Flavopiridol HCl

 

Distributor Tel: +886-2-2827-1197jimmy@new.abscience.com.tw

Biological Activity

Description	Flavopiridol competes with ATP to inhibit CDKs including CDK1, CDK2, CDK4 and CDK6 with IC50 of ~ 40 nM. It is 7.5-fold more selective for CDK1/2/4/6 than CDK7. Flavopiridol is initially found to inhibit EGFR and PKA. Phase 1/2.
Targets	CDK1	CDK2	CDK4	CDK6	CDK7
IC50	40 nM	40 nM	40 nM	40 nM	300 nM [1]
In vitro	Flavopiridol is initially found to inhibit the epidermal growth factor receptor and protein kinase A (IC50 = 21 and 122 μM). Flavopiridol is later shown to inhibit cell proliferation, at more physiologically relevant concentrations (IC50 = 66 nM) when Flavopiridol is tested in the National Cancer Institute Development Therapeutics Program panel of 60 human tumor cell lines. [1] Flavopiridol induces G1 arrest with inhibition of CDK2 and CDK4 in human breast carcinoma cells in a time and concentration dependent manner. [2] Short time treatment of Flavopiridol (~12 hours) induce apoptosis in hematopoietic cell lines including SUDHL4, SUDHL6 (B-cell lines), Jurkat and MOLT4 (T-cell lines ), and HL60 (myeloid). [3] In the clonogenic assay, Flavopiridol functions as a highly potent cytotoxic compound with a mean IC70 with 8 ng/mL in 23 human tumor models. [4] A recent study shows Flavopiridol treatment induces a substantial AKT-Ser473 phosphorylation in human glioblastoma T98G cell line. [5]
In vivo	At the maximal tolerated dose of 10 mg/kg/day administered p.o. on days 1-4 and 7-11, Flavopiridol effects tumor regression in PRXF1337 and tumor stasis lasting for 4 weeks in PRXF1369. [4] After treatment with 7.5 mg/kg Flavopiridol bolus intravenous (IV) or intraperitoneal on each of 5 consecutive days, 11 out of 12 advanced stage subcutaneous (s.c.) human HL-60 xenografts undergo complete regressions, and animals remain disease-free several months after one course of Flavopiridol treatment. SUDHL-4 s.c. lymphomas treated with flavopiridol at 7.5 mg/kg bolus IV for 5 days undergo either major (two out of eight mice) or complete (four out of eight mice) regression, with two animals remaining disease-free for more than 60 days. The overall growth delay is 73.2%. Daily IV or IP administration of flavopiridol results in peak plasma levels of about 7 µM, followed by a progressive decline to approximately 100 nM in 8 hours.[6]
Features

Protocol(Only for Reference)

Kinase Assay: ^[1]

Recombinant CDKs Kinase Reactions

CDKs activities are determined in microtiter plates as follows. Forty μg Gst-Rb are mixed with different amounts of Flavopiridol and unlabeled ATP. Reactions are then started by the addition of an ammonium sulfate cut of the S100 fraction obtained from insect cells expressing recombinant human CDKs. The final reaction conditions are 10 mM MgCl2, 50 mM Tris-HCl (pH 7.5), and 1 mM DTT. The final concentration of ATP is adjusted accordingly. Radiolabeled ATP is used as a phosphoryl donor. The reaction is carried out for 2.5 minutes at 30 °C after addition of enzyme and then terminated with the addition of EDTA. The Gst-Rb is then captured with glutathione-Sepharose and the incorporated radioactivity is determined by liquid scintillation counting.

Cell Assay: ^[2]

Cell lines	SUDHL4, SUDHL6, Jurkat, MOLT4, and HL60
Concentrations	0, 100 500, 5000 nM
Incubation Time	14 hours
Method	Cells grown at a density of 1 × 106 cells/mL are exposed to Flavopiridol for different concentrations and time periods. DNA is extracted. Briefly, cells are washed once with cold phosphate-buffered saline (PBS) and lysed with 3 mL lysis buffer (5 mM Tris-HCL [pH 7.5]; 20 mM EDTA; 0.5% Triton X-100) for 15 minutes at 4 °C. The chromatin of the cell lysates is isolated by centrifugation (20 minutes at 26,000g, 4 °C). The supernatants containing small DNA fragments are extracted sequentially with phenol, phenol:chloroform (1:1), and chloroform. Nucleic acids are precipitated in 0.5 M NaCl, 90% ethanol at -20 °C overnight. RNA is then digested by bovine RNAaseA (60 μg/mL). After sequential reextraction and reprecipitation, DNA is dissolved in 10 mM Tris-HCL (pH 7.5), 1 mM EDTA, 0.5% sodium dodecyl sulfate (SDS) before electrophoresis on 1.6% agarose gel.

