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US 2006/0216693 A1Published application
Method of screening compounds which inhibit the initiation of the retrotranscription of the rna of virus hiv-1 and means for implementing same
AssigneesCentre National de la Recherche Scientifique (CNRS), Universite Strasbourg 1 Louis Pasteur
InventorsFabienne Brule, Catherine Isel, Roland Marquet, Mickäel Rigourd
Life of this patent
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Abstract
The present invention relates to methods for screening compounds that can inhibit the early stage of an infection by the HIV-1 virus, that is to say the initiation step of viral genome reverse transcription, before integrating viral sequences into the cellular genome of the infected host organism.
Claims25
-24. (canceled)
An in vitro method for screening compounds inhibiting the initiation of HIV-1 virus RNA reverse transcription, comprising:
- a) contacting in a sample:
- (i) an RNA primer/RNA template complex formed between:
- (1) an RNA primer comprising from the 5′ end to the 3′ end:
- an intramolecular pairing sequence (109);
- a single-strand sequence (111);
- a stem-loop forming sequence (112);
- a single-strand sequence (113);
- an intermolecular pairing sequence (101) called anticodon sequence;
- an intermolecular pairing sequence (103);
- an intermolecular pairing sequence (105);
- an intramolecular pairing sequence (110);
- a single-strand sequence (114);
- an intermolecular pairing sequence (107) used as a primer;
- and
- (2) an RNA template comprising from the 5′ end to the 3′ end:
- an intramolecular pairing sequence (115);
- an intramolecular pairing sequence (116);
- an intermolecular pairing sequence (106);
- an intramolecular stem-loop forming sequence (117);
- an intramolecular pairing sequence (104);
- an intramolecular pairing sequence (102) called codon sequence;
- an intramolecular pairing sequence (118);
- a single-strand sequence (119);
- an intermolecular pairing sequence (108);
- an intramolecular stem-loop (8) forming sequence (120);
- a single-strand sequence (121);
- an intramolecular pairing sequence (122);
- wherein the RNA primer consists of:
- (a) either the in vitro transcription product of a nucleic acid encoding said RNA primer,
- (b) or the chemical synthesis or chemical hemisynthesis product of said RNA primer;
- and RNA primer being chosen amongst RNA primer sequence SEQ ID NO1 and RNA primer sequence SEQ ID NO3,
- being specified that:
- the paired sequences (101) and (102) form a helix (6C);
- the paired sequences (107) and (108) form a helix (7F);
- the paired sequences (109) and (110) form a helix (A);
- the paired sequences (107) and (108) form a helix (7F);
- the sequence (112) forms a stem-loop (B);
- the paired sequences (115) and (122) form a helix (1);
- the paired sequences (116) and (118) form a helix (2);
- the sequence (117) forms a stem-loop (4);
- the sequence (120) forms a stem-loop (8);
- with:
- (ii) a HIV-1 reverse transcriptase enzyme;
- and
- (iii) an appropriate dNTPs/ddNTP mixture;
- (iv) in the presence of a candidate compound to be tested;
- b) measuring the reverse transcription initiation activity in said sample;
- c) comparing the measured activity in step b) to the reverse transcription initiation activity measured in a control sample for which step a) is conducted without said candidate compound.
45 more elements
An RNA primer/RNA template complex formed between:
- (i) an RNA primer comprising from the 5′ end to the 3′ end:
- an intramolecular pairing sequence (109);
- a single-strand sequence (111);
- a stem-loop forming sequence (112);
- a single-strand sequence (113);
- an intermolecular pairing sequence (101) called anticodon sequence;
- an intermolecular pairing sequence (103);
- an intermolecular pairing sequence (105);
- an intramolecular pairing sequence (110);
- a single-strand sequence (114);
- an intermolecular pairing sequence (107) used as a primer;
- and
- (ii) an RNA template comprising from the 5′ end to the 3′ end:
- an intramolecular pairing sequence (115);
- an intramolecular pairing sequence (116);
- an intermolecular pairing sequence (106);
- an intramolecular stem-loop forming sequence (117);
- an intramolecular pairing sequence (104);
- an intramolecular pairing sequence (102) called codon sequence;
- an intramolecular pairing sequence (118);
- a single-strand sequence (119);
- an intermolecular pairing sequence (108);
- an intramolecular stem-loop (8) forming sequence (120);
- a single-strand sequence (121);
- an intramolecular pairing sequence (122);
- wherein the RNA primer consists of:
- (c) either the in vitro transcription product of a nucleic acid encoding said RNA primer,
- (d) or the chemical synthesis or chemical hemisynthesis product of said RNA primer;
- and RNA primer being chosen amongst RNA primer sequence SEQ ID NO:1 and RNA primer sequence SEQ ID NO:3,
- being specified that:
- the paired sequences (101) and (102) form a helix (6C);
- the paired sequences (107) and (108) form a helix (7F);
- the paired sequences (109) and (110) form a helix (A);
- the paired sequences (107) and (108) form a helix (7F);
- the sequence (112) forms a stem-loop (B);
- the paired sequences (115) and (122) form a helix (1);
- the paired sequences (116) and (118) form a helix (2);
- the sequence (117) forms a stem-loop (4);
- the sequence (120) forms a stem-loop (8).
36 more elements
Dependencies are inferred from the claim text.