Tofacitinib citrate (CP-690550 citrate) | Licensed by Pfizer

Validation & Quality Control

Product Citations(5)

J Exp Med, 2012, 209(2), 259-273

Blood, 2011, 118(14), 3911-3921

J Immunol, 2012, 189(6), 2784-2792

Int Immunol, 2011, 23(11), 701-712

Biochem Biophy Res Co, 2010, 402(3), 500-506

Customer Reviews(4) in vivo invitation

Quality Control & MSDS

Related Compound Libraries

Tofacitinib citrate (CP-690550 citrate) is available in the following compound libraries:

Chemical Library (96-well) Inhibitor Library (96-well) Kinase Inhibitor Library (96-well) Pfizer Licensed Library Tyrosine Kinase Inhibitor Library (96-Well)

Product Information

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Research Area
Research Area
Tofacitinib citrate (CP-690550 citrate) Mechanism
Tofacitinib citrate (CP-690550 citrate) Mechanism
The kinase domain of TYK2 consists of a small N-terminal domain and a larger C-terminal domain. The two domains are connected by a flexible hinge region. The substrate-binding site resides between the N- and the C-lobes. This hydrophobic cleft is competent in binding the adenine base of ATP, and the activation loop is expelled from the ATP-binding cavity. ^[1]
The crystal structure shows that Tofacitinib binds in the ATP-binding cavity of TYK2, and forms polar or non-polar interactions with a series of residues lining the substrate-binding cleft. The pyrrolopyrimidine ring is situated against the hinge region, and forms two hydrogen bonds with the main chain atoms of Glu979 and Val981. Besides, a hydrogen bond is formed between the N19 atom of the pyrrolopyrimidine ring and a water molecule. The piperidine ring is located in the TYK2 binding cavity, forming hydrophobic interactions with Leu903, Met978, Val911, Ser985, Leu1030 and Gly1040. Additionally, the methyl group of the piperidine ring is ideally situated in an underlying pocket at the binding cavity of TYK2. The terminal cyano-group packs against the glycine-rich loop, and forms polar interactions with main chain atoms spanning this flexible loop, including Gly909, Lys910 and Val911. Finally, Arg901 from the base of the glycine-rich loop and Tyr980 from the hinge produces a novel interaction by formation of a hydrogen bond. Such interaction stabilizes the hinge region for inhibitor binding. ^[1]
References
[1] Chrencik JE, et al. J Mol Biol. 2010, 400(3), 413-433.
Dr. Antonino Maria Sparta demonstrates a breakthrough in leukemia research by utilizing two inhibitors produced by Selleck Chemicals.

Cat.No.	Product Name	Solubility (25°C)
S1378	Ruxolitinib (INCB018424)	<1 mg/mL	61 mg/mL	61 mg/mL
S5001	Tofacitinib citrate (CP-690550 citrate)	<1 mg/mL	101 mg/mL	<1 mg/mL
S2736	TG101348 (SAR302503)	<1 mg/mL	105 mg/mL	21 mg/mL
S2789	Tofacitinib (CP-690550, Tasocitinib)	<1 mg/mL	63 mg/mL	<1 mg/mL
S2162	AZD1480	<1 mg/mL	70 mg/mL	5 mg/mL
S1143	AG-490	<1 mg/mL	59 mg/mL	6 mg/mL
S2219	Cyt387	<1 mg/mL	83 mg/mL	<1 mg/mL
S2686	NVP-BSK805 2HCl	3 mg/mL	113 mg/mL	15 mg/mL
S2692	TG101209	<1 mg/mL	102 mg/mL	<1 mg/mL
S2851	Baricitinib (LY3009104, INCB28050)	<1 mg/mL	74 mg/mL	<1 mg/mL
S2796	WP1066	<1 mg/mL	72 mg/mL	<1 mg/mL
S1134	AT9283	<1 mg/mL	76 mg/mL	38 mg/mL
S2806	CEP33779	<1 mg/mL	93 mg/mL	<1 mg/mL
S2214	AZ 960	<1 mg/mL	71 mg/mL	3 mg/mL
S2867	WHI-P154	<1 mg/mL	75 mg/mL	<1 mg/mL
S2179	LY2784544	<1 mg/mL	94 mg/mL	9 mg/mL
S2902	S-Ruxolitinib	5 mg/mL	61 mg/mL	61 mg/mL
S8004	ZM 39923 HCl	<1 mg/mL	30 mg/mL	8 mg/mL

Product Description

Biological Activity Protocol Chemical Information Customer Reviews Product Citations Tech Support & FAQs

