Sigma-Aldrich Nef Hck Activation Inhibit

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-]

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 andin vitrobinding 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 vitrobinding studies reveal two B9 K_Dvalues 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: yesPrimary Target
NefReversible: 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.CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany