Advances in the research of HIV-1 envelope glycoprotein gp120 inhibitors / 药学学报

From the process of human immunodeficiency virus-1 (HIV-1) invading cells, the combination of gp120 and CD4 is the first step for HIV-1 to invade cells. Interfering with this process can prevent HIV from recognizing target cells and inhibit virus replication. Therefore, HIV-1 gp120 is an important part of the HIV-1 life cycle. Fostesavir, a phosphatate prodrug derived from the gp120 inhibitor BMS-626529 modified by the prodrug strategy, was approved for the treatment of adult patients with multidrug resistant HIV-1 infection by the US FDA and the European Medicines Agency in 2020 and 2021, respectively. In this review, we focus on the research progress of small molecule inhibitors targeting the interaction of gp120-CD4 from the perspective of medicinal chemistry, in order to provide reference for the subsequent research of gp120 inhibitors.

Fostemsavir, a Drug with Novel Mechanism for the Treatment of HIV-1 Infection

HIV is a global problem with increased mortality and morbidity. The highly active antiretroviral therapy is effective in reducing the HIV RNA and improving the immune response. The drugs in the current regimen have certain disadvantages such as adverse effects, drug intolerance, and drug resistance. Since there is a demand for identifying the drugs with new mechanism of action, the compounds which target the viral gp120 receptor were screened and the most suitable drug among them was identified. In a Phase II and Phase III trial, the drug BMS-663068 fostemsavir was found to be efficacious in reducing the viral RNA levels. The drug is a prodrug that gets converted into metabolite temsavir BMS-626529. The preferred dose is 600 mg orally 12 hourly in patients who had undergone many treatment schedules with multidrug-resistant infection and those who cannot tolerate the drug regimen due to resistance and safety issues. The drug is metabolized by CYP3A4 and has drug interactions with CYP3A4 inducers and inhibitors. This review mainly comprises the mechanism of action, clinical trials, pharmacological properties, and adverse effects of the drug fostemsavir.

Small molecule HIV entry inhibitors targeting gp120 or gp41:research advances / 国际药学研究杂志

The envelope glycoprotein subunit gp120 and the transmembrane subunit gp41 of human immunodeficiency virus type 1(HIV-1)play important roles in the process of viral entry into target cells, and serve as key targets for developing HIV-1 entry inhibitors. Peptide HIV entry inhibitors, such as T20, are limited in clinical application due to the lack of oral properties. However, because of the possible oral administration and high bioavailability, small molecule compounds become more and more important in the research for HIV entry inhibitors. In this review, we summarize the recent advances in the development of small molecule HIV-1 entry inhibitors, including the NBD and BMS series of small-molecule inhibitors targeting gp120 as well as the NB-206 and ADS-J1 se- ries of small-molecule inhibitors targeting gp41. These small molecule inhibitors are promising compounds with research value, which lay a foundation for the design of more efficient and more reasonable small-molecule HIV-1 entry inhibitors.

Establishment of a vimentin knockout and HIV-1 gp120 transgenic mouse model / 南方医科大学学报

OBJECTIVE@#To construct a HIV-1 gp120 transgenic mice (gp120 Tg) with vimentin (VIM) gene knockout.@*METHODS@#Female HIV-1 gp120 Tg mice were mated to VIM heterozygote mice (F0). All the offspring mice were derived from these original founders so that both genotypes had the same mixed genetic background. The F1 mice were bred to generate of VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. PCR was performed for genotyping of the mice, and the expressions of VIM and gp120 in the brain tissues were examined using immunoblotting.@*RESULTS@#The results of PCR showed the presence of the target bands in VIM, VIM, VIM/gp120 Tg and VIM/gp120 Tg mice. In VIM/gp120 Tg mice, gp120 expression was detected throughout the brain regions while no VIM expression was detected.@*CONCLUSIONS@#We generated gp120 transgenic mouse models with VIM gene knockout, which facilitate the exploration of the role of VIM in gp120-induced neurotoxicity.

Role of HIV-1 envelope protein gp120 in neuronal injury-induced cognitive impairment / 中华微生物学和免疫学杂志

Objective To investigate the role of HIV-1 envelope protein gp120 in cognitive im-pairment induced by neuronal damage. Methods Western blot and immunofluorescence assay were used to detect microglia activation, inflammatory factor expression and neuronal damage after gp120 treatment. Neu-ronal damage and neurocognitive performance in gp120-transgenic mice were evaluated using immunohisto-chemical staining and behavioral analysis, respectively. Results In vivo and in vitro experiments showed that HIV-1 gp120 significantly induced the expression of caspase-1 and IL-1β, and indirectly caused neuro-nal synaptic shortening and neuronal damage (P<0. 05). Compared with wild-type mice, gp120-transgenic mice showed significant cortical and hippocampal glial activation, neuronal loss, dendritic damage and neu-rocognitive disorders. Conclusions HIV-1 gp120 might cause neuronal damage through activating the re-lease of inflammatory factor by microglia and involve in neurocognitive impairment.

