(3Z)-5-[(2,6-dichlorophenyl)methanesulfonyl]-3-({3,5-dimethyl-4-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidine-1-carbonyl]-1H-pyrrol-2-yl}methylidene)-2,3-dihydro-1H-indol-2-one

• ChemBase ID: 73171
• Molecular Formular: C32H34Cl2N4O4S
• Molecular Mass: 641.60776
• Monoisotopic Mass: 640.16778195
• SMILES: c1ccc(c(c1Cl)CS(=O)(=O)c1cc2c(cc1)NC(=O)/C/2=C\c1c(c(c([nH]1)C)C(=O)N1[C@H](CCC1)CN1CCCC1)C)Cl
• Canonical SMILES: O=C1Nc2c(/C/1=C/c1[nH]c(c(c1C)C(=O)N1CCC[C@@H]1CN1CCCC1)C)cc(cc2)S(=O)(=O)Cc1c(Cl)cccc1Cl
• InChI: InChI=1S/C32H34Cl2N4O4S/c1-19-29(35-20(2)30(19)32(40)38-14-6-7-21(38)17-37-12-3-4-13-37)16-24-23-15-22(10-11-28(23)36-31(24)39)43(41,42)18-25-26(33)8-5-9-27(25)34/h5,8-11,15-16,21,35H,3-4,6-7,12-14,17-18H2,1-2H3,(H,36,39)/b24-16-/t21-/m1/s1
• InChIKey: OYONTEXKYJZFHA-SSHUPFPWSA-N

NAMES AND DATABASE IDS

Names

• IUPAC name: (3Z)-5-[(2,6-dichlorophenyl)methanesulfonyl]-3-({3,5-dimethyl-4-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidine-1-carbonyl]-1H-pyrrol-2-yl}methylidene)-2,3-dihydro-1H-indol-2-one
• IUPAC Traditional name: (3Z)-5-[(2,6-dichlorophenyl)methanesulfonyl]-3-({3,5-dimethyl-4-[(2R)-2-(pyrrolidin-1-ylmethyl)pyrrolidine-1-carbonyl]-1H-pyrrol-2-yl}methylidene)-1H-indol-2-one
• Synonyms: PHA665752; PHA-665752; (3Z)-5-[[(2,6-dichlorophenyl)methyl]sulfonyl]-3-[[3,5-dimethyl-4-[[(2R)-2-(1-pyrrolidinylmethyl)-1-pyrrolidinyl]carbonyl]-1H-pyrrol-2-yl]methylene]-1,3-dihydro-2H-indol-2-one Hydrate; PHA 665752 Hydrate; PHA-665752 Hydrate

Database IDs

• CAS Number: 477575-56-7
• PubChem SID: 162038091
• PubChem CID: 10461815

CALCULATED PROPERTIES

• Acid pKa: 10.731995
• H Acceptors: 5
• H Donor: 2
• LogD (pH = 5.5): 1.901597
• LogD (pH = 7.4): 3.5050924
• Log P: 4.9807105
• Molar Refractivity: 174.3444 cm^3
• Polarizability: 65.87702 Å^3
• Polar Surface Area: 102.58 Å^2
• Rotatable Bonds: 7
• Lipinski's Rule of Five: false

