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Flavopiridol (Alvocidib, HMR-1275) CDK inhibidor

Name: Flavopiridol (Alvocidib, HMR-1275)
Brand: Selleck Chemicals
SKU: S1230
Rating: 5 (146 reviews)

Cat. No.S1230

Flavopiridol (Alvocidib) compite con el ATP para inhibir las CDK, incluyendo CDK1, CDK2, CDK4, CDK6 y CDK9 con valores de IC50 en el rango de 20-100 nM. Es más selectivo para CDK1, 2, 4, 6, 9 frente a CDK7. Inicialmente se encuentra que el Flavopiridol inhibe el EGFR y la PKA. El Flavopiridol induce autophagy y estrés del RE. El Flavopiridol bloquea la replicación del HIV-1. Fase 1/2.

Flavopiridol (Alvocidib, HMR-1275) CDK inhibidor Chemical Structure

Estructura química

Peso molecular: 401.84

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Control de calidad

Lote: Pureza: 99.92%

99.92

Dianas relacionadas	HSP PD-1/PD-L1 ROCK Wee1 DNA/RNA Synthesis Microtubule Associated Ras KRas Aurora Kinase Casein Kinase
Otros CDK Inhibidores	AS2863619 Ro-3306 SEL120 (SEL120-34A) hydrochloride INX-315 Tagtociclib (PF-07104091) PF-07220060 (CDK4/6-IN-6) AZD4573 ON123300 Enitociclib (BAY 1251152) Samuraciclib (ICEC0942) hydrochloride

