Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1995-04-10
2000-05-16
Stucker, Jeffrey
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 71, 435 72, 435 76, 435 792, 435 23, 435974, 435975, C12Q 170
Patent
active
060635622
ABSTRACT:
An in vitro method for predicting the identity of distinct, first-generation, drug-resistant, biologically-active, HIV protease mutants that may emerge in vivo in response to a drug targeted thereagainst. In a preferred embodiment, the in vitro method comprises the steps of (a) preparing, in the presence of the drug, a comprehensive library of all first-generation mutants of the protease differing therefrom by at least one and preferably no more than three amino acid substitutions, each of the protease mutants being generated as part of a polyprotein with the HIV reverse transcriptase protein; (b) isolating, in vitro, first-generation, drug-resistant, biologically-active, mutant proteases from said library by assaying for biological activity of the reverse transcriptase protein; and (c) identifying the distinct, first-generation, biologically-active, mutant proteases so isolated. The present invention also relates to an in vitro method for evaluating the efficacy of a drug against a biologically-active mutant or wild-type form of HIV protease, said method comprising the steps of (a) providing a mutant polyprotein, said mutant polyprotein including a biologically-inactive mutant form of the protease linked to HIV reverse transcriptase by one or more sites cleavable by the biologically-active or wild-type form of the protease; (b) adding the drug to the mutant polyprotein; (c) then, adding the biologically-active or wild-type form of the protease to the mutant polyprotein; and (d) then, assaying for the presence of biological activity for reverse transcriptase, whereby the presence of reverse transcriptase activity indicates that the drug is not efficacious against the biologically-active mutant or wild-type form of the protease tested. The present invention further relates to a kit for evaluating the efficacy of a drug against a biologically-active mutant or wild-type form of HIV protease.
REFERENCES:
patent: 5223408 (1993-06-01), Goeddel et al.
patent: 5223409 (1993-06-01), Ladner et al.
patent: 5258289 (1993-11-01), Davis et al.
patent: 5288514 (1994-02-01), Ellman
patent: 5300425 (1994-04-01), Kauvar
Chow et al., "Use of evolut. limit. of HIV-1 multidrug resist. to optimize therapy," Nature, 1993; 361:650-4.
Chow et al., "HIV-1 error revealed," Nature, 1993; 364:679.
Emini et al., "HIV and multidrug resistance," Nature, 1993; 364:679.
Tisdale et al., "Rapid in vitro select. of human immunodef. virus type 1 resistant to 3'-thiacytidine inhib. due to a mutation in the YMDD region of reverse transcript.," Proc. Natl. Acad. Sci. USA, 1993; 90:5653-6.
Taddie et al., "Genetic Charact. of the Vaccinia Virus DNA Polymerase: Identific. of Point Mutations Conferring Altered Drug Sensit. and Reduced Fidelity," Journal of Virology, 1991; 65(2):869-79.
Johnston et al., "Present Status and Future Prospects for HIV Therapies," Science, 1993; 260:1286-93.
Richman, "HIV Drug Resistance," Annu. Rev. Pharmacol. Toxicol., 1993; 32:149-64.
Meyerhans et al., "Temp. Fluctutations in HIV Quasispecies In Vivo Are Not Reflected by Sequen. HIV Isolations," Cell, 1989; 58:901-10.
Wain-Hobson, "The fastest genome evolution ever described: HIV variation in situ," Current Opinion in Genetics and Development, 1993; 3:878-83.
Honess et al., "Single Mutations at Many Sites within the DNA Polymerase Locus of Herpes Simplex Viruses Can Confer Hypersens. to Aphidicolin and Resistance to Phosponoacetic Acid," J. gen. Virol., 1984; 65:1-17.
Lacey et al., "Anal. of mutations in the thymidine kinase genes of drug-resist. varicella-zoster virus popul. using the polymerase chain reaction," Journal of General Virology, 1991; 72:623-30.
Alper, "Drug Discovery on the Assembly Line," Science, 264:1399-1401 (Jun. 3, 1994).
Matthews et al., "In Vitro Mutagen. of the Herpes Simplex Virus Type 1 DNA Polymerase Gene Results in Altered Drug Sensitiv. of the Enzyme," Journal of Virology, 1989; 63(11):4913-8.
Roberts et al., "Rational Design of Peptide-Based HIV Proteinase Inhibitors," Science, 1990; 248:358-61.
Handwerger et al., "Alterat. in Penicillin-Binding Proteins of Clinic. and Laboratory Isolated of Path. Streptococcus pneumoniae with Low Levels of Penicillin Resistance," The Journal of Infectious Diseases, 1986; 153(1):83-9.
Smith et al., "Resumption of Virus Production after Human Immunodef. Virus Infection of T Lymphocytes in the Presence of Azidothymidine," Journal of Virology, 1987; 61(12):3769-73.
