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Tofacitinib (CP-690550) JAK Inhibiteur

Name: Tofacitinib (CP-690550)
Brand: Selleck Chemicals
SKU: S2789
Rating: 5 (192 reviews)

Réf. Catalogue: S2789

Tofacitinib est un nouvel inhibiteur de JAK3 avec une IC50 de 1 nM dans les essais sans cellules, 20 à 100 fois moins puissant contre JAK2 et JAK1. Ce composé inhibe l'expression des protéines antiapoptotiques BCL-A1 et BCL-XL dans les cellules dendritiques plasmacytoïdes humaines (PDC) et induit l'apoptose des PDC.

Tofacitinib (CP-690550) JAK Inhibiteur Chemical Structure

Structure chimique

Poids moléculaire: 312.37

Aller à

Contrôle Qualité (Quality Control)

Lot : Pureté : 99.95%

99.95

Cibles apparentées	EGFR STAT Pim
Autre JAK Inhibiteurs	BMS-986165 (Deucravacitinib) AZD1480 WP1066 Momelotinib (CYT387) Filgotinib (GLPG0634) AT9283 Gandotinib (LY2784544) Pacritinib (SB1518) NVP-BSK805 2HCl TG101209

Culture cellulaire, traitement et concentration de travail
(Cell Culture, Treatment & Working Concentration)

