N-(2,6-diethylphenyl)-3-[4-(4-methylpiperazin-1-yl)benzamido]-1H,4H,5H,6H-pyrrolo[3,4-c]pyrazole-5-carboxamide

• ChemBase ID: 72687
• Molecular Formular: C28H35N7O2
• Molecular Mass: 501.6232
• Monoisotopic Mass: 501.2852234
• SMILES: c1(ccc(cc1)C(=O)Nc1c2c([nH]n1)CN(C2)C(=O)Nc1c(cccc1CC)CC)N1CCN(CC1)C
• Canonical SMILES: CCc1cccc(c1NC(=O)N1Cc2c(C1)[nH]nc2NC(=O)c1ccc(cc1)N1CCN(CC1)C)CC
• InChI: InChI=1S/C28H35N7O2/c1-4-19-7-6-8-20(5-2)25(19)29-28(37)35-17-23-24(18-35)31-32-26(23)30-27(36)21-9-11-22(12-10-21)34-15-13-33(3)14-16-34/h6-12H,4-5,13-18H2,1-3H3,(H,29,37)(H2,30,31,32,36)
• InChIKey: OBWNXGOQPLDDPS-UHFFFAOYSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: N-(2,6-diethylphenyl)-3-[4-(4-methylpiperazin-1-yl)benzamido]-1H,4H,5H,6H-pyrrolo[3,4-c]pyrazole-5-carboxamide
• IUPAC Traditional name: N-(2,6-diethylphenyl)-3-[4-(4-methylpiperazin-1-yl)benzamido]-1H,4H,6H-pyrrolo[3,4-c]pyrazole-5-carboxamide
• Synonyms: PHA-680632

Database IDs

• CAS Number: 398493-79-3
• PubChem SID: 162037608
• PubChem CID: 11249084

CALCULATED PROPERTIES

• Acid pKa: 10.493449
• H Acceptors: 5
• H Donor: 3
• LogD (pH = 5.5): 2.364922
• LogD (pH = 7.4): 4.0801616
• Log P: 4.5821342
• Molar Refractivity: 151.7333 cm^3
• Polarizability: 54.818478 Å^3
• Polar Surface Area: 96.6 Å^2
• Rotatable Bonds: 6
• Lipinski's Rule of Five: false

PROPERTIES

Physical Property

• Solubility: DMSO

Safety Information

• Storage Condition: -20°C

Pharmacology Properties

• Target: Aurora Kinase

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S1454

Research Area

• Description: Cancer

Biological Activity

• Description: PHA-680632 is potent inhibitor of Aurora A, Aurora B and Aurora C with IC50 of 27 nM, 135 nM and 120 nM, repectively.
• Targets: Aurora A; Aurora B; Aurora C
• IC50: 27 nM; 135 nM; 120 nM ^[1]
• In Vitro: PHA-680632 potently inhibits all three Aurora kinases (A, B, and C) with IC50 values of 27, 135, and 120 nM, respectively. PHA-680632 is selective for Aurora kinases, with 10- to 200-fold higher IC50 for FGFR1, FLT3, LCK, PLK1, STLK2, VEGFR2, and VEGFR3, and with IC50 higher than 10 μM for another 22 kinases. PHA-680632 shows potent anti-proliferative effects in a wide range of cell types with IC50 values of 0.06–7.15 μM, including HeLa, HCT116, HT29, LOVO, DU145, and NHDF cells. PHA-680632 (0.5 μM) causes polyploidy in tumor cells. The mechanism of action of PHA-680632 is in agreement with inhibition of Aurora kinases. ^[1]PHA680632 in association with radiation leads to additive effects in cancer cells, especially in the p53-deficient cells. Combined ionising radiation (IR) and treatment of PHA680632 (100–400 nM) prior to IR leads to an enhancement of radiation-induced Annexin V positive cells, micronuclei formation, and Brca1 foci formation only in HCT116 cells with deficient p53, other than the p53 wild-type counterparts. ^[2]
• In Vivo: HA-680632 (15–60 mg/kg) inhibits tumor growth in mice xenografts models of HL60, A2780, and HCT116 cells, by reducing tumor cell proliferation and increasing apoptosis. PHA-680632 (45 mg/kg) suppresses growth of activated ras-driven mammary tumors in mouse mammary tumor virus v-Ha-ras transgenic mice and results in complete tumor stabilization and partial regression. ^[1]

Protocol

• Protocol: Kinase Assay ^[1]
• Aurora Kinase Inhibition Assay: Inhibition of kinase activity by PHA-680632 is assessed using a scintillation proximity assay format. The biotinylated substrate is transphosphorylated by the kinase in presence of ATP traced with γ^33-ATP. The phosphorylated substrate is then captured using streptavidin-coated scintillation proximity assay beads and the extent of phosphorylation is evaluated by β-counter after a 4-hour rest for the floatation of the beads on a dense 5 M CsCl solution. In particular, a peptide derived from the Chocktide sequence (LRRWSLGL) is used as substrate for Aurora A, whereas the optimized peptide Auroratide is employed for Aurora B and C. The assay is run in a robotized format on 96-well plates. The potency of the compound toward Aurora kinases is evaluated and IC50 values are determined.
• Protocol: Cell Assay ^[1]
• Cell Lines: HeLa, HCT116, HT29, LOVO, DU145, and NHDF cells
• Concentrations: 0.001-1 μM, dissolved in DMSO as 10 mM stock solution
• Incubation Time: 72 hours
• Methods: Cells (5×10^3 to 1.5×10^4 per cm^2) are seeded in 24-well plate. After 24 hours, plates are treated with PHA-680632 and incubated for 72 hours. At the end of incubation time, cells are detached from each plate and counted using a cell counter. IC50s are calculated using percentage of growth versus untreated control.
• Protocol: Animal Study ^[2]
• Animal Models: Mice (female athymic nude) xenografts models of p53^?/? HCT116 cells
• Formulation: Dissolved in 20% Tween-80 in 5% glucose solution
• Doses: 40 mg/kg
• Administration: Intraperitoneal (i.p.) injection twice a day

References

• References: [1] Soncini C, et al. Clin Cancer Res, 2006, 12(13), 4080-4089.
• References: [2] Tao Y, et al. Br J Cancer, 2007, 97(12), 1664-1672.

REFERENCES

• Soncini C, et al. Clin Cancer Res, 2006, 12(13), 4080-4089.
• Tao Y, et al. Br J Cancer, 2007, 97(12), 1664-1672.