CHIR99021 HCl
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MedKoo CAT#: 407462

CAS#: 1797989-42-4 (HCl)

Description: CHIR99021, also known as CT99021, is a glycogen synthase kinase 3β (GSK3β) inhibitor that has antiproliferative activity in vitro and in vivo. CHIR-99021 inhibits GSK-3 with IC50 at 7 nM. In a series of carcinoma cell lines, the IC50 of CHIR99021 for proliferation is about 10μM. CHIR99021 promotes self-renewal of mouse embryonic stem cells by modulation of protein-encoding gene and long intergenic non-coding RNA expression.


Chemical Structure

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CHIR99021 HCl
CAS# 1797989-42-4 (HCl)

Theoretical Analysis

MedKoo Cat#: 407462
Name: CHIR99021 HCl
CAS#: 1797989-42-4 (HCl)
Chemical Formula: C22H19Cl3N8
Exact Mass:
Molecular Weight: 501.8
Elemental Analysis: C, 52.66; H, 3.82; Cl, 21.19; N, 22.33

Price and Availability

Size Price Availability Quantity
10.0mg USD 150.0 2 Weeks
25.0mg USD 250.0 2 Weeks
50.0mg USD 450.0 2 Weeks
100.0mg USD 650.0 2 Weeks
200.0mg USD 1050.0 2 Weeks
500.0mg USD 1650.0 2 Weeks
1.0g USD 2650.0 2 Weeks
2.0g USD 3850.0 2 Weeks
5.0g USD 5850.0 2 Weeks
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Related CAS #: 252917-06-9 (free base)   1782235-14-6 (3HCl)   2109414-84-6 (2HCl)   1797989-42-4 (HCl)    

Synonym: CHIR-911; CHIR911; CHIR 911; CT- 99021; CT-99021; CT- 99021; CHIR-73911; CHIR73911; CHIR 73911; GSK 3 inhibitor XVI; GSK 3IXV; CHIR99021; CHIR 99021;

IUPAC/Chemical Name: 6-((2-((4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)pyrimidin-2-yl)amino)ethyl)amino)nicotinonitrile hydrochloride

InChi Key: SCQDMKUZHIGAIB-UHFFFAOYSA-N

InChi Code: InChI=1S/C22H18Cl2N8.ClH/c1-13-10-29-21(31-13)17-12-30-22(32-20(17)16-4-3-15(23)8-18(16)24)27-7-6-26-19-5-2-14(9-25)11-28-19;/h2-5,8,10-12H,6-7H2,1H3,(H,26,28)(H,29,31)(H,27,30,32);1H

SMILES Code: N#CC1=CN=C(NCCNC2=NC=C(C3=NC(C)=CN3)C(C4=CC=C(Cl)C=C4Cl)=N2)C=C1.[H]Cl

Appearance: Solid powder

Purity: >98% (or refer to the Certificate of Analysis)

Shipping Condition: Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.

Storage Condition: Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).

Solubility: Soluble in DMSO

Shelf Life: >2 years if stored properly

Drug Formulation: This drug may be formulated in DMSO

Stock Solution Storage: 0 - 4 C for short term (days to weeks), or -20 C for long term (months).

HS Tariff Code: 2934.99.9001

Product Data:

Biological target: CHIR-99021 monohydrochloride (CT99021 monohydrochloride) is a potent and selective GSK-3α/β inhibitor with IC50s of 10 nM and 6.7 nM.
In vitro activity: Organoids treated with low dose of CHIR 99021 (1μM) showed increased levels of neural progenitor cell (NPC) markers SOX2 (WB, 1.5-fold increase) and PAX6 (IHC, 1.8-fold increase), and of radial glia marker BLBP (WB, 2.1-fold increase). No change was observed for levels of neuroepithelium marker E-cadherin and neuronal markers TUJ1 and DCX. In contrast, organoids treated with 10μM CHIR 99021 showed increased levels of E-cadherin (WB and IHC, 1.8-fold and 92-fold increase) and of intermediate progenitor marker TBR2 (2.9-fold increase), decreased levels of NPC markers SOX2 (WB, 1.9-fold decrease) and PAX6 (IHC, 2-fold decrease), and decreased levels of neuronal marker DCX (IHC, 7.8-fold decrease). This suggests that GSK3β signaling modulates neuroepithelium differentiation in a dose dependent manner. Reference: PLoS One. 2021; 16(5): e0251173. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099055/
In vivo activity: Phosphorylated GSK3β (p-GSK3β, Tyr216) protein levels were higher in ischemic regions of hearts from Lrp5-/- mice than in those from Wt mice (Fig 1A). Total GSK3β protein levels were also increased in the myocardium of Lrp5-/- mice compared to Wt mice. Phosphorylated GSK3β/Total- GSK3β ratios to analyze total myocardial content of activated GSK3β found no differences between genotypes (Fig 1A). CHIR99021 treatment significantly reduced p-GSK3β protein levels in Lrp5-/- mice but not in Wt mice. CHIR99021 treatment reduced p-GSK3β to the same level in both genotypes. Induction of myocardial injury (MI) causes a highly significant increase of p-GSK3β protein levels in Wt and Lrp5-/- mice, strongly suggesting that injury affects the canonical Wnt pathway (Fig 1B). Treatment with CHIR99021 before occlusion significantly reduced p-GSK3β levels in both genotypes (Fig 1C) further demonstrating efficacy of CHIR99021 treatments. Indeed, levels of p-GSK3β protein were significantly reduced after CHIR99021 treatments in damaged hearts leaving the canonical pathway open for activation and for β-catenin interaction with specific transcription factors. Reference: PLoS One. 2019; 14(6): e0218098. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6586285/