Biological Activity

Description	Tofacitinib citrate (CP-690550) is a potent JAK3 inhibitor with IC50 of 1 nM.
Targets	JAK3
IC50	1 nM ^[1]
In vitro	Tofacitinib citrate inhibits IL-2-mediated human T cell blast proliferation and IL-15-induced CD69 expression with IC50 of 11 nM and 48 nM, respectively. Tofacitinib citrate prevents mixed lymphocyte reaction with IC50 of 87 nM. Tofacitinib citrate treatment of murine factor-dependent cell Patersen–erythropoietin receptor (FDCP-EpoR) cells harboring human wild-type or V617F JAK2 leads to prevention of cell proliferation with IC50 of 2.1 µM and 0.25 µM, respectively. Tofacitinib citrate inhibits interleukin-6-induced phosphorylation of STAT1 and STAT3 with IC50 of 23 nM and 77 nM, respectively. Moreover, Tofacitinib citrate generates a significant pro-apoptotic effect on murine FDCP-EpoR cells carrying JAK2V^V617F, whereas a lesser effect is observed for cells carrying wild-type JAK2. This activity is coupled with the inhibition of phosphorylation of the key JAK2^V617F-dependent downstream signaling effectors signal transducer and activator of transcription (STAT)3, STAT5, and v-akt murine thymoma viral oncogene homolog (AKT). ^[2] Additionally, Tofacitinib citrate prevents IL-15-induced CD69 expression in human and cynomolgus monkey NK and CD8+ T cells in vitro. ^[3]
In vivo	Tofacitinib citrate decrease a delayed-type hyper-sensitivity response and extended cardiac allograft survival in murine models. Furthermore, Tofacitinib citrate treatment of ex-vivo-expanded erythroid progenitors from JAK2^V617F-positive PV patients results in specific, antiproliferative (IC50 = 0.2 μM) and pro-apoptotic activity. In contrast, expanded progenitors from healthy controls are less sensitive to Tofacitinib citrate in proliferation (IC50 > 1.0 μM), and apoptosis assays.^[2] During 2 weeks of Tofacitinib citrate dosing at 10 and 30 mg/kg/d, a significant, time-dependent decrease in NK cell numbers relative to vehicle treatment is observed. Effector memory CD8+ cell numbers in the Tofacitinib citrate-treated group are 55% less than those observed in animals treated with vehicle.^[3]
Clinical Trials	Tofacitinib citrate is under a Phase II clinical trial in the treatment of chronic plaque psoriasis.
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Enzyme assays	The JAK1, JAK2, and JAK3 kinase assays utilize a protein expressed in baculovirus-infected SF9 cells (a fusion protein of GST and the catalytic domain of human JAK enzyme) purified by affinity chromatography on glutathione−Sepharose. The substrate for the reaction is polyglutamic acid-tyrosine [PGT (4:1)], coated onto Nunc Maxi Sorp plates at 100 μg/mL overnight at 37 °C. The plates are washed three times, and JAK enzyme is added to the wells, which contained 100 μL of kinase buffer (50 mM HEPES, pH 7.3, 125 mM NaCl, 24 mM MgCl₂) + ATP + 1 mM sodium orthovanadate). For Tofacitinib citrate, it is also added for kinase assay at different doses. After incubation at room temperature for 30 min, the plates are washed three times. The level of phosphorylated tyrosine in a given well is determined by standard ELISA assay utilizing an anti-phosphotyrosine antibody.

Enzyme assays

The JAK1, JAK2, and JAK3 kinase assays utilize a protein expressed in baculovirus-infected SF9 cells (a fusion protein of GST and the catalytic domain of human JAK enzyme) purified by affinity chromatography on glutathione−Sepharose. The substrate for the reaction is polyglutamic acid-tyrosine [PGT (4:1)], coated onto Nunc Maxi Sorp plates at 100 μg/mL overnight at 37 °C. The plates are washed three times, and JAK enzyme is added to the wells, which contained 100 μL of kinase buffer (50 mM HEPES, pH 7.3, 125 mM NaCl, 24 mM MgCl₂) + ATP + 1 mM sodium orthovanadate). For Tofacitinib citrate, it is also added for kinase assay at different doses. After incubation at room temperature for 30 min, the plates are washed three times. The level of phosphorylated tyrosine in a given well is determined by standard ELISA assay utilizing an anti-phosphotyrosine antibody.

Cell Assay: ^[2]

Cell lines	FDCP-EpoR JAK2^WT and JAK2^V617F cell lines
Concentrations	0-4 μM
Incubation Time	72 hours
Method	Determination of growth inhibition by Tofacitinib citrate is performed using identical culture conditions for both FDCP-EpoR JAK2^WT and JAK2^V617F cell lines. Briefly, 1 × 10⁵ cells/mL are cultured in 96-well flat-bottom plates at 37 °C in a humidified 5% CO₂ atmosphere using RPMI 1640 supplemented with 1.25% FCS, and 5% WEHI supernatant. Decreased FCS concentration is necessary to prevent binding between Tofacitinib citrate and serum proteins. Growth inhibition assays are terminated by addition of 20 μL CellTiter96 One Solution Reagent. Flat-bottom plates are incubated for an additional 3 hours for MTT assay. Absorbance is determined at 595 nm on a BioTek Synergy-HT microplate reader. Results are the average standard deviation of three independent determinations.