Role of HIV-1 envelope protein gp120 in neuronal injury-induced cognitive impairment / 中华微生物学和免疫学杂志

Objective@#To investigate the role of HIV-1 envelope protein gp120 in cognitive impairment induced by neuronal damage.@*Methods@#Western blot and immunofluorescence assay were used to detect microglia activation, inflammatory factor expression and neuronal damage after gp120 treatment. Neuronal damage and neurocognitive performance in gp120-transgenic mice were evaluated using immunohistochemical staining and behavioral analysis, respectively.@*Results@#In vivo and in vitro experiments showed that HIV-1 gp120 significantly induced the expression of caspase-1 and IL-1β, and indirectly caused neuronal synaptic shortening and neuronal damage (P<0.05). Compared with wild-type mice, gp120-transgenic mice showed significant cortical and hippocampal glial activation, neuronal loss, dendritic damage and neurocognitive disorders.@*Conclusions@#HIV-1 gp120 might cause neuronal damage through activating the release of inflammatory factor by microglia and involve in neurocognitive impairment.

Compounds of traditional Chinese medicine and neuropathic pain / 中国天然药物

Neuropathic pain (NP) has become a serious global health issue and a huge clinical challenge without available effective treatment. P2 receptors family is involved in pain transmission and represents a promising target for pharmacological intervention. Traditional Chinese medicine (TCM) contains multiple components which are effective in targeting different pathological mechanisms involved in NP. Different from traditional analgesics, which target a single pathway, TCMs take the advantage of multiple components and multiple targets, and can significantly improve the efficacy of treatment and contribute to the prediction of the risks of NP. Compounds of TCM acting at nucleotide P2 receptors in neurons and glial cells could be considered as a potential research direction for moderating neuropathic pain. This review summarized the recently published data and highlighted several TCMs that relieved NP by acting at P2 receptors.

SBi4211 alleviates gp120-induced central nervous system injury / 南方医科大学学报

OBJECTIVE@#To explore the protective effect of SBi4211 (heptamidine), an inhibitor of S100B, against central nervous system injury induced by HIV-1 envelope protein gp120.@*METHODS@#In an @*RESULTS@#In the cell co-culture system, SBi4211 treatment significantly inhibited gp120-induced expression of S100B, RAGE and GFAP in U251 cells (@*CONCLUSIONS@#SBi4211 can protect neurons from gp120-induced neurotoxicity possibly by inhibiting the S100B/ RAGE-mediated signaling pathway.

Establishment of a gp120 transgenic mouse model with 7 nAChR knockout / 南方医科大学学报

OBJECTIVE@#To construct a HIV-1 gp120 transgenic mouse model (gp120) with 7 nicotinic acetylcholine receptor (7nAChR) gene knockout.@*METHODS@#The 7nAChR gene knockout mice (7R) were crossed with HIV-1gp120 transgenic mice (gp120) to generate F1 generation mice. We selected the F1 mice with the genotype of 7R/gp120 to mate to obtain the F2 mice. The genotypes of the F3 mice were identified by PCR, and the protein expressions in the double transgenic animal model was analyzed by immunohistochemistry. BV2 cells were treated with gp120 protein and 7nAChR inhibitor, and the expressions of IL-1β and TNF- were detected using ELISA.@*RESULTS@#The results of PCR showed the bands of the expected size in F3 mice. Two F3 mice with successful double gene editing (7R/gp120) were obtained, and immunohistochemistry showed that the brain tissue of the mice did not express 7 nAChR but with high gp120 protein expression. In the cell experiment, treatment with gp120 promoted the secretion of IL-1β and TNF- in BV2 cells, while inhibition of 7nAChR significantly decreased the expression of IL-1β and TNF- ( < 0.001).@*CONCLUSIONS@#By mating gp120 Tg mice with 7R mice, we obtained gp120 transgenic mice with 7nAChR gene deletion, which serve as a new animal model for exploring the role of 7nAChR in gp120-induced neurotoxicity.