Solubility Data

Solvent Max Conc. mg/mL Max Conc. mM
Solubility
DMSO 80.0 159.43
Ethanol 47.0 93.66
Water 7.14 14.23

Preparing Stock Solutions

The following data is based on the product molecular weight 501.8 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Recalculate based on batch purity %
Concentration / Solvent Volume / Mass 1 mg 5 mg 10 mg
1 mM 1.15 mL 5.76 mL 11.51 mL
5 mM 0.23 mL 1.15 mL 2.3 mL
10 mM 0.12 mL 0.58 mL 1.15 mL
50 mM 0.02 mL 0.12 mL 0.23 mL
Formulation protocol: 1. Delepine C, Pham VA, Tsang HWS, Sur M. GSK3ß inhibitor CHIR 99021 modulates cerebral organoid development through dose-dependent regulation of apoptosis, proliferation, differentiation and migration. PLoS One. 2021 May 5;16(5):e0251173. doi: 10.1371/journal.pone.0251173. PMID: 33951093; PMCID: PMC8099055. 2. Lee JS, Chae MK, Kikkawa DO, Lee EJ, Yoon JS. Glycogen Synthase Kinase-3β Mediates Proinflammatory Cytokine Secretion and Adipogenesis in Orbital Fibroblasts from Patients with Graves' Orbitopathy. Invest Ophthalmol Vis Sci. 2020 Jul 1;61(8):51. doi: 10.1167/iovs.61.8.51. PMID: 32735324; PMCID: PMC7426624. 3. Faccidomo S, Holstein SE, Santanam TS, Saunders BL, Swaim KS, Reid GT, O'Neill C, Eastman VR, Hodge CW. Pharmacological inhibition of glycogen synthase kinase 3 increases operant alcohol self-administration in a manner associated with altered pGSK-3β, protein interacting with C kinase and GluA2 protein expression in the reward pathway of male C57BL/6J mice. Behav Pharmacol. 2020 Feb;31(1):15-26. doi: 10.1097/FBP.0000000000000501. PMID: 31503067; PMCID: PMC6954298. 4. Badimon L, Casaní L, Camino-Lopez S, Juan-Babot O, Borrell-Pages M. GSK3β inhibition and canonical Wnt signaling in mice hearts after myocardial ischemic damage. PLoS One. 2019 Jun 20;14(6):e0218098. doi: 10.1371/journal.pone.0218098. PMID: 31220102; PMCID: PMC6586285.
In vitro protocol: 1. Delepine C, Pham VA, Tsang HWS, Sur M. GSK3ß inhibitor CHIR 99021 modulates cerebral organoid development through dose-dependent regulation of apoptosis, proliferation, differentiation and migration. PLoS One. 2021 May 5;16(5):e0251173. doi: 10.1371/journal.pone.0251173. PMID: 33951093; PMCID: PMC8099055. 2. Lee JS, Chae MK, Kikkawa DO, Lee EJ, Yoon JS. Glycogen Synthase Kinase-3β Mediates Proinflammatory Cytokine Secretion and Adipogenesis in Orbital Fibroblasts from Patients with Graves' Orbitopathy. Invest Ophthalmol Vis Sci. 2020 Jul 1;61(8):51. doi: 10.1167/iovs.61.8.51. PMID: 32735324; PMCID: PMC7426624.
In vivo protocol: 1. Faccidomo S, Holstein SE, Santanam TS, Saunders BL, Swaim KS, Reid GT, O'Neill C, Eastman VR, Hodge CW. Pharmacological inhibition of glycogen synthase kinase 3 increases operant alcohol self-administration in a manner associated with altered pGSK-3β, protein interacting with C kinase and GluA2 protein expression in the reward pathway of male C57BL/6J mice. Behav Pharmacol. 2020 Feb;31(1):15-26. doi: 10.1097/FBP.0000000000000501. PMID: 31503067; PMCID: PMC6954298. 2. Badimon L, Casaní L, Camino-Lopez S, Juan-Babot O, Borrell-Pages M. GSK3β inhibition and canonical Wnt signaling in mice hearts after myocardial ischemic damage. PLoS One. 2019 Jun 20;14(6):e0218098. doi: 10.1371/journal.pone.0218098. PMID: 31220102; PMCID: PMC6586285.

