NEF HCK ACTIVATION INHIBITOR 1PC X 10MG (C15-1304-101)

General description

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.

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.

Biochem/physiol Actions

Cell permeable: yes

Primary Target
Nef

Reversible: yes

Packaging

Packaged under inert gas

Warning

Toxicity: Standard Handling (A)

Reconstitution

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

Other Notes

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

Legal Information

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

Molecular Weight: 402.81. Empirical Formula: C₁₆H₁₁ClN₆O₃S. Assay: ≥. 98% (HPLC). Quality Level: 100. form: powder. manufacturer/tradename: Calbiochem^®. . storage condition: OK to freeze, protect from light. color: orange. solubility: DMSO: 2.5 . mg/mL. storage temp.: 2-8°C. SMILES string: C1: CC(: CC(: C1)Cl)N: NC2: C(NN(C2: O)C(: S)N)C3: CC: C(C: C3)[N+](: O)[O-]. Storage Class Code: 11 - Combustible Solids. WGK: WGK 2. Flash Point(F): Not applicable. Flash Point(C): Not applicable.