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SP600125

Name: SP600125
Brand: Selleck Chemicals
SKU: S1460
Rating: 5 (1016 reviews)

Synonyms: Nsc75890

Catalog No.S1460 Purity:99.94%

SP600125 (Nsc75890) is a broad-spectrum JNK inhibitor for JNK1, JNK2 and JNK3 with IC50 of 40 nM, 40 nM and 90 nM in cell-free assays, respectively; 10-fold greater selectivity against MKK4, 25-fold greater selectivity against MKK3, MKK6, PKB, and PKCα, and 100-fold selectivity against ERK2, p38, Chk1, EGFR etc. SP600125 is also a broad‐spectrum inhibitor of serine/threonine kinases including Aurora kinase A，FLT3 and TRKA with of IC50 of 60 nM, 90 nM and 70 nM. SP600125 inhibits autophagy and activates apoptosis.

SP600125 Chemical Structure

CAS No. 129-56-6

Purity & Quality Control

Batch: Purity: 99.94%

99.94

Products often used together with SP600125

Adezmapimod (SB203580)

SP600125 inhibits autophagy and activates apoptosis whereas, Adezmapimod induces mitophagy and autophagy.

PD98059

SP600125 and PD98059 are two kinases that possess pro-survival activities in TQ-induced cell death.

El-Najjar N, et al. Apoptosis. 2010 Feb;15(2):183-95.

SCH772984

SP600125 and SCH772984 significantly increase apoptosis of both PMN-MDSCs and M-MDSCs in vitro.

Yu J, et al. Front Oncol. 2021 Mar 18;11:647312.

LY294002

SP600125 and LY294002 suppress cell growth of SNU-216 and NCI-N87 cells.

Choi Y, et al. World J Gastroenterol. 2016 Nov 7; 22(41): 9141–9153.

Bentamapimod (AS602801)

SP600125 and Bentamapimod are pan-JNK inhibitors that cause a robust reduction in cell viability in a panel of GBM stem cells (GSCs) cultures.

MacLeod G, et al. Cell Rep. 2019 Apr 16;27(3):971-986.e9.

SP600125 Related Products

Related Targets	JNK1 JNK2 JNK3	Click to Expand
Related Products	JNK-IN-8 Anisomycin JNK Inhibitor IX Tanzisertib(CC-930) JNK Inhibitor VIII Bentamapimod (AS602801) BI-78D3 SP 600125, negative control DTP3 CC-401 Hydrochloride Polyphyllin I	Click to Expand
Related Compound Libraries	Kinase Inhibitor Library MAPK Inhibitor Library Cell Cycle compound library TGF-beta/Smad compound library Anti-alzheimer Disease Compound Library	Click to Expand

