Cat. No.: HY-15217

CHR-6494

Name: CHR-6494
Brand: MedChemExpress
SKU: HY-15217
Price: 66.00 USD
Availability: InStock
Rating: 5 (8 reviews)

8 Cited Publications

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Based on 8 publication(s) in Google Scholar

CHR-6494 is a potent inhibitor of haspin, with an IC₅₀ of 2 nM. CHR-6494 inhibits histone H3T3 phosphorylation. CHR-6494 can be used in the research of cancer.

For research use only. We do not sell to patients.

Purity : 99.28%
CAS No.: 1333377-65-3
Formula: C₁₆H₁₆N₆
Molecular Weight:292.34

Storage:

Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 1 year , -20°C, 6 months

CHR-6494 Chemical Structure

CAS No.: 1333377-65-3

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Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- IC₅₀ & Target
- Cellular Effect
- In Vitro
- In Vivo
Chemical Information
Publications (8)
Solvent & Solubility
- In Vitro
- In Vivo
Protocols (4)
Purity & Documentation
References
Help & FAQs
Classifications
Bioz

Biological Activity

Description

CHR-6494 is a potent inhibitor of haspin, with an IC₅₀ of 2 nM. CHR-6494 inhibits histone H3T3 phosphorylation. CHR-6494 can be used in the research of cancer^[1].

IC₅₀ & Target

^[1]

haspin

2 nM (IC₅₀)

Cellular Effect

Cell Line	Type	Value	Description	References
786-0	IC₅₀	1037 nM Compound: CHR-6494	Antiproliferative activity against human 786-0 cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay Antiproliferative activity against human 786-0 cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay	[PMID: 38354523]
A-375	EC₅₀	0.57 μM Compound: CHR-6494	Cytotoxicity against human A-375 cells incubated for 48 to 72 hrs by MTT assay Cytotoxicity against human A-375 cells incubated for 48 to 72 hrs by MTT assay	[PMID: 39038808]
ACHN	IC₅₀	937.2 nM Compound: CHR-6494	Antiproliferative activity against human ACHN cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay Antiproliferative activity against human ACHN cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay	[PMID: 38354523]
HBL-100	EC₅₀	25.7 μM Compound: CHR-6494	Antiproliferative activity against human HBL-100 cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay Antiproliferative activity against human HBL-100 cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay	[PMID: 38320389]
HCT-116	EC₅₀	2.2 μM Compound: CHR-6494	Antiproliferative activity against human HCT-116 cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay Antiproliferative activity against human HCT-116 cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay	[PMID: 38320389]
HeLa	EC₅₀	0.41 μM Compound: CHR-6494	Cytotoxicity against human HeLa cells incubated for 48 to 72 hrs by MTT assay Cytotoxicity against human HeLa cells incubated for 48 to 72 hrs by MTT assay	[PMID: 39038808]
HK-2	IC₅₀	1331 nM Compound: CHR-6494	Cytotoxicity against human HK-2 cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay Cytotoxicity against human HK-2 cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay	[PMID: 38354523]
IEC-6	EC₅₀	0.31 μM Compound: CHR-6494	Cytotoxicity against rat IEC-6 cells incubated for 48 to 72 hrs by MTT assay Cytotoxicity against rat IEC-6 cells incubated for 48 to 72 hrs by MTT assay	[PMID: 39038808]
MCF7	EC₅₀	0.86 μM Compound: CHR-6494	Cytotoxicity against human MCF7 cells incubated for 48 to 72 hrs by MTT assay Cytotoxicity against human MCF7 cells incubated for 48 to 72 hrs by MTT assay	[PMID: 39038808]
MIA PaCa-2	EC₅₀	0.48 μM Compound: CHR-6494	Cytotoxicity against human MIA PaCa-2 cells incubated for 48 to 72 hrs by MTT assay Cytotoxicity against human MIA PaCa-2 cells incubated for 48 to 72 hrs by MTT assay	[PMID: 39038808]
OS-RC-2	IC₅₀	1102 nM Compound: CHR-6494	Antiproliferative activity against human OS-RC-2 cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay Antiproliferative activity against human OS-RC-2 cells assessed as inhibition of cell growth incubated for 48 hrs by CCK-8 assay	[PMID: 38354523]
Raji	EC₅₀	0.82 μM Compound: CHR-6494	Cytotoxicity against human Raji cells incubated for 48 to 72 hrs by MTT assay Cytotoxicity against human Raji cells incubated for 48 to 72 hrs by MTT assay	[PMID: 39038808]
SH-SY5Y	EC₅₀	>50 μM Compound: CHR-6494	Antiproliferative activity against human SH-SY5Y cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay Antiproliferative activity against human SH-SY5Y cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay	[PMID: 38320389]
TERT-RPE1	EC₅₀	1.4 μM Compound: CHR-6494	Cytotoxicity against human TERT-RPE1 cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay Cytotoxicity against human TERT-RPE1 cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay	[PMID: 38320389]
U2OS	EC₅₀	0.9 μM Compound: CHR-6494	Antiproliferative activity against human U2OS cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay Antiproliferative activity against human U2OS cells assessed as reduction in cell viability incubated for 48 hrs by MTS assay	[PMID: 38320389]

