Fig. 6: Cyp-PROTACs exhibit improved anti-HIV-1 activity compared to parental Cyp ligand TWH106.

a Experimental design for (b–x): activated primary CD4+ T cells were pretreated for 48 h with TWH106 (orange), CG167 (blue), RJS308 (green) or DMSO (black), washed and infected with HIV-1 NL4.3 (2000 mU RT/10⁶ cells). Cells were treated with 1 μM or 5 μM Cyp inhibitor throughout the experiment (solid circles) or only pretreated with 5 μM Cyp inhibitor (dashed circle). Alternatively, cells were pretreated with 5 μM Cyp inhibitor and 50 μM VHL inhibitor VH298 (5 μM + VHLi, dashed circle). b–m Representative data from one donor at indicated days post infection (dpi), mean (n = 1 independent experiment performed in duplicate), (b–e) % Gag+ cells (f–i) virus levels in supernatant measured by SG-PERT (j–m) CypA mean fluorescence intensity (MFI), normalised to DMSO. n–x Data from 4 donors (1 μM, 5 μM and 5 μM pretreatment only) or 3 donors (5 μM pretreatment only + VHL inhibitor), all data normalised to DMSO, mean ± SD, (n–q) % Gag+ cells at 4 dpi, (r–u) virus levels in supernatant at 4 dpi measured by SG-PERT, (v–x) CypA MFI at time of infection (0 dpi). Statistical comparison using RM one-way ANOVA with Dunnett’s multiple comparisons test comparing to TWH106, * (P ≤ 0.05), ** (P ≤ 0.01), *** (P ≤ 0.001), **** (P ≤ 0.0001), P-values shown. Data from additional donors in Supplementary Fig. 14. w (right) histogram showing the spread of CypA-FITC fluorescence at 0 dpi, representative data from one donor. Gating strategies are shown in Supplementary Fig. 15a–c. Source data are provided in the Source Data file.