Tofacitinib (CP-690550) Citrate

For research use only.

Licensed by Pfizer Catalog No.S5001 Synonyms: Tasocitinib Citrate

147 publications

Tofacitinib (CP-690550) Citrate Chemical Structure

CAS No. 540737-29-9

Tofacitinib citrate (CP-690550, Tasocitinib) is a novel inhibitor of JAK with IC50 of 1 nM, 20 nM and 112 nM against JAK3, JAK2, and JAK1, respectively. Tofacitinib citrate has anti-infection activity.

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Selleck's Tofacitinib (CP-690550) Citrate has been cited by 147 publications

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Biological Activity

Description Tofacitinib citrate (CP-690550, Tasocitinib) is a novel inhibitor of JAK with IC50 of 1 nM, 20 nM and 112 nM against JAK3, JAK2, and JAK1, respectively. Tofacitinib citrate has anti-infection activity.
Targets
JAK3 [1]
(Cell-free assay)
JAK2 [4]
(Cell-free assay)
1 nM 20 nM
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 JAK2VV617F, whereas a lesser effect is observed for cells carrying wild-type JAK2. This activity is coupled with the inhibition of phosphorylation of the key JAK2V617F-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]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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T-cells M3\ENmZ2dmO2aX;uJIF{e2G7 NEL5NY42OCC2bzCzNFAhdk1? NVeyVIRwOSCqch?= NVX5cnNHUW6qaXLpeIlwdiCxZjDKRWsyN0qDS{OgbY4hcHWvYX6gR2Q1KHCxc3n0bZZmKFRiY3XscJMh[XO|ZYPz[YQh[XNiaX7obYJqfGmxbjDv[kBKVDJvc4TpcZVt[XSnZDDTWGFVPSCyaH;zdIhwenmuYYTpc44h[XRiVInyMVY6PSC{ZYPp[JVmKGG2IEWwJJRwKDNyMDDuUUBxemWrbnP1ZoF1\WRiZn;yJFEhcHJiZn;scI94\WRiYomgTWwuOiC|dHnteYxifGmxbjDt[YF{fXKnZDDh[pRmeiB|MDDtbY5{KGK7IHntcZVvd2Kub4SgZY5idHm|aYO= NFG1PFA9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OUi1NlA3QCd-Mkm4OVIxPjh:L3G+
T-cells MXvGeY5kfGmxbjDhd5NigQ>? NHjWVlQ,OzByIH7N MYKxJIhz M1L1RmlvcGmkaYTpc44hd2ZiSlHLNU9LSUt{L2TZT|IhcW5iaIXtZY4hS0R2IIDvd4l1cX[nIGSgZ4VtdHNiYYPz[ZN{\WRiYYOgbY5pcWKrdHnvckBw\iCLTE[td5RqdXWuYYTl[EBUXEGWMzDwbI9{eGixconsZZRqd25iYYSgQlMxOCCwTTDwdoVqdmO3YnH0[YQh\m:{IEGgbJIh\m:ubH;3[YQh[nliSVytOkB{fGmvdXzheIlwdiCvZXHzeZJm\CCjZoTldkA{OCCvaX7zJIJ6KGmvbYXuc4Jtd3RiYX7hcJl{cXN? NYLxe4pQRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkm4OVIxPjhpPkK5PFUzODZ6PD;hQi=>
TALL-1 Ml7LSpVv[3Srb36gZZN{[Xl? MoPiNUB2VQ>? NHfxcVM{KGi{cx?= NGHmb3pKdmirYnn0bY9vKG:oIFrBT|MhcW5iaIXtZY4hXEGOTD2xJINmdGy|IHHzd4V{e2WmIHHzJIlvcGmkaYTpc44hd2ZiSVytNkBqdmS3Y3XkJHNVSVR3IIDoc5NxcG:{eXzheIlwdiCjdDCxJJVOKHC{ZXnuZ5Vj[XSnZDDmc5IhOyCqcoOg[o9tdG:5ZXSgZpkhUUxvMjDpcoR2[3Srb36gcYVie3W{ZXSgZYZ1\XJiM{CgcYlveyCkeTDpcY12dm:kbH;0eIlv\w>? MlLqQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZ{NUi1NlEoRjJ4MkW4OVIyRC:jPh?=