Lignées cellulaires	Type d'essai	Concentration	Temps d'incubation	Description de l'activité	PMID
T cells	Function Assay	0.1/0.3/1 μM	24 h	disrupts γc-chain cytokine signaling	21383241
CD4+ T	Function Assay	100 nM	24/48 h	abrogates CD3-induced phosphorylation of STATs	21884580
CD4+ T	Growth Inhibition Assay	100/500 nM	48 h	inhibits anti-CD3-induced CD4+ T cell proliferation	21884580
CD4+ T	Function Assay	20/100/500 nM	48 h	inhibits the levels of IL-4, IFN-γ, IL-17A and IL-22	21884580
Sf9	Function assay		30 mins	Inhibition of human GST-fused JAK3 catalytic domain expressed in baculovirus-infected Sf9 cells using polyglutamic acid-tyrosine as substrate after 30 mins by ELISA, Ki = 0.0004 μM.	23668484
Sf9	Function assay		90 mins	Inhibition of human recombinant N-terminal GST-tagged JAK3 expressed in Sf9 cells using Biotin-LC-EQEDEPEGDYFEWLE as substrate after 90 mins by TR-FRET assay, IC50 = 0.0006 μM.	22087750
Sf9	Function assay		30 mins	Inhibition of human GST-fused JAK1 catalytic domain expressed in baculovirus-infected Sf9 cells using polyglutamic acid-tyrosine as substrate after 30 mins by ELISA, Ki = 0.0007 μM.	23668484
Sf9	Function assay		30 mins	Inhibition of human GST-fused JAK2 catalytic domain expressed in baculovirus-infected Sf9 cells using polyglutamic acid-tyrosine as substrate after 30 mins by ELISA, Ki = 0.0007 μM.	23668484
Sf9	Function assay			Inhibition of human recombinant GST-tagged JH1 domain (785 to1125) of JAK3 expressed in insect Sf9 cells by ELISA, IC50 = 0.001 μM.	14593182
Sf9	Function assay			Inhibition of GST-tagged human JAK3 catalytic domain expressed in Sf9 cells by ELISA, IC50 = 0.001 μM.	21105711
Sf9	Function assay		90 mins	Inhibition of human recombinant N-terminal GST-tagged JAK1 expressed in Sf9 cells using Biotin-LC-EQEDEPEGDYFEWLE as substrate after 90 mins by TR-FRET assay, IC50 = 0.003 μM.	22087750
Sf9	Function assay			Inhibition of GST-tagged human JAK3 catalytic domain expressed in Sf9 cells by ELISA, IC50 = 0.0033 μM.	21105711
Sf9	Function assay		90 mins	Inhibition of human recombinant N-terminal GST-tagged JAK2 expressed in Sf9 cells using Biotin-LC-EQEDEPEGDYFEWLE as substrate after 90 mins by TR-FRET assay, IC50 = 0.004 μM.	22087750
SF21	Function assay		10 mins	Inhibition of JAK2 (unknown origin) expressed in SF21 cells using Biotin-KAIETDKEYYTVKD as substrate and [33Pgamma]ATP incubated for 10 mins prior to substrate addition measured after 30 mins by Topcount analysis, IC50 = 0.004 μM.	23541670
insect cells	Function assay			Inhibition of GST-tagged Jak1 expressed in insect cells using 70 uM ATP, IC50 = 0.0061 μM.	21155605
insect cells	Function assay			Inhibition of GST-tagged Jak3 expressed in insect cells using 18 uM ATP, IC50 = 0.008 μM.	21155605
T-cells	Function assay		72 hrs	Inhibition of IL2-induced proliferation of human T cells assessed as [3H]thymidine incorporation after 72 hrs by scintillation counting, IC50 = 0.011 μM.	14593182
insect cells	Function assay			Inhibition of GST-tagged Jak2 expressed in insect cells using 20 uM ATP, IC50 = 0.012 μM.	21155605
MO7	Function assay			Inhibition of Jak3-mediated IL15-induced Stat5 phosphorylation in human MO7 cells by cell-based assay, IC50 = 0.024 μM.	21155605
Ba/F3	Function assay		60 mins	Inhibition of TEL-fused JAK1 expressed in Ba/F3 cells assessed as inhibition of STAT5 phosphorylation after 60 mins by AlphaScreen assay, IC50 = 0.026 μM.	22087750
T-cells	Function assay			Inhibition of JAK3/1 in human T cells expressing CD3 assessed as inhibition of IL2-stimulated STAT5a phosphorylation, IC50 = 0.028 μM.	23540648
PBMC	Function assay			Inhibition IL-7-indcued STAT5 phosphorylation in human PBMC cells by flow cytometry, IC50 = 0.039 μM.	26927423
