Adavosertib (MK-1775)

For research use only. Not for use in humans.

製品コードS1525 別名:AZD1775

Adavosertib (MK-1775)化学構造

分子量(MW):500.6

MK-1775 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.

サイズ 価格(税別) 在庫  
10mM (1mL in DMSO) JPY 41500 あり
JPY 30200 あり
JPY 46800 あり
JPY 80000 あり
JPY 129800 あり
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文献中Selleckの製品使用例(121)

製品安全説明書

Wee1阻害剤の選択性比較

生物活性

製品説明 MK-1775 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.
特性 The first reported Wee1 inhibitor.
ターゲット
Wee1 [1]
(Cell-free assay)
5.2 nM
体外試験

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]

細胞データ
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
ASPC-1 NGrYemNIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MYjJR|UxRTF|LkKgxtEhOS5zIN88US=> MVyyOVQ2QDl3NB?=
BxPC-3 NVr5T5RwT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M2fjdWlEPTB;MD64JOKyKDBwMEOg{txO NHW0ZWUzPTR3OEm1OC=>
CFPAC-1 MX;Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MmTLTWM2OD1|LkOgxtEhOC5{IN88US=> MnfhNlU1PTh7NUS=
HPAC NF70UWVIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M2\aZmlEPTB;MD61JOKyKDBwMEGg{txO M{Tad|I2PDV6OUW0
MIAPaCa-2 MWXHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MX\JR|UxRTBwNTFCtUAxNjB3IN88US=> NWSyNYhROjV2NUi5OVQ>
PANC-1 MX;Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MmLQTWM2OD1zMD62JOKyKDFwMTFOwG0> NITtTIczPTR3OEm1OC=>
SK-N-BE (2) NFTnb5NIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NVTjT21OUUN3ME2yMlTjiIoEsfMAjVAvOyEQvF2= NEf4ZWkzPTNyOEmxOi=>
SK-N-BE (2), PAN→MK M3\weWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MojSTWM2OD1{Nj625qCKyrIkgJm5MlYh|ryP M4XuWlI2OzB6OUG2
SK-N-BE (2), MK→PAN NEe0VpdIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MY\JR|UxRTJwNPMAjeKy6oDLMD6zJO69VQ>? M3rFdVI2OzB6OUG2
SK-N-AS NV6xflBvT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MnPkTWM2OD1yLkWw5qCKyrIkgJmwMlAzKM7:TR?= M1jCeFI2OzB6OUG2
SK-N-DZ NHviTFBIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NFT5OZFKSzVyPUCuN|bjiIoEsfMAjVAvODFizszN M2TDeVI2OzB6OUG2
SK-N-AS NVXveVg1SXCxcITvd4l{KEG|c3H5 MmLpOVAxKG6P MVm0PEBp NF34bJVqdmS3Y3XzJINmdGxiYYDvdJRwe2m| M4DxNFI2OzB6OUG2
SK-N-DZ MVPBdI9xfG:|aYOgRZN{[Xl? MUW1NFAhdk1? M3zH[VQ5KGh? MkTUbY5lfWOnczDj[YxtKGGyb4D0c5Nqew>? M2nWfFI2OzB6OUG2