Analysis on variation characteristics of HIV-1 Gp120 sequences in men who have sex with men in Guangzhou / 中华实验和临床病毒学杂志

Objective@#To analyze the variation characteristics of HIV-1 Gp120 sequences in men who have sex with men (MSM) in Guangzhou.@*Methods@#Plasma samples were collected from HIV-1 infected MSM before antiretroviral treatment. Viral RNA was extracted from plasma. Gp120 gene sequences were amplified by reverse transcription and nested-PCR using specific primers. Phylogenetic tree, length polymorphism, amino acid characteristics of V3 loop, co-receptors and signature amino acids were analyzed.@*Results@#The phylogenetic tree were divided into 4 clusters, and the most prevalent subtypes were CRF07_BC (34/61, 55.74%) and CRF01_AE (24/61, 39.34%). Majority of HIV-1 Gp120 sequences had 496-515 amino acids. Among five hypervariable regions, the V1 region had the highest levels of length polymorphism and V3 region had the lowest. The top four peptide of V3 loop were GPGQ (56/58, 96.55%). Most of the co-receptors HIV-1 strains used was CCR5(50/58, 86.21%)according to four methods of comprehensive prediction. There are four signature amino acids in CRF01_AE subtype strains, and the frequency of occurrence was 0.75-0.83; there are eight signature amino acids in CRF07_BC subtype strains, and the frequency was 0.74-0.94.@*Conclusions@#The length of Gp120 sequences in MSM in Guangzhou has a high polymorphism. The top four peptide of V3 loop, co-receptor and signature amino acid of V3 ring have formed unique patterns.

Osthole inhibits HIV gpl20 induced neuropathic pain by down-regulating P2X3 receptor in DRG / 中国药理学通报

Aim To investigate whether osthole can alleviate neuropathic pain induced by HIV gpl20 and its mechanism. Methods The paw withdrawal threshold and the paw withdrawal latency were observed to assess pain behaviors in four groups of the rats, including sham group, sham combined with osthole treatment group, gpl20 treatment group, and gpl20 combined with osthole treatment group. The protein expression levels of the P2X3 receptor, tumor necrosis factor-a receptor (TNF-aR), ERK, p-ERK in the L4-L6 dorsal root ganglia (DRG) were measured by Western blot. The mRNA expression level of P2X3 receptor was assessed by real-time quantitative polymerase chain reaction ( qPCR). The whole path clamp recording was used to measure HEK293 cell current activated by ATP. Results Osthole attenuated the mechanical and thermal hyperalgesia in gpl20 treated rats and down-regulated P2X3 receptor mRNA and protein expression in L4-L6 DRGs of gpl20 treated rats. Additionally, osthole simultaneously decreased the expression of TNF-ctR protein in L4-L6 DRGs and inhibited the phosphorylated ERK1/2 protein expression. Moreover, osthole reduced ATP activated current of HEK293 cells transfected with hP2X3R. Conclusion Osthole decreases the mechanical and thermal hyperalgesia induced by gpl20 through inhibiting P2X3R in DRG.

Expression, purification, characterization and immunogenicity of human immunodeficiency virus-1 glycoprotein gp120 derived from insect cells / 中华微生物学和免疫学杂志

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.

Expression, purification, characterization and immunogenicity of human immunodeficiency virus-1 glycoprotein gp120 derived from insect cells / 中华微生物学和免疫学杂志

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.

Role of P2X7 receptor in learning and memory dysfunction induced by gp120 in rats / 实用医学杂志

Objective To investigate the role of P2X7 receptor in learning and memory dysfunction induced by HIV-1 enveloped protein gp120 in rats. Methods The imitating HIV-1 associated dementia (HAD) animal models were established by intracerebroventricular (ICV) infusion of gp120 in rats. The effect of gp120 on the learning and memory dysfunction in rats was evaluated by Morris water maze (MWM) test. The role of P2X7 receptor (P2X7R) was studied by Western blot and PCR assay. Results The ICV infusion of gp120 for 3 days in rats could imitated the HAD animal model. Results of MWM test showed that the rats in the model group had longer escape latencies and errors compared with those in the control group (P < 0.01); Results of Western blot and PCR assay showed that the expressions of P2X7R and P2X7 mRNA in hippocampus of rats in the model group were significantly increased (P < 0.01). Conclusions The ICV infusion of gp120 in rats could imitate the HIV-1 associated dementia (HAD) animal models, and P2X7R may be involved in the pathophysiological process of learning and memory dysfunction caused by gp120.