Signaling Pathway

JNK Signaling Pathway

Cell Data

Cell Lines	Assay Type	Concentration	Incubation Time	Formulation	Activity Description	PMID
Hep3B	Function Assay	10 μM	1 h		Blocks autophagy and upregulation of Beclin 1 expression induced by ceramide	19060920
BV-2	Function Assay	2 μM	1 h		Inhibits the increase of sBAFF release in Gmix-treated BV-2 cells	19406831
RAW264.7	Function Assay	10 μM	12 h		Antiinflammatory activity assessed as inhibition of LPS-induced NO production with IC50 of 17μM	19497418
PC3	Function Assay	20 μM	1 h		Decreases the MMP2 and MMP9 expression	19633975
Plasmodium falciparum HB3	Antibacterial Assay		72 h	DMSO	Antiplasmodial activity with IC50 of 7.94328 μM	19734910
Plasmodium falciparum W2	Antibacterial Assay		72 h	DMSO	Antiplasmodial activity with IC50 of 7.94328 μM	19734910
Plasmodium falciparum 7G8	Antibacterial Assay		72 h	DMSO	Antiplasmodial activity with IC50 of 10 μM	19734910
Plasmodium falciparum 3D7	Antibacterial Assay		72 h	DMSO	Antiplasmodial activity with IC50 of 12.5893 μM	19734910
Plasmodium falciparum GB4	Antibacterial Assay		72 h	DMSO	Antiplasmodial activity with IC50 of 12.5893μM	19734910
HaCaT	Function Assay	20 μM	4 h	DMSO	Blocks the TNF-α-induced CYP4F11 transcription	19812349
HaCaT	Function Assay	20 μM	24 h	DMSO	Blocks the phosphorylation of c-Jun protein	19812349
A549	Function Assay	20 μM	1 h		Inhibition of TPA-induced MMP-2 and u-PA expression	20492175
PC12	Function Assay	10 μM	5 h	DMSO	Activation of Nrf2/ARE assessed as HO-1 protein induction pretreated with PD98059	21345685
PC12	Function Assay	10 μM	5 h	DMSO	Activation of Nrf2/ARE assessed as HO-1 protein induction pretreated with U0126	21345685
PC12	Function Assay	10 μM	5 h	DMSO	Activation of Nrf2/ARE assessed as HO-1 protein induction pretreated with SP600125	21345685
PC12	Function Assay	10 μM	5 h	DMSO	Activation of Nrf2/ARE assessed as HO-1 protein induction pretreated with SB203580	21345685
B16-F10	Function Assay		1 h		Inhibition of TNF-alpha-induced c-JUN phosphorylation	21815634
LoVo	Function Assay	1 μM	1 h		Inhibition of PGE2-induced expression of uPA and MMP-9 significantly	21859479
LoVo	Function Assay	1 μM	1 h		BlocksPGE2-induced cell migration significantly	21859479
THP-1	Function Assay	90 nM	30 min		Inhibition of tissue factor expression	22940059
PC3	Function Assay	25 μM	24 h		Inhibition of AP-1 and p21 luciferase activity induced by S179D PRL	23162652
SH-SY5Y	Function Assay	10 μM	1 h	DMSO	Neuroprotective activity assessed as reduction of anisomycin-induced cell death	23498914
SH-SY5Y	Kinase Assay	10 μM	1 h	DMSO	Inhibition of JNK3 assessed as blockade of anisomycin-induced c-jun phosphorylation at ser73	23498914
RAW264.7	Function Assay	10 μM	24 h		Antiinflammatory activity assessed as inhibition of IL-1beta release	23791078
RAW264.7	Function Assay	10 μM	24 h		Antiinflammatory activity assessed as inhibition of LPS-induced iNOS expression	23791078
RAW264.7	Function Assay	10 μM	2 h		Antiinflammatory activity assessed as inhibition of LPS-induced NO production	23791078
A549	Growth Inhibition Assay	20 μM	72 h	DMSO	Rapid and potent inhibition of cell proliferation	23912840
RAW264.7	Antiinflammatory assay				Antiinflammatory activity in mouse RAW264.7 cells assessed as inhibition of LPS-induced NO production relative to control, IC50 = 17 μM.	22831798
BMMC	Function assay	1 to 20 uM	7 days		Inhibition of RANKL/M-CSF-stimulated osteoclastogenesis in ICR mouse BMMC assessed as reduction in TRAP positive multinucleated cells at 1 to 20 uM incubated for 7 days by light microscopy	25397676
A673	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
BT-37	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
SK-N-SH	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
NB1643	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
OHS-50	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
SK-N-MC	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
NB-EBc1	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
LAN-5	qHTS assay				qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
Click to View More Cell Line Experimental Data

Biological Activity

Description

Targets

serine/threonine kinase ^[1]	JNK1 ^[1] (Cell-free assay)	JNK2 ^[1] (Cell-free assay)	Aurora A ^[4] (Cell-free assay)	TrkA ^[4] (Cell-free assay)	Click to View More Targets
	40 nM	40 nM	60 nM	70 nM	Click to View More Targets