BA/F3 NX\3WJl3TnWwY4Tpc44h[XO|YYm= MYWzNFAhdk1? MkjlN{BpenN? MkHwTY5pcWKrdHnvckBw\iCMQVuzJEh2dmuwb4fuJI9zcWerbjmg[ZhxemW|c3XkJIlvKG2xdYPlJGJCN0Z|IHPlcIx{KGG|c3Xzd4VlKGG|IILl[JVkfGmxbjDv[kBUXEGWNTDwbI9{eGixconsZZRqd25iYYSgN|AxKG6PIHHmeIVzKDNiaILzJIJ6KGmvbYXuc4Jtd3S2aX7nJIFv[Wy7c3nz NXvURXU5RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMk[yOVg2OjFpPkK2NlU5PTJzPD;hQi=>
OCL-AML5 M3m4XWZ2dmO2aX;uJIF{e2G7 NHvoXHgyKHWP NGPtR40{KGi{cx?= NG\jXHZKdmirYnn0bY9vKG:oIFrBT|IhcW5iaIXtZY4hV0OOLVHNUFUh[2WubIOgZZN{\XO|ZXSgZZMhcW6qaXLpeIlwdiCxZjDHUU1EW0ZiaX7keYNm\CCVVFHUOUBxcG:|cHjvdplt[XSrb36gZZQhOSC3TTDwdoVqdmO3YnH0[YQh\m:{IEOgbJJ{KG[xbHzve4VlKGK7IFfNMWNUTiCrbnT1Z5Rqd25ibXXhd5Vz\WRiYX\0[ZIhOzBibXnud{BjgSCrbX31co9jdG:2dHnu[y=> NEKxWJY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NkK1PFUzOSd-Mk[yOVg2OjF:L3G+
BA/F3 MlzaRY51cXC{b3zp[oVz[XSrdnWgZZN{[Xl? NIfxSlA4OiCqcoO= NVTUeIRwSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBud3W|ZTDCRU9HOyClZXzsd{BmgHC{ZYPzbY5oKFSHTD3KRWs{KGGodHXyJFczKGi{czDifUBk\WyuIITpeIVzKGeubzDhd5NigQ>? M3rqZ|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ4MkW4OVIyLz5{NkK1PFUzOTxxYU6=
BA/F3 MkXSSpVv[3Srb36gZZN{[Xl? MmftNUB2VQ>? MUKzJIhzew>? M2LpZWlvcGmkaYTpc44hd2ZiSlHLN{ApfW6tbn;3ckBwemmpaX6pJIV5eHKnc4Pl[EBqdiCvb4Xz[UBDSS:IMzDj[YxteyCjdDCxJJVOKHC{ZXnuZ5Vj[XSnZDDmc5IhOyCqcoOg[o9tdG:5ZXSgZpkheHWubHTve44hf2m2aDDzeJJmeHSjdnnkbY4h[mWjZIOgZpkhcW2vdX7vZoxwfHSrbnegZY5idHm|aYO= M3TLN|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ4MkW4OVIyLz5{NkK1PFUzOTxxYU6=
Huh7 NEDCPJZHfW6ldHnvckBie3OjeR?= NVT1fnVlOTBidV2= NGHvcJQ{OCCvaX7z MXXJcohq[mm2aX;uJI9nKFS7a{KgbY4hcHWvYX6gTJVpPyClZXzsd{Bie3Onc4Pl[EBieyC{ZXT1Z5Rqd25ib3[gTWZP[WyyaHG1MYlv\HWlZXSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKGG2IEGwJJVOKHC{ZT3pcoN2[mG2ZXSg[o9zKDNyIH3pcpMh[mWob4LlJGlHVmGucHjhOUB{fGmvdXzheIlwdiCob4KgN|AhdWmwczDtbY5{KGK7IHntcZVvd2Kub4T0bY5o MoHyQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjZ{M{GxOVkoRjJ4MkOxNVU6RC:jPh?=
Huh7 MX\GeY5kfGmxbjDhd5NigQ>? MmqyNVAhfU1? NFLIPZQ{OCCvaX7z MoTVTY5pcWKrdHnvckBw\iCWeXuyJIlvKGi3bXHuJGh2cDdiY3XscJMh[XO|ZYPz[YQh[XNicnXkeYN1cW:wIH;mJIJie2GuIHzleoVtKFOWQWSzJJBpd3OyaH;yfYxifGmxbjDheEAyOCC3TTDh[pRmeiB|MDDtbY5{KGK7IHntcZVvd2Kub4T0bY5o NIr3SVU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NkKzNVE2QSd-Mk[yN|EyPTl:L3G+
Huh7 NIfGXHRHfW6ldHnvckBie3OjeR?= M{nDWFEhfU1? MlzUN|AhdWmwcx?= MYDJcohq[mm2aX;uJI9nKFS7a{KgbY4hcHWvYX6gTJVpPyClZXzsd{Bie3Onc4Pl[EBieyC{ZXT1Z5Rqd25ib3[gTWZP[WyyaHG1MYlv\HWlZXSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKGG2IEGgeW0heHKnLXnuZ5Vj[XSnZDDmc5IhOzBibXnud{Bj\W[xcnWgTWZP[WyyaHG1JJN1cW23bHH0bY9vKG[xcjCzNEBucW6|IH3pcpMh[nliaX3teY5w[myxdITpcoc> NFu2Zlg9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NkKzNVE2QSd-Mk[yN|EyPTl:L3G+