Comparative analysis of Gp120 sequences of enterotropic and neurotropic SIV strains / 中国比较医学杂志

Objective To study the mutations of Env sequence of SIVmac239 after infection of Chinese rhesus monkeys, and compare the differences and characteristics of Gp120 sequences of enterotropic and neurotropic SIV strains. Methods Six strains of simian immunodeficiency virus were analyzed in this study: four separated from peripheral blood mononuclear cells of SIVmac239-infected monkeys and two neurotropic SIVmac251 strains.Isolated and cultured monoclonal virus was obtained by limiting dilution assay.Gp120 sequences were amplified after the RNA extraction and phylogenetic analysis was processed thereafter.So did the Gp120 amino acid sequence and N-glycosylation sites analysis of the enterotropic and neurotropic strains.Results SIVmac239 had different mutations in four rhesus monkeys.The diversity in amino acid sequences of the enterotropic and neurotropic strains concentrated in the V1 and V4 regions of Gp120.The enterotropic strains had an addition of glycosylation site in V4 but the glycosylation site changes of neurotropic strains were located in the conservative regions of C1, C2 and C3.Conclusions The addition of one glycosylation site in V4 region of GP120 and loss of one glycosylation site in C1 region are associated with enhanced enterotropism and neurotropism.The differences between the enterotropic and neurotropic strains are not dipicted in Gp120 V3 region which is closely related with the tropism of strains.

Eukaryotic expression and characterization of mouse TSLP and HIV-1 gp120 BAL V1/V2 fusion protein / 中国免疫学杂志

Objective:To express fusion protein of mouse thymic stromal lymphopoietin (TSLP) and HIV-1 gp120BAL V1/V2 subdomain in 293F cell.Methods:Full length of the V1V2 sequence from BAL isolate was fused with the C-terminus of mouse thymic stromal lymphopoietin (TSLP) and sub-cloned into pCEP-Pu vector to construct the eukaryotic expression plasmid-pCTV1V2BAL.The recombinant plasmid was confirmed by enzyme digestion and sequencing , then transfected into 293 F cells using PEI as a transfection reagent .The fusion protein was purified by metal chelate affinity chromatography and characterized by SDS -PAGE and Western blot . The epitopes of V1/V2 in fusion protein were identified by ELISA .Results:The SDS-PAGE and Western blot results showed that there were highly heterogeneous glycoprotein bands at the site between 35 kD and 55 kD, which reacted with anti-mTSLP rabbit polyclonal antibody and anti-His tag mouse monoclonal antibody .The ELISA analysis showed that antibodies to V 1/V2BAL existed in the sera of HIV-1 positive patients.Conclusion:The mTSLP-V1/V2 fusion protein was successfully expressed in 293F cells, which may be useful for HIV-1 vaccine research .

The Protective Effects of IGF-1 on Different Subpopulations of DRG Neurons with Neurotoxicity Induced by gp120 and Dideoxycytidine In Vitro

Peripheral neuropathy induced by human immunodeficiency virus (HIV) infection and antiretroviral therapy is not only difficult to distinguish in clinical practice, but also difficult to relieve the pain symptoms by analgesics because of the severity of the disease at the later stage. Hence, to explore the mechanisms of HIV-related neuropathy and find new therapeutic options are particularly important for relieving neuropathic pain symptoms of the patients. In the present study, primary cultured embryonic rat dorsal root ganglion (DRG) neurons were used to determine the neurotoxic effects of HIV-gp120 protein and/or antiretroviral drug dideoxycytidine (ddC) and the therapeutic actions of insulin-like growth factor-1 (IGF-1) on gp120- or ddC-induced neurotoxicity. DRG neurons were exposed to gp120 (500 pmol/L), ddC (50 micromol/L), gp120 (500 pmol/L) plus ddC (50 micromol/L), gp120 (500 pmol/L) plus IGF-1 (20 nmol/L), ddC (50 micromol/L) plus IGF-1 (20 nmol/L), gp120 (500 pmol/L) plus ddC (50 micromol/L) plus IGF-1 (20 nmol/L), respectively, for 72 hours. The results showed that gp120 and/or ddC caused neurotoxicity of primary cultured DRG neurons. Interestingly, the severity of neurotoxicity induced by gp120 and ddC was different in different subpopulation of DRG neurons. gp120 mainly affected large diameter DRG neurons (>25 microm), whereas ddC mainly affected small diameter DRG neurons (< or =25 microm). IGF-1 could reverse the neurotoxicity induced by gp120 and/or ddC on small, but not large, DRG neurons. These data provide new insights in elucidating the pathogenesis of HIV infection- or antiretroviral therapy-related peripheral neuropathy and facilitating the development of novel treatment strategies.

