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Adavosertib (AZD1775, MK-1775) Wee1 inhibitor

Name: Adavosertib (AZD1775, MK-1775)
Brand: Selleck Chemicals
SKU: S1525
Availability: InStock
Rating: 5 (239 reviews)

Cat.No.S1525

Adavosertib (MK-1775, AZD1775) is a potent and selective Wee1 inhibitor with IC50 of 5.2 nM in a cell-free assay; hinders G2 DNA damage checkpoint. Phase 2.

Chemical Structure

Molecular Weight: 500.6

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Quality Control

Batch: Purity: 99.99%

99.99

Related Targets	CDK HSP PD-1/PD-L1 ROCK DNA/RNA Synthesis Microtubule Associated Ras KRas Aurora Kinase Casein Kinase
Other Wee1 Inhibitors	PD0166285 Zedoresertib (Debio-0123, WEE1-IN-5) Azenosertib (Zn-C3) Potrasertib

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
ASPC-1	Growth Inhibition Assay			IC50=13.2 ± 1.1 μM	25458954
BxPC-3	Growth Inhibition Assay			IC50=0.8 ± 0.03 μM	25458954
CFPAC-1	Growth Inhibition Assay			IC50=3.3 ± 0.2 μM	25458954
HPAC	Growth Inhibition Assay			IC50=0.5 ± 0.01 μM	25458954
MIAPaCa-2	Growth Inhibition Assay			IC50=0.5 ± 0.05 μM	25458954
PANC-1	Growth Inhibition Assay			IC50=10.6 ± 1.1 μM	25458954
SK-N-BE (2)	Growth Inhibition Assay			IC50=2.4 ± 0.3 μM	25308916
SK-N-BE (2), PAN→MK	Growth Inhibition Assay			IC50=26.6 ± 9.6 μM	25308916
SK-N-BE (2), MK→PAN	Growth Inhibition Assay			IC50=2.4 ± 0.3 μM	25308916
SK-N-AS	Growth Inhibition Assay			IC50=0.50 ± 0.02 μM	25308916
SK-N-DZ	Growth Inhibition Assay			IC50=0.36 ± 0.01 μM	25308916
SK-N-AS	Apoptosis Assay	500 nM	48 h	induces cell apoptosis	25308916
SK-N-DZ	Apoptosis Assay	500 nM	48 h	induces cell apoptosis	25308916
THP-1	Growth Inhibition Assay	125/250/500 nM	48 h	increases cell death in a concentration-dependent manner	25084614
MV4-11	Growth Inhibition Assay	125/250/500 nM	48 h	increases cell death in a concentration-dependent manner	25084614
U937	Growth Inhibition Assay	125/250/500 nM	48 h	increases cell death in a concentration-dependent manner	25084614
HL-60	Growth Inhibition Assay	125/250/500 nM	48 h	increases cell death in a concentration-dependent manner	25084614
OCI-AML3	Growth Inhibition Assay	125/250/500 nM	48 h	increases cell death in a concentration-dependent manner	25084614
MOLM-13	Growth Inhibition Assay	125/250/500 nM	48 h	increases cell death in a concentration-dependent manner	25084614
CMK	Cell Viability Assay	10-10000 nM	72 h	reduces cell vialibity in a concentration-dependent manner	24962331
CMY	Cell Viability Assay	10-10000 nM	72 h	reduces cell vialibity in a concentration-dependent manner	24962331
Dayo	Growth Inhibition Assay			IC50=150 nM	24661910
UW228	Growth Inhibition Assay			IC50=232 nM	24661910
IST-MES1	Cell Viability Assay	150/250 nM	72 h	enhances the cisplatin cytotoxic effect in a concentration-dependent manner	24365782
IST-MES2	Cell Viability Assay	150/250 nM	72 h	enhances the cisplatin cytotoxic effect in a concentration-dependent manner	24365782
REN	Cell Viability Assay	150/250 nM	72 h	enhances the cisplatin cytotoxic effect in a concentration-dependent manner	24365782
NCI-H2452	Cell Viability Assay	150/250 nM	72 h	enhances the cisplatin cytotoxic effect in a concentration-dependent manner	24365782
MSTO-211H	Cell Viability Assay	150/250 nM	72 h	enhances the cisplatin cytotoxic effect in a concentration-dependent manner	24365782
NCI-H2052	Cell Viability Assay	150/250 nM	72 h	enhances the cisplatin cytotoxic effect in a concentration-dependent manner	24365782
WEE1	Growth Inhibition Assay			IC50=5.2 nM	23699655
CDC2	Growth Inhibition Assay			IC50＞1000 nM	23699655
CDK7	Growth Inhibition Assay			IC50＞1000 nM	23699655
MYT1	Growth Inhibition Assay			IC50=530 nM	23699655
T98G	Apoptosis Assay	100/250 nM	6 h	enhances radiation-induced cell killing	21992793
A549	Apoptosis Assay	200 nM	1 h	radiosensitizes NSCLC cells in a p53-dependent manner	21799033
H460	Apoptosis Assay	200 nM	1 h	radiosensitizes NSCLC cells in a p53-dependent manner	21799033
H1299	Apoptosis Assay	200 nM	1 h	radiosensitizes NSCLC cells in a p53-dependent manner	21799033
Calu-6	Apoptosis Assay	200 nM	1 h	radiosensitizes NSCLC cells in a p53-dependent manner	21799033
WiDr	Kinase Assays	10-10000 nM	8 h	inhibits phosphorylation of CDC2 at Tyr15 with an EC50 value of 85 nmol/L pretreated with gemcitabine	19887545
Function assay	Expi293F			Binding affinity to recombinant human full-length N-terminal His8-tagged Wee1 (1 to 646 residues) expressed in human Expi293F cells assessed as dessociation constant by quantitative real-time PCR method, Kd = 0.0032 μM.	28792760
Function assay	Expi293F			Binding affinity to recombinant human full-length N-terminal His8-tagged Wee2 (1 to 567 residues) expressed in human Expi293F cells assessed as dessociation constant by quantitative real-time PCR method, Kd = 0.0039 μM.	28792760
Antiproliferative assay	MDA-MB-231		72 hrs	Antiproliferative activity against human MDA-MB-231 cells measured after 72 hrs by CellTiter-Blue assay, IC50 = 0.26 μM.	28792760
Antiproliferative assay	HEK293T		72 hrs	Antiproliferative activity against HEK293T cells measured after 72 hrs by CellTiter-Blue assay, IC50 = 0.29 μM.	28792760
