Ispinesib (SB-715992)

Catalog No.S1452 Batch:S145201

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Technical Data

Formula

C30H33ClN4O2

Molecular Weight 517.06 CAS No. 336113-53-2
Solubility (25°C)* In vitro DMSO 103 mg/mL (199.2 mM)
Ethanol 103 mg/mL (199.2 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description Ispinesib (SB-715992, CK0238273) is a potent, specific and reversible inhibitor of kinesin spindle protein (KSP) with Ki app of 1.7 nM in a cell-free assay, no inhibition to CENP-E, RabK6, MCAK, MKLP1, KHC or Kif1A. Ispinesib induces mitotic arrest and apoptotic cell death.
Targets
KSP (HsEg5) [1]
(Cell-free assay)
1.7 nM(Ki app)
In vitro

Ispinesib is a potent, allosteric, reversible, and specific inhibitor of KSP, which changes the binding property of KSP to microtubules and disturbs its movement by inhibiting ADP release without altering the release of the KSP-ADP complex from the microtubule. [1] Ispinesib shows potent cytotoxic activity in a panel of tumor cell lines, including Colo205, Colo201, HT-29, M5076, Madison-109, and MX-1, with IC50 of 1.2 nM to 9.5 nM. [2] In PC-3 prostate cancer cells, Ispinesib (15 nM and 30 nM) blocks cell proliferation and induces apoptosis by regulating the expression levels of genes that controls apoptosis, cell proliferation, cell cycle, and cell signaling, such as EGFR, p27, p15, and IL-11. [3] In a panel of 53 breast cell lines, Ispinesib (7.4 nM–600 nM) demonstrates broad inhibitory activity. In BT-474 and MDA-MB-468 cells, Ispinesib (150 nM) induces apoptosis, as revealed by a higher proportion of apoptotic cells, lower antiapoptotic Bcl-XL level, and higher proapoptotic Bax and Bid levels. [4]

In vivo

Ispinesib (4.5 mg/kg–15 mg/kg) exhibits inhibitory effects against Colo205, Colo201, HT-29, but not MX-1 cells, in mouse xenograft models. SB-715992 (6 mg/kg–10 mg/kg ) also inhibits murine solid tumors, including Madison 109 lung carcinoma, M5076 sarcoma, as well as L1210 and P388 leukemias. [2] In mice xenograft models of breast cancer cells MCF-7, HCC1954, MDA-MB-468, and KPL4, Ispinesib (8 mg/kg–10 mg/kg) inhibits tumor growth. [4]

Features An allosteric, potent, specific, and reversible inhibitor of the mitotic kinesin spindle protein (KSP) (HsEg5).

Protocol (from reference)

Kinase Assay:

[1]

  • Steady-State Kinetic Analysis of Human KSP ATPase Activity and Inhibition by Ispinesib

    Kinesin specificity analysis is carried out using a pyruvate kinase-lactate dehydrogenase detection system that couples the production of ADP to oxidation of NADH. Absorbance changes are monitored at 340 nm. Steady-state studies using nanomolar concentrations of KSP are performed using a sensitive fluorescence-based assay utilizing a pyruvate kinase, pyruvate oxidase, and horseradish peroxidase (HRP) coupled detection system that couples the generation of ADP to oxidation of Amplex Red to fluorescent resorufin. Generation of resorufin is monitored by fluorescence (λexcitation = 520 nm and λemission = 580 nm). Steady-state biochemical experiments are performed in PEM25 buffer [25 mM Pipes-K+ (pH 6.8), 2 mM MgCl2, 1 mM EGTA] supplemented with 10 µM paclitaxel for experiments involving microtubules. The IC50 for steady-state inhibition is determined at 500 µM ATP, 5 µM Microtubules, and 1 nM KSP in PEM25 buffer. Ki app (apparent inhibitor dissociation constant) values of Ispinesib are extracted from the dose-response curves, with explicit correction for enzyme concentration by using the Morrison equation.Inhibitor modality (e.g., competitive, noncompetitive, uncompetitive, or mixed) under steady-state conditions is determined by measuring the effect of inhibitor concentration on initial velocity as a function of substrate concentrations. Data are fit using equations in GraFit to velocity equations for the various modes of inhibition.

Cell Assay:

[4]

  • Cell lines

    Breast cancer cells, including MCF-7, HCC1954, MDA-MB-468, and KPL4

  • Concentrations

    0.085 nM–33 µM

  • Incubation Time

    72 hours

  • Method

    Cells are plated in log phase of growth in 96-well plates and treated with Ispinesib for 72 hours. Then, cell growth is measured using CellTiter-Glo, and luminescence is detected using BioTek FLx800. Data are analyzed and the IC50 value, defined as the drug concentration that results in 50% growth inhibition relative to control, is calculated.

Animal Study:

[4]

  • Animal Models

    Nude (nu/nu) mice models of MCF7, KPL4, and HCC1954 cells; severe combined immunodeficient (SCID) mice model of MDA-MB-468 cells;

  • Dosages

    10 mg/kg for nude mice or 8 mg/kg for SCID mice

  • Administration

    Intraperitoneal injection on a q4d× schedule (3 doses, every 4 day

Customer Product Validation

Data from [Data independently produced by Mol Oncol, 2014, 8(8), 1548-60]

Data from [Data independently produced by J Biol Chem, 2014, 289(45), 31111-20]

Data from [PLoS One, 2013, 8(4), e60334]

Data from [PLoS One, 2013, 8(4), e60334]

Selleck's Ispinesib (SB-715992) has been cited by 27 publications

Small-molecule inhibition of kinesin KIF18A reveals a mitotic vulnerability enriched in chromosomally unstable cancers [ Nat Cancer, 2024, 5(1):66-84] PubMed: 38151625
E3 ubiquitin ligase ASB8 promotes selinexor-induced proteasomal degradation of XPO1 [ Biomed Pharmacother, 2023, 160:114305] PubMed: 36731340
Improving Localized Radiotherapy for Glioblastoma via Small Molecule Inhibition of KIF11 [ Cancers (Basel), 2023, 15(12)3173] PubMed: 37370783
The Global Phosphorylation Landscape of SARS-CoV-2 Infection [ Cell, 2020, 182(3):685-712.e19] PubMed: 32645325
The Global Phosphorylation Landscape of SARS-CoV-2 Infection [ Cell, 2020, 182(3):685-712.e19] PubMed: 32645325
Therapeutic targeting of KSP in preclinical models of high-risk neuroblastoma [ Sci Transl Med, 2020, 12(562)eaba4434] PubMed: 32967973
enAsCas12a Enables CRISPR-Directed Evolution to Screen for Functional Drug Resistance Mutations in Sequences Inaccessible to SpCas9 [ Mol Ther, 2020, S1525-0016(20)30482-2] PubMed: 33002419
Enhancing Brain Retention of a KIF11 Inhibitor Significantly Improves its Efficacy in a Mouse Model of Glioblastoma. [ Sci Rep, 2020, 16;10(1):6524] PubMed: 32300151
KIF11 and KIF15 mitotic kinesins are potential therapeutic vulnerabilities for malignant peripheral nerve sheath tumors [ Neurooncol Adv, 2020, 2(Suppl 1):i62-i74] PubMed: 32642733
Allele-selective lowering of mutant HTT protein by HTT-LC3 linker compounds [ Nature, 2019, 575(7781):203-209] PubMed: 31666698

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SHIPPING AND STORAGE
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