Cell lines

Jurkat cells; Human pulmonary artery endothelial cells (HPAECs)

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

20 μM, 16-18h

Applications

In Jurkat cells, GI 254023X inhibited cell proliferation and increased apoptosis in a concentration-dependent manner. Compared with control group, GI 254023X up-regulated the expression of Notch1 while down-regulated the expression of cleaved Notch1 in a time-dependent way. GI254023X also reduced the levels of MCL-1 and Hes-1 mRNA transcripts. In human pulmonary artery endothelial cells (HPAECs), GI254023X inhibited VE-cadherin cleavage and completely protected HPAECs from Hla-mediated barrier disruption.

Animal models

BALB/c mice injected with endotoxin-free recombinant Hla

Dosage form

200 mg/kg/day, dilution in 0.1 M carbonate buffer, 3-day period, intraperitoneal injection

Application

In BALB/c mice injected with endotoxin-free recombinant Hla, GI254023X enhanced vascular integrity, manifest by limited dye extravasation, suggested that GI254023X may afford protection against lethal infection. Mice were treated with either DMSO or GI254023X then infected with 5×107 CFUs S. aureus Newman. GI254023X-treated mice were less ill in appearance and demonstrated prolongation of time to death.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

Description:IC50: 5.3 nM (ADAM10)
A Disintegrin and metalloproteinase domain-containing protein 10, also known as ADAM10 or CDw156 or CD156c is a protein that in humans is encoded by the ADAM10 gene. ADAM10 (EC#: 3.4.24.81) is a sheddase, and has a broad specificity for peptide hydrolysis reactions (http://en.wikipedia.org/wiki/ADAM10). GI 254023X, synthesized by GSK, was reported to inhibit ADAM10 100-fold over ADAM17. GI 254023X could reduce constitutive cleavage of fractilkine from ECV-304 transfectants [1].
In vitro: Previusl study reported that compound GI254023X possessed comparable inhibitory potency for ADAM10 only and blocked TACE with more than 100-fold reduced potency [2].
In vivo: To examine the ability of GI254023X to inhibit Hla-mediated endothelial barrier disruption in vivo, mice treated for a 3-day period with GI254023X via intraperitoneal injection were subjected to a Miles assay following subcutaneous injection of recombinant toxin. Resutls showed that although all experimental animals succumbed to the lethal challenge, GI254023X-treated mice were less ill in appearance and demonstrated prolongation of time to death [3].
Clinical trial: GI254023X is currently in the preclinical development and no clinical trial is ongoing..
References
[1] Koichi Yokota, and Shin-Ichiro Nishimura. MMP/ADAM inhibitors: therapeutic potential for psoriasis. Expert Opin. Ther. Patents. 2005;15:421-435
[2] Hundhausen C, Misztela D, Berkhout TA, Broadway N, Saftig P, Reiss K, Hartmann D, Fahrenholz F, Postina R, Matthews V, Kallen KJ, Rose-John S, Ludwig A.The disintegrin-like metalloproteinase ADAM10 is involved in constitutive cleavage of CX3CL1 (fractalkine) and regulates CX3CL1-mediated cell-cell adhesion. Blood. 2003;102(4):1186-95.
[3] Powers ME, Kim HK, Wang Y, Bubeck Wardenburg J.ADAM10 mediates vascular injury induced by Staphylococcus aureus α-hemolysin. J Infect Dis. 2012;206(3):352-6.