PP2

別名：AG 1879,AGL 1879

製品コード：S7008 純度：99.5%

PP2 (AG 1879, AGL 1879), a Src family kinase inhibitor, potently inhibits Lck/Fyn with IC50 of 4 nM/5 nM in cell-free assays, ~100-fold less potent to EGFR, inactive for ZAP-70, JAK2 and PKA.

CAS No. 172889-27-9

サイズ	価格(税別)	在庫状況
10mM (1mL in DMSO)	JPY 55500	国内在庫あり
1mg	JPY 22000	国内在庫あり
5mg	JPY 44500	国内在庫あり
1g	JPY 448500	国内在庫なし(納期7~10日)

代表番号: 045-509-1970\|電子メール：[email protected] よく尋ねられる質問

文献中Selleckの製品使用例（113）

製品安全説明書

現在のバッチを見る：純度： 99.5%

99.5

PP2関連製品

関連ターゲット	Lck Fyn Lyn Yes Fgr	もっと見る
関連化合物ライブラリー	Tyrosine Kinase Inhibitor Library PI3K/Akt Inhibitor Library Angiogenesis Related compound Library HIF-1 Signaling Pathway Compound Library FDA-approved Anticancer Drug Library	もっと見る

シグナル伝達経路

Srcシグナル伝達経路

Src阻害剤の選択性比較

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	活性情報	PMID
A431	Function assay	10 uM		Inhibition of Src autophosphorylation of Y419 in A431 cells at 10 uM	16509573
8701-BC	Proapoptotic assay	10 uM		Proapoptotic activity against 8701-BC cells at 10 uM by PARP assay	16509573
Saos2	Cytotoxicity assay		48 hrs	Cytotoxicity against human Saos2 cells after 48 hrs by MTT assay, IC50 = 8.07 μM.	23932070
SH-SY5Y	Antiproliferative assay		72 hrs	Antiproliferative activity against human SH-SY5Y cells assessed as cell viability after 72 hrs by XTT assay, IC50 = 6.1 μM.	21856155
SaOS2	Antiproliferative assay			Antiproliferative activity against human SaOS2 cells assessed as cellular viability, IC50 = 8.1 μM.	17929792
T-cells	Function assay			Inhibition of tyrosine phosphorylation in human T cells, IC50 = 0.6 μM.	18077363
T-cells	Function assay			Inhibition of adhesion kinase in human T cells, IC50 = 0.6 μM.	18077363
A549	Growth inhibition assay			Growth inhibition of human A549 cells, IC50 = 0.01 μM.	28814374
A431	Function assay			Inhibitory effect on phospho-Src/nonphospho after EGF (100 uM) stimulation of A431 cells (21), IC50 = 17 μM.	15109642
MEG01	Antiproliferative assay			Antiproliferative activity against human MEG01 cells, IC50 = 17 μM.	18257513
A431	Function assay			Inhibitory effect on phospho-Src (Tyr416) after EGF (100 uM) stimulation of A431 cells (38), IC50 = 22 μM.	15109642
K562	Antiproliferative assay			Antiproliferative activity against human K562 cells, IC50 = 25 μM.	18257513
A431	Antiproliferative assay			Tested for antiproliferative activity against human A431 cells, IC50 = 32 μM.	15109642
A431	Antiproliferative assay			Antiproliferative activity against A431 cells, IC50 = 32.2 μM.	16509573
KU812	Antiproliferative assay			Antiproliferative activity against human KU812 cells, IC50 = 45 μM.	18257513
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生物活性

製品説明

PP2 (AG 1879, AGL 1879), a Src family kinase inhibitor, potently inhibits Lck/Fyn with IC50 of 4 nM/5 nM in cell-free assays, ~100-fold less potent to EGFR, inactive for ZAP-70, JAK2 and PKA.

Targets

LCK ^[1] (Cell-free assay)	Fyn ^[1] (Cell-free assay)
4 nM	5 nM

In Vitro
In vitro	PP2 inhibits Src by binding to an area of the molecule that does not overlap with the ATP binding domain. ^[2] PP2 (20 μM) induces 40-50% growth inhibition of HT29 cells, this concentration reduces the Src activity as early as 1 hour and maintains a 35% inhibition of Src activity for 2 days. PP2 (100 mM) decreases the Src activity of HT29 cells in a dose-dependent manner. PP2 (1 mM-100 mM) causes a dose-dependent growth inhibition of human colon cancer cells (HT29, SW480, and PMCO1), liver cancer cells (PLC/PRF/5, KYN-2, Li7, and HepG2), and breast cancer cells (MCF-7, MDA-MB-468, and BT-474). PP2 (20 μM) significantly increases aggregation in most of the cancer cells (HT29, SW480, PMCO1, PLC/PRF/5, KYN-2, Li7, MCF-7, and MDA-MB-468) in E-cadherin dependent manner. PP2 (20 μM) enhances E-cadherin expression and also strongly increases E-cadherin’s association with the actin cytoskeleton in cancer cells. PP2 (20 μM) increases the expression of α-catenin, β-catenin, and γ-catenin in HT29 cells, whereas in PLC/PRF/5 and MCF-7 cells, the total protein level of α-catenin does not change, but the levels of β- catenin and γ-catenin increases slightly. ^[3] PP2 inhibits proliferation of two cervical cancer cells (HeLa and SiHa) in a time- and dose-dependent manner. PP2 (10 μM) down-regulates pSrc-Y416, pEGFR-Y845, and -Y1173 expression levels in HeLa and SiHa cells. PP2 (10 μM) could modulate cell cycle arrest by up-regulating p21(Cip1) and p27(Kip1) in both HeLa and SiHa cells and down-regulating expression of cyclin A, and cyclin dependent kinase-2, -4 (Cdk-2, -4) in HeLa and of cyclin B and Cdk-2 in SiHa. ^[4]
Kinase Assay	Immune complex enzyme assays
Kinase Assay	The acid-treated enolase is diluted 1:20 with 1× PBS before aliquoting 100 mL/well into a Nunc 96-well high protein binding assay plate. Assay wells are then aspirated; blocked with 0.5% bovine serum, 1× PBS for 1 h at 37 ℃;and then washed five times with 300 mL of 1× PBS/well. The source of Lck is either LSTRA cells or Lck expressed in HeLa cells using a vaccinia expression system. FynT is expressed in HeLa cells using the vaccinia system. Cells (12.5× 10⁶/mL) are lysed in lysis buffer (20 mM Tris, pH 8.0, 150 mM NaCl, 0.5% Nonidet P-40, and 23 trypsin inhibitory units/mL aprotinin), and the lysates are clarified by centrifugation at 14,000 cpm for 15 min at 4 ℃ in an Eppendorf tube. The clarified lysates are then incubated with the appropriate anti-kinase antibody at 10 μg/mL for 2 h at 4 ℃. Protein A-Sepharose beads are added to the antibody/lysate mixture at 250 μL/mL and allowed to incubate for 30 min at 4 ℃. The beads are then washed twice in 1 mL of lysis buffer and twice in 1 mL of kinase buffer (25 mM HEPES, 3 mM MnCl₂, 5mM MgCl₂, and 100 μM sodium orthovanadate) and resuspended to 50% (w/v) in kinase buffer. Twenty-five microliters of the bead suspension is added to each well of the enolase-coated 96-well high protein binding plate together with an appropriate concentration of compound and [γ-³²P]ATP (25 μL/well of a 200 μCi/mL solution in kinase buffer). After incubation for 20 min at 20 ℃, 60 μLl of boiling 2× solubilization buffer containing 10 mM ATP is added to the assay wells to terminate the reactions. Thirty microliters of the samples is removed from the wells, boiled for 5 min, and run on a 7.5% SDS-polyacrylamide gel. The gels are subsequently dried and exposed to Kodak X-AR film. For quantitation, films are scanned using a Molecular Dynamics laser scanner, and the optical density of the major substrate band, enolase p46, is determined. In companion experiments for measuring the activity of compounds against Lck, the assay plate is washed with two wash cycles on a Skatron harvester using 50 mM EDTA, 1 mM ATP. Scintillation fluid (100 μL) is then added to the wells, and ³²P incorporation is measured using a micro-β-counter.
細胞実験	細胞株	HT29, SW480, PMCO1, PLC/PRF/5, KYN-2, Li7, HepG2, MCF-7, MDA-MB-468 and BT-474 cell lines
	濃度	~100 μM
	反応時間	2 days
	実験の流れ	Cell viability is determined using an in vitro toxicology assay kit following the manufacturer’s instructions. Cells are seeded in 96-well plates at day 0. Starting at day 1, cells are treated for 2 days with each of a series of increasing concentrations of PP2 (1 μM, 10 μM, and 100 μM). At the end of this period, cell proliferation is evaluated by a colorimetric assay based on the cleavage of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide by mitochondria dehydrogenase in viable cells, leading to formazan formation. This experiment is repeated three times with 10 determinations/tested concentration.
実験結果図	Methods	Biomarkers	結果図	PMID
	Western blot	p-PI3K / PI3K / p-AKT / AKT / Bcl-2 / Caspase-3 p-Src / Src / p-MAPK / MAPK		30250573
	Immunofluorescence	β-catenin FAK / p-FAK		18566211

In Vivo
In Vivo	PP2 (5 mg/kg/day) induces some slowing in the growth rate of the primary tumors relative to the control treated with vehicle in SCID mice inoculated HT29 cells in the spleen. PP2 (5 mg/kg/day) induces some slowing in the growth rate of the primary tumors relative to the control treated with vehicle in SCID mice inoculated HT29 cells in the spleen. PP2 (5 mg/kg/day) significantly reduces the relative liver weight and liver metastasis volume compared with the controls in SCID mice inoculated HT29 cells in the spleen. ^[3] PP2 (1.5 mg/kg i.p.) treated rats show approximately 50% reduction of infarct size on T2-weighted MRI and in TTC staining compared with controls in rats with focal ischemic brain injury. PP2 (1.5 mg/kg i.p.) results in better the neurological score than controls in rats with focal ischemic brain injury. ^[5]
動物実験	動物モデル	SCID mice inoculated HT29 cells in the spleen
	投与量	5 mg/kg/day
	投与経路	intraperitoneal injection

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
臨床試験 (data from https://clinicaltrials.gov, updated on 2024-01-11)
NCT03842371	Unknown status	Sepsis Syndrome	West China Hospital	February 11 2019	--
NCT02407626	Terminated	Myocardial Ischemia	Triemli Hospital\|University of Alberta	September 2015	Not Applicable
NCT02315287	Unknown status	Type 2 Diabetes	Seoul National University Bundang Hospital	September 2014	Phase 4

参考
[1] Hanke JH, et al. J Biol Chem, 1996, 271(2), 695-701. [2] Karni R, et al. FEBS Lett, 2003, 537(1-3), 47-52. [3] Nam JS, et al. Clin Cancer Res, 2002, 8(7), 2430-2436. [4] Kong L, et al. Mol Cell Biochem, 2011, 348(1-2), 11-19. [5] Lennmyr F, et al. Acta Neurol Scand, 2004, 110(3), 175-179. + 展開

参考

+ 展開

化学情報

分子量	301.77	化学式	C₁₅H₁₆ClN₅
CAS No.	172889-27-9	SDF	Download PP2 SDFをダウンロードする
Smiles	CC(C)(C)N1C2=NC=NC(=C2C(=N1)C3=CC=C(C=C3)Cl)N
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1年-80°Cin solvent
保管	3年-20°C粉	溶液の保管冷凍と解凍の繰り返しを避けるため小分けにしてください	1个月-20°Cin solvent

In vitro
Batch:

DMSO : 60 mg/mL ( (198.82 mM)；吸湿したDMSOは溶解度を減少させます。新しいDMSOをご使用ください。)

Ethanol : 2 mg/mL

Water : Insoluble

モル濃度計算器

in vivo
Batch:

Add solvents to the product individually and in order.

投与溶液組成計算機

実験計算

モル濃度計算器

質量	濃度	体積	分子量
=	×	×

希釈計算器分子量計算器

投与溶液組成計算機(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）μL 動物数匹

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

%DMSO+ %

計算結果：

投与溶媒濃度： mg/ml;

DMSOストック溶液調製方法： mg 試薬を μL DMSOに溶解する（濃度 mg/mL, 注：濃度が当該ロットのDMSO溶解度を超える場合はご連絡ください。 )

投与溶媒調製方法：Take μL DMSOストック溶液に μL PEG300，を加え、完全溶解後μL Tween 80，を加えて完全溶解させた後 μL ddH₂O，を加え完全に溶解させます。

投与溶媒調製方法：μL DMSOストック溶液に μL Corn oil，を加え、完全溶解。

注意：１.ストック溶液に沈殿、混濁などがないことをご確認ください；
２.順番通りに溶剤を加えてください。次のステップに進む前に溶液に沈殿、混濁などがないことを確認してから加えてください。ボルテックス、ソニケーション、水浴加熱など物理的な方法で溶解を早めることは可能です。

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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:
Could you please suggest me the in vivo details about the dilution to reduce the amount of DMSO to 1 to 5%?

回答
For in vivo study, we recommend to use 4% DMSO +Corn oil up to 2.5 mg/ml.

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):