Antiproliferative assay	MM1S		72 hrs	Antiproliferative activity against human MM1S cells measured after 72 hrs by CellTiter-Blue assay, IC50 = 0.31 μM.	28792760
Function assay	HEK293		1 hr	Inhibition of human full length PKMYT1 expressed in HEK293 cells using EFS (247 to 259 residues) as substrate after 1 hr by fluorescence polarization immunoasay, Ki = 0.47 μM.	29941193
Function assay	HEK293		1 hr	Inhibition of human full length PKMYT1 expressed in HEK293 cells using EFS (247 to 259 residues) as substrate after 1 hr by fluorescence polarization immunoasay, IC50 = 4.94 μM.	29941193
Function assay	MDA-MB-231	0.1 to 10 uM	6 hrs	Inhibition of Wee1 in human MDA-MB-231 cells assessed as decrease in CDK1 phosphorylation at Tyr 15 at 0.1 to 10 uM after 6 hrs by Western blot method	28792760
Function assay	HEK293T	0.1 to 10 uM	6 hrs	Inhibition of Wee1 in HEK293T cells assessed as decrease in CDK1 phosphorylation at Tyr15 at 0.1 to 10 uM after 6 hrs by Western blot method	28792760
Function assay	HEK293T	0.1 to 10 uM	6 hrs	Inhibition of PLK1 in HEK293T cells assessed as decrease in TCTP phosphorylation at 0.1 to 10 uM after 6 hrs by Western blot method	28792760
Function assay	MDA-MB-23	0.1 to 10 uM	6 hrs	Inhibition of PLK1 in human MDA-MB-23 cells assessed as decrease in TCTP phosphorylation at 0.1 to 10 uM after 6 hrs by Western blot method	28792760
qHTS assay	TC32			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for TC32 cells	29435139
qHTS assay	U-2 OS			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells	29435139
qHTS assay	A673			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
qHTS assay	DAOY			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells	29435139
qHTS assay	Saos-2			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells	29435139
qHTS assay	BT-37			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
qHTS assay	RD			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells	29435139
qHTS assay	SK-N-SH			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
qHTS assay	BT-12			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells	29435139
qHTS assay	NB1643			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
qHTS assay	OHS-50			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
qHTS assay	BT-12			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-12 cells	29435139
qHTS assay	DAOY			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for DAOY cells	29435139
qHTS assay	SK-N-SH			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-SH cells	29435139
qHTS assay	Rh41			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells	29435139
qHTS assay	A673			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for A673 cells)	29435139
qHTS assay	MG 63 (6-TG R)			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for MG 63 (6-TG R) cells	29435139
qHTS assay	U-2 OS			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for U-2 OS cells	29435139
qHTS assay	OHS-50			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for OHS-50 cells	29435139
qHTS assay	Rh41			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh41 cells	29435139
qHTS assay	RD			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for RD cells	29435139
qHTS assay	SJ-GBM2			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
qHTS assay	SK-N-MC			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
qHTS assay	NB-EBc1			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
qHTS assay	LAN-5			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
qHTS assay	Rh18			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells	29435139
qHTS assay	SJ-GBM2			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SJ-GBM2 cells	29435139
qHTS assay	Saos-2			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Saos-2 cells	29435139
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 100 mg/mL (199.76 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG 300 2 5%Tween80 3 50% ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	12.000mg/ml (23.97mM)	Taking the 1 mL working solution as an example, add 50 μL of 240 mg/mL clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify; add 50 μL of Tween-80 to the above system, mix evenly to clarify; Then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 95% Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	6.000mg/ml (11.99mM)	Taking the 1 mL working solution as an example, add 50 μL of 120 mg/mL clarified DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG 300 2 5%Tween80 3 50% ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	5.000mg/ml (9.99mM)	Taking the 1 mL working solution as an example, add 50 μL of 100 mg/mL clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify; add 50 μL of Tween-80 to the above system, mix evenly to clarify; Then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

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Average weight of animals: g

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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.