Cultivo celular, tratamiento y concentración de trabajo

Líneas celulares	Tipo de ensayo	Concentración	Tiempo de incubación	Descripción de la actividad	PMID
MCF-7 tumor cell	Proliferation assay			Inhibition of MCF-7 tumor cell proliferation	10843211
PC3 cell	Function assay			Inhibition of PC3 cell clonogenic assay, IC50=10 μM	11063609
HCT116 cell	Function assay			Inhibition of HCT116 cell clonogenic assay, IC50=13 μM	11063609
A2780 cell	Function assay			Inhibition of A2780 cell clonogenic assay， IC50=15 μM	11063609
Mia PaCa-2 cell	Function assay			Inhibition of Mia PaCa-2 cell clonogenic assay, IC50=36 μM	11063609
LNCaP human prostate carcinoma cell	Proliferation assay			Inhibition of LNCaP human prostate carcinoma cell proliferation	12190313
HCT116/VP35 human colon carcinoma cell	Proliferation assay			Inhibition of HCT116/VP35 human colon carcinoma cell proliferation, IC50=17 nM	12190313
HCT116 human colon carcinoma cell	Proliferation assay			Inhibition of HCT116 human colon carcinoma cell proliferation, IC50=18 nM	12190313
HCT116/VM46 human colon carcinoma cell	Proliferation assay			Inhibition of HCT116/VM46 human colon carcinoma cell proliferation, IC50=21 nM	12190313
A2780/DDP-R human ovarian carcinoma cell	Proliferation assay			Inhibition of A2780/DDP-R human ovarian carcinoma cell proliferation, IC50=38 nM	12190313
ABAE human fibroblast cell	Proliferation assay			Inhibition of ABAE human fibroblast cell proliferation, IC50=45 nM	12190313
HL60 human leukemia cell	Proliferation assay			Inhibition of HL60 human leukemia cell proliferation, IC50=46 nM	12190313
Hs 27 human fibroblast cell	Proliferation assay			Inhibition of Hs 27 human fibroblast cell proliferation, IC50=51 nM	12190313
CCRF-CEM human leukemia cell	Proliferation assay			Inhibition of CCRF-CEM human leukemia cell proliferation, IC50=52 nM	12190313
OVCAR-3 human ovarian carcinoma cell	Proliferation assay			Inhibition of OVCAR-3 human ovarian carcinoma cell proliferation, IC50=54 nM	12190313
A2780/DDP-S human ovarian carcinoma cell	Proliferation assay			Inhibition of A2780/DDP-S human ovarian carcinoma cell proliferation, IC50=56 nM	12190313
A2780/TAX-S human ovarian carcinoma cell	Proliferation assay			Inhibition of A2780/TAX-S human ovarian carcinoma cell proliferation, IC50=65 nM	12190313
LS174T human colon carcinoma cell	Proliferation assay			Inhibition of LS174T human colon carcinoma cell proliferation, IC50=65 nM	12190313
MCF-7 human breast carcinoma cell	Proliferation assay			Inhibition of MCF-7 human breast carcinoma cell proliferation, IC50=66 nM	12190313
PC3 human prostate carcinoma cell	Proliferation assay			Inhibition of PC3 human prostate carcinoma cell proliferation, IC50=66 nM	12190313
MLF mouse lung fibroblast cell	Proliferation assay			Inhibition of MLF mouse lung fibroblast cell proliferation, IC50=72 nM	12190313
LX-1 human lung carcinoma	Proliferation assay			Inhibition of LX-1 human lung carcinoma proliferation, IC50=75 nM	12190313
A431 human squamous cell	Proliferation assay			Inhibition of A431 human squamous cell carcinoma cell proliferation, IC50=75 nM	12190313
SKBR-3 human breast carcinoma cell	Proliferation assay			Inhibition of SKBR-3 human breast carcinoma cell proliferation, IC50=77 nM	12190313
A2780/TAX-R human ovarian carcinoma cell	Proliferation assay			Inhibition of A2780/TAX-R human ovarian carcinoma cell proliferation, IC50=78 nM	12190313
M109 mouse lung carcinoma cell	Proliferation assay			Inhibition of M109 mouse lung carcinoma cell proliferation, IC50=80 nM	12190313
CACO-2 human colon carcinoma cell	Proliferation assay			Inhibition of CACO-2 human colon carcinoma cell proliferation, IC50=86 nM	12190313
A549 human lung carcinoma cell	Proliferation assay			Inhibition of A549 human lung carcinoma cell proliferation, IC50=96 nM	12190313
MIP human colon carcinoma cell	Function assay			Inhibition of MIP human colon carcinoma cell line, IC50=0.12 μM	12190313
K562 human leukemia cell	Proliferation assay			Inhibition of K562 human leukemia cell proliferation, IC50=0.13 μM	12190313
human A2780 cell line	Proliferation assay		72 h	Antiproliferative effect against human A2780 cell line was determined in a whole cell 72 hr cytotoxicity assay, IC50=71 nM	15027863
human ovarian (A2780) cancer cell	Cytotoxic assay			Cytotoxic effect on human ovarian (A2780) cancer cell line, IC50=71 nM	15125971
