nur für Forschungszwecke
Kat.-Nr.S5243
| Verwandte Ziele | EGFR STAT Pim |
|---|---|
| Weitere JAK Inhibitoren | BMS-986165 (Deucravacitinib) AZD1480 WP1066 Momelotinib (CYT387) Filgotinib (GLPG0634) AT9283 Gandotinib (LY2784544) Pacritinib TG101209 Cerdulatinib (PRT062070) hydrochloride |
| Zelllinien | Assay-Typ | Konzentration | Inkubationszeit | Formulierung | Aktivitätsbeschreibung | PMID |
|---|---|---|---|---|---|---|
| Sf21 | Function assay | 1 hr | Inhibition of human JAK2 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.0028μM | 22591402 | ||
| Sf21 | Function assay | 1 hr | Inhibition of human JAK1 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.0033μM | 22591402 | ||
| Sf21 | Function assay | 1 hr | Inhibition of human TYK2 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.019μM | 22591402 | ||
| Sf21 | Function assay | 1 hr | Inhibition of human JAK3 kinase domain expressed in Sf21 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, IC50=0.428μM | 22591402 | ||
| 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=0.012μM | 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 prior to IL6-induction measured after 30 to 45 mins, EC50=0.024μM | 22698084 | ||
| SET2 | Function assay | Inhibition of JAK2 V617F mutant in human SET2 cells assessed as reduction in STAT5 phosphorylation, IC50=0.00184μM | 23061660 | |||
| TF1 | Function assay | 30 mins | Inhibition of JAK2 in human TF1 cells assessed as reduction in STAT5 phosphorylation incubated for 30 mins in presence of human recombinant EPO, IC50=0.00685μM | 23061660 | ||
| T-cells | Function assay | Inhibition of JAK3/1 in human T cells expressing CD3 assessed as inhibition of IL2-stimulated STAT5a phosphorylation, IC50=0.023μM | 23540648 | |||
| T-cells | Function assay | Inhibition of JAK2/1 in human T cells expressing CD3 assessed as inhibition of IFNgamma-stimulated STAT1 phosphorylation, IC50=0.031μM | 23540648 | |||
| Sf9 | Function assay | 1 hr | Inhibition of human JAK2 (828-1132) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0001μM | 23668484 | ||
| Sf9 | Function assay | 1 hr | Inhibition of human JAK1 (837-1142) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0002μM | 23668484 | ||
| Sf9 | Function assay | 1 hr | Inhibition of human TYK2 (873-1187) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0005μM | 23668484 | ||
| Sf9 | Function assay | 1 hr | Inhibition of human JAK3 (781-1124) expressed in baculovirus-infected Sf9 cells using EQEDEPEGDYFEWLE as substrate after 1 hr by HTRF assay, Ki=0.0032μM | 23668484 | ||
| 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=0.677μM | 24417533 | ||
| BA/F3 | Antiproliferative assay | 72 hrs | Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK1 after 72 hrs by cell titer glo assay | 26258521 | ||
| BA/F3 | Antiproliferative assay | 72 hrs | Antiproliferative activity against mouse BA/F3 cells expressing TEL-JAK2 after 72 hrs by cell titer glo assay | 26258521 | ||
| 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 | |
| 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 | |
| CD34+ | Function assay | Inhibition of JAK2 in human CD34+ cells assessed as inhibition of EPO-mediated cell proliferation, IC50=0.008μM | 26927423 | |||