TF1	Function assay		20 mins	Inhibition of JAK1 in human TF1 cells assessed as inhibition of IL6-induced STAT3 phosphorylation incubated for 20 mins followed by IL6 challenge for 30 mins in presence of whole blood, EC50 = 0.043 μM.	23659214
TF1	Function assay			Inhibition of JAK1 in human TF1 cells assessed as suppression of IL6-stimulated STAT3 phosphorylation by AlphaScreen assay, INH = 0.045 μM.	26372653
CTLL	Function assay			Inhibition of Jak3-mediated IL2-induced Stat5 phosphorylation in mouse CTLL cells by cell-based assay, IC50 = 0.048 μM.	21155605
Sf9	Function assay		90 mins	Inhibition of human recombinant N-terminal GST-tagged TYK2 expressed in Sf9 cells using Biotin-LC-EQEDEPEGDYFEWLE as substrate after 90 mins by TR-FRET assay, IC50 = 0.052 μM.	22087750
TF1	Function assay			Inhibition of JAK1 in human TF1 cells assessed as inhibition of IL2-induced STAT3 phosphorylation, EC50 = 0.053 μM.	22591402
TF1	Function assay		20 mins	Inhibition of JAK1 in human TF1 cells assessed as inhibition of IL6-induced STAT3 phosphorylation incubated for 20 mins prior to IL6-induction measured after 30 to 45 mins, EC50 = 0.053 μM.	22698084
TF1	Function assay		20 mins	Inhibition of JAK1 in human TF1 cells assessed as inhibition of IL6-induced STAT3 phosphorylation incubated for 20 mins followed by IL6 challenge for 30 mins, EC50 = 0.053 μM.	23659214
Ba/F3	Function assay		60 mins	Inhibition of TEL-fused JAK3 expressed in Ba/F3 cells assessed as inhibition of STAT5 phosphorylation after 60 mins by AlphaScreen assay, IC50 = 0.054 μM.	22087750
T-cells	Function assay			Inhibition of allogenic cells-stimulated proliferation in monkey T cells by mixed lymphocyte reaction method, IC50 = 0.057 μM.	14593182
CD34+	Function assay			Inhibition of JAK2 in human CD34+ cells assessed as inhibition of EPO-mediated cell proliferation, IC50 = 0.071 μM.	26927423
TF1	Function assay			Inhibition of IL4-induced proliferation of TF1 cells, IC50 = 0.08 μM.	16934457
T-cells	Function assay			Inhibition of allogenic cells-stimulated proliferation in human T cells by mixed lymphocyte reaction method, IC50 = 0.087 μM.	14593182
TF1	Function assay		20 mins	Inhibition of JAK2 in human TF1 cells assessed as inhibition of EPO-induced STAT5 phosphorylation incubated for 20 mins prior to EPO-induction measured after 30 to 45 mins, EC50 = 0.093 μM.	22698084
TF1	Function assay		20 mins	Inhibition of JAK2 in human TF1 cells assessed as inhibition of EPO-induced STAT5 phosphorylation incubated for 20 mins followed by IL6 challenge for 30 mins, EC50 = 0.093 μM.	23659214
PBMC	Function assay			Inhibition of JAK1 in human PBMC cells assessed as inhibition of IL-6-induced MCP1 secretion, IC50 = 0.095 μM.	26927423
TF1	Function assay		2 hrs	Inhibition of JAK2 in GMCSF-stimulated human TF1 cells assessed as suppression of STAT5 phosphorylation preincubated for 2 hrs followed by GMCSF stimulation for 50 mins by FACS reader analysis, IC50 = 0.095 μM.	27130359
TF1	Function assay			Inhibition of JAK2 in EPO-stimulated human TF1 cells expressing stably integrated beta-lactamase reporter gene under control of STAT5 response elements in interferon regulatory factor 1 gene promoter by fluorescence assay, IC50 = 0.107 μM.	29156136
Sf9	Function assay			Inhibition of GST-tagged human JAK1 catalytic domain expressed in Sf9 cells by ELISA, IC50 = 0.11 μM.	21105711
T-cells	Function assay			Inhibition of allogenic cells-stimulated proliferation in mouse T cells by mixed lymphocyte reaction method, IC50 = 0.115 μM.	14593182
ME180	Function assay			Inhibition of JAK1 in IL6-stimulated human ME180 cells expressing stably integrated beta-lactamase reporter gene under control of sis-inducible element by fluorescence assay, IC50 = 0.124 μM.	29156136