ID8 cells	Proliferation assay			Antiproliferative activity against ID8 cells, IC50=7 nM	17123821
MCF7 cells	Proliferation assay			Antiproliferative activity against MCF7 cells, IC50=26 nM	17123821
Sf9 cells	Function assay			Inhibition of recombinant cyclin A/CDK2 expressed in Sf9 cells, IC50=12 nM	17904366
human A2780 cells	Function assay			Inhibition of cdk-mediated NPM phosphorylation at thr199 in human A2780 cells	18469809
human A2780 cells	Function assay		24 h	Inhibition of cdk-mediated Rb phosphorylation at thr821 in human A2780 cells after 24 hrs	18469809
human NCI60 cells	Proliferation assay		72 h	Antiproliferative activity against human NCI60 cells after 72 hrs by sulforhodamine B assay, GI50=74.7 nM	21080703
human NCI60 cells	Proliferation assay		72 h	Antiproliferative activity against human NCI60 cells assessed as lethal effect after 72 hrs by sulforhodamine B assay, LC50=0.904 μM	21080703
human A2780 cells	Cytotoxic assay		24 h	Cytotoxicity against human A2780 cells after 24 hrs by MTT assay, GI50=23 nM	23301767
human MRC5 cells	Cytotoxic assay		72 h	Cytotoxicity against human MRC5 cells after 72 hrs by MTT assay, GI50=28 nM	23301767
human A2780 cells	Cytotoxic assay		72 h	Cytotoxicity against human A2780 cells after 72 hrs by MTT assay, GI50=29 nM	23301767
human A2780 cells	Cytotoxic assay		48 h	Cytotoxicity against human A2780 cells after 48 hrs by MTT assay, GI50=31 nM	23301767
human MRC5 cells	Cytotoxic assay		48 h	Cytotoxicity against human MRC5 cells after 48 hrs by MTT assay, GI50=39 nM	23301767
human MRC5 cells	Cytotoxic assay		24 h	Cytotoxicity against human MRC5 cells after 24 hrs by MTT assay, GI50=49 nM	23301767
human HMEC1 cells	Cytotoxic assay		24 h	Cytotoxicity against human HMEC1 cells after 24 hrs by MTT assay, GI50=61 nM	23301767
human HMEC1 cells	Cytotoxic assay		48 h	Cytotoxicity against human HMEC1 cells after 48 hrs by MTT assay, GI50=62 nM	23301767
human HMEC1 cells	Cytotoxic assay		72 h	Cytotoxicity against human HMEC1 cells after 72 hrs by MTT assay, GI50=66 nM	23301767
A2780	Function assay	330 nM	24 hr	Percentage A2780 cells in sb-G1 after 24 hr r at 330 nM (IC50), Cell cycle = 0.001 μM.	12190313
A2780	Apoptosis assay	330 nM	24 hr	Apoptotic A2780 cells after 24 hr r at 330 nM (IC50), Cell cycle = 0.001 μM.	12190313
A2780	Function assay	2980 nM	24 hr	Percentage A2780 cells in sub-G1 after 24 hr r at 2980 nM (IC90), Cell cycle = 0.002 μM.	12190313
A2780	Apoptosis assay	2980 nM	24 hr	Apoptotic A2780 cells after 24 hr r at 2980 nM (IC90), Cell cycle = 0.009 μM.	12190313
A2780	Function assay	2980 nM	24 hr	Percentage A2780 cells in S-phase after 24 hr r at 2980 nM (IC90), Cell cycle = 0.012 μM.	12190313
A2780	Function assay	330 nM	24 hr	Percentage A2780 cells in S-phase after 24 hr r at 330 nM (IC50), Cell cycle = 0.016 μM.	12190313
A2780	Function assay	330 nM	24 hr	Percentage A2780 cells in G2/M after 24 hr r at 330 nM (IC50), Cell cycle = 0.023 μM.	12190313
A2780	Function assay	2980 nM	24 hr	Percentage A2780 cells in G2/M after 24 hr r at 2980 nM (IC90), Cell cycle = 0.025 μM.	12190313
A2780	Function assay	330 nM	24 hr	Percentage A2780 cells in G1 after 24 hr r at 330 nM (IC50), Cell cycle = 0.06 μM.	12190313
A2780	Function assay	2980 nM	24 hr	Percentage A2780 cells in G1 after 24 hr r at 2980 nM (IC90), Cell cycle = 0.061 μM.	12190313
A2780	Function assay		24 hrs	Inhibition of cdk-mediated Rb phosphorylation at ser807/811 in human A2780 cells after 24 hrs	18469809
A2780	Function assay		24 hrs	Inhibition of cdk9-mediated RNA pol2 CTD phosphorylation at ser2 in human A2780 cells after 24 hrs	18469809
A2780	Function assay		24 hrs	Inhibition of cdk7-mediated RNA pol2 CTD phosphorylation at ser5 in human A2780 cells after 24 hrs	18469809
A2780	Apoptosis assay		24 hrs	Induction of apoptosis in human A2780 cells assessed as appearance of Mcl1 protein level after 24 hrs	18469809
DR-U2OS-GFP	Function assay	0.1 uM	56 hrs	Reduction of homologous recombination in human DR-U2OS-GFP cells expressing I-SceI nuclease assessed as reduction of RAD51 level at 0.1 uM after 56 hrs by immunoblotting	21417417
A2780	Function assay		24 hrs	Inhibition of CDK9 in human A2780 cells assessed as reduction of RNAPII CTD phosphorylation at Ser2 at GI50 concentration after 24 hrs by Western blotting analysis	23301767