| PBMC | Function assay | Inhibition of JAK1 in human PBMC cells assessed as inhibition of IL-6-induced MCP1 secretion, IC50=0.04μM | 26927423 | |||
| PBMC | Function assay | Inhibition IL-7-indcued STAT5 phosphorylation in human PBMC cells by flow cytometry, IC50=0.448μM | 26927423 | |||
| Sf21 | Function assay | 60 mins | Inhibition of human recombinant JAK2 expressed in Sf21 cells assessed as reduction in Ulight-CAGAGAIETDKEYYTVKD phosphorylation pre-incubated before substrate addition and measured after 60 mins by LANCE detection method, IC50=0.003μM | 27137359 | ||
| Sf21 | Function assay | 1 hr | Inhibition of recombinant human C-terminal 6His-tagged JAK2 (808 to end amino acids) expressed in Sf21 cells measured after 1 hr in presence of ATP by TR-FRET assay, IC50=0.0041μM | 27555284 | ||
| BaF3 | Function assay | Inhibition of JAK2 V617F mutant expressed in mouse BaF3 cells cells assessed as reduction in cell viability, EC50=0.186μM | 27555284 | |||
| HEL 92.1.7 | Antiproliferative assay | 3 days | Antiproliferative activity against HEL 92.1.7 cells assessed as viable cells measured after 3 days by WST-1 assay, IC50=14.7μM | 27555284 | ||
| HCC827 | Function assay | 30 uM | 24 hrs | Inhibition of JAK2 in human gefitinib-resistant HCC827 cells assessed as inhibition of STAT3 phosphorylation at Y705 site at 30 uM measured after 24 hrs by western blotting analysis | 27555284 | |
| HCC827 | Function assay | 30 uM | 24 hrs | Inhibition of JAK2 in wild-type human HCC827 cells assessed as inhibition of STAT3 phosphorylation at Y705 site at 30 uM measured after 24 hrs by western blotting analysis | 27555284 | |
| HEL | Antiproliferative assay | 48 hrs | Antiproliferative activity against HEL cells harboring JAK2 V617F mutant after 48 hrs by MTT assay, IC50=2.62μM | 27774135 | ||
| K562 | Antiproliferative assay | 48 hrs | Antiproliferative activity against human K562 cells after 48 hrs by MTT assay, IC50=10.3μM | 27774135 | ||
| MOLT4 | Antiproliferative assay | 48 hrs | Antiproliferative activity against human MOLT4 cells after 48 hrs by MTT assay, IC50=15.8μM | 27774135 | ||
| NCI-H23 | Antiproliferative assay | Antiproliferative activity against human NCI-H23 cells harboring KRAS G12C mutant at | 28038940 | |||
| NCI-H358 | Antiproliferative assay | Antiproliferative activity against human NCI-H358 cells harboring KRAS G12C mutant at | 28038940 | |||
| A673 | qHTS assay | qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells | 29435139 | |||
| SK-N-MC | qHTS assay | qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells | 29435139 | |||
| NB1643 | qHTS assay | qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells | 29435139 | |||
| LAN-5 | qHTS assay | qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells | 29435139 | |||
| HeLa | Function assay | 0.1 to 1 uM | 1 hr | Inhibition of JAK2 in human HeLa cells assessed as reduction in STAT5 phosphorylation at 0.1 to 1 uM after 1 hr by immunoblot analysis | 30243158 | |
| HeLa | Function assay | 0.1 to 1 uM | 1 hr | Inhibition of JAK2 in human HeLa cells assessed as increase in JAK2 phosphorylation at 0.1 to 1 uM after 1 hr by immunoblot analysis | 30243158 | |
| Sf9 | Function assay | 1 hr | Inhibition of human JAK2 (828 to 1132 residues) expressed in baculovirus infected Sf9 insect cells using EQEDEPEGDYFEWLE as substrate after 1 hr by fluorescence assay, IC50=0.0028μM | 30833158 | ||
