Characterization of human immunodeficiency virus type 1 variants with increased resistance to a C2-symmetric protease inhibitor

Inhibitors of the human immunodeficiency virus type 1 protease represent a promising class of antiviral drugs for the treatment of AIDS, and several are now in clinical trials. Here, we report the in vitro selection of viral variants with decreased sensitivity to a C2-symmetric protease inhibitor (A-77003). We show that a single amino acid substitution (Arg to Gln or Lys) at position 8 of the protease results in a substantial decrease in the inhibitory activity of the drug on the enzyme and a comparable increase in viral resistance. These findings, when analyzed by using the three-dimensional structure of the protease-drug complex, provide a strategic guide for the future development of inhibitors of the human immunodeficiency virus type 1 protease.

[1]  D. Richman.  Resistance of clinical isolates of human immunodeficiency virus to antiretroviral agents , 1993, Antimicrobial Agents and Chemotherapy.

[2]  D. Richman,et al.  HIV with reduced sensitivity to zidovudine (AZT) isolated during prolonged therapy. , 1989, Science.

[3]  G. Petsko,et al.  Weakly polar interactions in proteins. , 1988, Advances in protein chemistry.

[4]  B. Larder,et al.  Fifth mutation in human immunodeficiency virus type 1 reverse transcriptase contributes to the development of high-level resistance to zidovudine. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[5]  R. Gilcher.  Human retroviruses and AIDS. , 1988, The Journal of the Oklahoma State Medical Association.

[6]  A. Wlodawer,et al.  Different requirements for productive interaction between the active site of HIV-1 proteinase and substrates containing -hydrophobic*hydrophobic- or -aromatic*pro- cleavage sites. , 1992, Biochemistry.

[7]  John P. Overington,et al.  X-ray analysis of HIV-1 proteinase at 2.7 Å resolution confirms structural homology among retroviral enzymes , 1989, Nature.

[8]  T K Sawyer,et al.  A synthetic HIV-1 protease inhibitor with antiviral activity arrests HIV-like particle maturation. , 1990, Science.

[9]  D. Norbeck,et al.  Design, activity, and 2.8 A crystal structure of a C2 symmetric inhibitor complexed to HIV-1 protease. , 1990, Science.

[10]  S. Vasavanonda,et al.  Antiviral and pharmacokinetic properties of C2 symmetric inhibitors of the human immunodeficiency virus type 1 protease , 1991, Antimicrobial Agents and Chemotherapy.

[11]  J. Craig,et al.  Effects of a Specific Inhibitor of HIV Proteinase (Ro 31-8959) on Virus Maturation in a Chronically Infected Promonocytic Cell Line (U1) , 1991 .

[12]  P. Darke,et al.  Synthesis and antiviral activity of a series of HIV-1 protease inhibitors with functionality tethered to the P1 or P1' phenyl substituents: X-ray crystal structure assisted design. , 1992, Journal of Medicinal Chemistry.

[13]  D. Richman,et al.  Human immunodeficiency virus type 1 mutants resistant to nonnucleoside inhibitors of reverse transcriptase arise in tissue culture. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Jadhav,et al.  In vitro isolation and identification of human immunodeficiency virus (HIV) variants with reduced sensitivity to C-2 symmetrical inhibitors of HIV type 1 protease. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[15]  H. Gendelman,et al.  Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone , 1986, Journal of virology.

[16]  J. Fitzgibbon,et al.  Human immunodeficiency virus type 1 pol gene mutations which cause decreased susceptibility to 2',3'-dideoxycytidine , 1992, Antimicrobial Agents and Chemotherapy.

[17]  M. Jaskólski,et al.  Conserved folding in retroviral proteases: crystal structure of a synthetic HIV-1 protease. , 1989, Science.

[18]  E A Emini,et al.  Viral resistance to human immunodeficiency virus type 1-specific pyridinone reverse transcriptase inhibitors , 1991, Journal of virology.

[19]  D F Hoth,et al.  Present status and future prospects for HIV therapies. , 1993, Science.

[20]  M. Navia,et al.  Three-dimensional structure of aspartyl protease from human immunodeficiency virus HIV-1 , 1989, Nature.

[21]  C. Debouck,et al.  The HIV-1 protease as a therapeutic target for AIDS. , 1992, AIDS research and human retroviruses.

[22]  S D Kemp,et al.  Zidovudine-resistant human immunodeficiency virus selected by passage in cell culture , 1991, Journal of virology.

[23]  I B Duncan,et al.  Rational design of peptide-based HIV proteinase inhibitors. , 1990, Science.

[24]  S D Kemp,et al.  Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). , 1989, Science.

[25]  J. Robinson,et al.  PCR amplification of HIV-1 proteinase sequences directly from lab isolates allows determination of five conserved domains. , 1992, Virology.

[26]  M. Otto,et al.  HIV protease inhibitors. , 1995, AIDS.