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1: Wu Y, Liu F, Liu Y, Liu X, Ai Z, Guo Z, Zhang Y. GSK3 inhibitors CHIR99021 and 6-bromoindirubin-3'-oxime inhibit microRNA maturation in mouse embryonic stem cells. Sci Rep. 2015 Mar 2;5:8666. doi: 10.1038/srep08666. PubMed PMID: 25727520; PubMed Central PMCID: PMC4345320.

2: Bock AS, Leigh ND, Bryda EC. Effect of Gsk3 inhibitor CHIR99021 on aneuploidy levels in rat embryonic stem cells. In Vitro Cell Dev Biol Anim. 2014 Jun;50(6):572-9. doi: 10.1007/s11626-014-9734-5. Epub 2014 Feb 12. PubMed PMID: 24519175; PubMed Central PMCID: PMC4062835.

3: Li C, Zhang S, Lu Y, Zhang Y, Wang E, Cui Z. The roles of Notch3 on the cell proliferation and apoptosis induced by CHIR99021 in NSCLC cell lines: a functional link between Wnt and Notch signaling pathways. PLoS One. 2013 Dec 18;8(12):e84659. doi: 10.1371/journal.pone.0084659. eCollection 2013. PubMed PMID: 24367688; PubMed Central PMCID: PMC3867546.

4: Wu Y, Ai Z, Yao K, Cao L, Du J, Shi X, Guo Z, Zhang Y. CHIR99021 promotes self-renewal of mouse embryonic stem cells by modulation of protein-encoding gene and long intergenic non-coding RNA expression. Exp Cell Res. 2013 Oct 15;319(17):2684-99. doi: 10.1016/j.yexcr.2013.08.027. Epub 2013 Sep 7. PubMed PMID: 24021571.

5: Petkov S, Hyttel P, Niemann H. The small molecule inhibitors PD0325091 and CHIR99021 reduce expression of pluripotency-related genes in putative porcine induced pluripotent stem cells. Cell Reprogram. 2014 Aug;16(4):235-40. doi: 10.1089/cell.2014.0010. Epub 2014 Jun 24. PubMed PMID: 24960205.

6: Ai Z, Shao J, Wu Y, Yu M, Du J, Shi X, Shi X, Zhang Y, Guo Z. CHIR99021 enhances Klf4 Expression through β-Catenin Signaling and miR-7a Regulation in J1 Mouse Embryonic Stem Cells. PLoS One. 2016 Mar 3;11(3):e0150936. doi: 10.1371/journal.pone.0150936. eCollection 2016. PubMed PMID: 26938105; PubMed Central PMCID: PMC4777400.

7: Yin X, Farin HF, van Es JH, Clevers H, Langer R, Karp JM. Niche-independent high-purity cultures of Lgr5+ intestinal stem cells and their progeny. Nat Methods. 2014 Jan;11(1):106-12. doi: 10.1038/nmeth.2737. Epub 2013 Dec 1. PubMed PMID: 24292484; PubMed Central PMCID: PMC3951815.

8: Ye S, Tan L, Yang R, Fang B, Qu S, Schulze EN, Song H, Ying Q, Li P. Pleiotropy of glycogen synthase kinase-3 inhibition by CHIR99021 promotes self-renewal of embryonic stem cells from refractory mouse strains. PLoS One. 2012;7(4):e35892. doi: 10.1371/journal.pone.0035892. Epub 2012 Apr 23. PubMed PMID: 22540008; PubMed Central PMCID: PMC3335080.

9: Wang X, Wei L, Cramer JM, Leibowitz BJ, Judge C, Epperly M, Greenberger J, Wang F, Li L, Stelzner MG, Dunn JC, Martin MG, Lagasse E, Zhang L, Yu J. Pharmacologically blocking p53-dependent apoptosis protects intestinal stem cells and mice from radiation. Sci Rep. 2015 Apr 10;5:8566. doi: 10.1038/srep08566. PubMed PMID: 25858503; PubMed Central PMCID: PMC4392360.

10: Lam AQ, Freedman BS, Morizane R, Lerou PH, Valerius MT, Bonventre JV. Rapid and efficient differentiation of human pluripotent stem cells into intermediate mesoderm that forms tubules expressing kidney proximal tubular markers. J Am Soc Nephrol. 2014 Jun;25(6):1211-25. doi: 10.1681/ASN.2013080831. Epub 2013 Dec 19. PubMed PMID: 24357672; PubMed Central PMCID: PMC4033376.

