solo para uso en investigación
Cat. No.S5001
| Dianas relacionadas | EGFR STAT Pim |
|---|---|
| Otros JAK Inhibidores | BMS-986165 (Deucravacitinib) AZD1480 WP1066 Momelotinib (CYT387) Filgotinib (GLPG0634) AT9283 Gandotinib (LY2784544) Pacritinib TG101209 Cerdulatinib (PRT062070) hydrochloride |
| Líneas celulares | Tipo de ensayo | Concentración | Tiempo de incubación | Formulación | Descripción de la actividad | PMID |
|---|---|---|---|---|---|---|
| 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 | 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 cells | 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 | 45 mins | 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 | ||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 14593182 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 21105711 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 19762238 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 30460842 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 23659214 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 16934457 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 26372653 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 22087750 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 30139575 | |||
| A2780 | Function assay | Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells | 27771180 | |||
| 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 | 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 | |
| Haga clic para ver más datos experimentales de líneas celulares | ||||||
| Peso molecular | 504.49 | Fórmula | C16H20N6O.C6H8O7 |
Almacenamiento (Desde la fecha de recepción) | |
|---|---|---|---|---|---|
| Nº CAS | 540737-29-9 | Descargar SDF | Almacenamiento de soluciones madre |
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| Sinónimos | Tasocitinib Citrate,CP-690550 | 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 | ||
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In vitro |
DMSO
: 100 mg/mL
(198.21 mM)
Water : Insoluble Ethanol : Insoluble |
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In vivo |
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Paso 1: Introduzca la información a continuación (Recomendado: Un animal adicional para tener en cuenta la pérdida durante el experimento)
Paso 2: Introduzca la formulación in vivo (Esto es solo la calculadora, no la formulación. Por favor, contáctenos primero si no hay una formulación in vivo en la sección de Solubilidad.)
Resultados del cálculo:
Concentración de trabajo: mg/ml;
Método para preparar el líquido maestro de DMSO: mg fármaco predissuelto en μL DMSO ( Concentración del líquido maestro mg/mL, Por favor, contáctenos primero si la concentración excede la solubilidad del DMSO del lote del fármaco. )
Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadirμL PEG300, mezclar y clarificar, luego añadirμL Tween 80, mezclar y clarificar, luego añadir μL ddH2O, mezclar y clarificar.
Método para preparar la formulación in vivo: Tomar μL DMSO líquido maestro, luego añadir μL Aceite de maíz, mezclar y clarificar.
Nota: 1. Por favor, asegúrese de que el líquido esté claro antes de añadir el siguiente disolvente.
2. Asegúrese de añadir el (los) disolvente(s) en orden. Debe asegurarse de que la solución obtenida, en la adición anterior, sea una solución clara antes de proceder a añadir el siguiente disolvente. Se pueden utilizar métodos físicos como el vórtice, el ultrasonido o el baño de agua caliente para ayudar a la disolución.
| Targets/IC50/Ki |
JAK3
(Cell-free assay) 1 nM
JAK2
(Cell-free assay) 20 nM
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|---|---|
| In vitro |
El citrato de Tofacitinib inhibe la proliferación de blastos de células T humanas mediada por IL-2 y la expresión de CD69 inducida por IL-15 con IC50 de 11 nM y 48 nM, respectivamente. El citrato de Tofacitinib previene la reacción linfocitaria mixta con una IC50 de 87 nM. El tratamiento de células Patersen-receptor de eritropoyetina (FDCP-EpoR) murinas dependientes de factores que albergan JAK2 humano de tipo salvaje o V617F con citrato de Tofacitinib conduce a la prevención de la proliferación celular con IC50 de 2,1 µM y 0,25 µM, respectivamente. El citrato de Tofacitinib inhibe la fosforilación de STAT1 y STAT3 inducida por interleucina-6 con IC50 de 23 nM y 77 nM, respectivamente. Además, el citrato de Tofacitinib genera un efecto proapoptótico significativo en células FDCP-EpoR murinas que portan JAK2VV617F, mientras que se observa un efecto menor para las células que portan JAK2 de tipo salvaje. Esta actividad está acoplada con la inhibición de la fosforilación de los efectores de señalización descendentes clave dependientes de JAK2V617F, el transductor de señal y activador de transcripción (STAT)3, STAT5, y el homólogo del oncogén viral del timoma murino v-akt (AKT). Además, el citrato de Tofacitinib previene la expresión de CD69 inducida por IL-15 en células NK y células T CD8+ humanas y de macaco cangrejero in vitro.
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| Ensayo de quinasa |
Ensayos enzimáticos
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Los ensayos de quinasa JAK1, JAK2 y JAK3 utilizan una proteína expresada en células SF9 infectadas con baculovirus (una proteína de fusión de GST y el dominio catalítico de la enzima JAK humana) purificada por cromatografía de afinidad en glutatión-Sepharose. El sustrato para la reacción es ácido poliglutámico-tirosina [PGT (4:1)], recubierto en placas Nunc Maxi Sorp a 100 μg/mL durante la noche a 37 °C. Las placas se lavan tres veces y se añade la enzima JAK a los pocillos, que contenían 100 μL de tampón de quinasa (50 mM HEPES, pH 7,3, 125 mM NaCl, 24 mM MgCl2) + ATP + 1 mM de ortovanadato de sodio. Para CP-690550 (Tofacitinib) Citrate, también se añade para el ensayo de quinasa a diferentes dosis. Después de la incubación a temperatura ambiente durante 30 min, las placas se lavan tres veces. El nivel de tirosina fosforilada en un pocillo dado se determina mediante un ensayo ELISA estándar utilizando un anticuerpo anti-fosfotirosina.
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| In vivo |
El citrato de Tofacitinib disminuye una respuesta de hipersensibilidad de tipo retardado y prolonga la supervivencia del aloinjerto cardíaco en modelos murinos. Además, el tratamiento con citrato de Tofacitinib de progenitores eritroides expandidos ex vivo de pacientes con PV positivos para JAK2V617F da como resultado una actividad antiproliferativa específica (IC50 = 0,2 μM) y proapoptótica. En contraste, los progenitores expandidos de controles sanos son menos sensibles al citrato de Tofacitinib en ensayos de proliferación (IC50 > 1,0 μM) y apoptosis. Durante 2 semanas de dosificación con citrato de Tofacitinib a 10 y 30 mg/kg/d, se observa una disminución significativa y dependiente del tiempo en el número de células NK en relación con el tratamiento con el vehículo. El número de células CD8+ de memoria efectora en el grupo tratado con citrato de Tofacitinib es un 55% menor que el observado en animales tratados con el vehículo.
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Referencias |
|
| Métodos | Biomarcadores | Imágenes | PMID |
|---|---|---|---|
| Western blot | JAK3 / p-JAK3 / STAT3 LMP1 / EBNA1 / BZLF1 JAK3 / STAT5 / p-STAT5 |
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26082451 |
| Growth inhibition assay | Cell number |
|
27732937 |
(datos de https://clinicaltrials.gov, actualizado el 2024-05-22)
| Número NCT | Reclutamiento | Condiciones | Patrocinador/Colaboradores | Fecha de inicio | Fases |
|---|---|---|---|---|---|
| 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 |
Tel: +1-832-582-8158 Ext:3
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Pregunta 1:
What is the difference between the two products (S5001, S2789)?
Respuesta:
Tofacitinib (S2789) is the base form of this compound. The biological activity of these two compounds is the same. However, S5001 is better than S2789 for oral gavage.