A2780	Cell cycle assay		24 hrs	Cell cycle arrest in human A2780 cells assessed as accumulation at G2/M phase at less than GI50 after 24 hrs by flow cytometric analysis	23301767
A2780	Function assay		24 hrs	Inhibition of CDK9 in human A2780 cells assessed as downregulation of MCL1 at GI50 to 5XGI50 concentration after 24 hrs by Western blotting analysis	23301767
A2780	Apoptosis assay		24 hrs	Induction of apoptosis in human A2780 cells assessed as induction of PARP cleavage at GI50 to 5XGI50 concentration after 24 hrs by Western blotting analysis	23301767
A2780	Function assay		24 hrs	Inhibition of CDK9 in human A2780 cells assessed as downregulation of HDM2 at GI50 to 5XGI50 concentration after 24 hrs by Western blotting analysis	23301767
MT4	Antiproliferative assay			Antiviral activity against Human immunodeficiency virus 1 NL 4-3 infected in MT4 cells measured on day 4 post infection by p24 assay, EC50 = 0.015 μM.	25914804
MT4	Cytotoxicity assay			Cytotoxicity against human MT4 cells, IC50 = 0.067 μM.	25914804
Sf9	Function assay		15 mins	Inhibition of CDK2/cyclin E1 (unknown origin) expressed in Sf9 insect cells using UlightCFFKNIVTPRTPPPSQGK-amide substrate after 15 mins by autoradiography, IC50 = 0.13 μM.	25914804
A549	Antiproliferative assay		3 days	Antiproliferative activity against human A549 cells after 3 days by SRB method, GI50 = 0.14 μM.	25914804
DU145	Antiproliferative assay		3 days	Antiproliferative activity against human DU145 cells after 3 days by SRB method, GI50 = 0.15 μM.	25914804
KB	Antiproliferative assay		3 days	Antiproliferative activity against human KB cells after 3 days by SRB method, GI50 = 0.16 μM.	25914804
KBVIN	Antiproliferative assay		3 days	Antiproliferative activity against human KBVIN cells after 3 days by SRB method, GI50 = 0.18 μM.	25914804
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by Celltiter-Glo reagent based assay in presence of 10% fetal bovine serum, EC50 = 0.034 μM.	26985305
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by Celltiter-Glo reagent based assay in presence of 0.625% fetal bovine serum, EC50 = 0.059 μM.	26985305
Sf9	Function assay		10 mins	Inhibition of human His6-tagged CDK9/cyclin T1 expressed in baculovirus infected sf9 cells using GST-CTD as substrate after 10 mins in presence of [gamma-32P]ATP by SDS-PAGE analysis, IC50 = 0.0025 μM.	27171036
Sf21	Function assay			Inhibition of recombinant human full length C-terminal His6-tagged CDK9/cyclin T1 expressed in baculovirus infected Sf21 insect cells using PDKtide as substrate, IC50 = 0.011 μM.	27171036
HeLa	Cytotoxicity assay		72 hrs	Cytotoxicity against human HeLa cells assessed as decrease in cell viability after 72 hrs by MTT assay, CC50 = 0.12 μM.	27171036
sf21	Function assay			Inhibition of full length human N-terminal His6-tagged CDK6/N-terminal GST-tagged cyclin D3 expressed in sf21 cells using histone H1 substrate, IC50 = 0.395 μM.	27171036
Sf21	Function assay			Inhibition of recombinant human full length C-terminal His6-tagged CDK7/cyclin H/N-terminal GST-tagged MAT1 expressed in baculovirus infected Sf21 insect cells using cdk7 substrate peptide, IC50 = 0.514 μM.	27171036
HepG2	Antiproliferative assay		72 hrs	Antiproliferative activity against human HepG2 cells after 72 hrs by CelTiter-Glo assay, EC50 = 0.1464 μM.	29407975
KOPN8	Antiproliferative assay		72 hrs	Antiproliferative activity against human KOPN8 cells after 72 hrs by CelTiter-Glo assay, EC50 = 0.1926 μM.	29407975
SEM	Antiproliferative assay		72 hrs	Antiproliferative activity against human SEM cells after 72 hrs by CelTiter-Glo assay, EC50 = 0.2043 μM.	29407975
UOCB1	Antiproliferative assay		72 hrs	Antiproliferative activity against human UOCB1 cells after 72 hrs by CelTiter-Glo assay, EC50 = 0.2084 μM.	29407975
KOPN8	Apoptosis assay	0.5 uM	3 to 24 hrs	Induction of apoptosis in human KOPN8 cells assessed as upregulation of cleaved PARP level at 0.5 uM after 3 to 24 hrs by Western blot analysis	29407975
KOPN8	Apoptosis assay	0.5 uM	1 hr	Induction of apoptosis in human KOPN8 cells assessed as upregulation of cleaved PARP level at 0.5 uM pre-treated with NAC for 1 hr and measured after 3 to 24 hrs by Western blot analysis	29407975
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Información química, almacenamiento y estabilidad

