Kinesin spindle protein (KSP) inhibitors. Part 1: The discovery of 3,5-diaryl-4,5-dihydropyrazoles as potent and selective inhibitors of the mitotic kinesin KSP

Bioorganic and Medicinal Chemistry Letters

Volumn 15, Issue 8, 2005, Pages 2041-2045

  Cox, Christopher D ^a   Breslin, Michael J ^a   Mariano, Brenda J ^a   Coleman, Paul J ^a   Buser, Carolyn A ^a   Walsh, Eileen S ^a   Hamilton, Kelly ^a   Huber, Hans E ^a   Kohl, Nancy E ^a   Torrent, Maricel ^a   Yan, Youwei ^a   Kuo, Laurence C ^a   Hartman, George D ^a  

^a MERCK RESEARCH LABORATORIES   (United States)

Author keywords

Anti mitotics; Dihydropyrazoles; KSP; Mitotic kinesins

Indexed keywords

KINESIN SPINDLE PROTEIN; NUCLEOTIDE; PROTEIN INHIBITOR; UNCLASSIFIED DRUG;

ALLOSTERISM; APOPTOSIS; ARTICLE; BINDING SITE; CANCER CELL CULTURE; CONCENTRATION (PARAMETERS); DRUG ACTIVITY; DRUG BINDING; DRUG POTENCY; DRUG SCREENING; DRUG SELECTIVITY; MICROTUBULE; MITOSIS; X RAY CRYSTALLOGRAPHY;

EID: 20144388363     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2005.02.055     Document Type: Article

References (22)

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15. Dihydropyrazole 13f can be easily accessed by heating chalcone in the presence of methylhydrazine in EtOH at 80°C for 2 h
16. 2: 317.1096. Found 317.1105. See Ref. 15 for assignment of absolute stereochemistry as S
17. 50's of KSPi 15 when tested against the motor domains of eight additional kinesins (CENP-E, MKLP-1, Kif3A, Kif1B, uKHC, nKHC, KIF14, and MCAK) were greater than 50 μM in ATPase assays similar to that described for KSP. The selection of these counter-screens was based on considerations of homology and function
18. 2, pH 6.8) containing 1 mM GTP. The assay was initiated by adding cold tubulin at 2.5 mg/mL to each well (to a total volume of 200 μL; 1% final DMSO)
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20. 2+) was determined by the use of the difference Fourier method and refined to an R-factor of 0.25. The coordinates have been deposited with RCSB Protein Data Bank under the accession code 1YRS
21. We have solved the crystal structure of the ternary complex of KSP, ADP and over 20 structurally-related inhibitors. In each case, only the S-antipode was found to bind in the active site. The absolute stereochemistry of 14 was assigned by analogy
22. Caspase-3 activity in cell lysates was determined using the ApoAlert Caspase-3 fluorescent assay kit (Clontech, Palo Alto, CA), which measures the release of the 7-amino-4-trifluoromethylcoumarin (AFC) fluorophore from the substrate DEVD-AFC