In vitro
In vitro	SP600125 is originally characterized as a selective ATP-competitive inhibitor of c-Jun N-terminal kinase JNK. In Jurkat T cells, SP600125 inhibits the phosphorylation of c-Jun with IC50 of 5 μM to 10 μM. In CD4+ cells, such as Th0 cells isolated from either human cord or peripheral blood, SP600125 blocks cell activation and differentiation and inhibits the expression of inflammatory genes COX-2, IL-2, IL-10, IFN-γ, and TNF-α, with IC50 of 5 μM to 12 μM. ^[1] However, later studies reveal that SP600125 also suppresses aryl hydrocarbon receptor (AhR) ^[2], Mps1 ^[3], and a panel of other serine/threonine kinases, including Aurora kinase A, FLT3, MELK, and TRKA ^[4]. In a mouse beta cells MIN6, SP600125 (20 μM) induces the phosphorylation of p38 MAPK and its downstream CREB-dependent promoter activation. ^[5] In HCT116 cells, SP600125 (20 μM) blocks the G2 phase to mitosis transition and induces endoreplication. This ability of SP600125 is independent of JNK inhibition, but due to its inhibition of CDK1-cyclin B activation upstream of Aurora A and Polo-like kinase 1. ^[6]
Kinase Assay	In Vitro Kinase Assays
Kinase Assay	The potency of SP600125 towards kinases, including MPS1, JNK, and Aurora kinase A, is determined based on the specific measurement of radioactive phosphotransfer to the substrate. For each enzyme, the absolute K_m values for ATP and the specific substrate are initially determined and each assay is then run at optimized [ATP] (2·αK_m) and [substrate] (5·K_m) concentrations. MPS1 activity is measured using 5 nM of MPS1 recombinant protein in 50 mM HEPES pH 7.5, 2.5 mM MgCl₂, 1 mM MnCl₂, 1 mM DTT, 3 μM NaVO₃, 2 mM β-glycerophosphate, 0.2 mg/mL BSA, 200 μM P38-βtide substrate-peptide (KRQADEEMTGYVATRWYRAE), and 8 μM ATP with 1.5 nM ³³P-γ-ATP. Ten serial 1:3 dilutions (from 30 μM to 1.5 nM) of SP600125 are tested and IC50 determined.
Cell Research	Cell lines	HCT116, A2780, and U2OS cells
	Concentrations	0–5 μM
	Incubation Time	72 hours
	Method	Cells are seeded in 384 well-plates. One day after seeding, the cells are treated with SP600125 for 72 hours and the plates are then processed using a CellTiter-Glo assay. Inhibitory activity is evaluated comparing treated versus control data and IC50 value of proliferation is calculated.
Experimental Result Images	Methods	Biomarkers	Images	PMID
	Western blot	p-JNK p-IGF1R / IGF1R / p-Akt / Akt / p-ERK / ERK p-Src / Src p-c-Jun / c-Jun / pJNK / JNK Survivin / Bcl-2 / PARP p-FADD / FADD / p-c-Jun / c-Jun		25226534
	Immunofluorescence	AIF / Endo G E-cadherin / β-catenin α-catenin / Actin		21738692
	Growth inhibition assay	Cell viability (U-87 MG) Cell viability (A549)		27176481

In Vivo
In vivo	In mice, SP600125 (15 mg/kg or 30 mg/kg) significantly inhibits lipopolysaccharide (LPS)-induced TNF-α expression and anti-CD3-induced apoptosis of CD4⁺ CD8⁺ thymocytes. ^[1]
Animal Research	Animal Models	Mouse LPS/TNF model (female CD-1)
	Dosages	15 or 30 mg/kg
	Administration	Administered via intravenous injection or orally

References

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Chemical Information & Solubility

Molecular Weight	220.23	Formula	C₁₄H₈N₂O
CAS No.	129-56-6	SDF	Download SP600125 SDF
Smiles	C1=CC=C2C(=C1)C3=NNC4=CC=CC(=C43)C2=O
Storage (From the date of receipt)	3 years-20°Cpowder	Storage of Stock Solutions Aliquot the stock solutions to avoid repeated freeze-thaw cycles	1 year-80°Cin solvent
Storage (From the date of receipt)	3 years-20°Cpowder		1 month-20°Cin solvent

In vitro
Batch:

DMSO : 60 mg/mL ( (272.44 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molecular Weight Calculator

In vivo Batch: Add solvents to the product individually and in order.					In vivo Formulation Calculator
	Clear solution	5% DMSO 1 95% Corn oil	0.3mg/ml (1.36mM)	Taking the 1 mL working solution as an example, add 50 μL of 6 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.	In vivo Formulation Calculator
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	2.0mg/ml (9.08mM)	Taking the 1 mL working solution as an example, add 50 μL of 40 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	10%DMSO 1 90%Corn oil	6.0mg/ml (27.24mM)	Taking the 1 mL working solution as an example, add 100 μL of 60 mg/ml clear DMSO stock solution to 900 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	2.2mg/ml (9.99mM)	Taking the 1 mL working solution as an example, add 50 μL of 44 mg/ml clarified DMSO stock solution to 400 μL PEG300, mix evenly to clarify it; add 50 μL Tween80 to the above system, mix evenly to clarify it; then continue to add 500 μL ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

Preparing Stock Solutions

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
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Dilution Calculator Molecular Weight Calculator

In vivo Formulation Calculator (Clear solution)

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

Dosage: mg/kg Average weight of animals: g Dosing volume per animal:μL Number of animals:

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Calculation results:

Working concentration: mg/ml;

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.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

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.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Frequently Asked Questions

Question 1:
how to reconstitute the inhibitor for in vivo studies?

Answer:
S1460 can be dissolved in 5% DMSO/corn oil at 5 mg/ml as a clear solution for injection. The inhibitor dissolved in vehicle 30% PEG400/0.5% Tween80/5%Propylene glycol, at 30mg/ml is a suspension and can be used for oral administration.

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