±42.4)×109/L,两组比较差异有统计学意义(P<0.01)。结论 PIs或NNRTIs为主治疗HCV/HIV合并感染的患者对血细胞没有影响。

新型HIV逆转录酶和整合酶双靶点抑制剂的虚拟筛选

目的构建3-OH HEPT类针对HIV逆转录酶非核苷类抑制剂作用靶点和整合酶的双靶点抑制剂[RT(NNRTI)/IN]药效团模型,对中药化学数据库(TCMD)进行搜索,寻找潜在的新型HIV双靶点抑制剂。方法从已知的3-OH HEPT类HIVRT(NNRTI)/IN双靶点抑制剂出发,构建药效团模型,基于药效团模型和Lipinski五规则对TCMD进行筛选,发现新的抗HIV双靶点抑制剂。结果构建的药效团模型包括6个药效特征基团,以符合6个药效特征基团中任意5个为条件,在测试集数据库中验证性搜索命中全部16个化合物,检出率达到了100%。利用所建的六点药效团模型和Lipinski五规则对TCMD筛选,得到符合条件的化合物229个,其中包含已知的抗HIV活性化合物。结论建立的药效团模型和筛选策略为后续研究打下了较好的基础。

环糊精/非核苷类逆转录酶抑制剂包合物研究

非核苷类逆转录酶抑制剂(NNRTIs)是抗艾滋病毒的一线药物,具有低毒高效的显著疗效。但其溶水性差、生物利用度低等缺点限制了NNRTIs抗病毒潜力的发挥。环糊精包合物能达到提高生物利用度、增加水溶性等目的。本文概述了环糊精与非核苷逆转录酶抑制剂包合物研究进展,结果表明环糊精包合物能提高其水溶性、生物利用度及临床应用效果。

Regioselective synthesis and anti-HIV activity of the novel 2- and 4-substituted pyrazolo[4,5-e] [ 1,2,4]thiadiazines

The new regioisomer 7-methyl-pyrazolo[4,5-e][1,2,4]thiadiazine nucleus (5) was synthesized, and its novel mono-N2- or N4-substituted pyrazolo[4,5-e][1,2,4]thiadiazines (6, 7) were regioselectively prepared by deprotonation of N2 or/and N4 atoms with different molar ratio of NaH and alkyl halides. Anti-HIV-1 screening tests showed some compounds to be potent as HIV-1 non-nucleoside reverse transcriptase inhibitors (HIV-1 NNRTIs).

Receptor-based Molecular Designs of DABO Derivatives as HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors

A series of 46 dihydro-alkoxy-benzyl-oxopyrimidines （DABOs）, a class of highly potent non-nucleoside reverse transcriptase inhibitors （NNRTIs）, was studied by molecular docking followed by comparative molecular field analysis （CoMFA） and comparative molecular similarity index analysis （CoMSIA）. The results showed that the H-bonding interactions between the C=O and NH of the pyrimidine ring and Lys101, hydrophobic interactions between R, R1, X sites of ligands and neighboring amino acid residuals, and the electrostatic interactions between ligands and His235 and Lys101 residues were the dominant factors affecting the binding affinities. Based on an optimal docking conformation, 3D-QSAR models of 46 DABO derivatives were developed. The r^2 and cross-validated r^2 （q^2） of an optimal CoMSIA model were 0.862 and 0.532, respectively. Based on the QSAR studies, 9 new compounds were designed by the method of LeapFrog. The binding energies and docking scores （GScore） of 9 new compounds were better than that of a template molecule with the highest observed activity. The results showed that the molecular designs of DABOs should be focused on the hydrophobic interactions with the bottom of the binding pocket as well as van der Waals interactions with the entrance of binding pocket.

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.

Computational modeling studies on anti-HIV-1 non-nucleoside reverse transcriptase inhibition by dihydroalkoxybenzyloxopyrimidines analogues: an electrotopological atomistic approach

For the first time we report quantitative structure activity relationship (QSAR) studies based on Kier-Hall Electrotopological State (E-State) Indices for Dihydroalkoxybenzyloxopyrimidines (DABO) derivatives acting as NNRTIs of HIV-1. A dataset of 74 compounds was compiled from published studies and randomly subdivided into training and test sets. To understand the pharmacophoric effect, Kier-Hall Electrotopological State descriptors namely SN1, SN3, SF, SAr, SS, SO, SNO2, SCl, SY (Y = S-alkyl and NH-alkyl), SX (X = Me) and biological activity were used as independent and dependent variable respectively. Statistical results were highly encouraging for the training set multiple linear regression [(MLR): r2 = 0.961, F = 100.41 and q2 = 0.926, neural networks (NN): r2 = 0.966, F = 115.594, degrees of freedom = 40 and k-nearest neighbour (k-NN): r2 = 0.770, q2 = 0.757, degrees of freedom = 40]. Results of validation using a test set showed the same trend as training set (NN > MLR > kNN). The above results suggest that of various functional groups present in DABO such as SN3, SO, SCl, SAr and SNO2 contribute more significantly towards activity. On the other hand SN1, SS, and SF do not play any role in enhancing the activity. The substitution of S-alkyl and NH-alkyl at C2 position is essential though it does not contribute much towards the activity. The substitution of methyl group at C5 position is unfavorable and exhibit negative impact on inhibitory activity. Therefore, it seems reasonable to choose E-state indices as suitable and significant descriptors for exploring the relationship between the pIC50 and the pharmacological properties of the compounds.

抗艾滋病药物研究进展

过去十多年里,艾滋病(AIDS)已形成世界范围大流行,被称为“二十世纪的瘟疫”.近年来,亚洲地区流行速度急剧上升,到本世纪末将有上千万HIV感染者.为了研制治疗AIDS的药物,人们做出了广泛的努力.到1997年,已有十多种药物获得美国FDA批准用于HIV感染的治疗,按作用机制可把它们分为三大类;1.核苷类似物抑制病毒的逆转录酶;2.非核苷类似物抑制病毒的逆转录酶;3.HIV蛋白酶的抑制剂.然而。