In Vitro

CHR-6494 (0-10^-5 nM; 72 hours) dose-dependently inhibits the growth of cancer cells, such as HCT-116, HeLa, MDA-MB-231, and Wi-38 cells, with IC₅₀s of 500 nM, 473 nM, 752 nM and 1059 nM, respectively^[1].
CHR-6494 (500 nM) produces a mitotic catastrophe with abnormal morphology of the mitotic spindle and centrosome amplification, and upregulates the spindle assembly checkpoint protein BUB1 and the marker of mitotic arrest cyclin B1^[1].
CHR-6494 exhibits inhibitory activities against melanoma cell lines, including BRAFV600E mutants, NRAS mutants, and wild type cells, with IC₅₀s ranging from 396 nM to 1229 nM^[2].
CHR-6494 (300 nM and 600 nM; 72 hours) induces apoptosis, increases caspase 3/7 activity by 3- and 6-fold, respectively in COLO-792 cells, and to 8.5- and 16-fold in RPMI-7951 cells^[2].
CHR-6494 in combination with MEK inhibitors synergistically inhibits viability of melanoma cells, enhances apoptosis in melanoma cells, modulates cell cycle progression independently by arresting melanoma cells at different phases, and suppresses migration of melanoma cells^[2].
CHR-6494 (50, 200 nM; 1 week) enhances the antiproliferative effects of MLN8237 in MDA-MB-231, SKBR3 breast cancer cells^[3].
CHR-6494 (200 nM; 72 hours) enhances the apoptosis of MDA-MB-231 and SKBR3 cells when combined with MLN8237^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.

In Vivo

CHR-6494 (50 mg/kg; i.p. in two cycles of five consecutive days for 15 days) inhibits the growth of tumor and cuases no obvious body weight change in nude mice bearing HCT-116 human colorectal cancer cells^[1].
CHR-6494 (20 mg/kg; intraperitoneal injection for 15 consecutive days) inhibits the tumor volume and weight compared with the control group in nude mice bearing MDA-MB-231 xenograft tumors^[3].
CHR-6494 (20 mg/kg; intraperitoneal injection for 15 consecutive days) enhances the tumor volume and weight inhibition of MLN8237 (20 mg/kg; p.o.) in vivo^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Chemical Information

CAS No. 1333377-65-3
Appearance Solid
Molecular Weight 292.34
Formula C₁₆H₁₆N₆
Color White to yellow
SMILES

CCCNC(C=C1)=NN2C1=NC=C2C3=CC(C=NN4)=C4C=C3
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	1 year
	-20°C	6 months

Publications (8)

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Journal Impact Factor

Cancer Commun (Lond)

CRISPR/Cas9 screening identifies a kinetochore-microtubule dependent mechanism for Aurora-A inhibitor resistance in breast cancer. [Abstract]

2021 Feb;41(2):121-139. PMID: 33471959
Cell Death Discov

Haspin balances the ratio of asymmetric cell division through Wnt5a and regulates cell fate decisions in mouse embryonic stem cells. [Abstract]

2023 Aug 23;9(1):307. PMID: 37612272
Eur J Med Chem

Novel inhibitors targeting the PGK1 metabolic enzyme in glycolysis exhibit effective antitumor activity against kidney renal clear cell carcinoma in vitro and in vivo. [Abstract]

2024 Mar 5:267:116209. PMID: 38354523
Mol Syst Biol

Unraveling mitotic protein networks by 3D multiplexed epitope drug screening. [Abstract]

2018 Aug 13;14(8):e8238. PMID: 30104419
J Mol Med (Berl)

Combination of PARP inhibitor and temozolomide to suppress chordoma progression. [Abstract]

2019 Aug;97(8):1183-1193. PMID: 31201471
Transl Oncol

Combined HASPIN and mTOR inhibition is synergistic against KRAS-driven carcinomas. [Abstract]

2022 Dec:26:101540. PMID: 36115073
Mol Cell Endocrinol

The N-terminal region of HASPIN regulates phosphorylation of AURKA and meiotic progression in spermatocytes. [Abstract]

2025 Aug 28:609:112645. PMID: 40885374
Research Square Preprint

2024 Jan 31.

Solvent & Solubility

In Vitro:

DMSO : 50 mg/mL (171.03 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

3.4207 mL

17.1034 mL

34.2067 mL

5 mM

0.6841 mL

3.4207 mL

6.8413 mL

10 mM

0.3421 mL

1.7103 mL

3.4207 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

Molarity Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass Mass

Concentration Concentration

Volume Volume

Molecular Weight Molecular Weight

Dilution Calculator

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

Stock Solution Working Solution

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline

Solubility: ≥ 2.5 mg/mL (8.55 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Protocol 2