Animal Study: ^[2]

Animal Models	Mauritius-origin adult cynomolgus monkeys
Formulation	0.5% methylcellulose in distilled water
Dosages	10, 30 mg/kg/d
Administration	Oral gavage

1

References

Chemical Information

Download Tofacitinib citrate (CP-690550 citrate) SDF

Molecular Weight (MW)	504.49
Formula	C₁₆H₂₀N₆O.C₆H₈O₇
CAS No.	540737-29-9
Synonyms	N/A

Solubility (25°C) DMSO 101 mg/mL Water <1 mg/mL Ethanol <1 mg/mL
Storage	2 years -20°CPowder
	2 weeks4°Cin DMSO
	6 months-80°Cin DMSO

Chemical Name	3-((3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile citrate

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Research Area

Customer Reviews (4)

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Rating

Source

Dr. Akihiko Yoshimura of Akihiko Yoshimura. Tofacitinib citrate (CP-690550 citrate) purchased from Selleck

Method

real-time PCR, ELISA, Western blot

Cell Lines

hind paws cells of mice

Concentrations

15mg/kg/day

Incubation Time

2 weeks

Results

STAT3-inhibition antagonizes arthritis effects in vivo.

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Rating

Source

Dr. Akihiko Yoshimura of Akihiko Yoshimura. Tofacitinib citrate (CP-690550 citrate) purchased from Selleck

Method

Histopathology, ﬂuorescent immunohistochemistry

Cell Lines

DBA/J1 male mice

Concentrations

15mg/kg/day

Incubation Time

2 weeks

Results

Joint destruction seen in the CIA model was rescued by CP690,550 treatment

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Rating

Source

Dr. Akihiko Yoshimura of Akihiko Yoshimura. Tofacitinib citrate (CP-690550 citrate) purchased from Selleck

Method

Immunoblotting, real-time PCR

Cell Lines

MC3T3-E1 cells

Concentrations

0-1000 nM

Incubation Time

24 h

Results

The STAT3 inhibitor CP690,550 inhibits arthritis in vivo and the expression of IL-6 cytokine family in vitro.

Click to enlarge

Rating

Source

Biochem Biophy Res Commun, 2010, 402, 500–506. Tofacitinib citrate (CP-690550 citrate) purchased from Selleck

Method

RT-PCR, ELISA, TTC staining, Neurological assessment

Cell Lines

γδT cells and activated memory T cells

Concentrations

10 μM

Incubation Time

Results

CP-690550 efﬁciently suppressed IL-17 production by γδT cells as well as memory CD4+T cells.We investigated the effect of CP-690550 in ischemic brain damage in mice. CP-690550 or vehicle was injected intra-peritoneally after I/R. The degree of CBF reduction was similar in the two groups (Fig. B). As shown in Fig. 1C, the functional neurological deﬁcit induced by I/R surgery was signiﬁcantly improved in mice receiving CP-690550 7 days after MCAO compared to control mice. Infarct size was smaller in mice treated with CP-690550 3 days after MCAO than in untreated mice, as shown in Fig. D

Product Citations (5)

Genetic resistance to JAK2 enzymatic inhibitors is overcome by HSP90 inhibition. [Weigert O, et al. J Exp Med 2012;209(2), 259-273]
PubMed: 22271575
FERM domain mutations induce gain of function in JAK3 in adult T-cell leukemia/lymphoma. [Elliott NE, et al. Blood 2011;118(14), 3911-3921]
PubMed: 21821710
Inhibition of JAKs in macrophages increases lipopolysaccharide-induced cytokine production by blocking IL-10-mediated feedback. [Pattison MJ, et al. J Immunol 2012;189(6), 2784-2792]
PubMed: 22904308

IL-1b and TNFa-initiated IL-6-STAT3 pathway is critical in mediating inflammatory cytokines and RANKL expression in inflammatory arthritis. [Mori T, et al. Int Immunol 2011;23(11), 701-712]
PubMed: 21937456
Therapeutic effect of IL-12/23 and their signaling pathway blockade on brain ischemia model. [Konoeda F, et al. Biochem Biophy Res Co 2010;402(3), 500-506]
PubMed: 20965150

Tech Support & FAQs

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

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AT9283

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AG-490
AG-490 (Tyrphostin B42) is an inhibitor of EGFR, ErbB2 and JAK2 with IC50 of 0.1 μM, 13.5 μM, and ~10 μM, respectively.
AZD1480

AZD1480 is a novel ATP-competitive inhibitor of JAK1 and JAK2 with IC50 of 1.3 nM and 0.26 nM, respectively.
LY2784544