THP-1 NXTrUGhST3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MljoNVI2NzJ3MD:1NFAhdk1? Ml\pOFghcA>? Mo\WbY5kemWjc3XzJINmdGxiZHXheIghcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> NETyVmszPTB6NE[xOC=>
MV4-11 M{LZR2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MnrHNVI2NzJ3MD:1NFAhdk1? M{DoVFQ5KGh? Mk\KbY5kemWjc3XzJINmdGxiZHXheIghcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> MWiyOVA5PDZzNB?=
U937 MmiyS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NUP3VnFUOTJ3L{K1NE82ODBibl2= MX20PEBp NH\jPG9qdmO{ZXHz[ZMh[2WubDDk[YF1cCCrbjDhJINwdmOnboTyZZRqd25vZHXw[Y5l\W62IH3hco5meg>? NF3ERWUzPTB6NE[xOC=>
HL-60 NGfiT21Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NGrjeXMyOjVxMkWwM|UxOCCwTR?= MYO0PEBp NFXqUmVqdmO{ZXHz[ZMh[2WubDDk[YF1cCCrbjDhJINwdmOnboTyZZRqd25vZHXw[Y5l\W62IH3hco5meg>? NY\PZmdLOjVyOES2NVQ>
OCI-AML3 MYnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MUKxNlUwOjVyL{WwNEBvVQ>? MXe0PEBp NUHjcpNZcW6lcnXhd4V{KGOnbHyg[IVifGhiaX6gZUBkd26lZX70doF1cW:wLXTldIVv\GWwdDDtZY5v\XJ? NV7tW3lxOjVyOES2NVQ>
MOLM-13 NVrOXWprT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MlHXNVI2NzJ3MD:1NFAhdk1? MVi0PEBp NGfiRZhqdmO{ZXHz[ZMh[2WubDDk[YF1cCCrbjDhJINwdmOnboTyZZRqd25vZHXw[Y5l\W62IH3hco5meg>? NFrxfVQzPTB6NE[xOC=>
CMK M{f4O2NmdGxiVnnhZoltcXS7IFHzd4F6 MWqxNE0yODByMDDuUS=> Mn\hO|IhcA>? MVjy[YR2[2W|IHPlcIwhfmmjbHnibZR6KGmwIHGgZ49v[2WwdILheIlwdi2mZYDlcoRmdnRibXHucoVz M1KxWFI1QTZ{M{Ox
CMY NWPYNHJNS2WubDDWbYFjcWyrdImgRZN{[Xl? MWixNE0yODByMDDuUS=> MUW3NkBp MkewdoVlfWOnczDj[YxtKH[rYXzpZol1gSCrbjDhJINwdmOnboTyZZRqd25vZHXw[Y5l\W62IH3hco5meg>? NYLhdJBYOjR7NkKzN|E>
Dayo MXzHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MXTJR|UxRTF3MDDuUS=> NXq1ZY95OjR4NkG5NVA>
UW228 MlvtS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MljNTWM2OD1{M{Kgcm0> M1rFVFI1PjZzOUGw
IST-MES1 NVz2e2c5S2WubDDWbYFjcWyrdImgRZN{[Xl? NILEeFAyPTBxMkWwJI5O MVS3NkBp MoGx[Y5p[W6lZYOgeIhmKGOrc4DsZZRqdiCleYTveI95cWNiZX\m[YN1KGmwIHGgZ49v[2WwdILheIlwdi2mZYDlcoRmdnRibXHucoVz NIPhSVIzPDN4NUe4Ni=>
IST-MES2 MYHD[YxtKF[rYXLpcIl1gSCDc4PhfS=> NF;FTGsyPTBxMkWwJI5O Mk\BO|IhcA>? MV\lcohidmOnczD0bIUh[2m|cHzheIlvKGO7dH;0c5hq[yCnZn\lZ5QhcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> NGHIUZozPDN4NUe4Ni=>
