Nef Hck Activation Inhibitor ; B9

Biochem/physiol Actions

Reversible: yes

Cell permeable: yes

Primary TargetNef

General description

A cell-permeable diphenylpyrazolo compound that is shown to disrupt HIV accessary protein Nef dimerization (IC₅₀ = 3 µM in HEK293T cells) and prevent Nef-mediated Src family kinase (SFK) Hck activation (IC₅₀ = 2.8 µM) without directly affecting the catalytic activity of c-Src, Hck, Lck, or Lyn (IC₅₀ >20 µM). Shown to effectively suppress HIV and SIV viral replication (IC₅₀﹤0.3 µM in 9 d HIV-1 NL4-3-infected CEM-T4 cultures; IC₅₀ = 1 µM in 5 d SIV δB670-infected CEM-174 cultures) and infectivity (45% and 50% inhibition against HIV-1 NL4-3 and SIV δB670, respectively, following 48 h 3 µM B9 treatment in TZM-bl reporter cells) in a Nef-dependent manner, being ineffective against the replication of Nef-defective HIV-1 mutant. Molecular docking studies predict a high-affinity B9-targeting site formed at the Nef dimerization interface and a low-affinity B9-binding site on each Nef monomer at the dimer interface periphery, with Asn126 contributing to both modes of binding. Consistently, in vitro binding studies reveal two B9 K_D values of 1.72 nM and 860 nM toward full-length HIV-1 Nef and a complete loss of B9 binding toward Nef N126A/L/Q mutants.

A cell-permeable diphenylpyrazolo compound that disrupts HIV accessary protein Nef dimerization (IC₅₀ = 3 µM in HEK293T cells) and prevents Nef-mediated Src family kinase Hck activation (IC₅₀ = 2.8 µM) without directly affecting the catalytic activity of c-Src, Hck, Lck, or Lyn. Shown to suppress HIV and SIV viral replication (IC₅₀/strain/culture = 1 µM/SIV δB670/CEM-174 and ﹤0.3 µM/HIV-1 NL4-3/CEM-T4) and infectivity, being ineffective against the replication of Nef-defective HIV-1 mutant. Molecular docking and in vitro binding studies reveal a high-affinity B9-targeting site formed at the Nef dimerization interface and a low-affinity binding site on each Nef monomer at the dimer interface periphery.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Emert-Sedlak, L.A., et al. 2013. Chem. Biol.20, 82.

Packaging

Packaged under inert gas

Reconstitution

Following reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 6 months at -20°C.

Warning

Toxicity: Standard Handling (A)