Atrasentan 173937-91-2_Endothelin Receptor_GPCR & G Protein_Signaling Pathways_Products

Atrasentan (ABT-627; NSC720763; A-147627; trade name: Xinlay) is a novel, selective and potent endothelin receptor/ET antagonist with anticancer activity. It suppresses ETA with an IC50 of 0.0551 nM. It's an experimental medication being researched to treat different cancers, including non-small cell lung cancer. Additionally, it's being looked into as a treatment for diabetic kidney disease. In patients who were not responding to hormone therapy, trasentan was not successful in a phase 3 trial for prostate cancer. Atrasentan is a medication that blocks the function of endothelin-1 and may be used to treat hormone-resistant prostate cancer that has metastasized.

Physicochemical Properties

Molecular Formula	C29H38N2O6
Molecular Weight	510.62182
Exact Mass	510.273
Elemental Analysis	C, 68.21; H, 7.50; N, 5.49; O, 18.80
CAS #	173937-91-2
Related CAS #	Atrasentan hydrochloride; 195733-43-8; 178738-96-0 (sodium); 195704-72-4
PubChem CID	159594
Appearance	White to light yellow solid powder
Density	1.188g/cm3
Boiling Point	659.4ºC at 760mmHg
Flash Point	352.6ºC
Vapour Pressure	2.76E-18mmHg at 25°C
LogP	4.631
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	12
Heavy Atom Count	37
Complexity	734
Defined Atom Stereocenter Count	3
SMILES	O=C([C@H]1[C@H](C2=CC=C(OC)C=C2)N(CC(N(CCCC)CCCC)=O)C[C@@H]1C3=CC=C(OCO4)C4=C3)O
InChi Key	MOTJMGVDPWRKOC-QPVYNBJUSA-N
InChi Code	InChI=1S/C29H38N2O6/c1-4-6-14-30(15-7-5-2)26(32)18-31-17-23(21-10-13-24-25(16-21)37-19-36-24)27(29(33)34)28(31)20-8-11-22(35-3)12-9-20/h8-13,16,23,27-28H,4-7,14-15,17-19H2,1-3H3,(H,33,34)/t23-,27-,28+/m1/s1
Chemical Name	(2R,3R,4S)-4-(1,3-benzodioxol-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid
Synonyms	ABT-627; (+)-A 127722; ABT627; ABT 627; ABT-627; NSC720763; A147627; Abbott 147627; trade name: Xinlay; A-147627
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

ETA ( IC50 = 0.055 nM )

ln Vitro

Atrasentan (ABT-627, 0-50 μM) strongly inhibits the growth of LNCaP and C4-2b prostate cancer cells. When combined with Taxotere, ABT-627 causes a notably higher reduction in viable prostate cancer cells compared to when either drug is used alone. It also exhibits a higher degree of NF-κB DNA binding activity down-regulation[2]. Atrasentan significantly induces a number of CYPs and drug transporters (CYP3A4 is 12-fold induced at 50 μM, for example). It is a weak BCRP inhibitor (IC50 in MDCKII-BCRP cells = 59.8±11 μM) and a moderate P-gp inhibitor (IC50 in P388/dx cells = 15.1±1.6 μM)[3].

ln Vivo

Atrasentan (3 mg/kg, p.o.) suppresses the pressor response brought on by big endothelin-1 (1 nmol/kg) in pithed rats[1]. Aatrasentan (ABT-627, 10 mg/kg, i.p.) and Taxotere alone partially inhibited the growth of C4-2b tumors within the bone environment in the SCID-hu model[2].