Add each solvent one by one: 10% DMSO 90% Corn Oil

Solubility: 1 mg/mL (3.42 mM); Suspended solution; Need ultrasonic

This protocol yields a suspended solution of 1 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (10.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage Dosage
mg/kg
Animal Weight (Per Animal) Animal Weight (Per Animal)
g
Dosing Volume (Per Animal) Dosing Volume (Per Animal)
μL
Number of Animals Number of Animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

%
DMSO +

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
%
+
+
%
Tween-80 + +
%
Saline +

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Working solution concentration: 0.22 mg/mL

Protocols

All Signal Pathway Related Molecular Biology Cell Biology

Kinase activity and phosphorylation assays
Kinase activity assays measure the ability of kinases to transfer phosphate groups from ATP to specific substrates, while phosphorylation assays detect the presence and levels of phosphorylated proteins. Common methods include radiolabeled ATP incorporation (e. g. ,), ADP release detection via bioluminescence (e. g. ,[3]), enzyme-linked immunosorbent assays (ELISA) for phospho-specific epitopes (e. g. ,[6]), and microtiter-based formats for high-throughput screening (e. g. ,[8]). The ADP-Glo assay quantifies kinase activity by measuring ADP produced during phosphorylation using a luciferase-based system. Radiometric assays involve autoradiography or scintillation counting after incorporation of ³²P-labeled ATP into substrate proteins. ELISA-based approaches rely on phospho-specific antibodies to detect activated kinases in cell lysates or purified samples.
Western Blot
Western blotting (WB) is a commonly used experimental method in molecular biology, biochemistry, and immunogenetics for identifying and quantifying target proteins. It combines gel electrophoresis with immunoassay, enabling researchers to analyze protein expression, post-translational modifications, and molecular weight.
Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
Protocol for Kinase activity and phosphorylation assays
Kinase activity assays measure transfer of phosphate from ATP to a protein or peptide substrate, generating phosphorylated substrate, ADP, or incorporated radiolabeled phosphate as the readout; phosphorylation assays measure site-specific phosphorylation in cells or tissues as a proxy for kinase-pathway activation, inhibition, or substrate regulation. Phosphorylation can be detected by phospho-specific Western blot, immunoprecipitation kinase assay, phospho-immunofluorescence, phospho-flow cytometry, luminescent ADP detection, radiolabeled ATP incorporation, or reporter-based pathway assays, and these readouts can be applied to cancer cells, primary neurons, mouse tumors, organoids, inflammatory macrophages, ferroptosis studies, and mitophagy studies when the kinase target is biologically relevant.

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (285 KB)
SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Huertas D, et al. Antitumor activity of a small-molecule inhibitor of the histone kinase Haspin. Oncogene. 2012 Mar 15;31(11):1408-18. [Content Brief]

[2]. Han L, et al. Anti-Melanoma Activities of Haspin Inhibitor CHR-6494 Deployed as a Single Agent or in a Synergistic Combination with MEK Inhibitor. J Cancer. 2017 Aug 25;8(15):2933-2943. [Content Brief]

[3]. Chen A, et al. CRISPR/Cas9 screening identifies a kinetochore-microtubule dependent mechanism for Aurora-A inhibitor resistance in breast cancer. Cancer Commun (Lond). 2021 Feb;41(2):121-139. [Content Brief]

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Storage method and period of stock solution: -80°C, 1 year; -20°C, 6 months. When stored at -80°C, please use it within 1 year. When stored at -20°C, please use it within 6 months.

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
DMSO	1 mM	3.4207 mL	17.1034 mL	34.2067 mL	85.5169 mL
	5 mM	0.6841 mL	3.4207 mL	6.8413 mL	17.1034 mL
	10 mM	0.3421 mL	1.7103 mL	3.4207 mL	8.5517 mL
	15 mM	0.2280 mL	1.1402 mL	2.2804 mL	5.7011 mL
	20 mM	0.1710 mL	0.8552 mL	1.7103 mL	4.2758 mL
	25 mM	0.1368 mL	0.6841 mL	1.3683 mL	3.4207 mL
	30 mM	0.1140 mL	0.5701 mL	1.1402 mL	2.8506 mL
	40 mM	0.0855 mL	0.4276 mL	0.8552 mL	2.1379 mL
	50 mM	0.0684 mL	0.3421 mL	0.6841 mL	1.7103 mL
	60 mM	0.0570 mL	0.2851 mL	0.5701 mL	1.4253 mL
	80 mM	0.0428 mL	0.2138 mL	0.4276 mL	1.0690 mL
	100 mM	0.0342 mL	0.1710 mL	0.3421 mL	0.8552 mL

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CHR-6494

Publications Citing Use of MedChemExpress (MCE) CHR-6494