| Sf9 | Function assay | 1 hr | Inhibition of human JAK1 (837 to 1142 residues) expressed in baculovirus infected Sf9 insect cells using EQEDEPEGDYFEWLE as substrate after 1 hr by fluorescence assay, IC50=0.0033μM | 30833158 | ||
| HEL | Antiproliferative assay | 48 hrs | Synergistic antiproliferative activity against HEL cells harboring JAK2 V617F mutant assessed as reduction in cell viability after 48 hrs in presence of SAHA by MTT assay, IC50=0.3μM | 30901208 | ||
| K562 | Antiproliferative assay | 48 hrs | Synergistic antiproliferative activity against human K562 cells assessed as reduction in cell viability after 48 hrs in presence of SAHA by MTT assay, IC50=1.03μM | 30901208 | ||
| MOLT4 | Antiproliferative assay | 48 hrs | Antiproliferative activity against human MOLT4 cells assessed as reduction in cell viability after 48 hrs by MTT assay, IC50=15.8μM | 30901208 | ||
| HEL | Antiproliferative assay | 48 hrs | Antiproliferative activity against HEL cells harboring JAK2 V617F mutant assessed as reduction in cell viability after 48 hrs by MTT assay, IC50=18.6μM | 30901208 | ||
| K562 | Antiproliferative assay | 48 hrs | Antiproliferative activity against human K562 cells assessed as reduction in cell viability after 48 hrs by MTT assay, IC50=23.2μM | 30901208 | ||
| A549 | Function assay | 80 to 400 nM | 6 hrs | Inhibition of HDAC2 in human A549 cells assessed as increase in acetyl histone H4 level at 80 to 400 nM after 6 hrs by Western blot analysis | 30901208 | |
| A549 | Function assay | 80 to 400 nM | 6 hrs | Inhibition of HDAC1 in human A549 cells assessed as increase in acetyl histone H4 level at 80 to 400 nM after 6 hrs by Western blot analysis | 30901208 | |
| A549 | Function assay | 80 to 400 nM | 6 hrs | Inhibition of HDAC3 in human A549 cells assessed as increase in acetyl histone H4 level at 80 to 400 nM after 6 hrs by Western blot analysis | 30901208 | |
| A549 | Function assay | 80 to 400 nM | 6 hrs | Inhibition of HDAC6 in human A549 cells assessed as increase in acetyl alpha tubulin level at 80 to 400 nM after 6 hrs by Western blot analysis | 30901208 | |
| HEL | Function assay | 100 mg/kg | 5 days | Drug level in tumor of BALB/c nu mouse xenografted with HEL cells at 100 mg/kg/day, ip administered for 5 days and measured 1 hr post-last dose by LC-MS/MS analysis | 30901208 | |
| Sf9 | Function assay | 30 secs | Inhibition of human recombinant N-terminal hexahistidine tagged JAK2 JH1 catalytic domain (835 to 1132 residues) expressed in baculovirus infected Sf9 cells using Tyr6 peptide as substrate incubated for 30 secs under shaking condition measured after 1 hr , IC50=0.0006μM | 30981578 | ||
| Sf21 | Function assay | 1 hr | Inhibition of recombinant human N-terminal epitope-tagged JAK2 (828 to 1132 residues) expressed in baculovirus infected Sf21 insect cells using EQEDEPEGDYFEWLE as substrate after 1 hr by homogeneous time-resolved fluorescence assay, IC50=0.0028μM | 30981578 | ||
| Sf9 | Function assay | 30 secs | Inhibition of human recombinant N-terminal hexahistidine tagged JAK1 JH1 catalytic domain (854 to 1154 residues) expressed in baculovirus infected Sf9 cells using Tyr6 peptide as substrate incubated for 30 secs under shaking condition measured after 1 hr , IC50=0.004μM | 30981578 | ||
