Zibotentan (ZD4054) 186497-07-4_Endothelin Receptor_GPCR & G Protein_Signaling Pathways_Products

Zibotentan (formerly known as ZD-4054; ZD4054) is a specific and orally bioavailable Endothelin (ET)A antagonist with potential antitumor activity. It shows no action against ETB and has an IC50 of 21 nM for inhibiting the endothelin receptor. The FDA approved zibotentan for prostate cancer treatment on a fast track basis; however, the drug did not pass clinical trials. By selectively binding to the ET-A receptor, zibotentan blocks the actions of endothelin that stimulate the growth of tumor cells. By blocking AKT and p42/44MAPK phosphorylation, ZD4054 successfully reduced basal and ET-1-induced cell proliferation. It also enhanced apoptosis by blocking Bcl-2, activating caspase-3, and poly(ADP-ribose) polymerase protein.

Physicochemical Properties

Molecular Formula

C19H16N6O4S

Molecular Weight

424.43

Exact Mass

424.095

Elemental Analysis

C, 53.77; H, 3.80; N, 19.80; O, 15.08; S, 7.55

CAS #

186497-07-4

Related CAS #

186497-07-4

PubChem CID

9910224

Appearance

White to off-white solid powder

Density

1.4±0.1 g/cm3

Boiling Point

637.0±65.0 °C at 760 mmHg

Flash Point

339.0±34.3 °C

Vapour Pressure

0.0±1.9 mmHg at 25°C

Index of Refraction

1.628

LogP

2.38

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

654

Defined Atom Stereocenter Count

SMILES

O=S(NC1=NC=C(C)N=C1OC)(C2=CC=CN=C2C3=CC=C(C4=NN=CO4)C=C3)=O

InChi Key

FJHHZXWJVIEFGJ-UHFFFAOYSA-N

InChi Code

InChI=1S/C19H16N6O4S/c1-12-10-21-17(19(23-12)28-2)25-30(26,27)15-4-3-9-20-16(15)13-5-7-14(8-6-13)18-24-22-11-29-18/h3-11H,1-2H3,(H,21,25)

Chemical Name

N-(3-methoxy-5-methylpyrazin-2-yl)-2-[4-(1,3,4-oxadiazol-2-yl)phenyl]pyridine-3-sulfonamide

Synonyms

ZD4054; ZD-4054; ZD 4054

HS Tariff Code

2934.99.9001

Storage

Powder-20°C 3 years

4°C 2 years

In solvent -80°C 6 months

-20°C 1 month

Shipping Condition

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets

ET-A ( IC50 = 21 nM )

ln Vitro

In vitro activity: Zibotentan binds to endothelin A receptor (ET_A) with high affinity (Ki of 13 nM) and has no affinity for endothelin B receptor (ET_B) (IC50 of >10 μM). This is because it specifically inhibits ET_A-mediated antiapoptotic effects but not ET_B-mediated proapoptotic effects in human and rat smooth muscle cells.[1] Treatment with zibotentan at a concentration of 1 μM inhibits the mitogenic activity induced by ET-1 in the ovarian carcinoma cell lines HEY and OVCA 433, which secrete ET-1 and express ET_A and ET_B mRNA.[2] In HEY and OVCA 433 cells, ZD4054 (1 μM) inhibits ET-1-induced EGFR transactivation. Zibotentan (1 μM) reverses the epithelial-mesenchymal transition (EMT) mediated by ET-1 by upregulating the expression and promoter activity of E-cadherin and blocking the secretion of vascular endothelial growth factor (VEGF) and invasiveness in HEY and OVCA 433 cells.[3] Additionally, zibotentan significantly reduces the basal and ET-1-induced proliferation of cells in SKOV-3 and A-2780 cells. This is linked to phosphorylation of AKT and p42/44MAPK as well as increased apoptosis via bcl-2 inhibition and activation of caspase-3 and poly(ADP-ribose) polymerase proteins.[4]

ln Vivo

Zibotentan potently suppresses the growth of HEY ovarian carcinoma xenografts in mice by 69% while posing no toxicity, when administered at a dose of 10 mg/kg/day for 21 days. This effect is correlated with a 37% reduction in Ki-67 expression, a 62% inhibition of tumor-induced vascularization, and a blocking of cell proliferation. Zibotentan treatment consistently and potently increases the expression of E-cadherin while significantly inhibiting the expression of matrix mET_Alloproteinase-2 (MMP-2) and VEGF, as well as the activation of p42/44 MAPK and EGFR. [3]

Enzyme Assay

The inhibition by Zibotentan (varying concentrations) of ¹²⁵iodine-ET-1 binding to cloned human ET_A is assessed using standard radioligand-binding techniques. Mice erythroleukaemic cells are used to express human recombinant ET_A, and cell membranes are prepared for competitive binding experiments using ¹²⁵iodine-ET-1 as the radioligand. In vitro experiments with Zibotentan are conducted in triplicate, ranging from 100 pM to 100 μM in half-log increments. The expression used to quantify the inhibition of ET-1 binding is the geometric mean pIC50 value, which represents the concentration required to inhibit 50% of binding, along with a 95% confidence interval (CI). Zibotentan's affinity for cloned human ET_A is evaluated by applying Cheng and Prusoff's equation to find the equilibrium dissociation constant (Ki) in an additional receptor-binding screen that makes use of more concentration-response curves from three different studies.

Cell Assay

After a 24-hour incubation period in serum-free DMEM, cells are subjected to a 48-hour Zibotentan exposure. Following therapy, cells are lysed, the supernatant is extracted, and using a microplate reader, the assay is performed to look for histone-associated DNA fragments at 405 nm. Adherent and floating cells are collected for the purpose of detecting early apoptotic events. Using the Vybrant Apoptosis Kit, cells are double stained with propidium iodide and FITC-conjugated Annexin V. Their cytofluorometric analysis is then performed right away.

Animal Protocol

Dissolved in DMSO, and diluted in PBS; 10 mg/kg; i.p. injection

Female athymic (nu+/nu+) mice bearing established HEY human ovarian carcinoma xenografts

References

[1]. Br J Cancer . 2005 Jun 20;92(12):2148-52.

[2]. Exp Biol Med (Maywood) . 2006 Jun;231(6):1132-5.

[3]. Cancer Res . 2007 Jul 1;67(13):6351-9.

[4]. Mol Cancer Ther . 2007 Jul;6(7):2003-11.

Additional Infomation

N-(3-methoxy-5-methyl-2-pyrazinyl)-2-[4-(1,3,4-oxadiazol-2-yl)phenyl]-3-pyridinesulfonamide is a phenylpyridine.
Zibotentan is an orally available selective antagonist of the endothelin-A (ET-A) receptor with potential antineoplastic activity. Zibotentan binds selectively to the ET-A receptor, thereby inhibiting endothelin-mediated mechanisms that promote tumor cell proliferation.

Drug Indication
Investigated for use/treatment in prostate cancer.

Solubility Data

Solubility (In Vitro)

DMSO: 24~25 mg/mL (56.5~58.9 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.89 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

Solubility in Formulation 2: 1% DMSO +30% polyethylene glycol+1% Tween 80 : 30 mg/mL

(Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3561 mL	11.7805 mL	23.5610 mL
	5 mM	0.4712 mL	2.3561 mL	4.7122 mL
	10 mM	0.2356 mL	1.1781 mL	2.3561 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

PeptideDB

Zibotentan (ZD4054) 186497-07-4

CAS No.: 186497-07-4

Physicochemical Properties

Biological Activity

Solubility Data