Peso molecular	401.84	Fórmula	C₂₁H₂₀ClNO₅	Almacenamiento (Desde la fecha de recepción)	3 años-20°Cpolvo
Nº CAS	146426-40-6	Descargar SDF		Almacenamiento de soluciones madre	1 año-80°Cen disolvente 1 mes-20°Cen disolvente
Sinónimos	NSC 649890, HMR-1275, L86-8275			Smiles	CN1CCC(C(C1)O)C2=C(C=C(C3=C2OC(=CC3=O)C4=CC=CC=C4Cl)O)O

Solubilidad

In vitro
Lote:

DMSO : 80 mg/mL (199.08 mM)
(El DMSO contaminado con humedad puede reducir la solubilidad. Usar DMSO fresco y anhidro.)

Water : Insoluble

Ethanol : Insoluble

Calculadora de Molaridad

Masa	Concentración	Volumen	Peso molecular
=	×	×

Calculadora de Dilución Calculadora de Peso Molecular

In vivo Lote:
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 30%PEG300 2 65%ddH2O Validado por los laboratorios Selleck. Si necesita ajustes en esta formulación, contacte con nuestro equipo de ventas para pruebas personalizadas.	2.500mg/ml (6.22mM)	Taking the 1 mL working solution as an example, add 50 μL of 50 mg/ml clarified DMSO stock solution to 300 μL of PEG 300, mix evenly to clarify it; then continue to add 650 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.
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.

Calculadora de formulación in vivo (Solución clara)

Paso 1: Introduzca la información a continuación (Recomendado: Un animal adicional para tener en cuenta la pérdida durante el experimento)

Dosis: mg/kg Peso promedio de los animales: g Volumen de dosificación por animal:μL Número de animales:

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.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

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.

