Adavosertib (MK-1775)

For research use only.

Catalog No.S1525 Synonyms: AZD1775

123 publications

Adavosertib (MK-1775) Chemical Structure

CAS No. 955365-80-7

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.

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Selleck's Adavosertib (MK-1775) has been cited by 123 publications

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Choose Selective Wee1 Inhibitors

Biological Activity

Description 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.
Features The first reported Wee1 inhibitor.
Targets
Wee1 [1]
(Cell-free assay)
5.2 nM
In vitro

MK-1775 inhibits Wee1 kinase in an ATP-competitive manner. Compared to Wee1, MK-1775 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, MK-1775 treatment inhibits the basal phosphorylation of CDC2 at Tyr15 (CDC2Y15) with EC50 of 49 nM, and suppresses gemcitabine-, carboplatin- or cisplatin-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. MK-1775 treatment alone at 30-100 nM has no significant antiproliferative effect in WiDr and H1299 cells, whereas MK-1775 at 300 nM, sufficient to inhibit Wee1 by >80%, displays moderate but significant antiproliferative effects by 34.1% in WiDr cells and 28.4% in H1299 cells. [1]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
ASPC-1 MlnaS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NFrCW5NKSzVyPUGzMlIhyrFiMT6xJO69VQ>? M2XrWlI2PDV6OUW0
BxPC-3 Mn\pS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NFP0cXZKSzVyPUCuPEDDuSByLkCzJO69VQ>? MkXZNlU1PTh7NUS=
CFPAC-1 NHHiUXRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MoHqTWM2OD1|LkOgxtEhOC5{IN88US=> MoXCNlU1PTh7NUS=
HPAC NHHLZnhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NF7BbXBKSzVyPUCuOUDDuSByLkCxJO69VQ>? NWC4NmZxOjV2NUi5OVQ>
MIAPaCa-2 NHzDS4VIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NHvObXJKSzVyPUCuOUDDuSByLkC1JO69VQ>? NUHEVGJFOjV2NUi5OVQ>
PANC-1 MmPIS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M2PJVmlEPTB;MUCuOkDDuSBzLkGg{txO M4jEUlI2PDV6OUW0
SK-N-BE (2) NGiyXoZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= Mlf0TWM2OD1{LkVihKnDueLCiUCuN{DPxE1? NHHIcJAzPTNyOEmxOi=>
SK-N-BE (2), PAN→MK NUC5WZhqT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M4XaV2lEPTB;Mk[uOwKBkcLz4pEJPU43KM7:TR?= NWfRNJdROjV|MEi5NVY>
SK-N-BE (2), MK→PAN MnfES5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NYX3N|FzUUN3ME2yMlTjiIoEsfMAjVAvOyEQvF2= MXWyOVMxQDlzNh?=
