W0 2011/000721 A1 I||||||||||||||||||||||||||||||||||||||||||||||||||||||||| - Questel

unable to persist on the cell surface, even when used at 5 mM concentration,
leading to no antiinfective activity in the trans-infection assay (Figure 3).
Dose-response curves were obtained for 12 using CD4+ T lymphocytes from
healthy donors who are either genetically resistant (P1 ) or genetically
susceptible (P2) to HIV infection. B-THP-1/DC-SIGN cells were challenged
with HIV-1 (BaL strain) in the presence of increasing concentrations of 12.
After washing and co-culture with the CD4+ T lymphocytes analysis of p24
concentration in the supematants allowed to assess viral infection. Each point
was obtained in duplicate. At 100 μM the inhibition of infection was complete
for both donors and a 5 μM IC 50 could be estimated (Figure 4).
Infection studies were performed not only on laboratory adapted HIV strains
but also on viral strains isolated from patients. In particular clades V6 (CCR5
tropic) (Figure 5) and V17 (CXCR4 tropic) (Figure 6) were tested: 12 showed
>99% inhibition at 50 μM concentration.
7-aminoactinomicin D (7-AAD) labelling of the cells after the incubation period
and cytofluori metric analysis showed that the anti-infective properties of the
inhibitors are real and not an epiphenomenon, due to cell death. Thus, the data
for 12 shown in Figure 7 indicate that the percentage of 7-AAD positive cells
(apoptotic cells) did not change significantly in the absence of the compound or
in its presence up to 250 μM (the highest concentration tested in the infection
studies). Similar data were obtained after incubating the cells with 12 for 24
hours (not shown).
Figure 10 shows the anti-infective activity of a group of multivalent compounds.
12 and 12a are within the present invention, Man4 is a tetravalent presentation
of mannose on the same scaffold; G3PS-di24 is a fully succinylated BH30
dendrimer (which is a third generation dendrimer based o n 2,2'-
bis(methoxyol)propanoic acid (bis-MPA)) presenting 24-26 copies of the amine