11: Qin H, Hejna M, Liu Y, Percharde M, Wossidlo M, Blouin L, Durruthy-Durruthy J, Wong P, Qi Z, Yu J, Qi LS, Sebastiano V, Song JS, Ramalho-Santos M. YAP Induces Human Naive Pluripotency. Cell Rep. 2016 Mar 15;14(10):2301-12. doi: 10.1016/j.celrep.2016.02.036. Epub 2016 Mar 3. PubMed PMID: 26947063; PubMed Central PMCID: PMC4807727.

12: Fu Y, Huang C, Xu X, Gu H, Ye Y, Jiang C, Qiu Z, Xie X. Direct reprogramming of mouse fibroblasts into cardiomyocytes with chemical cocktails. Cell Res. 2015 Sep;25(9):1013-24. doi: 10.1038/cr.2015.99. Epub 2015 Aug 21. PubMed PMID: 26292833; PubMed Central PMCID: PMC4559819.

13: Ninomiya H, Mizuno K, Terada R, Miura T, Ohnuma K, Takahashi S, Asashima M, Michiue T. Improved efficiency of definitive endoderm induction from human induced pluripotent stem cells in feeder and serum-free culture system. In Vitro Cell Dev Biol Anim. 2015 Jan;51(1):1-8. doi: 10.1007/s11626-014-9801-y. Epub 2014 Aug 15. PubMed PMID: 25124871.

14: Cheng T, Zhai K, Chang Y, Yao G, He J, Wang F, Kong H, Xin H, Wang H, Jin M, Gong B, Gu L, Yang Z, Wu Y, Ji G, Sun Y. CHIR99021 combined with retinoic acid promotes the differentiation of primordial germ cells from human embryonic stem cells. Oncotarget. 2017 Jan 31;8(5):7814-7826. doi: 10.18632/oncotarget.13958. PubMed PMID: 27999196; PubMed Central PMCID: PMC5352363.

15: Cheng L, Hu W, Qiu B, Zhao J, Yu Y, Guan W, Wang M, Yang W, Pei G. Generation of neural progenitor cells by chemical cocktails and hypoxia. Cell Res. 2014 Jun;24(6):665-79. doi: 10.1038/cr.2014.32. Epub 2014 Mar 18. Erratum in: Cell Res. 2015 May;25(5):645-6. PubMed PMID: 24638034; PubMed Central PMCID: PMC4042166.

16: Besing RC, Paul JR, Hablitz LM, Rogers CO, Johnson RL, Young ME, Gamble KL. Circadian rhythmicity of active GSK3 isoforms modulates molecular clock gene rhythms in the suprachiasmatic nucleus. J Biol Rhythms. 2015 Apr;30(2):155-60. doi: 10.1177/0748730415573167. Epub 2015 Feb 27. PubMed PMID: 25724980; PubMed Central PMCID: PMC4586074.

17: Roccio M, Hahnewald S, Perny M, Senn P. Cell cycle reactivation of cochlear progenitor cells in neonatal FUCCI mice by a GSK3 small molecule inhibitor. Sci Rep. 2015 Dec 8;5:17886. doi: 10.1038/srep17886. PubMed PMID: 26643939; PubMed Central PMCID: PMC4672274.

18: van den Berg CW, Elliott DA, Braam SR, Mummery CL, Davis RP. Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes Under Defined Conditions. Methods Mol Biol. 2016;1353:163-80. doi: 10.1007/7651_2014_178. PubMed PMID: 25626427.

19: Briolotti P, Chaloin L, Balaguer P, Da Silva F, Tománková V, Pascussi JM, Duret C, Fabre JM, Ramos J, Klieber S, Maurel P, Daujat-Chavanieu M, Gerbal-Chaloin S. Analysis of Glycogen Synthase Kinase Inhibitors That Regulate Cytochrome P450 Expression in Primary Human Hepatocytes by Activation of β-Catenin, Aryl Hydrocarbon Receptor and Pregnane X Receptor Signaling. Toxicol Sci. 2015 Nov;148(1):261-75. doi: 10.1093/toxsci/kfv177. Epub 2015 Aug 10. PubMed PMID: 26259606.

20: Preston GC, Sinclair LV, Kaskar A, Hukelmann JL, Navarro MN, Ferrero I, MacDonald HR, Cowling VH, Cantrell DA. Single cell tuning of Myc expression by antigen receptor signal strength and interleukin-2 in T lymphocytes. EMBO J. 2015 Aug 4;34(15):2008-24. doi: 10.15252/embj.201490252. Epub 2015 Jul 1. PubMed PMID: 26136212; PubMed Central PMCID: PMC4551349.