Animal Study: ^[4] [6]

Animal Models	Human prostate cancer xenografts, PRXFI337 and PRXFI369, grown s.c. in nude mice [4] Human promyelocytic leukemia HL-60, human B-cell follicular lymphoma SUDHL-4, and acquired immunodeficiency syndrome (AIDS)-related human B-cell lymphoma AS283 xenografts in mice. [6]
Dosages	10 mg/kg/d [4]; 7.5 mg/kg/d [6]
Administration	p.o.[4]; i.p. or i.v. [6]
Solubility	30% propylene glycol, 5% Tween 80, 65% D5W, 30 mg/mL

1

References

Clinical Trial Information( data from http://clinicaltrials.gov)

NCT Number	Recruitment	Conditions	Sponsor /Collaborators	Start Date	Phases
NCT00991952	Active, not recruiting	Adenocarcinoma of the Gastroesophageal Junction|Diffuse Adenocarcinoma of the Stomach|Intestinal Adenocarcinoma of the Stomach|Mixed Adenocarcinoma of the Stomach|Recurrent Gastric Cancer|Stage IIIA G	National Cancer Institute (NCI)	2009-09	Phase 2
NCT01076556	Active, not recruiting	B-cell Chronic Lymphocytic Leukemia|Contiguous Stage II Small Lymphocytic Lymphoma|Noncontiguous Stage II Small Lymphocytic Lymphoma|Prolymphocytic Leukemia|Recurrent Small Lymphocytic Lymphoma|Refrac	National Cancer Institute (NCI)	2010-08	Phase 1
NCT01413880	Active, not recruiting	Acute Myelogenous Leukemia	Sidney Kimmel Comprehensive Cancer Center	2011-05	Phase 2
NCT01349972	Recruiting	Acute Myeloid Leukemia With Multilineage Dysplasia Following Myelodysplastic Syndrome|Adult Acute Minimally Differentiated Myeloid Leukemia (M0)|Adult Acute Monoblastic Leukemia (M5a)|Adult Acute Mono	National Cancer Institute (NCI)	2011-08	Phase 2
NCT00957905	Active, not recruiting	Recurrent Extragonadal Germ Cell Tumor|Recurrent Extragonadal Non-seminomatous Germ Cell Tumor|Recurrent Extragonadal Seminoma|Recurrent Malignant Testicular Germ Cell Tumor|Recurrent Ovarian Germ Cel	National Cancer Institute (NCI)	2013-05	Phase 2

Chemical Information

Download Flavopiridol HCl SDF

Chemical Name	2-(2-chlorophenyl)-5,7-dihydroxy-8-((3S,4R)-3-hydroxy-1-methylpiperidin-4-yl)-4H-chromen-4-one hydrochloride

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.4383	4.383	8.766	13.149

Customer Reviews (1)

Click to enlarge	Rating
	Source	PNAS 2011.May 108:8417. Flavopiridol HCl purchased from Selleck
	Method	Fluorescent Microscopy/Confocal Imaging
	Cell Lines	Tg:Pomc-Pttg embryo, Pomc-eGFP embryo
	Concentrations	50 μM
	Incubation Time	18-48 h
	Results	Although flavopiridol retarded early embryonic development before corticotroph ontogeny occurred, in vivo treatment of zebrafish embryos with R-roscovitine, olomoucine, PD-0332991, and CAY10572 starting at 18 hpf caused no apparent growth defect by 40 hpf. Strikingly, R-roscovitine-treated embryos exhibited approximately 40% reduction in pituitary POMC-eGFP expression compared with controls.

Product Citations (1)

• 
  

  Comparative drug screening in NUT midline carcinoma [Beesley AH Br J Cancer 2014;10.1038/bjc.2014.54]