| Sf9 | Function assay | Binding affinity to human recombinant N-terminal hexahistidine tagged JAK2 JH1 catalytic domain (835 to 1132 residues) expressed in baculovirus infected Sf9 cells assessed as dissociation constant by surface plasmon resonance assay, Kd=0.0282μM | 30981578 | |||
| Sf9 | Function assay | 30 secs | Inhibition of human recombinant C-terminal hexahistidine tagged JAK3 JH1 catalytic domain (811 to 1124 residues) expressed in baculovirus infected Sf9 cells using Tyr6 peptide as substrate incubated for 30 secs under shaking condition measured after 1 hr , IC50=0.051μM | 30981578 | ||
| HEL | Antiproliferative assay | 3 days | Antiproliferative activity against HEL cells harboring JAK2 V617F mutant measured after 3 days by CCK8 assay, IC50=7.639μM | 30981578 | ||
| insect cells | Function assay | 10 mins | Inhibition of recombinant human N-terminal GST-tagged JAK1 (866 to 1154 residues) expressed in insect cells using FITC-labeled C6-KKHTDDGYMPMSPGVA-NH peptide as substrate after 10 mins in presence of 5 mM ATP by caliper mobility shift assay, IC50=0.02μM | 32297743 | ||
| insect cells | Function assay | 10 mins | Inhibition of recombinant human N-terminal GST-tagged JAK2 (831 to 1132 residues) expressed in insect cells using 5FAM-labeled GEEPLYWSFPAKKK-NH2 peptide as substrate after 10 mins in presence of 5 mM ATP by caliper mobility shift assay, IC50=0.02μM | 32297743 | ||
| BAF3 | Cytotoxicity assay | 48 hrs | Cytotoxicity against mouse BAF3 cells expressing JAK2 V617F mutant after 48 hrs by CellTiterGlo assay, IC50=0.126μM | ChEMBL | ||
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| Molekulargewicht | 404.36 | Formel | C17H18N6.H3O4P |
Lagerung (Ab dem Eingangsdatum) | |
|---|---|---|---|---|---|
| CAS-Nr. | 1092939-17-7 | -- | Lagerung von Stammlösungen |
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| Synonyme | INCB018424, INC424 | Smiles | C1CCC(C1)C(CC#N)N2C=C(C=N2)C3=C4C=CNC4=NC=N3.OP(=O)(O)O | ||
|
In vitro |
DMSO
: 81 mg/mL
(200.31 mM)
Ethanol : 9 mg/mL Water : Insoluble |
|
In vivo |
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Schritt 1: Geben Sie die untenstehenden Informationen ein (Empfohlen: Ein zusätzliches Tier zur Berücksichtigung von Verlusten während des Experiments)
Schritt 2: Geben Sie die In-vivo-Formulierung ein (Dies ist nur der Rechner, keine Formulierung. Bitte kontaktieren Sie uns zuerst, wenn es im Abschnitt "Löslichkeit" keine In-vivo-Formulierung gibt.)
Berechnungsergebnisse:
Arbeitskonzentration: mg/ml;
Methode zur Herstellung der DMSO-Stammlösung: mg Wirkstoff vorgelöst in μL DMSO ( Konzentration der Stammlösung mg/mL, Bitte kontaktieren Sie uns zuerst, wenn die Konzentration die DMSO-Löslichkeit der Wirkstoffcharge überschreitet. )
Methode zur Herstellung der In-vivo-Formulierung: Nehmen Sie μL DMSO Stammlösung, dann hinzufügenμL PEG300, mischen und klären, dann hinzufügenμL Tween 80, mischen und klären, dann hinzufügen μL ddH2O, mischen und klären.
Methode zur Herstellung der In-vivo-Formulierung: Nehmen Sie μL DMSO Stammlösung, dann hinzufügen μL Maisöl, mischen und klären.
Hinweis: 1. Bitte stellen Sie sicher, dass die Flüssigkeit klar ist, bevor Sie das nächste Lösungsmittel hinzufügen.
