In vitro screening of compounds that inhibit reverse transcription, useful for treatment and prevention of human immune deficiency virus infection, from their effect on a primer-matrix complex

摘要

In vitro method of screening for compounds (A) that inhibit the initiation of reverse transcription of RNA of HIV-1. In vitro method of screening for compounds (A) that inhibit the initiation of reverse transcription of RNA of HIV-1 comprises: (a) incubating together a complex (C) of primer RNA (P) and matrix RNA (M); HIV-1 reverse transcriptase (RT); mixture of (di)deoxynucleotidfe triphosphates ((d)dNTP) and test compound; (b) measuring the activity of initiation of reverse transcription; and (c) comparing this activity with that in a control sample that does not contain test compound. (P), which is either an in vitro transcript from the appropriate nucleic acid or a (semi-)synthetic product, comprises, in the 5' to 3' direction, an intramolecular pairing sequence (109); single-stranded sequence (111); sequence (112) that forms a stem-loop; single-stranded region (113); intermolecular pairing sequence (101) ('anticodon'); intermolecular pairing sequences (103) and (105); intramolecular pairing sequence (110); single-stranded sequence (114) and intermolecular pairing sequence (107), functioning as primer. (M) comprises, in the 5' to 3' direction, intramolecular pairing sequences (115) and (116); intermolecular pairing sequences (106); sequence (117) that forms an intramolecular stem-loop; intramolecular pairing sequence (104); intramolecular pairing sequence (102) ('codon'); intramolecular pairing sequence (118); single-stranded sequence (119); intermolecular pairing sequence (108); sequence (120) forming an intramolecular stem-loop; single-stranded sequence (121) and intramolecular pairing sequence (122). When paired, (101) and (102) form helix 6C; (107) and (108) form helix 7F; (109) and (110) form helix A; (115) and (122) form helix 1; (116) and (118) form helix 2; while (112), (117) and (120) form stem-loops B, 4 and 8. Independent claims are also included for the following: (1) (C) as a new product; and (2) kit for the process that contains (C). ACTIVITY : Anti-HIV. No details of tests for anti-HIV activity are given. MECHANISM OF ACTION : Inhibiting reverse transcription of HIV-1 RNA.