Caracterização molecular da região do envelope do HIV-1 em crianças e adolescentes da cidade de São Paulo, Brasil / Molecular characterization of HIV-1 envelope in children and teenager in São Paulo city, Brazil

Introdução: O manejo de crianças vivendo com HIV/AIDS é um desafio e pode ser beneficiado com informações adicionais para subsidiar as decisões clínicas. Foram avaliados dados clínicos e laboratoriais a partir de uma coorte pediátrica em São Paulo, Brasil, visando a identificação de marcadores de prognóstico da doença. Métodos: Todas as crianças foram acompanhadas em uma unidade pediátrica de referência em São Paulo, Brasil. Todas as crianças com consentimento do estudo e informações de acompanhamento foram incluídas. Foi realizado sequenciamento parcial do gene do envelope do HIV-1 das amostra disponíveis (plasma ou PBMC). As amostras foram amplificadas por nested PCR, sequenciados com ABI3100 usando Big Dye e as sequências foram editadas manualmente. As taxas de falso positivo (FPRs, predição genotípica do tropismo viral) foram avaliadas pelo website geno2-pheno[co-receptores]. Os desfechos de evolução da doença avaliados foram: 1- aviremia (CV < 1.7cópias/mL versus viremia detectável); e 2- Progressão da doença (TCD4> 350cels/mm3 e clinicamente assintomáticos versus sintomas ou TCD4 <350cels/mm3). EpiInfo6 e Stata 8 foram utilizados para análise estatística e valores de p<0.05 foram considerados.Resultados: Das 87 crianças incluídas, 71 tiveram acompanhamento por mais de um ano. Foi observado que 45% do sexo masculino, com idade mediana de 8 anos (1-15). Na entrada, 86% apresentavam sintomas, com mediana de TCD4 e carga viral respectivamente de 454 células/mm3 e 5.1 log10. Na última avaliação, 75% eram assintomáticos, 37% avirêmicos, com mediana de linfócitos TCD4 470cels/mm3. O tropismo viral R5 na entrada foi observado em 57% dos pacientes e esteve associada a valores de TCD4 mais elevados durante o seguimento e a sintomatologia, quando comparados com pacientes X4. Aviremia foi associado ao uso de HAART como primeiro esquema, CV indetectável durante o seguimento e adesão boa ao TARV. A progressão boa da doença foi...

The Role of HIV Replicative Fitness in Perinatal Transmission of HIV / 中国病毒学·英文版

Perinatal transmission of Human immunodeficiency virus(HIV),also called mother-to-child transmission(MTCT),accounts for 90% of infections in infants worldwide and occurs in 30%-45% of children born to untreated HIV-1 infected mothers.Among HIV-1 infected mothers,some viruses are transmitted from mothers to their infants while others are not.The relationship between virologic properties and the pathogenesis caused by HIV-1 remains unclear.Previous studies have demonstrated that one obvious source of selective pressure in the perinatal transmission of HIV-1 is maternal neutralizing antibodies.Recent studies have shown that viruses which are successfully transmitted to the child have growth advantages over those not transmitted,when those two viruses are grown together.Furthermore,the higher fitness is determined by the gp120 protein of the virus envelope.This suggests that the selective transmission of viruses with higher fitness occurred exclusively,regardless of transmission routes.There are many factors contributing to the selective transmission and HIV replicative fitness is an important one that should not be neglected.This review summarizes current knowledge of the role of HIV replicative fitness in HIV MTCT transmission and the determinants of viral fitness upon MTCT.

Use of Mutual Information Arrays to Predict Coevolving Sites in the Full Length HIV gp120 Protein for Subtypes B and C / 中国病毒学·英文版

It is well established that different sites within a protein evolve at different rates according to their role within the protein; identification of these correlated mutations can aid in tasks such as ab initio protein structure,structure function analysis or sequence alignment.Mutual Information is a standard measure for coevolution between two sites but its application is limited by signal to noise ratio.In this work we report a preliminary study to investigate whether larger sequence sets could circumvent this problem by calculating mutual information arrays for two sets of drug naive sequences from the HIV gp120 protein for the B and C subtypes.Our results suggest that while the larger sequences sets can improve the signal to noise ratio,the gain is offset by the high mutation rate of the HIV virus which makes it more difficult to achieve consistent alignments.Nevertheless,we were able to predict a number of coevolving sites that were supported by previous experimental studies as well as a region close to the C terminal of the protein that was highly variable in the C subtype but highly conserved in the B subtype.