2. Achten Sie darauf, das/die Lösungsmittel der Reihe nach hinzuzufügen. Sie müssen sicherstellen, dass die bei der vorherigen Zugabe erhaltene Lösung eine klare Lösung ist, bevor Sie mit der Zugabe des nächsten Lösungsmittels fortfahren. Physikalische Methoden wie Vortex, Ultraschall oder ein heißes Wasserbad können zur Unterstützung des Lösens verwendet werden.
| Targets/IC50/Ki |
JAK2
(Cell-free assay) 2.8 nM
JAK1
(Cell-free assay) 3.3 nM
|
|---|---|
| In vitro |
INCB018424 hemmt potent und selektiv die JAK2V617F-vermittelte Signalgebung und Proliferation in Ba/F3-Zellen und HEL-Zellen. INCB018424 erhöht die Apoptose in Ba/F3-Zellen dosisabhängig signifikant. INCB018424 (64 nM) führt zu einer Verdopplung der Zellen mit depolarisierten Mitochondrien in Ba/F3-Zellen. INCB018424 hemmt die Proliferation von erythroiden Progenitoren von normalen Spendern und Polycythaemia vera-Patienten mit einer IC50 von 407 nM bzw. 223 nM. INCB018424 zeigt eine bemerkenswerte Potenz gegen die erythroidale Koloniebildung mit einer IC50 von 67 nM. |
| Kinase-Assay |
Bindungsassay
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Rekombinante Proteine werden mittels Sf21-Zellen und Baculovirus-Vektoren exprimiert und mittels Affinitätschromatographie gereinigt. Für JAK-Kinase-Assays wird ein homogenes zeitaufgelöstes Fluoreszenz-Assay mit dem Peptidsubstrat (-EQEDEPEGDYFEWLE) verwendet. Jede Enzymreaktion wird mit Ruxolitinib oder Kontrolle, JAK-Enzym, 500 nM Peptid, Adenosintriphosphat (ATP; 1mM) und 2% Dimethylsulfoxid (DMSO) für 1 Stunde durchgeführt. Die 50%ige Hemmkonzentration (IC50) wird als die INCB018424-Konzentration berechnet, die für die Hemmung von 50% des Fluoreszenzsignals erforderlich ist.
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| In vivo |
INCB018424 (180 mg/kg, oral, zweimal täglich) führt in einem JAK2V617F-getriebenen Mausmodell zu einer Überlebensrate von über 90% bis Tag 22. INCB018424 (180 mg/kg, oral, zweimal täglich) reduziert signifikant die Splenomegalie und die zirkulierenden Spiegel entzündlicher Zytokine und eliminiert bevorzugt neoplastische Zellen, was zu einem signifikant verlängerten Überleben ohne myelosuppressive oder immunsuppressive Effekte in einem JAK2V617F-getriebenen Mausmodell führt. Der primäre Endpunkt wird bei 41,9% der Patienten in der Ruxolitinib-Gruppe im Vergleich zu 0,7% in der Placebo-Gruppe in der doppelblinden Studie zur Myelofibrose erreicht. Ruxolitinib führt zur Aufrechterhaltung der Milzvolumenreduktion und einer Verbesserung von 50% oder mehr im gesamten Symptom-Score. Insgesamt 28% der Patienten in der Ruxolitinib-Gruppe (15 mg zweimal täglich) weisen in Woche 48 bei Patienten mit Myelofibrose eine Milzvolumenreduktion von mindestens 35% auf, verglichen mit 0% in der Gruppe, die die beste verfügbare Therapie erhielt. Die mittlere palpable Milzlänge hat sich mit Ruxolitinib um 56% verringert, aber mit der besten verfügbaren Therapie in Woche 48 um 4% erhöht. Patienten in der Ruxolitinib-Gruppe zeigten eine Verbesserung der allgemeinen Lebensqualitätsmessungen und eine Reduktion der mit Myelofibrose verbundenen Symptome. |
Literatur |
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(Daten von https://clinicaltrials.gov, aktualisiert am 2024-05-22)
| NCT-Nummer | Rekrutierung | Erkrankungen | Sponsor/Kooperationspartner | Startdatum | Phasen |
|---|---|---|---|---|---|
| NCT06310304 | Active not recruiting | Healthy Participants |
Incyte Corporation |
March 26 2024 | Phase 1 |
| NCT02596347 | Completed | Chronic Beryllium Disease (CBD)|Beryllium Sensitization (BeS) |
National Jewish Health |
April 2015 | -- |
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