<Suppliers Price>

Ispinesib

Names

[ CAS No. ]:
336113-53-2

[ Name ]:
Ispinesib

[Synonym ]:
N-(3-aminopropyl)-N-[(1R)-1-(3-benzyl-7-chloro-4-oxo-3,4-dihydroquinazolin-2-yl)-2-methylpropyl]-4-methylbenzamide
N-(3-Aminopropyl)-N-[(1R)-1-(3-benzyl-7-chloro-4-oxo-3,4-dihydro-2-quinazolinyl)-2-methylpropyl]-4-methylbenzamide
Benzamide, N-(3-aminopropyl)-N-[(1R)-1-[7-chloro-3,4-dihydro-4-oxo-3-(phenylmethyl)-2-quinazolinyl]-2-methylpropyl]-4-methyl-
Ispinesib mesylate
S1452_Selleck
Ispinesib mesilate
Ispinesib
UNII-BKT5F9C2NI
SB-715992

Biological Activity

[Description]:

Ispinesib is a specific inhibitor of KSP, with a Ki app of 1.7 nM.

[Related Catalog]:

Signaling Pathways >> Cell Cycle/DNA Damage >> Kinesin
Signaling Pathways >> Cytoskeleton >> Kinesin
Research Areas >> Cancer

[Target]

KSP:1.7 nM (Ki app)


[In Vitro]

Ispinesib is a potent, highly specific inhibitor of KSP, with a Ki app of 1.7 nM[1]. Ispinesib (150 nM) inhibits BT-474 and MDA-MB-468 cell lines, with GI50s of 45 and 19 nM, respectively[2]. Ispinesib (SB715992, 15 and 30 nM) suppresses the proliferation of PC-3 prostate cancer cell by 48.65% and 52.16%, and induces apoptosis of prostate cancer cell by 1094.88% and 1516.70%, respectively. Ispinesib up regulates genes responsible for apoptosis and cell cycle arrest, and down regulates genes responsible for cell proliferation and survival. The anti-proliferation and pro-apoptotic activities of Ispinesib can be enhanced by genistein[3].

[In Vivo]

Ispinesib (SCID, 8 mg/kg; nude, 10 mg/kg, q4d × 3) reduces tumor volume in mice bearing tumor xenografts of ER-positive (MCF7), HER2-positive (KPL4, HCC1954, and BT-474), and triple-negative (MDA-MB-468) breast cancer cells via i.p. one dose every 4 days repeated three times[2].

[Kinase Assay]

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 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 MTs, and 1 nM KSP in PEM25 buffer. Ki app (apparent inhibitor dissociation constant) estimates of Ispinesib are extracted from the concentration-response curves, with explicit correction for enzyme concentration[1].

[Cell Assay]

PC-3 prostate cancer cells are seeded in 96 well plates at a density of 4 × 103 cells/well. PC-3 cells are incubated for 24 hours to allow attachment to the surface of each well of the tissue culture plate. Then, the cells are treated with varying concentration of reagents and incubated for 1 to 3 days. First, PC-3 cells are treated with 15 and 30 nM of Ispinesib, respectively. Second, PC-3 cells are subjected to combinational treatments with 7.5 or 10 nM of Ispinesib plus 30 μM of genistein. Finally, PC-3 cells are pre-treated with 30 μM of genistein for 24 hours followed by treatment with 15 nM of Ispinesib. Control cells are treated with 0.3 mM Na2CO3 (vehicle control). After treatment, PC3 cells are incubated at 37°C with MTT (0.5 mg/mL) for 2 hours and isopropyl alcohol at room temperature for 1 hour. The spectrophotometric absorbance of each sample is then determined by using ULTRA Multifunctional Micro Plate Reader at 595 nm[3].

[Animal admin]

Mice with a tumor volume of ∼250 mm3 receive a single dose of Ispinesib (10 mg/kg). Tumors are dissected, fixed in 10% buffered formalin, and embedded in paraffin, and 5-μm tissue sections are prepared. Antigen retrieval is done by boiling in 50 mM citrate buffer (pH 5.5), and sections are then incubated in 3% hydrogen peroxide, washed in PBS-0.1% Tween, and blocked in 10% goat serum. Phospho-histone H3 (PH3) antibody is detected using Alexa Fluor 488 secondary antibody. Images are taken with a microscope at ×10 magnification and captured using MetaMorph software to quantify PH3 expression by computing the area ratio of PH3-positive cells per total cells. Ki67/cleaved caspase-3 staining is done. Nonfluorescent images are taken on an Olympus BX41 microscope at ×20 magnification[2].

[References]

[1]. Lad L, et al. Mechanism of inhibition of human KSP by ispinesib. Biochemistry. 2008 Mar 18;47(11):3576-85.

[2]. Purcell JW, et al. Activity of the kinesin spindle protein inhibitor ispinesib (SB-715992) in models of breast cancer. Clin Cancer Res. 2010 Jan 15;16(2):566-76.

[3]. Davis DA, et al. Increased therapeutic potential of an experimental anti-mitotic inhibitor SB715992 by genistein in PC-3 human prostate cancer cell line. BMC Cancer. 2006 Jan 24;6:22.


[Related Small Molecules]

ARRY-520 | GSK-923295 | Kif15-IN-1 | Monastrol | Kif15-IN-2 | Litronesib | CW-069 | K 858 | SB743921 | AZ82 | Eg5 Inhibitor V, trans-24 | EMD534085 | Paprotrain

Chemical & Physical Properties

[ Density]:
1.2±0.1 g/cm3

[ Boiling Point ]:
708.0±70.0 °C at 760 mmHg

[ Molecular Formula ]:
C30H33ClN4O2

[ Molecular Weight ]:
517.062

[ Flash Point ]:
382.0±35.7 °C

[ Exact Mass ]:
516.229187

[ PSA ]:
81.22000

[ LogP ]:
5.20

[ Vapour Pressure ]:
0.0±2.3 mmHg at 25°C

[ Index of Refraction ]:
1.619

[ Storage condition ]:
-20℃

Related Compounds