CTLL-2	Antiproliferative assay		72 hrs	Antiproliferative activity against IL-2-stimulated mouse CTLL-2 cells expressing JAK1/JAK3 after 72 hrs by alamar blue assay, IC50 = 0.132 μM.	19762238
HT2	Function assay			Inhibition of JAK3 in mouse HT2 cells assessed as suppression of cell growth, IC50 = 0.156 μM.	27771180
DND/L12	Function assay		30 mins	Inhibition of JAK3 in human DND/L12 cells after 30 mins by luciferase assay in presence of human serum albumin, IC90 = 0.16 μM.	14593182
T-cells	Function assay			Inhibition of JAK2/1 in human T cells expressing CD3 assessed as inhibition of IFNgamma-stimulated STAT1 phosphorylation, IC50 = 0.17 μM.	23540648
insect	Function assay			Inhibition of GST-tagged Tyk2 expressed in insect cells using 35 uM ATP, IC50 = 0.176 μM.	21155605
Ba/F3	Function assay		60 mins	Inhibition of TEL-fused JAK2 expressed in Ba/F3 cells assessed as inhibition of STAT5 phosphorylation after 60 mins by AlphaScreen assay, IC50 = 0.265 μM.	22087750
TF1	Function assay			Inhibition of JAK2 in human TF1 cells assessed as suppression of cell growth, IC50 = 0.2751 μM.	27771180
CD34+	Function assay			Inhibition of JAK2 homodimer in human CD34+ cells spiked into human whole blood assessed as inhibition of EPO-induced STAT-5 phosphorylation preincubated for 45 mins followed by EPO addition measured after 15 mins by FACS analysis, IC50 = 0.302 μM.	24417533
HUO3	Function assay		4 days	Inhibition of human GM-CSF-induced proliferation in human HUO3 cells assessed as [3H]thymidine incorporation after 4 days by scintillation counting, IC50 = 0.324 μM.	14593182
HU03	Function assay		72 hrs	Inhibition of GM-CSF-induced human HU03 cells proliferation after 72 hrs by [3H]thymidine incorporation assay, IC50 = 0.324 μM.	21105711
BAF3	Antiproliferative assay		72 hrs	Antiproliferative activity against mouse BAF3 cells expressing TEL-JAK3 kinase after 72 hrs by alamar blue assay, IC50 = 0.57 μM.	19762238
T-cells	Function assay		3 days	Inhibition of cell proliferation of IL2-activated and CD3 and CD28 antibody-stimulated mouse CD4-positive T cells after 3 days by XTT assay, IC50 = 0.63 μM.	30460842
NK92	Function assay		20 mins	Inhibition of TYK2 in human NK92 cells assessed as inhibition of IL12-induced STAT4 phosphorylation incubated for 20 mins followed by IL12 challenge for 45 mins, EC50 = 0.71 μM.	23659214
TF1	Function assay			Inhibition of IL3-induced proliferation of TF1 cells, IC50 = 0.8 μM.	16934457
UT7	Function assay			Inhibition of JAK2 in human UT7 cells assessed as suppression of EPO-stimulated STAT5 phosphorylation by AlphaScreen assay, INH = 1.12 μM.	26372653
Ba/F3	Function assay		60 mins	Inhibition of TEL-fused TYK2 expressed in Ba/F3 cells assessed as inhibition of STAT5 phosphorylation after 60 mins by AlphaScreen assay, IC50 = 1.2 μM.	22087750
T-cells	Function assay		72 hrs	Inhibition of JAK3 in rat splenocytic T cells assessed as reduction in IL-2-induced cell proliferation after 72 hrs by MTS assay, IC50 = 1.2 μM.	30139575
SZ4	Function assay			Inhibition of JAK3 in human SZ4 cells assessed as reduction in IL2-stimulated STAT5 phosphorylation preincubated for 1 hr followed by IL2 stimulation and measured after 15 mins by MSD assay, IC50 = 1.21 μM.	30460842
BAF3	Antiproliferative assay		72 hrs	Antiproliferative activity against mouse BAF3 cells expressing TEL-JAK2 kinase after 72 hrs by alamar blue assay, IC50 = 1.574 μM.	19762238
HEL	Function assay		1 hr	Inhibition of JAK2 V617F mutant in human HEL cells after 1 hr, IC50 = 4.724 μM.	27771180
Jurkat	Function assay		24 hrs	Inhibition of anti-CD3/anti-CD28-induced IL2 production in human Jurkat cells after 24 hrs by scintillation counting, IC50 = 7.84 μM.	14593182
YT	Function assay	30 ng/ml		Inhibition of IL2-induced JAK3 phosphorylation in human YT cells at 30 ng/ml by immunoblotting analysis	14593182