PROPERTIES

Safety Information

• Storage Condition: -20°C

Pharmacology Properties

• Target: c-Met

Product Information

• Salt Data: Free Base

DETAILS

Selleck Chemicals - S1070

Biological Activity

• Description: PHA-665752 is a potent, selective and ATP-competitive c-Met inhibitor with IC50 of 9 nM.
• Targets: c-Met
• IC50: 9 nM ^[1]
• In Vitro: PHA-665752 significantly inhibits c-Met kinase activity with K_i of 4 nM, and exhibits >50-fold selectivity for c-Met compared with various tyrosine and serine-threonine kinases. PHA-665752 potently inhibits the HGF-stimulated c-Met autophosphorylation with IC50 of 25-50 nM. PHA-665752 also significantly blocks HGF- and c-Met-dependent functions such as cell motility and cell proliferation with IC50 of 40-50 nM and 18-42 nM, respectively. In addition, PHA-665752 potently inhibits HGF-stimulated or constitutive phosphorylation of mediators of downstream of c-Met such as Gab-1, ERK, Akt, STAT3, PLC-γ, and FAK in multiple tumor cell lines. ^[1] PHA-665752 inhibits cell growth in TPR-MET-transformed BaF3 cells with IC50 of <60 nm,="" and="" inhibits="" constitutive="" cell="" motility="" and="" migration="" by="" 92.5%="" at="" 0.2="" μm.="" inhibition="" of="" c-met="" by="" pha665752="" (0.2="" μm)="" also="" induces="" cell="" apoptosis="" of="" 33.1%="" and="" g1="" cell="" cycle="" arrest="" with="" cells="" in="" g1="" phase="" increasing="" from="" 42.4%="" to="" 77.0%.="" pha665752="" can="" cooperate="" with="" rapamycin="" to="" inhibit="" cell="" growth="" of="" tpr-met-transformed="" baf3="" cells="" and="" non-small="" cell="" lung="" cancer="" h441="" cells.="">^[2]
• In Vivo: Administration of PHA-665752 induces a dose-dependent tumor growth inhibition of S114 xenografts by 20 %, 39% and 68%, at dose of 7.5, 15, and 30 mg/kg/day, respectively. ^[1] PHA665752 treatment significantly reduces the tumor growth of NCI-H69, NCI-H441 and A549 in mouse xenografts by 99%, 75%, and 59%, respectively. PHA665752 also significantly inhibits angiogenesis by >85%, due to decreasing the production of vascular endothelial growth factor and increasing the production of the angiogenesis inhibitor thrombospondin-1. ^[3]

Protocol

• Protocol: Kinase Assay ^[1]
• In vitro enzyme assay: The c-Met kinase domain GST-fusion protein is used for the c-Met assay. The IC50 value of PHA-665752 for the inhibition of c-Met is based on phosphorylation of kinase peptide substrates or poly-glu-tyr in the presence of ATP and divalent cation (MgCl₂ or MnCl₂ 10-20 mM). The linear range (i.e., the time period over which the rate remains equivalent to the initial rate) is determined for c-Met, and the kinetic measurement and IC50 determination are performed within this range.
• Protocol: Cell Assay ^[1]
• Cell Lines: S114, GTL-16, NCI-H441, and BxPC-3
• Concentrations: Dissolved in DMSO, final concentrations ~10 μM
• Incubation Time: 18, or 72 hours
• Methods: For proliferation assays, cells are grown in medium with 0.1% FBS for 48 hours after which they are treated with various concentrations of PHA-665752 in HGF (50 ng/mL) in a medium containing 2% FBS. After 18 hours, cells are incubated with BrdUrd for 1 hour, fixed, and stained with anti-BrdUrd peroxidase-conjugated antibody, and plates are read at 630 nm. For apoptosis assays, cells are grown in medium with 2% FBS in presence and absence of HGF (50 ng/mL) and various concentrations of PHA-665752 for 72 hours. After 72 hours, a mixture containing ethidium bromide and acridine orange is added, and apoptotic cells (bright orange cells or cell fragments) are counted by fluorescence microscopy.
• Protocol: Animal Study ^[1]
• Animal Models: Female athymic mice (nu/nu) bearing S114 or GTL-16 tumor xenografts
• Formulation: Formulated in vehicle (L-lactate (pH 4.8) and 10% polyethylene glycol)
• Doses: ~30 mg/kg/day
• Administration: Injection via bolus i.v.

References

• References: [1] Christensen JG, et al. Cancer Res, 2003, 63(21), 7345-7355.
• References: [2] Ma PC, et al. Clin Cancer Res, 2005, 11(6), 2312-2319.
• References: [3] Puri N, et al. Cancer Res, 2007, 67(8), 3529-3534.

Toronto Research Chemicals - P294425

PHA-665752 ia a c-Met kinase inhibitor. PHA-665752 is ATP-competitive, an active-site inhibitor with greater than 50-fold selectivity for c-Met vs a panel of tyrosine and serine-threonine kinases.

REFERENCES

• Christensen JG, et al. Cancer Res, 2003, 63(21), 7345-7355.
• Ma PC, et al. Clin Cancer Res, 2005, 11(6), 2312-2319.
• Puri N, et al. Cancer Res, 2007, 67(8), 3529-3534.
• Cheng, H., et al.: J. Clin. Oncol., 20, 1544 (2002)
• Ma, P., et al.: Cancer Metastasis Rev., 22, 309 (2002)
• Christensen, J., et al.: Cancer Lett., 225, 1 (2002)
• Lo Muzio, L., et al.: Tumor Biol., 27, 116 (2002)