Mecanismo de acción

Características	First CDK inhibitor to be used in human clinical trials.
Targets/IC₅₀/Ki	CDK9 (Cell-free assay) 20 nM CDK1 (Cell-free assay) 30 nM CDK4 (Cell-free assay) 20-40 nM CDK2 (Cell-free assay) 40 nM CDK6 (Cell-free assay) 60 nM CDK7 (Cell-free assay) 875 nM
In vitro	Flavopiridol (Alvocidib) muestra menor actividad contra quinasas no relacionadas como MAP, PAK, PKC y EGFR con un IC50 de >14 μM. Inhibe significativamente el crecimiento de colonias de células HCT116, A2780, PC3 y Mia PaCa-2 con IC50 de 13 nM, 15 nM, 10 nM y 36 nM, respectivamente. Este compuesto también inhibe potentemente la actividad de la glucógeno sintasa quinasa-3 (GSK-3) con un IC50 de 280 nM. En comparación con otras CDKs, inhibe la actividad de CDK7 con menor potencia, con un IC50 de 875 nM. A 0.5 μM, inhibe tanto pSer807/811 Rb como pThr199 NPM, mientras que se observan cambios leves en pThr821 Rb. También disminuye el nivel general de ARN polimerasa II, así como la fosforilación de la ARN polimerasa II en los repetidos CTD en Ser2 Ser5. Como un inhibidor de CDK de amplio espectro, puede inhibir la progresión del Cell Cycle tanto en G1 como en G2. A 0.3 μM, induce la detención en G1 en células MCF-7 o MDA-MB-468 mediante la inhibición de la actividad quinasa de CDK4 o CDK2. Exhibe una potente citotoxicidad contra una amplia variedad de líneas celulares tumorales con valores de IC50 que van desde 16 nM para LNCAP hasta 130 nM para K562.
Ensayo de quinasa	Ensayo de quinasa CDK
Ensayo de quinasa	Para el ensayo de quinasa CDK1/ciclina B1, las reacciones de quinasa consisten en 100 ng del complejo GST-CDK1/ciclina B1 (humano) expresado por baculovirus, 1 μg de histona HI, 0.2 μCi de [γ-³³P]ATP, 25 μM de ATP en 50 μL de tampón de quinasa (50 mM Tris, pH 8.0, 10 mM MgCl₂, 1 mM EGTA, 0.5 mM DTT). Para el ensayo de quinasa CDK2/ciclina E, las reacciones de quinasa consisten en 5 ng del complejo GST-CDK2/ciclina E (humano) expresado por baculovirus, 0.5 μg de proteína de fusión GST-RB (aminoácidos 776-928 de la proteína del retinoblastoma), 0.2 μCi de [γ-³³P]ATP, 25 μM de ATP en 50 μL de tampón de quinasa (50 mM Hepes, pH 8.0, 10 mM MgCl₂, 1 mM EGTA, 2 mM DTT). Para el ensayo de quinasa CDK4/ciclina D1, las reacciones de quinasa consisten en 150 ng de GST-CDK4/ciclina D1 (humano) expresado por baculovirus, 280 ng de Stag-ciclina D1, 0.5 μg de proteína de fusión GST-RB (aminoácidos 776-928 de la proteína del retinoblastoma), 0.2 μCi de [γ-³³P]ATP, 25 μM de ATP en 50 μL de tampón de quinasa (50 mM Hepes, pH 8.0, 10 mM MgCl₂, 1 mM EGTA, 2 mM DTT). Las reacciones se incuban durante 45 minutos para CDK1 y CDK2, o 1 hora para CDK4 a 30 °C y se detienen con la adición de ácido tricloroacético (TCA) frío a una concentración final del 15 %. Los precipitados de TCA se recogen en placas unifilter GF/C utilizando un recolector universal Filtermate y los filtros se cuantifican utilizando un contador de centelleo líquido de 96 pocillos TopCount. Flavopiridol (Alvocidib) se disuelve a 10 mM en dimetilformamida (DMF) y se evalúa en seis concentraciones, cada una por triplicado. La concentración final de DMF en el ensayo = 2 %. Los valores de IC50 se derivan mediante análisis de regresión no lineal y tienen un coeficiente de variación = 16 %. Para ensayar su actividad sobre CDK6, se establece un ensayo de unión por filtro. Los siguientes componentes se combinan en la mezcla de reacción: 2 μL de CDK6 (0.7 mg/μL), 5 μL de histona H1 (6 mg/mL), 14 μL de tampón de quinasa (60 mM β-glicerofosfato, 30 mM p-nitrofenil