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Chemical Information, Storage & Stability

Molecular Weight	500.6	Formula	C₂₇H₃₂N₈O₂	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	955365-80-7	Download SDF		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	AZD1775			Smiles	CC(C)(C1=NC(=CC=C1)N2C3=NC(=NC=C3C(=O)N2CC=C)NC4=CC=C(C=C4)N5CCN(CC5)C)O

Mechanism of Action

Features	The first reported Wee1 inhibitor.
Targets/IC₅₀/Ki	Wee1 (Cell-free assay) 5.2 nM
In vitro	Adavosertib (MK-1775) inhibits Wee1 kinase in an ATP-competitive manner. Compared to Wee1, it displays 2- to 3-fold less potency against Yes with IC50 of 14 nM, 10-fold less potency against seven other kinases with >80% inhibition at 1 μM, and >100-fold selectivity over human Myt 1, another kinase that inhibits cyclin-dependent kinase 1 (CDC2) by phosphorylation at an alternative site (Thr14). By abrogating the DNA damage checkpoint via blockade of Wee1 activity in WiDr cells bearing mutated p53, this compound treatment inhibits the basal phosphorylation of CDC2 at Tyr15 (CDC2Y15) with EC50 of 49 nM, and suppresses induced phosphorylation of CDC2 and cell cycle arrest in a dose-dependent manner, with EC50 of 82 nM and 81 nM, 180 nM and 163 nM, as well as 159 nM and 160 nM, respectively. Its treatment alone at 30-100 nM has no significant antiproliferative effect in WiDr and H1299 cells, whereas at 300 nM, sufficient to inhibit Wee1 by >80%, it displays moderate but significant antiproliferative effects by 34.1% in WiDr cells and 28.4% in H1299 cells.
Kinase Assay	In vitro kinase assays
Kinase Assay	Recombinant human Wee1 is used. Kinase reaction is conducted with 10 μM ATP, 1.0 μCi of [γ-³³P]ATP, and 2.5 μg of poly(Lys, Tyr) as a substrate in the presence of increasing concentrations of Adavosertib (MK-1775) at 30°C for 30 minutes. Radioactivity incorporated into the substrate is trapped on MultiScreen-PH plates and is counted on a liquid scintillation counter.
In vivo	Adavosertib (MK-1775) treatment alone at ~20 mg/kg displays minimal antitumor effects against WiDr xenografts in rats with T/C of 69% at day 3. Its antitumor efficacy in the nude rat HeLa-luc and TOV21G-shp53 xenograft models is also moderate.
References	[1] https://pubmed.ncbi.nlm.nih.gov/19887545/

Applications

Methods	Biomarkers	PMID
Western blot	p-Cdk1(Y15) / Cdk1 p-KAP1(S824) / p-Chk2(T68) / p-Chk1(S345) PARP / CF-PARP / pH3(S10) / p-CDC25c(S216) / p-CDK2(Y15) WEE1	25609063
Immunofluorescence	tubulin / p-HH3(S10) γH2AX Cleaved caspase-3 / pH3	30755439
Growth inhibition assay	Cell viability IC50	25458954

Clinical Trial Information

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

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT03253679	Completed	Advanced Malignant Solid Neoplasm\|Refractory Malignant Solid Neoplasm	National Cancer Institute (NCI)	January 16 2019	Phase 2
NCT03668340	Active not recruiting	Uterine Cancer	Dana-Farber Cancer Institute\|AstraZeneca	October 22 2018	Phase 2
NCT03028766	Completed	Hypopharynx Squamous Cell Carcinoma\|Oral Cavity Squamous Cell Carcinoma\|Larynx Cancer	University of Birmingham\|AstraZeneca\|Cancer Research UK	June 22 2017	Phase 1

Tech Support

Handling Instructions

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Frequently Asked Questions

Question 1:
How to prepare its methylcellulose solution? and how to prepare methylcellulose itself? Once make the solution, how should i keep it?

Answer:
It is a suspension or emulsion in 0.5% methylcellulose, and it is ok to treat mice orally. It is recommended to dissolve methylcellulose in saline. It will take some time to dissolve methylcellulose, and you can vortex it for a while. The methylcellulose solution of this compound can be stored at 4°C for a week.

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C8=C7/X	C8:	LOG(C8):