Cilengitide

 

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Biological Activity

Description	Cilengitide is a potent integrin inhibitor for αvβ3 receptor and αvβ5 receptor with IC50 of 4.1 nM and 79 nM, respectively; ~10-fold selectivity against gpIIbIIIa. Phase 2.
Targets	αvβ3 receptor	αvβ5 receptor
IC50	4.1 nM	79 nM [2]
In vitro	Cilengitide is a cyclized pentapeptide peptidomimetic designed to compete for the arginine-glycine-aspartic acid (RGD) peptide sequence that regulates integrin-ligand binding. Cilengitide selectively and potently blocks the ligation of theαvβ3 andαvβ5 integrins to provisional matrix proteins such as vitronectin, fibronectin, fibrinogen, von Willebrand factor, osteopontin, and others. [1] Cilegitide inhibits angiogenesis in vitro. 10 μM Cilengitide completely inhibits attachment of BAE, BME and HUVE cells on vitronectin and fibronectin. Cilengitide inhibits in vitro angiogenesis of BAE cells on three-dimensional collagen and fibrin gels pretreated with FGF-2(or VEGF-A) with IC50 of 15 μM and 8 μM, 4 μM and 3 μM, respectively. [2] Cilengitide blocks proliferation and induces apoptosis of endothelial cells as well as differentiation of human endothelial precursor cells (EPCs). 50 μg/mL Cilengitide completely inhibits the proliferation of human microvascular endothelial cell line HMEC-1 and leads to apoptosis in ~30% cells. [3] 1.0 μM Cilengitide treating for 9 days inhibits the proliferation of EPCs by nearly 40%. 1 μM Cilengitide inhibits the differentiation of EPCs by more than 80% at 14 days. [4] Cilengitide inhibits adhesion and induces apoptosis of tumor cells. 25 μg/mL Cilengitide causes detachment of DAOY cells (medulloblastoma) and U87MG cells (glioblastoma) from vitronectin and tenascin by more than 60%. 25 μg/mL Cilengitide induces a nearly 50% apoptosis rate of these cells. [5]
In vivo	Cilengitide is activity against tumor growth and angiogenesis as single-agent. 100 μg Cilengitide induces a significant decrease in the number of CD 31+ vessels seen in tumors (2/high-power field) compared with control tumors (56/high-power field). 100 μg Cilengitide increases cellular apoptosis in the brain tumors of animals (2.2% apoptotic cells/high-power field) compared with those receiving the inactive peptide (1.7% cells/high-power field). Cilengitide treatment results in prolonged survival of the mice bearing melanoma xenografts M21 compared with control treatment group. (36.5 vs 17.3 days). [5] Cilengitide can augment the therapeutic benefit associated with cytotoxic agents including chemotherapy and radiation therapy in tumor models. Cilengitide (250 mg/dose) alone does not alter tumor growth of breast cancer xenografts when compared with untreated mice, but combined modality RIT (CMRIT) using RIT and six doses of Cilengitide (250 mg/dose) increases efficacy of treatment, with the cure rate for mice that receives only RIT increasing from 15 to 53%. CMRIT significantly increases apoptosis of tumor and endothelial cells 5 days, and decreases tumor proliferation. [6]
Features

Protocol(Only for Reference)

Kinase Assay: ^[2]

Integrin-binding competition assay

Recombinant soluble integrins are immobilized, and peptides, which are serially diluted in Tris-buffered saline (TBS++) (0.1% (w/v) BSA, 150 mM NaCl, 1 mM CaCl2, 1 mM MgCl2 10 μM MnCl2, 20 mM Tris-HCl; pH 7.4), are added in parallel with biotinylated vitronectin (to 1μg/mL). After a 3-h incubation at 37℃ and washing with Tris–buffered saline, bound ligand is detected by incubation with an antibiotin alkaline phosphatase-conjugated antibody (BioRad) followed by development with p-nitrophenyl phosphatase substrate. The reaction is stopped by the addition of NaOH and the color intensity read at 405 nm.

Cell Assay: ^[3]

Cell lines	Human microvascular endothelial cell line HMEC-1
Concentrations	1-50 μg/mL
Incubation Time	3 days
Method	HMEC-1 (1×104 per well) are seeded on uncoated 48 well plates and incubated in medium containing 4% FCS with Cilengitide. After incubation for 72 hours at 37℃, cells are trypsinized and counted.

Animal Study: ^[5]

Animal Models	Human glioblastoma xenografts U87 MG
Dosages	100μg
Administration	Daily i.p.
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
NCT01118676	Recruiting	Locally Advanced Non Small Cell Lung Cancer (NSCLC)*	Institut Claudius Regaud|Merck KGaA	2010-03	Phase 1
NCT01504165	Completed	Renal Impairment	Merck KGaA	2012-01	Phase 1
NCT01517776	Recruiting	Gliomas	Martin-Luther-Universit t Halle-Wittenberg|Merck KGaA	2012-01	Phase 2
NCT01558687	Terminated	Supratentorial Newly Diagnosed Inoperable Gliobastoma	Merck KGaA	2012-08	Phase 1
NCT01782976	Not yet recruiting	Glioblastoma	M.D. Anderson Cancer Center|BRAIN TUMOR TRIALS COLLABORATIVE (BTTC)|EMD Serono	2013-06	Phase 2

Chemical Information

Download Cilengitide SDF

Chemical Name	Cyclo(L-arginylglycyl-L-α-aspartyl-D-phenylalanyl-N-methyl-L-valyl)

Preparing Stock Solutions

Stock Solution (1ml DMSO)	1mM	10mM	20mM	30mM
Mass(mg)	0.58866	5.8866	11.7732	17.6598

Customer Reviews (1)

Click to enlarge	Rating
	Source	Dr.Milica Pesic from Institute for Biological Research .Cilengitide purchased from Selleck
	Method	MTT
	Cell Lines	HTB 183, A549
	Concentrations	0-50 uM
	Incubation Time	72 h
	Results	Cell growth inhibition of non-small cell lung carcinoma (NSCLC) by Cilengitide.