SK-N-AS M4X5d2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NV3KVIV6UUN3ME2wMlUx6oDLwsJihKkxNjB{IN88US=> MnzwNlU{ODh7MU[=
SK-N-DZ NIiyUGJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MWLJR|UxRTBwM{dihKnDueLCiUCuNFEh|ryP Mmf1NlU{ODh7MU[=
SK-N-AS Mn34RZBweHSxc3nzJGF{e2G7 NVTTN5VFPTByIH7N NWi0[FJGPDhiaB?= NFG5[XlqdmS3Y3XzJINmdGxiYYDvdJRwe2m| NGTibokzPTNyOEmxOi=>
SK-N-DZ MVnBdI9xfG:|aYOgRZN{[Xl? NVHLNppVPTByIH7N NH7Z[lc1QCCq M{DDNYlv\HWlZYOgZ4VtdCCjcH;weI9{cXN? MWWyOVMxQDlzNh?=
THP-1 M4LiR2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NWO3V41IOTJ3L{K1NE82ODBibl2= M4r6SVQ5KGh? MlnDbY5kemWjc3XzJINmdGxiZHXheIghcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> NUi1PZpLOjVyOES2NVQ>
MV4-11 M3G0V2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MkmzNVI2NzJ3MD:1NFAhdk1? NG\icIM1QCCq Mof2bY5kemWjc3XzJINmdGxiZHXheIghcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> M4LaZlI2ODh2NkG0
U937 NYTVTmFmT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NI\p[VUyOjVxMkWwM|UxOCCwTR?= NIf3SXc1QCCq M1;Yb4lv[3KnYYPld{Bk\WyuIHTlZZRpKGmwIHGgZ49v[2WwdILheIlwdi2mZYDlcoRmdnRibXHucoVz NHn2[VIzPTB6NE[xOC=>
HL-60 NIX4TVhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NITybmYyOjVxMkWwM|UxOCCwTR?= NH7k[ZQ1QCCq MkmwbY5kemWjc3XzJINmdGxiZHXheIghcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> NIrLSGkzPTB6NE[xOC=>
OCI-AML3 M{Hv[2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NVXNRlR4OTJ3L{K1NE82ODBibl2= MofFOFghcA>? NXH1PWd3cW6lcnXhd4V{KGOnbHyg[IVifGhiaX6gZUBkd26lZX70doF1cW:wLXTldIVv\GWwdDDtZY5v\XJ? M1jjdFI2ODh2NkG0
MOLM-13 M2XHRmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NVv0[JNJOTJ3L{K1NE82ODBibl2= MXe0PEBp NXPGO291cW6lcnXhd4V{KGOnbHyg[IVifGhiaX6gZUBkd26lZX70doF1cW:wLXTldIVv\GWwdDDtZY5v\XJ? MlXjNlUxQDR4MUS=
CMK MXLD[YxtKF[rYXLpcIl1gSCDc4PhfS=> MYKxNE0yODByMDDuUS=> M2DJRVczKGh? Mln4doVlfWOnczDj[YxtKH[rYXzpZol1gSCrbjDhJINwdmOnboTyZZRqd25vZHXw[Y5l\W62IH3hco5meg>? NFnCSY0zPDl4MkOzNS=>
CMY MYfD[YxtKF[rYXLpcIl1gSCDc4PhfS=> MYexNE0yODByMDDuUS=> NWLy[Y5IPzJiaB?= MWTy[YR2[2W|IHPlcIwhfmmjbHnibZR6KGmwIHGgZ49v[2WwdILheIlwdi2mZYDlcoRmdnRibXHucoVz M3nXRlI1QTZ{M{Ox
Dayo NIrrbJRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= Ml;qTWM2OD1zNUCgcm0> M4PQW|I1PjZzOUGw
UW228 NH;NRVJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MoDITWM2OD1{M{Kgcm0> M{TOWVI1PjZzOUGw
IST-MES1 MnjsR4VtdCCYaXHibYxqfHliQYPzZZk> NHnqPYcyPTBxMkWwJI5O MoXRO|IhcA>? NEHtPXNmdmijbnPld{B1cGViY3nzdIxifGmwIHP5eI91d3irYzDl[oZm[3RiaX6gZUBkd26lZX70doF1cW:wLXTldIVv\GWwdDDtZY5v\XJ? MXGyOFM3PTd6Mh?=
