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Flavopiridol (Alvocidib, HMR-1275) CDK Inhibiteur

Name: Flavopiridol (Alvocidib, HMR-1275)
Brand: Selleck Chemicals
SKU: S1230
Rating: 5 (148 reviews)

Réf. Catalogue: S1230

Le Flavopiridol (Alvocidib) entre en compétition avec l'ATP pour inhiber les CDK, y compris CDK1, CDK2, CDK4, CDK6 et CDK9 avec des valeurs d'IC50 comprises entre 20 et 100 nM. Il est plus sélectif pour CDK1, 2, 4, 6, 9 par rapport à CDK7. Le Flavopiridol est initialement découvert pour inhiber l'EGFR et la PKA. Le Flavopiridol induit l'autophagy et le stress du RE. Le Flavopiridol bloque la réplication du HIV-1. Phase 1/2.

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

Structure chimique

Poids moléculaire: 401.84

Aller à

Contrôle Qualité (Quality Control)

Lot : Pureté : 99.92%

99.92

Cibles apparentées	HSP PD-1/PD-L1 ROCK Wee1 DNA/RNA Synthesis Microtubule Associated Ras KRas Aurora Kinase Casein Kinase
Autre CDK Inhibiteurs	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

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
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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Informations chimiques, stockage et stabilité (Chemical Information, Storage & Stability)

Poids moléculaire	401.84	Formule	C₂₁H₂₀ClNO₅	Stockage (À compter de la date de réception)	3 ans-20°Cpoudre
N° CAS	146426-40-6	Télécharger le SDF		Stockage des solutions mères	1 an-80°Cen solvant 1 mois-20°Cen solvant
Synonymes	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

Solubilité (Solubility)

In vitro
Lot:

DMSO : 80 mg/mL (199.08 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 30%PEG300 2 65%ddH2O Validé par les laboratoires Selleck. Si vous avez besoin d'ajustements à cette formulation, contactez notre équipe commerciale pour des tests personnalisés.	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.

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)

