TABLE 7.
Rates of HIV-1 protease cleavage of p25, half-lives of dissociation of MIs, and MI affinities from Gag polymorphisms
| Virus or VLP | Innate p25 cleavage |
GSK’254 relative cleavage rates vs wild-type | Single-cycle antiviral assay, FCEC50 vs wild-type |
MI dissociation from HIV-1 VLPs,a half-life ± SD, min |
Relative GSK’254/GSK’795 dissociative half-lives | MI affinity to HIV-1 VLPs,a
Kd ± SD, nM |
Relative GSK’254/GSK’795 affinity | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Kclv min−1 ± SD | Half-life, min | GSK’254 | GSK’795 | GSK’254 | GSK’795 | GSK’254 ± SD | GSK’795 ± SD | ||||
| Wild type | 0.0017 ± 0.0003 | 418 | 1.0 | 1 | 1 | 361 ± 65 | 51 ± 18 | 7.1 | 2.3 ± 2.3 | 4.8 ± 1.5 | 2.1 |
| V370A/ΔT371 | 0.0030 ± 0.0003 | 235 | 1.8 | ND | ND | 459 ± 155 | 48 ± 17 | 9.6 | ND | 26 | ND |
| ΔV370 | 0.0044 ± 0.0007 | 158 | 2.6 | 2.7 | 2.2 | 315 ± 125 | 46 ± 13 | 6.8 | 3.0 ± 3.1 | 27 ± 17 | 9.0 |
| V362I | 0.0047 ± 0.0005 | 148 | 2.8 | 1.0 | 2.8 | 269 ± 89 | 45 ± 17 | 6.0 | 0.6 ± 0.4 | 4.3 ± 2.7 | 7.2 |
| V370A | 0.0046 ± 0.0011 | 156 | 2.7 | 0.7 | 1.2 | 434 ± 139 | 36 ± 16 | 12.1 | 0.7 ± 0.3 | 6.5 ± 3.7 | 9.3 |
| V362I/V370A | 0.0097 ± 0.0022 | 73 | 5.7 | ND | ND | 227 ± 83 | 35 ± 13 | 6.5 | 0.5 ± 0.5 | 5.0 ± 3.6 | 10.0 |
| A364V | 0.0157 ± 0.0013 | 44 | 9.2 | >1000 | >600 | ≤1 | ≤1 | 1.0 | 15 ± 1 | 27 ± 10 | 1.8 |
a
Tritium-labeled GSK’254 and GSK’795 surrogates with the C-20/C-29 double bond reduced with 3[H] were used to determine MI dissociation half-life and binding affinity.