17 research outputs found
6‑(1-Benzyl‑1<i>H</i>‑pyrrol-2-yl)-2,4-dioxo-5-hexenoic Acids as Dual Inhibitors of Recombinant HIV‑1 Integrase and Ribonuclease H, Synthesized by a Parallel Synthesis Approach
The
increasing efficiency of HAART has helped to transform HIV/AIDS
into a chronic disease. Still, resistance and drug–drug interactions
warrant the development of new anti-HIV agents. We previously discovered
hit <b>6</b>, active against HIV-1 replication and targeting
RNase H in vitro. Because of its diketo-acid moiety, we speculated
that this chemotype could serve to develop dual inhibitors of both
RNase H and integrase. Here, we describe a new series of 1-benzyl-pyrrolyl
diketohexenoic derivatives, <b>7a</b>–<b>y</b> and <b>8a</b>–<b>y</b>, synthesized following a parallel
solution-phase approach. Those 50 analogues have been tested on recombinant
enzymes (RNase H and integrase) and in cell-based assays. Approximately
half (22) exibited inhibition of HIV replication. Compounds <b>7b</b>, <b>7u</b>, and <b>8g</b> were the most active
against the RNase H activity of reverse-transcriptase, with IC<sub>50</sub> values of 3, 3, and 2.5 μM, respectively. Compound <b>8g</b> was also the most potent integrase inhibitor with an IC<sub>50</sub> value of 26 nM
Cell cycle.
<p>HEK293 cells stably expressing Pdcd4<sub>(39–91)</sub>luc were incubated for 24 h with DMSO (blue) or increasing concentrations of compounds <b>1</b>, <b>2</b>, <b>3</b>, <b>4</b>, <b>5</b>, and <b>6</b> (3 μM (violet), 10 μM (yellow), and 30 μM (red)). Cell cycle analysis was performed after permeabilization using 7-AAD. Data are presented as means ± SEM of at least 3 independent experiments (* p<0.05, ** p<0.01, *** p<0.01).</p
Nonspecific activity.
<p>HEK293 cells stably expressing Pdcd4<sub>(mut39–91)</sub>luc were incubated for 8 h with TPA [10 nM] in combination with increasing concentrations (0.1–30 μM) of compounds <b>1</b>, <b>2</b>, <b>3</b>, <b>4</b>, <b>5</b>, and <b>6</b>. Luciferase activity is given relative to TPA-treated controls. Data are presented as means ± SEM of at least 3 independent experiments (* p<0.05, ** p<0.01, *** p<0.01).</p
Proliferation.
<p>HEK293 cells stably expressing Pdcd4<sub>(39–91)</sub>luc were incubated for 5 days with DMSO (blue) or increasing concentrations of compounds <b>1</b>, <b>2</b>, <b>3</b>, <b>4</b>, <b>5</b>, and <b>6</b> (1 μM (green), 3 μM (violet), 10 μM (yellow), and 30 μM (red)). Confluency was assessed every 4 h in an IncuCyte instrument. Proliferation kinetics shown are representative for at least 3 independent experiments and represent means of 3 wells treated in parallel (* p<0.05, ** p<0.01, *** p<0.01).</p
Conformational analysis of compounds 1, 2 and 4.
<p>(A) A systematic approach was chosen to explore the conformational space around the three rotable bonds. (B-D) Global minimum energy conformations (<i>left panel</i>) and energy plots (<i>right panel</i>) of compounds <b>1</b> (B), <b>2</b> (C), and <b>4</b> (D).</p
Pdcd4 stabilizing activity.
<p>HEK293 cells stably expressing the Pdcd4 stability reporter Pdcd4<sub>(39–91)</sub>luc were incubated for 8 h with TPA [10 nM] in combination with increasing concentrations (0.1–30 μM) of compounds <b>1</b>, <b>2</b>, <b>3</b>, <b>4</b>, <b>5</b>, and <b>6</b>. Pdcd4 stabilizing activity was determined relative to Δ(RLU<sub>DMSO</sub>–RLU<sub>TPA</sub>). Data are presented as means ± SEM of at least 3 independent experiments (* p<0.05, ** p<0.01, *** p<0.01).</p
Chemical structures.
<p>Structures of compounds <b>1</b> (1,2-bis(4-chlorophenyl)disulfide), <b>2</b> (4-chloro-<i>N</i>-(4-chlorophenyl)benzamide), <b>3</b> ((<i>E</i>)-1,2-bis(4-chlorophenyl)diazene), <b>4</b> (1,2-bis(4-chlorophenyl)hydrazine), <b>5</b> (1,2-bis(4-methoxyphenyl)disulfide), and <b>6</b> (1,2-bis(4-nitrophenyl)disulfide).</p
Viability.
<p>HEK293 cells stably expressing Pdcd4<sub>(39–91)</sub>luc were incubated for 24 h with DMSO (blue) or increasing concentrations of compounds <b>1</b>, <b>2</b>, <b>3</b>, <b>4</b>, <b>5</b>, and <b>6</b> (3 μM (violet), 10 μM (yellow), and 30 μM (red)). Cell death analysis was performed using Annexin V and 7-AAD co-staining. Data are presented as means ± SEM of at least 3 independent experiments (* p<0.05, ** p<0.01, *** p<0.01).</p
Design, Synthesis, and Structure–Activity Relationship of <i>N</i>‑Arylnaphthylamine Derivatives as Amyloid Aggregation Inhibitors
Dyes like CR are able to inhibit the aggregation of Aβ
fibrils. Thus, a screening of a series of dyes including ABBB (<b>1</b>) was performed. Its main component <b>2</b> tested
in an in vitro assay (i.e., ThT assay)
showed good potency at inhibiting fibrils association. Congeners <b>4</b>–<b>9</b> have been designed and synthesized
as inhibitors of Aβ aggregation. A number of these newly synthesized
compounds have been found to be active in the ThT assay with IC<sub>50</sub> of 1–57.4 μM. The most potent compound of this
series, <b>4k</b>, showed micromolar activity in this test.
Another potent derivative <b>4q</b> (IC<sub>50</sub> = 5.6 μM)
rapidly crossed the blood–brain barrier, achieving whole brain
concentrations higher than in plasma. So <b>4q</b> could be
developed to find novel potent antiaggregating βA agents useful
in Alzheimer disease as well as other neurological diseases characterized
by deposits of amyloid aggregates
New Nucleotide-Competitive Non-Nucleoside Inhibitors of Terminal Deoxynucleotidyl Transferase: Discovery, Characterization, and Crystal Structure in Complex with the Target
Terminal
deoxynucletidyl transferase (TdT) is overexpressed in some cancer
types, where it might compete with pol μ during the mutagenic
repair of double strand breaks (DSBs) through the nonhomologous end
joining (NHEJ) pathway. Here we report the discovery and characterization
of pyrrolyl and indolyl diketo acids that specifically target TdT
and behave as nucleotide-competitive inhibitors. These compounds show
a selective toxicity toward MOLT-4 compared to HeLa cells that correlate
well with in vitro selectivity for TdT. The binding site of two of
these inhibitors was determined by cocrystallization with TdT, explaining
why these compounds are competitive inhibitors of the deoxynucleotide
triphosphate (dNTP). In addition, because of the observed dual localization
of the phenyl substituent, these studies open the possibility of rationally
designing more potent compounds