YT	Function assay	30 ng/ml		Inhibition of IL2-induced STAT5A phosphorylation in human YT cells at 30 ng/ml by immunoblotting analysis	14593182
YT	Function assay	30 ng/ml		Inhibition of IL2-induced STAT5B phosphorylation in human YT cells at 30 ng/ml by immunoblotting analysis	14593182
NK-cells	Immunosuppressive assay	1 to 5 mg/kg	4 weeks	Immunosuppressive activity in naive cynomolgus monkey assessed as effect on CD3-CD16+ NK cells at 1 to 5 mg/kg, po for 4 weeks	14593182
T-cells	Function assay	5 to 500 nM	1 hr	Inhibition of IL2-induced Stat5 phosphorylation in human CD4+ T cells at 5 to 500 nM after 1 hr by Western blot	19053756
SUM149PT	Function assay	3 uM	16 hrs	Induction of PTPN6 in human SUM149PT cells assessed as inhibition of STAT3 phosphorylation at 3 uM after 16 hrs by Western blotting analysis in presence of pervanadate	24978112
Hs578T	Function assay	3 uM	16 hrs	Induction of PTPN6 in human Hs578T cells assessed as inhibition of STAT3 phosphorylation at 3 uM after 16 hrs by Western blotting analysis in presence of pervanadate	24978112
T-cells	Function assay	50 to 300 nM	1 hr	Inhibition of JAK1/JAK3 in human CD4 positive T cells assessed as inhibition of IL4-stimulated STAT6 phosphorylation at 50 to 300 nM preincubated for 1 hr followed by IL-4 stimulation measured after 30 mins by immunoblot analysis	29852068
T-cells	Function assay	50 to 300 nM	1 hr	Inhibition of JAK1/JAK3 in human CD4 positive T cells assessed as inhibition of IL2-stimulated STAT5 phosphorylation at Tyr-695 residue at 50 to 300 nM preincubated for 1 hr followed by IL-2 stimulation measured after 30 mins by immunoblot analysis	29852068
T-cells	Function assay	>300 nM	1 hr	Inhibition of JAK1/JAK2/TYK2 in human CD4 positive T cells assessed as inhibition of IL6-stimulated STAT3 phosphorylation at >300 nM preincubated for 1 hr followed by IL-6 stimulation measured after 30 mins by immunoblot analysis	29852068
TALL-1	Function assay	1 uM	3 hrs	Inhibition of JAK3 in human TALL-1 cells assessed as inhibition of IL-2 induced STAT5 phosphorylation at 1 uM preincubated for 3 hrs followed by IL-2 induction measured after 30 mins by immunoblotting	26258521
BA/F3	Function assay	300 nM	3 hrs	Inhibition of JAK3 (unknown origin) expressed in mouse BA/F3 cells assessed as reduction of STAT5 phosphorylation at 300 nM after 3 hrs by immunoblotting analysis	26258521
OCL-AML5	Function assay	1 uM	3 hrs	Inhibition of JAK2 in human OCL-AML5 cells assessed as inhibition of GM-CSF induced STAT5 phosphorylation at 1 uM preincubated for 3 hrs followed by GM-CSF induction measured after 30 mins by immunoblotting	26258521
BA/F3	Antiproliferative assay		72 hrs	Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK3 after 72 hrs by cell titer glo assay	26258521
BA/F3	Function assay	1 uM	3 hrs	Inhibition of JAK3 (unknown origin) expressed in mouse BA/F3 cells at 1 uM preincubated for 3 hrs followed by pulldown with streptavidin beads by immunoblotting analysis	26258521
Huh7	Function assay	10 uM	30 mins	Inhibition of Tyk2 in human Huh7 cells assessed as reduction of IFNalpha5-induced STAT3 phosphorylation at 10 uM pre-incubated for 30 mins before IFNalpha5 stimulation for 30 mins mins by immunoblotting	26231159
Huh7	Function assay	10 uM	30 mins	Inhibition of Tyk2 in human Huh7 cells assessed as reduction of basal level STAT3 phosphorylation at 10 uM after 30 mins by immunoblotting	26231159
Huh7	Function assay	1 uM	30 mins	Inhibition of Tyk2 in human Huh7 cells assessed as reduction of IFNalpha5-induced STAT3 phosphorylation at 1 uM pre-incubated for 30 mins before IFNalpha5 stimulation for 30 mins mins by immunoblotting	26231159
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Informations chimiques, stockage et stabilité (Chemical Information, Storage & Stability)