REN NInQSGxE\WyuIG\pZYJqdGm2eTDBd5NigQ>? NYDoeYhzOTVyL{K1NEBvVQ>? MWG3NkBp MYjlcohidmOnczD0bIUh[2m|cHzheIlvKGO7dH;0c5hq[yCnZn\lZ5QhcW5iYTDjc45k\W62cnH0bY9vNWSncHXu[IVvfCCvYX7u[ZI> NHXa[ZUzPDN4NUe4Ni=>
NCI-H2452 NFnIb2JE\WyuIG\pZYJqdGm2eTDBd5NigQ>? MX6xOVAwOjVyIH7N MW[3NkBp NUXLWoNb\W6qYX7j[ZMhfGinIHPpd5Bt[XSrbjDjfZRwfG:6aXOg[YZn\WO2IHnuJIEh[2:wY3XueJJifGmxbj3k[ZBmdmSnboSgcYFvdmW{ NWCyPXkxOjR|NkW3PFI>
MSTO-211H NFrhRpBE\WyuIG\pZYJqdGm2eTDBd5NigQ>? MnHmNVUxNzJ3MDDuUS=> NXu3cHBVPzJiaB?= NX;1THdX\W6qYX7j[ZMhfGinIHPpd5Bt[XSrbjDjfZRwfG:6aXOg[YZn\WO2IHnuJIEh[2:wY3XueJJifGmxbj3k[ZBmdmSnboSgcYFvdmW{ M{m1clI1OzZ3N{iy
NCI-H2052 NVnJeWhPS2WubDDWbYFjcWyrdImgRZN{[Xl? NWP1bnpvOTVyL{K1NEBvVQ>? MoLUO|IhcA>? NG\UTJFmdmijbnPld{B1cGViY3nzdIxifGmwIHP5eI91d3irYzDl[oZm[3RiaX6gZUBkd26lZX70doF1cW:wLXTldIVv\GWwdDDtZY5v\XJ? MoKxNlQ{PjV5OEK=
WEE1 NHzWOmZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NE\zZXBKSzVyPUWuNkBvVQ>? MWiyN|Y6QTZ3NR?=
CDC2 MWHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MX;JR|Ux97zgMUCwNEBvVQ>? NWfYfphoOjN4OUm2OVU>
CDK7 NX7nZZdwT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MYnJR|Ux97zgMUCwNEBvVQ>? Mor1NlM3QTl4NUW=
MYT1 M4LRT2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NEPRXmpKSzVyPUWzNEBvVQ>? NX3QcGZ[OjN4OUm2OVU>
T98G  MYfBdI9xfG:|aYOgRZN{[Xl? MVyxNFAwOjVyIH7N MWi2JIg> M4jaSYVvcGGwY3XzJJJi\GmjdHnvck1qdmS3Y3XkJINmdGxia3nscIlv\w>? MnLaNlE6QTJ5OUO=
A549 NUfJdmNwSXCxcITvd4l{KEG|c3H5 Ml\rNlAxKG6P MoDwNUBp M1HNcJJi\Gmxc3Xud4l1cXqnczDOV2NNSyClZXzsd{BqdiCjIIC1N{1l\XCnbnTlcpQhdWGwbnXy M2[4NFIyPzl7MEOz
H460 NI\Xd4FCeG:ydH;zbZMhSXO|YYm= MmLXNlAxKG6P M4TRPFEhcA>? M1z2e5Ji\Gmxc3Xud4l1cXqnczDOV2NNSyClZXzsd{BqdiCjIIC1N{1l\XCnbnTlcpQhdWGwbnXy MXKyNVc6QTB|Mx?=
H1299 MXvBdI9xfG:|aYOgRZN{[Xl? MoC4NlAxKG6P MWCxJIg> MWPyZYRqd3OnboPpeIl7\XNiTmPDUGMh[2WubIOgbY4h[SCyNUOt[IVx\W6mZX70JI1idm6nch?= NIT6O2czOTd7OUCzNy=>
Calu-6  MlPyRZBweHSxc3nzJGF{e2G7 MX6yNFAhdk1? MmK4NUBp MX7yZYRqd3OnboPpeIl7\XNiTmPDUGMh[2WubIOgbY4h[SCyNUOt[IVx\W6mZX70JI1idm6nch?= NYrjcVRCOjF5OUmwN|M>
WiDr NWHNS2xqU2mwYYPlJGF{e2G7cx?= MUixNE0yODByMDDuUS=> NFSyNog5KGh? NGX0NWlqdmirYnn0d{BxcG:|cHjvdplt[XSrb36gc4YhS0SFMjDheEBVgXJzNTD3bZRpKGGwIFXDOVDDqH[jbIXlJI9nKDh3IH7tc4wwVCCycnX0doVifGWmIIfpeIgh\2WvY3n0ZYJqdmV? NVywUXI5OTl6OEe1OFU>