Enzyme Assay

Atrasentan is used to treat and incubate the cells. After two PBS washes, they are lysed in an ice-cold lysis buffer [1 mM EGTA, 1 mM EDTA, 1.5 mM Tris (pH 7.4), 150 mM NaCl, 1% Triton X-100, 2.5 mM sodium PPi, 1 mM β-glycerophosphate, 1 mM sodium orthovanadate, 1 μg/mL leupeptin, and 1 mM PMSF]. After centrifuging the extracts to get rid of any remaining cell debris, the bicinchoninic acid (BCA) protein assay reagent is used to measure the amount of protein in the supernatants. Proteins (150 μg) cross-linked to agarose hydrazide beads and incubated with mild rocking at 4°C for the entire night. The lysis buffer and kinase assay buffer (25 mM Tris (pH 7.5), 10 mM MgCl₂, 5 mM β-glycerol phosphate, 0.1 mM sodium orthovanadate, 2 mM DTT) are used to wash the immunoprecipitated products twice after Akt is selectively immunoprecipitated from the cell lysates. The products are then resuspended in 40 μL of kinase assay buffer that contains 200 μM ATP and 1 μg GSK-3α/β fusion protein. The addition of Lamelli SDS sample buffer stops the kinase assay reaction after it has been running for 30 minutes at 30°C. 10% SDS-PAGE is used to resolve the reaction products, and then an antiphosphorylated GSK-3α/β antibody is used for Western blotting. After separating 40 μg of protein from the lysate samples using 10% SDS-PAGE, anti-Akt antibody is used for Western blotting in order to determine the total amount of Akt.

Cell Assay

In 96-well microtiter culture plates, the three prostate cancer cell lines (LNCaP, C4-2b, and PC-3) are seeded at a density of 3 × 10³ cells per well. Following an overnight incubation period, the medium is taken out and replaced with a new one that has been diluted from a 10-mM stock to contain varying concentrations of ABT-627 (0-50 μM). 20 μL of MTT solution (5 mg/mL in PBS) is added to each well after the drug has been incubated for 72 hours, and the wells are then incubated for an additional two hours. After the process is finished, the MTT formazan that was produced by metabolically viable cells is dissolved in 100 μL of isopropanol and the supernatant is aspirated. The absorbance is measured at 595 nm on a plate reader after the plates are mixed for 30 minutes on a gyratory shaker.

Animal Protocol

Rats are orally given YM598 (0.3, 1, and 3 mg/kg), atrasentan (0.3, 1, and 3 mg/kg), or 0.5% methyl cellulose as a vehicle using a dosing cannula. 5 mL/kg is the dosage volume for both the test material and the vehicle. The rats are anesthetized with sodium pentobarbital about 20 minutes after the compounds are administered, and they are then pithed and ventilated 30 minutes after the dosage. Big endothelin-1 (1 nmol/kg) is injected intravenously and blood pressure is recorded about an hour after the compounds are taken orally. In these two experiments, linear regression analysis is used to determine the dose of test compound that causes 50% inhibition (ID50) of the big endothelin-1-induced increase in diastolic blood pressure.

References

[1]. Superiority of YM598 over atrasentan as a selective endothelin ETA receptor antagonist. Eur J Pharmacol. 2004 Sep 13;498(1-3):171-7.

[2]. In vitro and in vivo molecular evidence for better therapeutic efficacy of ABT-627 combination in prostate cancer. Cancer Res. 2007 Apr 15;67(8):3818-26.

[3]. Interaction potential of the endothelin-A receptor antagonist atrasentan with drug transporters and drug-metabolising enzymes assessed in vitro. Cancer Chemother Pharmacol. 2011 Oct;68(4):1093-8.

Additional Infomation

Atrasentan is a member of pyrrolidines.
Atrasentan is a substance that is being studied in the treatment of cancer. It belongs to the family of drugs called endothelin-1 protein receptor antagonists. It is a novel, selective endothelin A receptor antagonist (SERA).
A pyrrolidine and benzodioxole derivative that acts a RECEPTOR, ENDOTHELIN A antagonist. It has therapeutic potential as an antineoplastic agent and for the treatment of DIABETIC NEPHROPATHIES.

Drug Indication
Investigated for use/treatment in prostate cancer and cancer/tumors (unspecified).

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

Injection Formulations
(e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).
Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.9584 mL	9.7920 mL	19.5840 mL
	5 mM	0.3917 mL	1.9584 mL	3.9168 mL
	10 mM	0.1958 mL	0.9792 mL	1.9584 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

Atrasentan 173937-91-2

CAS No.: 173937-91-2

Physicochemical Properties

Biological Activity

Solubility Data