Poids moléculaire	312.37	Formule	C₁₆H₂₀N₆O	Stockage (À compter de la date de réception)	3 ans-20°Cpoudre
N° CAS	477600-75-2	Télécharger le SDF		Stockage des solutions mères	1 an-80°Cen solvant 1 mois-20°Cen solvant
Synonymes	CP-690550, Tasocitinib			Smiles	CC1CCN(CC1N(C)C2=NC=NC3=C2C=CN3)C(=O)CC#N

Solubilité (Solubility)

In vitro
Lot:

DMSO : 62 mg/mL (198.48 mM)
(Le DMSO contaminé par l'humidité peut réduire la solubilité. Utiliser du DMSO frais et anhydre.)

Water : Insoluble

Ethanol : Insoluble

Calculateur de molarité

Masse	Concentration	Volume	Poids moléculaire
=	×	×

Calculateur de dilution Calculateur de poids moléculaire

In vivo Lot:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validé par les laboratoires Selleck. Si vous avez besoin d'ajustements à cette formulation, contactez notre équipe commerciale pour des tests personnalisés.	3.100mg/ml (9.92mM)	Taking the 1 mL working solution as an example, add 50 μL of 62 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 clarify; 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.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

Calculateur de formulation in vivo (Solution claire)

Étape 1 : Saisir les informations ci-dessous (Recommandé : Un animal supplémentaire pour tenir compte des pertes pendant l'expérience)

Dosage : mg/kg Poids moyen des animaux : g Volume de dosage par animal :μL Nombre d'animaux :

Étape 2 : Saisir la formulation in vivo (Ceci est seulement le calculateur, pas la formulation. Veuillez nous contacter d'abord s'il n'y a pas de formulation in vivo dans la section Solubilité.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Résultats du calcul :

Concentration de travail : mg/ml;

Méthode de préparation du liquide maître DMSO : mg médicament prédissous dans μL DMSO ( Concentration du liquide maître mg/mL, Veuillez nous contacter d'abord si la concentration dépasse la solubilité du DMSO du lot de médicament. )

Méthode de préparation de la formulation in vivo : Prendre μL DMSO liquide maître, puis ajouterμL PEG300, mélanger et clarifier, puis ajouterμL Tween 80, mélanger et clarifier, puis ajouter μL ddH₂O, mélanger et clarifier.

Méthode de préparation de la formulation in vivo : Prendre μL DMSO liquide maître, puis ajouter μL Huile de maïs, mélanger et clarifier.

Note : 1. Veuillez vous assurer que le liquide est clair avant d'ajouter le solvant suivant.
2. Assurez-vous d'ajouter le(s) solvant(s) dans l'ordre. Vous devez vous assurer que la solution obtenue, lors de l'ajout précédent, est une solution claire avant de procéder à l'ajout du solvant suivant. Des méthodes physiques telles que le vortex, les ultrasons ou le bain-marie chaud peuvent être utilisées pour faciliter la dissolution.

Mécanisme d'action (Mechanism of Action)

