Home Cell Cycle Chk inhibitor MK-8776 (SCH 900776)

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MK-8776 (SCH 900776) Chk inhibitor

Cat.No.S2735

MK-8776 (SCH 900776) is a selective Chk1 inhibitor with IC50 of 3 nM in a cell-free assay, showing 500-fold selectivity against Chk2. It is in Phase 2.
MK-8776 (SCH 900776) Chk inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 376.25

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Quality Control

Batch: Purity: 99.98%
99.98

Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
SKOV3 Growth Inhibition Assay 0.3 µM 8 d sensitizes the cell lines to 23548269
BxPC-3 Growth Inhibition Assay 10-1000 nM 24-48h enhances the chemosensitization to 23804422
MiaPaCa-2 Growth Inhibition Assay 10-1000 nM 24-48h enhances the chemosensitization to 23804422
AsPC-1 Growth Inhibition Assay 10-1000 nM 24-48h enhances the chemosensitization to 23804422
H1299 Growth Inhibition Assay 500 nM 24 h DMSO enhances the chemosensitization to PMX 24113549
H1993 Growth Inhibition Assay 500 nM 24 h DMSO enhances the chemosensitization to PMX 24113549
H1437 Growth Inhibition Assay 500 nM 24 h DMSO enhances the chemosensitization to PMX 24113549
H23 Growth Inhibition Assay 500 nM 24 h DMSO enhances the chemosensitization to PMX 24113549
AsPC-1 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
TK10 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
A498 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
U20S Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
SNB19 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
MDA-MB-435 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
U87 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
HCC2998 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
MDA-MB-231 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
MiaPaCa-2 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
MCF10A Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
HCT116 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
IGROV-1 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
SW620 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
HCT115 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
U251 Growth Inhibition Assay 200/2000 nM 24 h decreases the IC50 of 24359526
OVCAR-8 Growth Inhibition Assay 0.3 µM 8 d sensitizes the cell lines to 23548269
MV-4-11 Apoptosis Assay 100-700 nM 48 h induces apoptosis dose dependently 23536721
U937 Apoptosis Assay 100-700 nM 48 h induces apoptosis dose dependently 23536721
MOLM-13  Apoptosis Assay 100-700 nM 48 h induces apoptosis dose dependently 23536721
A2058  Cell Viability Assay 37.5-300 nM 72 h DMSO reduces the MK-1775 EC50 by 5-fold to an average of 45 nM 23148684
H2009 Cell Viability Assay 500 nM 72 h DMSO results in G1/S-phase accumulation combined with MK-1775 23148684
Su.86.86 Cell Viability Assay 500 nM 72 h DMSO results in G1/S-phase accumulation combined with MK-1775 23148684
HRE Cell Viability Assay 500 nM 72 h DMSO results in G1/S-phase accumulation combined with MK-1775 23148684
HMEC Cell Viability Assay 500 nM 72 h DMSO results in G1/S-phase accumulation combined with MK-1775 23148684
U2OS  Function Assay 2 µM 0-24 h induces phosphorylation of Chk1 at serine 345 at both concentrations as early as 2 h after administration 22937147
U2OS  Growth Inhibition Assay 0-10 µM 24/48 h inhibits cell growth dose dependently 22937147
U937 Function Assay 100-500 nM 4 h  decreases the cytarabine-induced Chk1 autophosphorylation at Ser296 and prevents Cdc25A downregulation 22869869
U937 Function Assay 100 nM 4 h  reverses the cytarabine-induced inhibition of 3H-thymidine incorporation into DNA 22869869
U937 Function Assay 100-500 nM 4 h  induces increased phosphorylation of H2AX 22869869
HL-60 Apoptosis Assay 30/100/300 nM 24 h DMSO enhances cytarabine-induced apoptosis 22869869
ML-1 Apoptosis Assay 25/50/100 nM 24 h DMSO enhances cytarabine-induced apoptosis 22869869
HCT116 Function Assay 1 µM 24 h abrogates of cell cycle arrest  22510560
U2OS Function Assay 1 µM 24 h abrogates of cell cycle arrest  22510560
Sf9 Function assay Inhibition of recombinant CDK2/Cyclin A expressed in insect Sf9 cells assessed as inhibition of [33P]-ATP incorporation into biotinylated histone H1 after 1 hr by liquid scintillation counting, IC50 = 0.16 μM. 21094607
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 75 mg/mL (199.33 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