Larder et al., "Zidovudine-Resistant Human Immunodef. Virus Selected by Pass. in Cell Cult.," Journal of Virology, 1991; 65(12):5232-6.
Tanouye, "Pharmaceutical Consortium to Begin Clinical Trials of Comb. AIDS Drugs," Wall Street Journal (Apr. 14, 1994).
Larder et al., "Infect. potential of human immunodef. virus type 1 reverse transcriptase mutants with altered inhib. sensit.,"Proc. Natl. Acad. Sci. USA, 1989; 86:4803-7.
Kageyama et al., "In Vitro Inhib. of Human Immunodef. Virus (HIV) Type 1 Replication by C.sub.2 Symmetry-Based HIV Protease Inhib. as Single Agents or in Combin.," Antimicrobial Agents and Chemotherapy, 1992; 36(5):926-33.
Ho et al., "Characterization of Human Immunodef. Virus Type 1 Variants with Increased Resist. to a C.sub.2 -Symmetric Protease Inhib.," Journal of Virology, 68(3):2016-20 (Mar. 1994).
Neu, "The Crisis in Antibiotic Resistance," Science, 1992; 257:1064-73.
Mohri et al., "Quantitation of zidovudine-resistant human immunodef. virus type 1 in the blood of treated and untreated patients," Proc. Natl. Acad. Sci. USA, 1993; 90:25-9.
Barbas III et al., "Semisynth. combinatorial antibody libraries:A chem soln. to the diversity prob.," Proc.Natl.Acad.Sci.USA,1992;89:4457-61.
Amberg et al., "SurfZAP.TM. Vector*:Linking Phenotype to Genotype for Phagemid Display Libraries," Strategies in molecular biology, 6:2-4, published prior to the filing of the present application.
McCafferty et al., "Phage antibod.:filamentous phage displaying antibody variable domains," Nature, 1990; 348:552-4.
Matthews et al., "Substrate Phage:Selection of Protease Substrates by Monovalent Phage Display," Science, 1993; 260:1113-7.
Fields et al., "A novel genetic system to detect protein-protein interactions," Nature, 1989; 340:245-6.
Saag et al., "Extensive variation of human immunodeficiency virus type-1 in vivo," Nature, 1988; 334:440-4.
Nashed et al., "Continuous Spectrophotometric Assay for Retroviral Proteases of HIV-1 and AMV," Biochemical and Biophysical Research Communications, 1989; 163(2):1079-85.
Richards et al., "Sensitive, Soluble Chromogenic Substrates for HIV-1 Proteinase," The Journal of Biological Chemistry, 1990;265(14):7733-6.
Neu, "Quinolone Antimicrobial Agents," Annu. Rev. Med., 1992; 43:465-86.
Fasching et al., "gyrA Mutations in Ciprofloxacin-Resistant, Methicillin-Resistant Staphylococcus aureus from Indiana, Minnesota, and Tennessee," The Journal of Infectious Diseases, 1991; 164:976-9.
Larder et al., "Zidovudine resistance predicted by direct detection of mutations in DNA from HIV-infected lymphocytes," AIDS, 1991, 5:137-44.
Boucher et al., "Ordered Appearance of Zidovudine Resistance Mutations during Treatment of 18 Human Immunodeficiency Virus-Positive Subj.," The Journal of Infectious Diseases, 1992; 165:105-10.
Lopez-Galindez et al., "Charact. of genetic variation and 3'-azido-3'-deoxythymidine-resistance mutations of human immunodef. virus by the RNase A mismatch cleavage method," Proc. Natl. Acad. Sci. USA, 1991; 88:4280-4.
Nunberg et al., "Viral. Resist. to Human Immunodef. Virus Type 1-Spec. Pyrid. Rev. Transcr. Inhib.," Journal of Virology,1991;65(9):4887-92.
Dueweke et al., "A mutation in reverse transcriptase of bis(heteroaryl)piperazine-resistant human immunodef. virus type 1 that confers increased sensitivity to other nonnucleoside inhib.," Proc. Natl. Acad. Sci. USA, 1993; 90:4713-7.
Banerjee et al., "inhA, a Gene Encoding a Target for Isoniazid and Ethionamide in Mycobacterium tuberculosis," Science, 263:227-30 (Jan. 14, 1994).
Richman et al., "Detection of Mutations Associated with Zidovudine Resist. in Human Immunodef. Virus by Use of the Polymerase Chain Reaction," The Journ
Heefner Donald L.
Melnick Laurence M.
Sepracor Inc.
Stucker Jeffrey
LandOfFree
In vitro method for predicting the evolutionary response of HIV does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with In vitro method for predicting the evolutionary response of HIV , we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and In vitro method for predicting the evolutionary response of HIV will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-257293