Caractéristiques	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) displays less activity against unrelated kinases such as MAP, PAK, PKC, and EGFR with IC50 of >14 μM. It significantly inhibits the colony growth of HCT116, A2780, PC3, and Mia PaCa-2 cells with IC50 of 13 nM, 15 nM, 10 nM and 36 nM, respecitively. This compound also potently inhibits the activity of Glycogen synthase kinase-3 (GSK-3) with an IC50 of 280 nm. Compared with other CDKs, it inhibits the activity of CDK7 less potently with IC50 of 875 nM. At 0.5 μM, it inhibits both pSer807/811 Rb and pThr199 NPM, whereas mild changes are observed at pThr821 Rb. It also decreases the overall RNA polymerase II level, as well as the phosphorylation of RNA polymerase II on the CTD repeats at Ser2 Ser5. As a broad spectrum CDK inhibitor, it can inhibit cell cycle progression in either G1 or G2. At 0.3 μM, it induces G1 arrest in either MCF-7 or MDA-MB-468 cells by inhibition of the CDK4 or CDK2 kinase activity. It exhibits potent cytotoxicity against a wide variety of tumor cell lines with IC50 values ranging form 16 nM for LNCAP to 130 nM for K562.
Essai kinase	Essai kinase CDK
Essai kinase	Pour l'essai kinase CDK1/cycline B1, les réactions de kinase consistent en 100 ng de complexe GST-CDK1/cycline B1 (humain) exprimé par baculovirus, 1 μg d'histone HI, 0,2 μCi de [γ-³³P]ATP, 25 μM d'ATP dans 50 μL de tampon kinase (50 mM Tris, pH 8,0, 10 mM MgCl₂, 1 mM EGTA, 0,5 mM DTT). Pour l'essai kinase CDK2/cycline E, les réactions de kinase consistent en 5 ng de complexe GST-CDK2/cycline E (humain) exprimé par baculovirus, 0,5 μg de protéine de fusion GST-RB (acides aminés 776-928 de la protéine du rétinoblastome), 0,2 μCi de [γ-³³P]ATP, 25 μM d'ATP dans 50 μL de tampon kinase (50 mM Hepes, pH 8,0, 10 mM MgCl₂, 1 mM EGTA, 2 mM DTT). Pour l'essai kinase CDK4/cycline D1, les réactions de kinase consistent en 150 ng de GST-CDK4/cycline D1 (humain) exprimé par baculovirus, 280 ng de Stag-cycline D1, 0,5 μg de protéine de fusion GST-RB (acides aminés 776-928 de la protéine du rétinoblastome), 0,2 μCi de [γ-³³P]ATP, 25 μM d'ATP dans 50 μL de tampon kinase (50 mM Hepes, pH 8,0, 10 mM MgCl₂, 1 mM EGTA, 2 mM DTT). Les réactions sont incubées pendant 45 minutes pour CDK1 et CDK2, ou 1 heure pour CDK4 à 30 °C et arrêtées par l'addition d'acide trichloroacétique froid (TCA) à une concentration finale de 15 %. Les précipités de TCA sont recueillis sur des plaques unifilter GF/C à l'aide d'un moissonneur universel Filtermate et les filtres sont quantifiés à l'aide d'un compteur à scintillation liquide TopCount à 96 puits. Le Flavopiridol (Alvocidib) est dissous à 10 mM dans du diméthylformamide (DMF) et évalué à six concentrations, chacune en triple. La concentration finale de DMF dans l'essai = 2 %. Les valeurs d'IC50 sont dérivées par analyse de régression non linéaire et ont un coefficient de variation = 16 %. Pour évaluer son activité sur CDK6, un essai de liaison sur filtre est établi. Les éléments suivants sont combinés dans le mélange réactionnel : 2 μL de CDK6 (0,7 mg/μL), 5 μL d'histone H1 (6 mg/mL), 14 μL de tampon kinase (60 mM β-glycérophosphate, 30 mM p-nitrophényl phosphate, 25 mM MOPS (pH 7,0), 5 mM EGTA, 15 mM MgCl₂, 1 mM DTT, 0,1 mM Na-vanadate), 3 μL de concentrations croissantes de ce composé diluées dans 50 % de DMSO, et 6 μL de ³³P-ATP (1 mCi/mL) dans de l'ATP non radioactif à une concentration de 90 μM (concentration finale : 15 μM). L'essai est initié par l'addition de ³³P-ATP. La réaction est incubée pendant 20 minutes à 30°C. Une aliquote de 25 μL du surnageant est ensuite déposée sur du papier phosphocellulose Whatman P81. Les filtres sont lavés 5 fois avec une solution d'acide phosphorique à 1 %. Les filtres humides sont comptés en présence de 1 mL de liquide de scintillation. L'activité de Cdk9 est mesurée en utilisant 50 nM de Cdk9/cycline T recombinant dans 50 mM HEPES pH 7,5, 10 mM MgCl₂, 1 mM DTT, 3 μM Na₃VO₄, 150 μM de peptide CDT de l'ARN polymérase et 80 μM d'ATP. L'essai Cdk7 est réalisé dans le même tampon en utilisant 37 nM de kinase purifiée en présence de 200 μM d'ATP et 10 μM de protéine de liaison à la myéline comme substrat. La puissance de ce composé envers CDK9 et CDK7 est déterminée en utilisant soit un essai basé sur un échangeur d'anions fort (résine Dowex 1-X8, forme formate) soit un essai de scintillation de proximité. Les valeurs d'IC50 sont calculées à partir des courbes dose-réponse.
In vivo	Administration of S1230 at 7.5 mg/kg for 7 days displays slight antitumor activity against P388 murine leukemia, resulting in %T/C value of 110, and active against the human A2780 ovarian carcinoma implanted sc in nude mice, producing 1.5 log cell kill (LCK). S1230 treatment at 1-2.5 mg/kg for 10 days significantly suppresses collagen-induced arthritis in mice in a dose-dependent manner, by inhibiting synovial hyperplasia and joint destruction, whereas serum concentrations of anti-collagen type II (CII) Abs and proliferative responses to CII are maintained. In the p21-intact Hct116 xenografts in nude mice, administration of CPT-11 (100 mg/kg) followed by S1230 (3 mg/kg) 7 and 16 hours later significantly inhibits tumor regression by 86% and 82%, respectively, displaying >2 fold inhibition compared with CPT-11 alone by 40 %. The combination produces ~30% complete response rate (CR) in contrast to CPT-11 alone where no CR is found.
Références	[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/

Applications (Applications)

Méthodes	Biomarqueurs	Images	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

Informations sur l'essai clinique (Clinical Trial Information)

(données du https://clinicaltrials.gov, mis à jour le 2023-11-09)

Numéro NCT	Recrutement	Conditions	Promoteur/Collaborateurs	Date de début	Phases
NCT03593915	TERMINATED	Myelodysplastic Syndromes (MDS)	Sumitomo Pharma America, Inc.	2018-08-29	PHASE1; PHASE2
NCT03969420	TERMINATED	Acute Myeloid Leukemia (AML)	Sumitomo Pharma America, Inc.	2020-01-15	PHASE2
NCT03441555	COMPLETED	Acute Myeloid Leukemia (AML)	AbbVie	2018-05-30	PHASE1
NCT04493099	WITHDRAWN	Recurrent Acute Myeloid Leukemia; Refractory Acute Myeloid Leukemia	M.D. Anderson Cancer Center	2020-10	PHASE1; PHASE2
NCT03563560	COMPLETED	Acute Myeloid Leukemia	Sumitomo Pharma Co., Ltd.	2018-05-15	PHASE1
NCT03298984	COMPLETED	Acute Myeloid Leukemia	Sumitomo Pharma America, Inc.	2017-09-25	PHASE1

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