Prexasertib HCl (LY2606368)

Catalog No.S7178 Batch:S7178

Technical Data

Formula	C₁₈H₁₉N₇O₂.2HCl
Molecular Weight	438.31	CAS No.	1234015-54-3
Solubility (25°C)*	In vitro	DMSO	2 mg/mL (4.56 mM)
		Water	Insoluble
		Ethanol	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

Prexasertib(LY2606368) is an ATP-competitive CHK1 inhibitor with a Ki value of 0.9 nmol/L. For CHK2 and RSK, its IC50 values are 8 nM and 9 nM respectively in cell-free assay.

Targets

Chk1 ^[1] (Cell-free assay)	Chk2 ^[1] (Cell-free assay)	RSK ^[1] (Cell-free assay)
0.9 nM(Ki)	8 nM	9 nM

In vitro

In nonclinical studies, LY2606368 induced DNA damage as measured by replication catastrophe and increases in pH2A.X, a marker of double-stranded DNA breaks^[1]. Treatment of cells with LY2606368 results in the rapid appearance of TUNEL and pH2AX-positive double-stranded DNA breaks in the S-phase cell population. In a functional assay, LY2606368 potently abrogated the G2–M checkpoint activated by doxorubicin in p53-deficient HeLa cells with an EC50 of 9 nmol/L. LY2606368 was broadly antiproliferative with IC50 values typically <50 nmol/L in the most sensitive cell lines with a minority of cell lines showing considerable resistance with IC50's >1,000 nmol/L. LY2606368 requires CDC25A and CDK2 to cause DNA damage^[2].

In vivo

LY2606368 inhibited tumor growth in cancer xenografts as monotherapy and in combination with other agents^[1]. In an orthotopic SKOV3 ovarian cancer model, LY2606368 was shown to inhibit the growth of primary tumors and significantly reduce the incidence of metastases and ascites accumulation. LY2606368 also demonstrated efficacy in an SW1990 orthotopic pancreatic cancer model resulting in a 92% inhibition of primary tumor growth and the elimination of metastases to the lymphnode, spleen, and intestine^[3].

Protocol (from reference)

Cell Assay:^{^[2]}	Cell lines HeLa cells Concentrations 33 or 100 nmol/L Incubation Time 12 h Method HeLa cells were plated onto T25 flasks and allowed to recover for 24 hours. LY2606368 was then added to give final concentrations of 33 or 100 nmol/L. In some experiments, 20μmol/L Z-VAD-FMK was included during the drug treatment. Cells were treated for 12 hours, and during the last 2 hours, colchicine was added to 1 μg/mL. Fixation of nuclei for metaphase spreads was done following the method of Bayani and Squire. Chromosome spreads were done. A 12-μL volume of cell suspension in 3:1 methanol/acetic acid fixative was dropped from a height of 3 cm onto dry glass slides or coverslips. The slides were then heated for 45 seconds on a 43°C metal block, before being removed to allow drying to complete at room temperature. Coverslips were mounted on slides with Vectashield Hard Set mounting medium with DAPI. Slides were examined with a Leica DMR fluorescence microscope and images were captured using a SPOT RT3 Slider camera.
Animal Study:^{^[2]}	Animal Models Female CD-1 nu-/nu- mice Dosages 15 mg/kg Administration s.c.

Cell Assay:^{^[2]}

Cell lines
HeLa cells
Concentrations
33 or 100 nmol/L
Incubation Time
12 h
Method
HeLa cells were plated onto T25 flasks and allowed to recover for 24 hours. LY2606368 was then added to give final concentrations of 33 or 100 nmol/L. In some experiments, 20μmol/L Z-VAD-FMK was included during the drug treatment. Cells were treated for 12 hours, and during the last 2 hours, colchicine was added to 1 μg/mL. Fixation of nuclei for metaphase spreads was done following the method of Bayani and Squire. Chromosome spreads were done. A 12-μL volume of cell suspension in 3:1 methanol/acetic acid fixative was dropped from a height of 3 cm onto dry glass slides or coverslips. The slides were then heated for 45 seconds on a 43°C metal block, before being removed to allow drying to complete at room temperature. Coverslips were mounted on slides with Vectashield Hard Set mounting medium with DAPI. Slides were examined with a Leica DMR fluorescence microscope and images were captured using a SPOT RT3 Slider camera.

Animal Study:^{^[2]}

Animal Models
Female CD-1 nu-/nu- mice
Dosages
15 mg/kg
Administration
s.c.

References

Customer Product Validation

: Data from [Data independently produced by , , Mol Cancer Ther, 2018, 17(12):2676-2688]

Selleck's Prexasertib HCl (LY2606368) has been cited by 39 publications

Comprehensive multi-omics analysis reveals WEE1 as a synergistic lethal target with hyperthermia through CDK1 super-activation [ Nat Commun, 2024, 15(1):2089]	PubMed: 38453961
Transcriptional Differential Analysis of Nitazoxanide-Mediated Anticanine Parvovirus Effect in F81 Cells [ Viruses, 2024, 16(2)282]	PubMed: 38400057
Actionable loss of SLF2 drives B-cell lymphomagenesis and impairs the DNA damage response [ EMBO Mol Med, 2023, e16431.]	PubMed: 37485814
Single-cell trajectory analysis reveals a CD9 positive state to contribute to exit from stem cell-like and embryonic diapause states and transit to drug-resistant states [ Cell Death Discov, 2023, 9(1):285]	PubMed: 37542044
Combination therapy with WEE1 inhibition and trifluridine/tipiracil against esophageal squamous cell carcinoma [ Cancer Sci, 2023, 10.1111/cas.15966]	PubMed: 37724648
Replicative stress in gastroesophageal cancer is associated with chromosomal instability and sensitivity to DNA damage response inhibitors [ iScience, 2023, 10.1016/j.isci.2023.108169]	PubMed: 37965133
Identification of therapeutic sensitivities in a spheroid drug combination screen of Neurofibromatosis Type I associated High Grade Gliomas [ PLoS One, 2023, 18(2):e0277315]	PubMed: 36730269
CHEK2 signaling is the key regulator of oocyte survival after chemotherapy [ Sci Adv, 2023, 9(42):eadg0898]	PubMed: 37862420
Replicative stress in gastroesophageal adenocarcinoma is associated with chromosomal instability and sensitivity to DNA damage response inhibitors [ bioRxiv, 2023, 2023.03.27.534412]	PubMed: 37034740
Clinically relevant CHK1 inhibitors abrogate wild-type and Y537S mutant ERα expression and proliferation in luminal primary and metastatic breast cancer cells [ J Exp Clin Cancer Res, 2022, 41(1):141]	PubMed: 35418303

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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