fosfato, 25 mM MOPS (pH 7.0), 5 mM EGTA, 15 mM MgCl₂, 1 mM DTT, 0.1 mM Na-vanadato), 3 μL de concentraciones crecientes de este compuesto diluidas en 50 % de DMSO, y 6 μL de ³³P-ATP (1 mCi/mL) en ATP no radiactivo a una concentración de 90 μM (concentración final: 15 μM). El ensayo se inicia con la adición de ³³P-ATP. La reacción se incuba durante 20 minutos a 30 °C. Una alícuota de 25 μL del sobrenadante se aplica luego sobre papel de fosfocelulosa Whatman P81. Los filtros se lavan 5 veces con una solución de ácido fosfórico al 1 %. Los filtros húmedos se cuentan en presencia de 1 mL de líquido de centelleo. La actividad de Cdk9 se mide utilizando 50 nM de Cdk9/ciclina T recombinante en 50 mM HEPES pH 7.5, 10 mM MgCl₂, 1 mM DTT, 3 μM Na₃VO₄, 150 μM de péptido CDT de ARN polimerasa y 80 μM de ATP. El ensayo de Cdk7 se realiza en el mismo tampón utilizando 37 nM de quinasa purificada en presencia de 200 μM de ATP y 10 μM de proteína de unión a mielina como sustrato. La potencia de este compuesto hacia CDK9 y CDK7 se determina utilizando un ensayo basado en un intercambiador aniónico fuerte (resina Dowex 1-X8, forma formato) o un ensayo de proximidad por centelleo. Los valores de IC50 se calculan a partir de las curvas de dosis-respuesta.
In vivo	La administración de S1230 a 7.5 mg/kg durante 7 días muestra una ligera actividad antitumoral contra la leucemia murina P388, resultando en un valor %T/C de 110, y es activa contra el carcinoma ovárico humano A2780 implantado subcutáneamente en ratones nude, produciendo una eliminación de 1.5 log de células (LCK). El tratamiento con S1230 a 1-2.5 mg/kg durante 10 días suprime significativamente la artritis inducida por colágeno en ratones de manera dosis-dependiente, al inhibir la hiperplasia sinovial y la destrucción articular, mientras que las concentraciones séricas de anticuerpos anti-colágeno tipo II (CII) y las respuestas proliferativas al CII se mantienen. En los xenoinjertos de Hct116 con p21 intacta en ratones nude, la administración de CPT-11 (100 mg/kg) seguida de S1230 (3 mg/kg) 7 y 16 horas después inhibe significativamente la regresión tumoral en un 86 % y 82 %, respectivamente, mostrando una inhibición > 2 veces en comparación con CPT-11 solo en un 40 %. La combinación produce una tasa de respuesta completa (CR) de aproximadamente el 30 %, en contraste con CPT-11 solo, donde no se encuentra CR.
Referencias	[1] https://pubmed.ncbi.nlm.nih.gov/11063609/ [2] https://pubmed.ncbi.nlm.nih.gov/15689157/ [3] https://pubmed.ncbi.nlm.nih.gov/18469809/ [4] https://pubmed.ncbi.nlm.nih.gov/8674031/ [5] https://pubmed.ncbi.nlm.nih.gov/12190313/ [6] https://pubmed.ncbi.nlm.nih.gov/18209094/ [7] https://pubmed.ncbi.nlm.nih.gov/11751522/ [8] https://pubmed.ncbi.nlm.nih.gov/18469809/ [9] https://pubmed.ncbi.nlm.nih.gov/11311660/

Aplicaciones

Métodos	Biomarcadores	Imágenes	PMID
Western blot	Cleaved caspase-8 / Cleaved caspase-9 / Cleaved caspase-3 p-RNAPII / p-eIF4E / Mnk1 p-ERK / ERK / p-p38 / p-4EBP1 / 4EBP1 / p-S6 CDK2 / CDK4 / Cyclin A / p21 / p27 / Rb		31193061
Growth inhibition assay	Cell viability		31193061

Información del ensayo clínico

(datos de https://clinicaltrials.gov, actualizado el 2024-05-22)

Número NCT	Reclutamiento	Condiciones	Patrocinador/Colaboradores	Fecha de inicio	Fases
NCT03441555	Completed	Acute Myeloid Leukemia (AML)	AbbVie\|Sumitomo Pharma America Inc.	May 30 2018	Phase 1
NCT03298984	Completed	Acute Myeloid Leukemia	Sumitomo Pharma America Inc.	September 25 2017	Phase 1

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