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Mass Concentration Volume Molecular Weight
Dilution Calculator Molecular Weight Calculator

In vivo
Batch:

In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
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Chemical Information, Storage & Stability

Molecular Weight 376.25 Formula

C15H18BrN7

 Storage (From the date of receipt)
CAS No. 891494-63-6 Download SDF Storage of Stock Solutions

Synonyms N/A Smiles CN1C=C(C=N1)C2=C3N=C(C(=C(N3N=C2)N)Br)C4CCCNC4

Mechanism of Action

Targets/IC50/Ki
Chk1
(Cell-free assay)
3 nM
CDK2
(Cell-free assay)
0.16 μM
In vitro

MK-8776 (SCH 900776) is a less potent inhibitor of Chk2 and CDK2 with IC50 of 1.5 μM and 0.16 μM, respectively. It shows no significant inhibition of cytochrome P450 human liver microsomal isoforms 1A2, 2C9, 2C19, 2D6, and 3A4. In combination with an antimetabolite, this compound induces accumulation of γ-H2AX within 2 hours, indicative of replication fork collapse and double stranded DNA breaks. Additionally, it suppresses accumulation of the Chk1 pS296 autophosphorylation in a dose-dependent manner. Exposure of proliferating WS1 cells to SCH 900776 is associated with rapid, dose-dependent accumulation of Chk1 pS345, indicating that cycling populations of normal cells induce Chk1 pS345 following exposure to it as part of a futile cycle, perhaps driven by AT-family kinases and DNA-PK.
Kinase Assay
Chk1 SPA assay
An in vitro assay utilizing recombinant His-Chk1 expressed in the baculovirus expression system as an enzyme source and biotinylated peptide based upon CDC25C as substrate. His-Chk1 is diluted to 32 nM in kinase buffer containing 50 mM Tris pH 8.0, 10 mM MgCl2, and 1 mM DTT. CDC25C (CDC25 Ser216 C-term biotinylated peptide) peptide is diluted to 1.93 μM in kinase buffer. For each kinase reaction, 20 μL of 32 nM Chk1 enzyme solution and 20 μL of 1.926 μM CDC25C are mixed and combined with 10 μL of MK-8776 (SCH 900776) diluted in 10% DMSO, making final reaction concentrations of 6.2 nM Chk1, 385 nM CDC25C and 1% DMSO after addition of start solution. The reaction is started by addition of 50 μL of start solution consisting of 2 μM ATP and 0.2 μCi of 33P-ATP, making a final reaction concentration of 1 μM ATP, with 0.2 μCi of 33P-ATP per reaction. Kinase reactions run for 2 hours at room temperature and are stopped by the addition of 100 μL of stop solution consisting of 2 M NaCl, 1% H3PO4, and 5 mg/mL Streptavidin-coated SPA beads. SPA beads are captured using a 96-well GF/B filter plate and a Filtermate universal harvester. Beads are washed twice with 2 M NaCl and twice with 2 M NaCl with 1% phosphoric acid. Signal is then assayed using a TopCount 96-well liquid scintillation counter. Dose-response curves are generated from duplicate 8 point serial dilutions of this compound. IC50 values are derived by nonlinear regression analysis.
In vivo

Dose escalation of MK-8776 (SCH 900776) (16 mg/kg and 32 mg/kg) induces incremental improvements in tumor response. Importantly, doses of this compound associate with robust biomarker activation and improved tumor response are not associated with enhanced toxicity on hematological parameters in BALB/c mice.

References
  • [4] https://pubmed.ncbi.nlm.nih.gov/34267371/

Applications

Methods Biomarkers Images PMID
Western blot Cyclin E / pY15-CDK / γH2AX p-chk1(ser345) / CDC25A
S2735-WB2
26595527

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT00779584 Completed
Hodgkin Disease|Lymphoma Non-Hodgkin|Neoplasms
Merck Sharp & Dohme LLC
 October 17 2008 Phase 1

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Frequently Asked Questions

Question 1:
I would like to know whether your product S2735 is the optically pure R enantiomer or whether it is a racemic mix.

Answer:
It is the R enantiomer of our S2735.

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