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
JAK3 / STAT5 / p-STAT5; 

PubMed: 27732937     


The following cell lines: EBV-negative B cell line (BJAB), EBV-transformed lymphoblastoid cell line (LCL), EBV-negative T cell lines (Jurkat and MOLT4), EBV-positive T cell lines (SNT15 and SNT16), EBV-negative NK cell line (KHYG1), and EBV-positive NK cell lines (KAI3 and SNK6), were treated without (−) or with (+) 1 μM tofacitinib for 24 h and cell lysates were then immunoblotted for the indicated proteins.

LMP1 / EBNA1 / BZLF1; 

PubMed: 27732937     


SNT13, SNT15, SNT16, KAI3 and SNK6 cell lines were treated with 5 μM tofacitinib for 48 h, and cell lysates were immunoblotted for LMP1, EBNA1 and BZLF1. Actin was blotted as a loading control.

JAK3 / p-JAK3 / STAT3; 

PubMed: 26082451     


CTCL cell lines were subjected to western blot (tofacitinib 10, 100, 200 nM) (G) after 48 hours of treatment.

27732937 26082451
Growth inhibition assay
Cell number; 

PubMed: 27732937     


BJAB, LCL, Jurkat, SNT15, KHYG1 and KAI3 cells were treated with the indicated concentrations of tofacitinib, and viable cells were counted at the indicated times using the trypan blue exclusion test. Values are means ± SE of the results from triplicate experiments. *P < 0.05 as compared with DMSO-treated cells.

27732937
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 JAK2V617F-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]

Protocol

Kinase Assay:

[1]

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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 Research:

[2]

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  • Cell lines: FDCP-EpoR JAK2WT and JAK2V617F 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 JAK2WT and JAK2V617F 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.


    (Only for Reference)
Animal Research:

[2]

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  • Animal Models: Mauritius-origin adult cynomolgus monkeys
  • Dosages: 10, 30 mg/kg/d
  • Administration: Oral gavage
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 100 mg/mL warmed (198.21 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
0.5% methylcellulose
For best results, use promptly after mixing.
30 mg/mL

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 504.49
Formula

C16H20N6O.C6H8O7

CAS No. 540737-29-9
Storage powder
in solvent
Synonyms Tasocitinib Citrate
Smiles CC1CCN(CC1N(C)C2=NC=NC3=C2C=CN3)C(=O)CC#N.C(C(=O)O)C(CC(=O)O)(C(=O)O)O

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 ul Number of animals
Step 2: Enter the in vivo formulation ()
% DMSO % % Tween 80 % ddH2O
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Bio Calculators

Molarity Calculator

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

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*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

Dilution Calculator

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration (start) x Volume (start) = Concentration (final) x Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

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    V2

* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / COA (available online).

The Serial Dilution Calculator Equation

  • Serial Dilutions

  • Computed Result

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
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Molecular Weight Calculator

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Total Molecular Weight: g/mol

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (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.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04496960 Not yet recruiting Drug: tofacitinib|Other: Placebo Sjogren''s Syndrome National Institute of Dental and Craniofacial Research (NIDCR)|National Institutes of Health Clinical Center (CC) January 5 2021 Phase 1|Phase 2
NCT04624230 Not yet recruiting Drug: tofacitinib Ulcerative Colitis Pfizer November 20 2020 Phase 3
NCT04468425 Not yet recruiting Drug: Tofacitinib Citrate Healthy Subjects TWi Biotechnology Inc. November 11 2020 Phase 1
NCT04424303 Not yet recruiting Drug: Tofacitinib Ulcerative Colitis Pfizer September 30 2020 --
NCT04576000 Recruiting Drug: Tofacitinib Ulcerative Colitis Robarts Clinical Trials Inc. August 25 2020 --
NCT04141904 Suspended Drug: Tofacitinib 5 MG [Xeljanz]|Other: Placebo Depression|Inflammation University of Oxford|Wellcome Trust|Medical Research Council February 10 2020 Phase 1|Phase 2

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

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

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Frequently Asked Questions

  • Question 1:

    What is the difference between the two products (S5001, S2789)?

  • Answer:

    Tofacitinib (S2789) is the base form of Tofacitinib citrate (S5001). The biological activity of these two compound are same. S5001 is better than S2789 for Oral gavage.

JAK Signaling Pathway Map

JAK Inhibitors with Unique Features

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID