Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Exploring Inhibitors of HIV-1 Protease: Interaction Studies with Applications for Drug Discovery
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry.
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A variety of HIV-1 protease inhibitors and their interactions with the enzyme have been characterized in order to identify novel and improved drugs against AIDS. The investigated inhibitors were represented by clinical and non-clinical inhibitors, active site and allosteric inhibitors, transition-state analogues and metal-ions. In addition, different enzyme variants were used to investigate the contribution of different amino acid residues to the interaction with different ligands. The problem of resistance has been addressed by exploring novel types of inhibitors, and resistant mutants of HIV-1 protease. A study resolving the inhibition of HIV-1 protease by Cu2+ showed that the enzyme can be allosterically inhibited and that copper inhibition is a result of an interaction with His-69 and a subsequent conformational change. Several types of transition-state analogues were analyzed with respect to their inhibition of wild-type and resistant mutants of HIV-1 protease. Unfortunately cyclic compounds were not found to be better than linear compounds. Moreover, it was not possible to identify structure-activity relationships that clearly correlated with efficacy towards mutants and a biosensor based method for more detailed kinetic studies was therefore adopted. By cross-linking the immobilized enzyme on the biosensor matrix, a stable surface was obtained and kinetic rate constants could be determined for the interaction between the enzyme and inhibitors. Additional improvements in the methodology involved identification of a more representative interaction model, allowing more detailed studies of interactions with resistant mutants and varying conditions. Finally, absorption to lipid membranes and interaction with human serum albumin and α1-glycoprotein by clinical drugs were studied in a simplified ADME model system for improvement of the earlier stages of drug development. These studies have revealed important characteristics of these drugs that can potentially be modeled into new compounds that have improved efficacy of both wild-type and resistant mutants of HIV-1 protease.

Place, publisher, year, edition, pages
Uppsala: Institutionen för naturvetenskaplig biokemi , 2004. , p. 47
Series
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 1038
Keywords [en]
Biochemistry, kinetics, biosensors, ADME, drug discovery, SPR, resistance, inhibition, copper
Keywords [sv]
Biokemi
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-4655ISBN: 91-554-6087-9 (print)OAI: oai:DiVA.org:uu-4655DiVA, id: diva2:165402
Public defence
2004-12-03, B41, BMC, Husargatan 3, Uppsala, 13:30 (English)
Opponent
Supervisors
Available from: 2004-11-12 Created: 2004-11-12 Last updated: 2017-05-04Bibliographically approved
List of papers
1. Inhibition of HIV-1 protease by Cu2+ occurs by a reversible allosteric mechanism involving His-69
Open this publication in new window or tab >>Inhibition of HIV-1 protease by Cu2+ occurs by a reversible allosteric mechanism involving His-69
(English)Article in journal (Refereed) Submitted
Identifiers
urn:nbn:se:uu:diva-92354 (URN)
Available from: 2004-11-12 Created: 2004-11-12 Last updated: 2009-11-24Bibliographically approved
2. Resistance profiles of cyclic and linear inhibitors of HIV-1 protease
Open this publication in new window or tab >>Resistance profiles of cyclic and linear inhibitors of HIV-1 protease
Show others...
2002 (English)In: Antiviral Chemistry & Chemotherapy, ISSN 0956-3202, E-ISSN 2040-2066, Vol. 13, no 1, p. 27-37Article in journal (Refereed) Published
Abstract [en]

Resistance to anti-HIV protease drugs is a major problem in the design of AIDS drugs with long-term efficacy. To identify structural features associated with a certain resistance profile, the inhibitory properties of a series of symmetric and asymmetric cyclic sulfamide, cyclic urea and linear transition-state analogue inhibitors of HIV-1 protease were investigated using wild-type and mutant enzyme. To allow a detailed structure-inhibition analysis, enzyme with single, double, triple and quadruple combinations of G48V, V82A, 184V and L90M substitutions was used. Kinetic analysis of the mutants revealed that catalytic efficiency was 1-30% of that for the wild-type enzyme, a consequence of reduced kcat in all cases and an increased KM for all mutants except for the G48V enzyme. The overall structure-inhibitory profiles of the cyclic compounds were similar, and the inhibition of the V82A, 184V and G48V/L90M mutants were less efficient than of the wild-type enzyme. The greatest increase in Ki was generally observed for the 184V mutant and least for the G48V/L90M mutant, and additional combinations of mutations did not result in improved inhibition profiles for the cyclic compounds. An extended analysis of additional mutants, and including a set of linear compounds, showed that the profile was unique for each compound, and did not reveal any general structural features associated with a certain inhibition profile. The effects of structural modifications in the inhibitors, or of mutations, were not additive and they differed depending on their context. The results demonstrate the difficulties in predicting resistance, even for closely related compounds, and designing compounds with improved resistance profiles.

National Category
Pharmaceutical Sciences Natural Sciences
Identifiers
urn:nbn:se:uu:diva-64332 (URN)12180647 (PubMedID)
Available from: 2006-03-04 Created: 2006-03-04 Last updated: 2018-01-10Bibliographically approved
3. Determination of interaction kinetic constants for HIV-1 protease inhibitors using optical biosensor technology
Open this publication in new window or tab >>Determination of interaction kinetic constants for HIV-1 protease inhibitors using optical biosensor technology
Show others...
2001 (English)In: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 291, no 2, p. 207-218Article in journal (Refereed) Published
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-92356 (URN)10.1006/abio.2001.5025 (DOI)
Available from: 2004-11-12 Created: 2004-11-12 Last updated: 2017-12-14Bibliographically approved
4. Identification of complexities in HIV-1 protease - ligand interactions using biosensor technology
Open this publication in new window or tab >>Identification of complexities in HIV-1 protease - ligand interactions using biosensor technology
Show others...
(English)Manuscript (Other (popular science, discussion, etc.))
Identifiers
urn:nbn:se:uu:diva-92357 (URN)
Available from: 2004-11-12 Created: 2004-11-12 Last updated: 2009-11-27Bibliographically approved
5. Early absorption and distribution analysis of antitumor and anti-AIDS drugs: lipid membrane and plasma protein interactions
Open this publication in new window or tab >>Early absorption and distribution analysis of antitumor and anti-AIDS drugs: lipid membrane and plasma protein interactions
2005 (English)In: Journal of Medicinal Chemistry, ISSN 0022-2623, E-ISSN 1520-4804, Vol. 48, no 10, p. 3536-46Article in journal (Refereed) Published
Keywords
Absorption, Anti-HIV Agents/*chemistry, Antineoplastic Agents/*chemistry, Binding Sites, Biosensing Techniques, Blood Proteins/*chemistry, HIV Protease/chemistry, HIV Protease Inhibitors/chemistry, Humans, Kinetics, Lipids/*chemistry, Membranes; Artificial, Orosomucoid/chemistry, Research Support; Non-U.S. Gov't, Serum Albumin/chemistry, Surface Plasmon Resonance, Taxoids/chemistry
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-76584 (URN)10.1021/jm049343o (DOI)15887962 (PubMedID)
Available from: 2006-03-04 Created: 2006-03-04 Last updated: 2017-12-14Bibliographically approved

Open Access in DiVA

fulltext(896 kB)1551 downloads
File information
File name FULLTEXT01.pdfFile size 896 kBChecksum MD5
35c48f0d16dd878600bf9dc657e707cf155195c0e5ea406ddf1b68b07ad88240bf995231
Type fulltextMimetype application/pdf
Buy this publication >>

By organisation
Department of Biochemistry
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 1551 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 1328 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf