GSK 2830371

Alias: GSK 2830371; GSK-2830371; GSK2830371.

Cat No.:V3535 Purity: ≥98%

COA Product Data Instructions for use SDS

GSK 2830371 Chemical Structure CAS No.: 1404456-53-6
Product category: Phosphatase

This product is for research use only, not for human use. We do not sell to patients.

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Nature 2021, 597(7874):119-125.

Cell 2020,183:1202-1218.e25.

Science Adv 2022: 8, eabm9427.

Nature 597, 119–125 (2021)

Cell Stem Cell 2020,26(6):845-861.e12.

Science Trans Med 2023,15(701):eadd7872.

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J Am Chem Soc 2022,144:21831-21836.

Cell 2020,183:1202-1218.e25.

Science Adv 2022:8,eabm9427.

Nature 2021,597(7874):119-125.

Cell 2020,183:1202-1218.e25.

PNAS Nexus 2022,1(3):pgac063.

ACS Nano 2023,17(10):9110-9125.

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Purity & Quality Control Documentation

Current batch：

Purity: ≥98%

COA

MSDS

Instructions

Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Biological Data

Product Description

GSK-2830371 (GSK2830371) is a first-in-class, potent, orally bioactive and highly selective allosteric inhibitor of Wip1 (wild-type p53-induced phosphatas) phosphatase with potential anticancer activity. It inhibits Wip1 with an IC50 of 6 nM. Treatment of tumor cells with the inhibitor GSK2830371 increases phosphorylation of Wip1 substrates and causes growth inhibition in both hematopoietic tumor cell lines and Wip1-amplified breast tumor cells harboring wild-type TP53. Oral administration of Wip1 inhibitors in mice results in expected pharmacodynamic effects and causes inhibition of lymphoma xenograft growth. To our knowledge, GSK2830371 is the first orally active, allosteric inhibitor of Wip1 phosphatase.

Biological Activity I Assay Protocols (From Reference)

Targets

IC50: 6 nM (Wip1 phosphatase)[1]

ln Vitro

With IC50 values of 6 nM and 13 nM, respectively, GSK 2830371 potently inhibits the dephosphorylation of Wip1 (2-420) by FDP and the endogenous substrate phospho-p38 MAPK (T180). Treatment with GSK 2830371 (0.04, 0.11, 0.33, 1, 3, and 9 μM) enhanced substrate phosphorylation in PPM1D-amplified MCF7 breast cancer cells in a concentration-dependent manner. GSK 2830371 (0.001, 0.01, 0.1, 1 and 10 μM) treatment of MX-1 and MCF7 (Wip1 amplified, p53 wild-type) cells resulted in concentration-dependent effects in cell growth experiments [1]. In MCF-7 cells, GSK2830371's 50% growth inhibitory concentration (GI50) is 2.65 μM±0.54 (SEM). When 2.5μM GSK2830371 is added to MCF-7 cells, both isoforms of WIP1 undergo considerable time-dependent degradation within 8 hours. This phenomenon is correlated with the stability of p53 and the phosphorylation of p53Ser15 (pp53Ser15) [2].

ln Vivo

GSK 2830371 was administered orally to DOHH2 tumors in pharmacodynamic experiments, which resulted in enhanced phosphorylation of Chk2 (T68) and p53 (S15) and decreased Wip1 protein concentration. Following 14 days of oral dosing at 150 mg/kg body weight BID (twice daily) and TID (three times daily), respectively, GSK 2830371 reduced the development of DOHH2 tumor xenografts by 41% and 68%. Similar suppression of tumor growth was noted in mice given 75 or 150 mg/kg body weight on a BID basis. The short half-life of GSK 2830371 in mice is consistent with the TID regimen's stronger reduction of tumor growth, indicating that prolonged inhibition of Wip1 may be necessary for the greatest anticancer effects [1].

Cell Assay

Cells are seeded into 96 well plates at 200-400 cells per well and treated with GSK 2830371 dilution series on day 1. After 7 d, we used the CellTiter-Glo cell viability assay to determine effects on cell growth. Luminescent signal is detected on an EnVision 2104. For clonogenic assays, cells are seeded in 12-well tissue culture plates at 2,000 cells per well. Cells are treated with a compound dilution series on day 1 and again on day 7. After 14 d, cells are washed with 1× PBS, stained with 1 mL of Coomassie Brilliant Blue R-250, and colonies are quantitated with the Optomax Sorcerer colony counter[1].

Animal Protocol

Mice[1] Female SCID mice are subcutaneously inoculated with 1×107 DOHH2 cells and tumor growth is monitored with electronic calipers. When tumors reached 100-200 mm3, animals are treated orally twice (BID) or thrice (TID) daily with vehicle (2% DMSO and 40% Captisol, pH 4.0) or GSK 2830371. For efficacy studies, eight mice per group are administered with GSK 2830371 at 75 or 150 mg per kg body weight BID or 150 mg per kg body weight TID. Tumor growth inhibition (% difference in tumor growth compared to control) is calculated when tumors for vehicle-control mice exceeded 1,000 mm3 (day 11). For pharmacodynamic biomarker analysis, DOHH2 tumors from three mice are harvested 2 or 4 h after final dose following 14 d of treatment with either vehicle or GSK 2830371 at 75 or 150 mg per kg body weight, BID; tumors are frozen in liquid nitrogen for subsequent lysis and western blot analysis.

References

[1]. Gilmartin AG, et al. Allosteric Wip1 phosphatase inhibition through flap-subdomain interaction. Nat Chem Biol. 2014 Mar;10(3):181-7.
[2]. Esfandiari A, et al. Chemical Inhibition of Wild-Type p53-Induced Phosphatase 1 (WIP1/PPM1D) by GSK2830371 Potentiates the Sensitivity to MDM2 Inhibitors in a p53-Dependent Manner. Mol Cancer Ther. 2016 Mar;15(3):379-91

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula	C23H29CLN4O2S
Molecular Weight	461.0200
CAS #	1404456-53-6
SMILES	O=C(C1=CC=C(CNC2=CC(Cl)=CN=C2C)S1)N[C@@H](CC3CCCC3)C(NC4CC4)=O
InChi Key	IVDUVEGCMXCMSO-FQEVSTJZSA-N
InChi Code	InChI=1S/C23H29ClN4O2S/c1-14-19(11-16(24)12-25-14)26-13-18-8-9-21(31-18)23(30)28-20(10-15-4-2-3-5-15)22(29)27-17-6-7-17/h8-9,11-12,15,17,20,26H,2-7,10,13H2,1H3,(H,27,29)(H,28,30)/t20-/m0/s1
Chemical Name	(S)-5-(((5-chloro-2-methylpyridin-3-yl)amino)methyl)-N-(3-cyclopentyl-1-(cyclopropylamino)-1-oxopropan-2-yl)thiophene-2-carboxamide
Synonyms	GSK 2830371; GSK-2830371; GSK2830371.
Storage	Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility Data

Solubility (In Vitro)	DMSO : ≥ 51 mg/mL (~110.62 mM)
Solubility (In Vivo)	Solubility in Formulation 1: ≥ 2.5 mg/mL (5.42 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (5.42 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More Solubility in Formulation 3: ≥ 2.5 mg/mL (5.42 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.)

Solubility (In Vitro)

DMSO : ≥ 51 mg/mL (~110.62 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.42 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: 2.5 mg/mL (5.42 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

View More

Solubility in Formulation 3: ≥ 2.5 mg/mL (5.42 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

(Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1691 mL	10.8455 mL	21.6910 mL
	5 mM	0.4338 mL	2.1691 mL	4.3382 mL
	10 mM	0.2169 mL	1.0846 mL	2.1691 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

Calculator

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Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?

Enter 350.26 in the Molecular Weight (MW) box
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The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

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Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:

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The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box

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Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4 c12h18n3o4
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Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
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The answer appears in the Volume (to add to vial) box

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)

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Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)

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Calculation results

Working concentration： mg/mL；

Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O，mix and clarify.

Note： (1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.

Biological Data

A) The effect on growth of a panel of p53 wild-type (Green) and Mutant/null (Maroon) cell line pairs with differentPPM1Dgenetic status to 0.08-10μM GSK2830371 exposure for 168hr, using sulforhodamine (SRB) growth inhibition assays. B) Basal expression of WIP1 and p53 in cell lines (SE-short film exposure, LE-long exposure). C) The sensitivity of a panel of p53 Wt (Green) and Mt/null (Maroon) cell line pairs with differentPPM1Dgenetic status to 0.08-10μM Nutlin-3 and 0.008-1μM RG7388 in 168hr SRB growth inhibition assays in the presence and absence of the highest non-growth inhibitory dose of GSK2830371 (2.5μM).Mol Cancer Ther.2016 Mar;15(3):379-91.

A) GSK2830371 (2.5μM) treatment of MCF-7 cells over 8 hours shows WIP1 degradation over time, p53 stabilisation and Phospho-p53Ser15(pp53Ser15) accumulation. B) p53Ser15phosphorylation in MCF-7 cells 30min post 2Gy ionising radiation in the presence or absence of 2.5μM. GSK2831371 inhibits WIP1 catalytic activity independent of WIP1 protein levels. C) 4 hours treatment with 2.5μM GSK2830371 results in degradation of full-length and truncated WIP1 in HCT116+/+and U2OS cells. D) Lysates obtained from HCT116+/+cells treated with 20μM proteasome inhibitor MG132 and 2.5μM GSK2830371 ± 3.0μM Nutlin-3 overnight underwent immunoprecipitation (IP) with anti-ubiquitin antibody (Ub-Ab) and the precipitates probed for WIP1 and p53 by western blot. Input samples (left panel) are western blots of total lysate before IP for comparison with the IP results on the right hand panel.Mol Cancer Ther.2016 Mar;15(3):379-91.

A) Dose-dependent increased in caspase 3/7 activity of NGP cells after 24 hours treatment with Nutlin-3 (Nut-3 GI50 ≈ 3.0μM) alone or in combination with 2.5μM GSK2830371. B) Increase in caspase 3/7 activity in NGP and SJSA-1 cells and theirTP53mutant daughter cell lines 48 hours after treatment with Nutlin-3 ± 2.5μM GSK2830371. C) Reduction in clonogenic efficiency of HCT116+/+cells following exposure to 0.5 × GI50 concentration of Nut-3 in the presence of 2.5μM GSK2830371 compared to either inhibitor alone over 10 days. D) Immunoblot of NGP cells showing Nut-3 dependent phosphorylation of p53 at Ser15 is markedly enhanced by GSK2830371 at 4 & 24hrs exposure and leads to increased caspase 3 cleavage at 48 hours.Mol Cancer Ther.2016 Mar;15(3):379-91.