LY2784544 is selective to V617F mutant JAK2 with IC50 of 55 nM in Ba/F3 cells.
AZ 960

AZ 960 is a novel ATP competitive JAK2 inhibitor with IC50 and K_i of <3 nM and 0.45 nM, respectively.
Cyt387
CYT387 is an ATP-competitive inhibitor of JAK1 and JAK2 with IC50 of 11 nM and 18 nM, respectively.
NVP-BSK805 2HCl

NVP-BSK805 is a potent and selective ATP-competitive JAK2 inhibitor with IC50 of 0.5 nM.
TG101209

TG101209 is a potent inhibitor of JAK2, Flt3 and RET with IC50 of 6 nM, 25 nM and 17 nM, respectively.
TG101348 (SAR302503)

TG-101348 (SAR302503) is a selective inhibitor of JAK2 with IC50 of 3 nM.
Tofacitinib (CP-690550, Tasocitinib)

CP-690550 is a novel inhibitor of JAK3 with IC50 of 1 nM.

JAK/STAT Pathway Products

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Tofacitinib citrate (CP-690550 citrate) Licensed by Pfizer

Validation & Quality Control

Product Citations(5)

Customer Reviews(4) in vivo invitation

Quality Control & MSDS

Related Compound Libraries

Tofacitinib citrate (CP-690550 citrate) is available in the following compound libraries:

Product Information

Compare JAK Inhibitors

Research Area

Tofacitinib citrate (CP-690550 citrate) Mechanism

Product Description

Biological Activity

Protocol(Only for Reference)

Kinase Assay: ^[1]

Cell Assay: ^[2]

Animal Study: ^[2]

References

Chemical Information

Research Area

Customer Reviews (4)

Product Citations (5)

Tech Support & FAQs

If you have any other enquiries, please leave a message.

Related JAK Inhibitors

JAK/STAT Pathway Products

Recently Viewed Items

Cat.No.	Product Name	Solubility (25°C)
Cat.No.	Product Name	Water	DMSO	Ethanol
S1378	Ruxolitinib (INCB018424)	<1 mg/mL	61 mg/mL	61 mg/mL
S5001	Tofacitinib citrate (CP-690550 citrate)	<1 mg/mL	101 mg/mL	<1 mg/mL
S2736	TG101348 (SAR302503)	<1 mg/mL	105 mg/mL	21 mg/mL
S2789	Tofacitinib (CP-690550, Tasocitinib)	<1 mg/mL	63 mg/mL	<1 mg/mL
S2162	AZD1480	<1 mg/mL	70 mg/mL	5 mg/mL
S1143	AG-490	<1 mg/mL	59 mg/mL	6 mg/mL
S2219	Cyt387	<1 mg/mL	83 mg/mL	<1 mg/mL
S2686	NVP-BSK805 2HCl	3 mg/mL	113 mg/mL	15 mg/mL
S2692	TG101209	<1 mg/mL	102 mg/mL	<1 mg/mL
S2851	Baricitinib (LY3009104, INCB28050)	<1 mg/mL	74 mg/mL	<1 mg/mL
S2796	WP1066	<1 mg/mL	72 mg/mL	<1 mg/mL
S1134	AT9283	<1 mg/mL	76 mg/mL	38 mg/mL
S2806	CEP33779	<1 mg/mL	93 mg/mL	<1 mg/mL
S2214	AZ 960	<1 mg/mL	71 mg/mL	3 mg/mL
S2867	WHI-P154	<1 mg/mL	75 mg/mL	<1 mg/mL
S2179	LY2784544	<1 mg/mL	94 mg/mL	9 mg/mL
S2902	S-Ruxolitinib	5 mg/mL	61 mg/mL	61 mg/mL
S8004	ZM 39923 HCl	<1 mg/mL	30 mg/mL	8 mg/mL

Choose Your Country or Region

Product Categories

Tofacitinib citrate (CP-690550 citrate) Licensed by Pfizer

Validation & Quality Control

Product Citations(5)

Customer Reviews(4) in vivo invitation

Quality Control & MSDS

Related Compound Libraries

Tofacitinib citrate (CP-690550 citrate) is available in the following compound libraries:

Product Information

Compare JAK Inhibitors

Research Area

Tofacitinib citrate (CP-690550 citrate) Mechanism

Product Description

Biological Activity

Protocol(Only for Reference)

Kinase Assay: [1]

Cell Assay: [2]

Animal Study: [2]

References

Chemical Information

Research Area

Customer Reviews (4)

Product Citations (5)

Tech Support & FAQs

If you have any other enquiries, please leave a message.

Related JAK Inhibitors

JAK/STAT Pathway Products

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Kinase Assay: ^[1]

Cell Assay: ^[2]

Animal Study: ^[2]