他の多くの細胞株試験データをご覧になる場合はこちらをクリックして下さい

アッセイ
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
体内試験 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]

お薦めの試験操作(参考用のみ)

キナーゼ試験:

[1]

- 合併

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.
細胞試験:

[1]

- 合併
  • 細胞株: WiDr, NCI-H1299, TOV21G, and HeLa
  • 濃度: Dissolved in DMSO, final concentrations ~10 μM
  • 反応時間: 24 hours
  • 実験の流れ:

    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.


    (参考用のみ)
動物試験:

[1]

- 合併
  • 動物モデル: Immunodeficient nude rats (F344/NJcl-rnu) bearing WiDr, HeLa-luc, or TOV21G-shp53 tumors
  • 投薬量: ~20 mg/kg/day
  • 投与方法: Orally
    (参考用のみ)

溶解度 (25°C)

体外 DMSO 80 mg/mL (159.8 mM)
Ethanol 10 mg/mL (19.97 mM)
Water 0.0001 mg/mL (0.0 mM)
体内 左から(NMPから)右の順に溶剤を製品に加えます(文献ではなく、Selleckの実験によるデータ):
2% DMSO+30% PEG 300+5% Tween 80+ddH2O
混合させたのち直ちに使用することを推奨します。
5mg/mL

* 溶解度測定はSelleck技術部門によって行われており、その他文献に示されている溶解度と差異がある可能性がありますが、同一ロットの生産工程で起きる正常な現象ですからご安心ください。

化学情報

分子量 500.6
化学式

C27H32N8O2

CAS No. 955365-80-7
Storage powder
in solvent
別名 AZD1775
Smiles CN1CCN(CC1)C2=CC=C(NC3=NC4=C(C=N3)C(=O)N(CC=C)N4C5=NC(=CC=C5)C(C)(C)O)C=C2

投与溶媒組成計算器(クリア溶液)

ステップ1:実験データを入力してください。(実験操作によるロスを考慮し、動物数を1匹分多くして計算・調製することを推奨します)
投与量 mg/kg 動物平均体重 g 投与体積(動物毎) ul 動物数
ステップ2:投与溶媒の組成を入力してください。(ロット毎に適した溶解組成が異なる場合があります。詳細については弊社までお問い合わせください)
% DMSO % % Tween 80 % ddH2O
計算リセット

便利ツール

モル濃度計算器

モル濃度計算器

求めたい質量、体積または濃度を計算してください。

質量 (mg) = 濃度 (mM) x 体積 (mL) x 分子量 (g/mol)

モル濃度計算器方程式

  • 質量
    濃度
    体積
    分子量

*貯蔵液を準備するとき、常に、オンであるとわかる製品のバッチに特有の分子量を使って、を通してラベルとMSDS/COA(製品ページで利用可能な)。

希釈計算器

希釈計算器

貯蔵液を準備するために必要な希釈率を計算してください。Selleck希釈計算器は、以下の方程式に基づきます:

開始濃度 x 開始体積 = 最終濃度 x 最終体積

希釈の計算式

この方程式は、一般に略語を使われます:C1V1 = C2V2 ( 入力 出力 )

  • C1
    V1
    C2
    V2

常に貯蔵液を準備するとき、小びんラベルとMSDS/COA(オンラインで利用できる)で見つかる製品のバッチに特有の分子量を使ってください。

連続希釈計算器方程式

  • 連続希釈剤

  • 計算結果

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
    C6=C5/X C6: LOG(C6):
    C7=C6/X C7: LOG(C7):
    C8=C7/X C8: LOG(C8):
分子量計算器

分子量计算器

そのモル質量と元素組成を計算するために、合成物の化学式を入力してください:

総分子量:g/mol

チップス: 化学式は大文字と小文字の区別ができます。C10H16N2O2 c10h16n2o2

モル濃度計算器

質量 濃度 体積 分子量

臨床試験

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

技術サポート

ストックの作り方、阻害剤の保管方法、細胞実験や動物実験の際に注意すべき点など、製品を取扱う時に問い合わせが多かった質問に対しては取扱説明書でお答えしています。

Handling Instructions

他に質問がある場合は、お気軽にお問い合わせください。

  • * 必須

よくある質問(FAQ)

  • 質問1:

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

  • 回答:

    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.

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