Targets/IC₅₀/Ki	JAK3 (Cell-free assay) 1 nM JAK2 (Cell-free assay) 20 nM JAK1 (Cell-free assay) 112 nM
In vitro	CP-690550 is a specific, orally inhibitor of JAK3, it is 20- to 100-fold less potent for JAK2 and JAK1 with IC50 of 20 nM and 112 nM, respectively. This compound doesn't have potent activity against 30 other kinases (all median IC50 > 3000 nM). It inhibits IL-2–induced proliferation with 30-fold greater potency than its effects on GM-CSF–induced proliferation. This chemical effectively inhibits a murine mixed lymphocyte reaction (MLR) (IC50 = 91 nM). It potently inhibits IL-4 induced upregulation of CD23 (IC50=57 nM) and class II major histocompatibility complex (MHCII) expression (IC50=71 nM) on murine B cells. A recent research indicates low dose of this compound accelerates the onset of experimental autoimmune encephalomyelitis by potentiating Th17 differentiation.
Essai kinase	Essai de la kinase JAK3
Essai kinase	Un fragment codant pour le domaine catalytique de la JAK3 humaine (785aa à 1125aa, domaine catalytique JH1) est amplifié par PCR à partir de l'ADNc pleine longueur et cloné dans le site EcoRI du vecteur de transfert de baculovirus pVL1393. Le baculovirus recombinant est utilisé pour infecter les cellules Sf9 (Spodoptera frugipedra) et la protéine de fusion GSTJAK3 recombinante est isolée sur de la sépharose glutathion. La protéine de fusion est éluée avec du glutathion réduit et stockée dans un tampon contenant 50 mM Tris, pH 7,5, 10 mM DTT et 10% de glycérol. L'activité de la kinase JAK3 est mesurée par ELISA comme suit : Les plaques sont recouvertes pendant une nuit d'un copolymère aléatoire d'acide L-glutamique et de tyrosine (4:1) (100 ug/mL). Les plaques sont lavées et la JAK3 JH1:GST recombinante (100 ng/puits) avec ou sans inhibiteurs est incubée à température ambiante pendant 30 minutes, après quoi l'anticorps anti-phosphotyrosine PY20 conjugué à la HRP (ICN) est ajouté et développé par TMB (3,3',5,5'-tétraméthylbenzidine). D'autres kinases (tableau 1) sont produites dans E. coli ou dans des cellules d'insectes, selon ce qui est jugé optimal pour la kinase donnée. L'activité catalytique des tyrosines kinases est mesurée à l'aide de l'ELISA susmentionné, tandis que les sérine/thréonine kinases sont testées à l'aide d'essais enzymatiques radioactifs.
In vivo	In a murine model of heterotopic heart transplantation (DBA2 donor heart into C57/BL6 host), Tofacitinib results in a dose-dependent increase in survival of transplanted hearts.The EC50 (drug concentration in blood at which 50% of mice will maintain their graft for >28 days) to be ~60 ng/mL.This compound prevents rejection of allogeneic kidneys in nonhuman primate (NHPs, macaca fascicularis) (MST of 62 and 83 days for the 50 to 100 ng/ml groups and 200 to 400 ng/ml groups, respectively). Mice chronically dosed with this compound (1.5-15 mg/kg/day) demonstrates dose and time-dependent alterations in lymphocyte subsets when examined by flow cytometry. The most dramatic change observed is a 96% reduction in splenic NK1.1+TCRb-cell numbers following 21 days of treatment. Delayed-type hypersensitivity (DTH) responses in sensitized mice are reduced in a dose-dependent manner following treatment with this chemical (1.87–30 mg/kg, s.c.). Extended survival of neonatal Balb/c hearts implanted into the ear pinna of MHC mismatched C3H/HEN mice is observed with this compound monotherapy (10–30 mg/kg/day), but improved upon combination with cyclosporin (10 mg/kg/day).
Références	[1] https://pubmed.ncbi.nlm.nih.gov/14593182/ [2] https://pubmed.ncbi.nlm.nih.gov/14678034/ [3] https://pubmed.ncbi.nlm.nih.gov/18242596/ [4] https://pubmed.ncbi.nlm.nih.gov/22252297/ [5] https://pubmed.ncbi.nlm.nih.gov/16378072/ [6] https://pubmed.ncbi.nlm.nih.gov/14678034/

Applications (Applications)

Méthodes	Biomarqueurs	Images	PMID
Western blot	JAK3 / STAT5 / p-STAT5 LMP1 / EBNA1 / BZLF1 JAK3 / p-JAK3 / STAT3		27732937
Growth inhibition assay	Cell number		27732937

Informations sur l'essai clinique (Clinical Trial Information)

(données du https://clinicaltrials.gov, mis à jour le 2024-05-22)

Numéro NCT	Recrutement	Conditions	Promoteur/Collaborateurs	Date de début	Phases
NCT06202560	Enrolling by invitation	Frontal Fibrosing Alopecia\|Lichen Planopilaris	Institute of Dermatology Thailand	November 29 2023	Not Applicable
NCT05487703	Completed	Arthritis Rheumatoid	Pfizer	November 14 2022	--
NCT05082428	Completed	Ulcerative Colitis	Pfizer	May 30 2022	--
NCT05728008	Completed	Ulcerative Colitis	IRCCS San Raffaele	April 5 2022	--
NCT04768504	Recruiting	Immune-Mediated Colitis	Khashayar Esfahani\|Sir Mortimer B. Davis - Jewish General Hospital	March 22 2022	Phase 2

Foire aux questions (Frequently Asked Questions)

Question 1:
I was just wondering what the main differences between S2789 and S5001?

Réponse :
S2789 is the base form of S5001 (Citrate). The biological activity of these two compounds are same. The S5001 (Citrate) is more suitable for oral administration.

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C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):