IST-MES2 M2fkTWNmdGxiVnnhZoltcXS7IFHzd4F6 Mm\uNVUxNzJ3MDDuUS=> M3TMZlczKGh? NV[wVFUz\W6qYX7j[ZMhfGinIHPpd5Bt[XSrbjDjfZRwfG:6aXOg[YZn\WO2IHnuJIEh[2:wY3XueJJifGmxbj3k[ZBmdmSnboSgcYFvdmW{ NFrJ[mwzPDN4NUe4Ni=>
REN NF64d25E\WyuIG\pZYJqdGm2eTDBd5NigQ>? MlLpNVUxNzJ3MDDuUS=> NXj6RZpOPzJiaB?= NGSwOlZmdmijbnPld{B1cGViY3nzdIxifGmwIHP5eI91d3irYzDl[oZm[3RiaX6gZUBkd26lZX70doF1cW:wLXTldIVv\GWwdDDtZY5v\XJ? MoTNNlQ{PjV5OEK=
NCI-H2452 NETIUnBE\WyuIG\pZYJqdGm2eTDBd5NigQ>? NHm5NoEyPTBxMkWwJI5O NUX3WlNGPzJiaB?= M2nkfYVvcGGwY3XzJJRp\SClaYPwcIF1cW5iY4n0c5RwgGmlIHXm[oVkfCCrbjDhJINwdmOnboTyZZRqd25vZHXw[Y5l\W62IH3hco5meg>? Mn7QNlQ{PjV5OEK=
MSTO-211H NH\IVoNE\WyuIG\pZYJqdGm2eTDBd5NigQ>? MnfpNVUxNzJ3MDDuUS=> M33MSFczKGh? MkK2[Y5p[W6lZYOgeIhmKGOrc4DsZZRqdiCleYTveI95cWNiZX\m[YN1KGmwIHGgZ49v[2WwdILheIlwdi2mZYDlcoRmdnRibXHucoVz M{HVNFI1OzZ3N{iy
NCI-H2052 M3\KXmNmdGxiVnnhZoltcXS7IFHzd4F6 MWSxOVAwOjVyIH7N Mkn6O|IhcA>? MVrlcohidmOnczD0bIUh[2m|cHzheIlvKGO7dH;0c5hq[yCnZn\lZ5QhcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> NYTyfo9tOjR|NkW3PFI>
WEE1 NYO5PWxzT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MmL1TWM2OD13LkKgcm0> Mo\uNlM3QTl4NUW=
CDC2 NF7CT21Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MVLJR|Ux97zgMUCwNEBvVQ>? MX2yN|Y6QTZ3NR?=
CDK7 MlvLS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MlXTTWM2OO,:nkGwNFAhdk1? MkfMNlM3QTl4NUW=
MYT1 M4PtT2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NUTJUJBRUUN3ME21N|Ahdk1? M{nGRVI{Pjl7NkW1
T98G  NXnCPHlQSXCxcITvd4l{KEG|c3H5 NGrGW|IyODBxMkWwJI5O NGf3S2M3KGh? MUTlcohidmOnczDyZYRq[XSrb36tbY5lfWOnZDDj[YxtKGurbHzpcoc> NEO2SlMzOTl7Mke5Ny=>
A549 NHXZbIlCeG:ydH;zbZMhSXO|YYm= MVyyNFAhdk1? NXzxVYlbOSCq M{jl[ZJi\Gmxc3Xud4l1cXqnczDOV2NNSyClZXzsd{BqdiCjIIC1N{1l\XCnbnTlcpQhdWGwbnXy MlPNNlE4QTlyM{O=
H460 MXHBdI9xfG:|aYOgRZN{[Xl? MUSyNFAhdk1? MVOxJIg> MmDtdoFlcW:|ZX7zbZRqgmW|IF7TR2xEKGOnbHzzJIlvKGFicEWzMYRmeGWwZHXueEBu[W6wZYK= NF33c24zOTd7OUCzNy=>
H1299 NGrrUIVCeG:ydH;zbZMhSXO|YYm= NFP5b5YzODBibl2= NIL3O24yKGh? M3PaZZJi\Gmxc3Xud4l1cXqnczDOV2NNSyClZXzsd{BqdiCjIIC1N{1l\XCnbnTlcpQhdWGwbnXy MonDNlE4QTlyM{O=
Calu-6  NHrJNFVCeG:ydH;zbZMhSXO|YYm= MWmyNFAhdk1? NXLXbFhWOSCq NITSbWxz[WSrb4PlcpNqfGm8ZYOgUnNEVENiY3XscJMhcW5iYTDwOVMu\GWyZX7k[Y51KG2jbn7ldi=> MV[yNVc6QTB|Mx?=
WiDr NH3aRnhMcW6jc3WgRZN{[Xm| Mmi1NVAuOTByMECgcm0> M1[5dVghcA>? NUX5TGlLcW6qaXLpeJMheGixc4Doc5J6dGG2aX;uJI9nKEOGQ{KgZZQhXHm{MUWge4l1cCCjbjDFR|UxyqC4YXz1[UBw\iB6NTDucY9tN0xicILleJJm[XSnZDD3bZRpKGenbXPpeIFjcW6n NFjoeJUyQTh6N{W0OS=>

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
p-Cdk1(Y15) / Cdk1; 

PubMed: 25609063     


The impact of a 24 hour pre-treatment of MK-1775 on p-Cdk1 was assessed by western blotting. Because of low basal phosphorylation, GBM22 was irradiated (RT) with 10 Gy.

p-KAP1(S824) / p-Chk2(T68) / p-Chk1(S345); 

PubMed: 25609063     


Western blot evaluation of GBM6 and GBM22 short-term explant cultures 24 hours after treatment with either MK-1775 or TMZ or the combination. 

PARP / CF-PARP / pH3(S10) / p-CDC25c(S216) / p-CDK2(Y15); 

PubMed: 25458954     


Pancreatic cancer cells were treated with vehicle control or 500 nM MK-1775 for 48 h. Whole cell lysates were subjected to Western blotting and probed with anti-PARP, -p-H3, -γH2AX, -p-CHK1, -CHK1, -p-CDC25C, -p-CDK1, - CDK1, -p-CDK2, -CDK2, or -β-actin antibody.

WEE1; 

PubMed: 27616351     


MIA PaCa2, PANC-1, Hs 766T, Capan-1 and PL11 cells were treated with MK-1775 (400 nM/L) and/or MMC (150 nM/L) for 24 hours. Whole cell lysates were prepared using RIPA and western blot was performed to assess the protein expression of WEE1 (90 kDa), pCDK1(y15) (34 kDa), CDK1 (34 kDa) and GAPDH (36 kDa) as loading control. 

25609063 25458954 27616351
Immunofluorescence
tubulin / p-HH3(S10); 

PubMed: 30755439     


FaDu and UNC7 cells were treated with adavosertib (500 nM), alisertib (250 nM), or adavosertib + alisertib for 24 hours and followed by immunofluorescent staining with anti-tubulin (Green) and anti-pHH3 (S10; Red). Nucleus was stained with DAPI. (A) Representative images of mitotic cells were captured by confocal microscopy. Scale bar: 10 µm.

γH2AX; 

PubMed: 25609063     


A) γH2AX foci formation in GBM6 and GBM22 were assessed 24 h after a single treatment of 300 nM MK-1775. 

Cleaved caspase-3 / pH3; 

PubMed: 27616351     


MIA PaCa2 cells were treated and dual stained with pH3 to observe mitotic entry; and cleaved caspase 3 (CL-CSP3) to observe caspase 3 activity.

30755439 25609063 27616351
Growth inhibition assay
Cell viability; 

PubMed: 25458954     


Pancreatic cancer cell lines were cultured in 96-well plates at 37℃ for 48 h in complete medium with variable concentrations of MK-1775 and viable cell numbers were determined using MTT reagent and a microplate reader. The data are presented as means ± standard errors from at least 3 independent experiments.

IC50; 

PubMed: 25084614     


AML cell lines were cultured for 72 h in complete medium with variable concentrations of MK-1775 (MK) and viable cell numbers were determined using MTT assays. IC50 values were calculated as drug concentration necessary to inhibit 50% growth compared to untreated control cells.

25458954 25084614
In vivo 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. Antitumor efficacy by MK-1775 alone in the nude rat HeLa-luc and TOV21G-shp53 xenograft models is also moderate. [1]

Protocol

Kinase Assay:

[1]

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In vitro kinase assays:

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 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.
Cell Research:

[1]

- Collapse
  • Cell lines: WiDr, NCI-H1299, TOV21G, and HeLa
  • Concentrations: Dissolved in DMSO, final concentrations ~10 μM
  • Incubation Time: 24 hours
  • Method:

    Cells are treated with or without gemcitabine for 24 hours, then with MK-1775 for an additional 24 hours. Cell viability is determined with a WST-8 kit using SpectraMax. Cellular caspase-3/7 activities are determined with a Caspase-3/7 Glo kit.


    (Only for Reference)
Animal Research:

[1]

- Collapse
  • Animal Models: Immunodeficient nude rats (F344/NJcl-rnu) bearing WiDr, HeLa-luc, or TOV21G-shp53 tumors
  • Dosages: ~20 mg/kg/day
  • Administration: Orally
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 80 mg/mL (159.8 mM)
Water 0.0001 mg/mL (0.0 mM)
Ethanol ''10 mg/mL
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+5% Tween 80+ddH2O
For best results, use promptly after mixing.
5mg/mL

* 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.

Chemical Information

Molecular Weight 500.6
Formula

C27H32N8O2

CAS No. 955365-80-7
Storage powder
in 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

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Clinical Trial Information

NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT03668340 Recruiting Drug: AZD1775 Uterine Cancer Dana-Farber Cancer Institute|AstraZeneca October 22 2018 Phase 2
NCT03028766 Active not recruiting Drug: AZD1775|Drug: Cisplatin|Radiation: Radiotherapy 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

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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Frequently Asked Questions

  • Question 1:

    How to prepare MK1775 methylcellulose solution? and how to prepare methylcellulose itself? Once make the MK1775 methylcellulose solution, how should i keep it?

  • Answer:

    MK1775 in 0.5% methylcellulose is a suspension or emulsion, 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 MK1775 methylcellulose solution can be stored at 4°C for a week.

Wee1 Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID