Peptidic HIV fusion inhibitors targeting envelope glycoprotein transmembrane subunit gp41 / 中国药理学通报

HIV envelope glycoprotein transmembrane subunit gp41 plays a major role in the fusion of viral and target cell membranes. The extracellular region of gp41 consists of N-terminal fusion peptide and downstream N- and C-heptad repeat (NHR and CHR) regions. The peptidesderived from the NHR and CHR regions, designated N- and C-peptides, respectively, have potent inhibitory activity on the HIV mediated cell fusion. C-peptide T-20 has just got the approval of U.S. FDA, which became the first success of one new class anti-HIV agents, named HIV-fusion inhibitors. However, a relatively long peptide such as T-20 suffers from several limitations including proteolytic sensitivity, large dosage, therefore it is unable to produced by gene engineering. Alternately, shorter peptidic fusion inhibitors and active peptides suitable for gene engineering are pursued. In the recent years, this kind of peptide modifications are hot spots in HIV research field and contribute a lot to the inhibitory mechanism of N- and C-peptide.

Screening and identification of HIV-1 gp41 core structure epitope from random phage display peptide library / 中国免疫学杂志

Objective:To identify and characterize the epitopes on core structure of HIV 1 gp41.Methods:A random phage displayed dodecapeptide library was screened with a conformation specific monoclonal antibody NC 1 specifically against the core structure of HIV 1 gp41.The positive clones were identified by sandwich ELISA,soluble NC 1 blocking assay and competitive inhibition assay.Results:After three rounds of screening,10 of 24 phage clones were identified as positive clones which can bind to NC 1.Amino acid sequences deduced from DNA sequences showed five different sequences:HDVHHRWVYLLS?ITVNEWLYTSEQ?HGRSHGMFKPKR?MGPIARPHWHLN?DMYRSPRPKPDT.The binding between phage clones(displayed HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN, respectively)and NC 1 could be inhibited by N36 C34 complex.Soluble NC 1 could block the binding between phage clones and NC 1.Conclusion:The results indicate that HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN are the mimotopes which could mimic the core structrue epitopes of six helix bundle of HIV 1 gp41.

Studies on a high-throughput screening method for identification of HIV fusion inhibitors targeting gp41 / 中国药理学通报

AIM To modify and improve a screening assay so that it becomes more convenient, economic and adaptable in China for high throughput screening of HIV fusion inhibitors targeting gp41. METHODS The original screening method reported by Jiang et al (J Virol. Methods 1999;80:85 96) was modified by: ① using a conformation specific monoclonal antibody to replace a polyclonal antibody for coating plates; ②simplifying the procedures; ③using parts of the reagents produced in China. RESULTS The modified screening assay is simpler, more convenient, and more economic than the original assay, but its sensitivity is comparable to and specificity is a little better than the original method. CONCLUSIONS The modified screening assay is more convenient and economic and can be used in China for high throughput screening of HIV fusion inhibitors from complex sample, such as phage display peptide libraries, microorganism fermentation liquids, herbs and other natural products.