AVE0991 sodium, the sodium salt of AVE-0991, is a novel, orally bioactive non-peptide mimic and high affinity agonist of angiotensin-(1-7) with an IC50 of 21 nM.

Physicochemical Properties

Molecular Formula	C29H31N4O5S2-.NA+
Molecular Weight	602.7
Exact Mass	602.16
Elemental Analysis	C, 57.79; H, 5.18; N, 9.30; Na, 3.81; O, 13.27; S, 10.64
CAS #	306288-04-0
Related CAS #	AVE 0991; 304462-19-9
PubChem CID	78357809
Appearance	White to yellow solid powder
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	11
Heavy Atom Count	41
Complexity	938
Defined Atom Stereocenter Count	0
InChi Key	GABSTAFOMFJFOI-UHFFFAOYSA-M
InChi Code	InChI=1S/C29H32N4O5S2.Na/c1-5-30-29(35)32-40(36,37)28-24(16-23(39-28)15-19(2)3)21-13-11-20(12-14-21)17-33-25(18-34)27(38-4)31-26(33)22-9-7-6-8-10-22;/h6-14,16,18-19H,5,15,17H2,1-4H3,(H2,30,32,35);/q;+1/p-1
Chemical Name	sodium;ethylcarbamoyl-[3-[4-[(5-formyl-4-methoxy-2-phenylimidazol-1-yl)methyl]phenyl]-5-(2-methylpropyl)thiophen-2-yl]sulfonylazanide
Synonyms	AVE-0991 sodium salt; AVE 0991; AVE-0991; AVE0991
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	Ang-(1-7) receptor ( IC50 = 21±35 nM )
ln Vitro	AVE 0991 is a nonpeptide compound that acts on the endothelium in a manner akin to that of Ang-(1-3). The high-affinity binding of [125I]-Ang-(1-7) to bovine aortic endothelial cell membranes is competitive between AVE 0991 and unlabeled Ang-(1–7), with IC50s of 220±280 nM and 21±35 nM, respectively. The release of NO and O2-at peak concentrations by AVE 0991 sodium salt and Ang-(1–7) (both 10 μM) do not differ significantly (NO: 295±20 and 270±25 nM; O2-: 18±2 and 20±4 nM). In contrast to Ang-(1-7)[1], the amount of bioactive NO released by AVE 0991 is approximately five times greater.
ln Vivo	AVE 0991 (0.58 nmol/g) results in a significant reduction of water diuresis in WT mice as compared to animals treated with the vehicle (0.06±0.03 mL versus 0.27±0.05; n = 9 for each group; P<0.01). The antidiuretic action of AVE 0991 (AVE) is linked to a rise in urine osmolality (1669±231.0 mOsm/KgH2O) in comparison to 681.1±165.8 mOsm/KgH2O in mice treated with vehicle; P<0.01). When water is added, the antidiuretic effect of AVE 0991 is eliminated due to the genetic deletion of Mas (0.37±0.10 mL [n=9] versus 0.27±0.03 mL [n=11] in mice treated with AVE 0991). Administration of AVE 0991 (0.58 nmol/g) to water-loaded Swiss mice results in a significantly lower urinary volume than animals treated with a vehicle (0.13±0.05 mL [n=16] versus 0.51±0.04 mL [n=40]; P<0.01)[2]. This observation was also made with C57BL/6 mice on the same subject. AVE-0991 treatment for one week results in a significant decrease in perfusion pressure (56.55±0.86 vs. 68.73±0.69 mmHg in vehicle-treated rats) and an increase in systolic tension (11.40±0.05 vs. 9.84±0.15 g in vehicle-treated rats).It also causes a rise in the rate of tension (+dT/dt; 184.30±0.50 vs. 155.20±1.97 g/s in vehicle-treated rats), a fall in the rate of tension (dT/dt; 179.60±1.39 vs. 150.80±2.42 g/s in vehicle-treated rats). Notably, rats treated with vehicles showed a marginally higher heart rate (HR) of 220.40±0.71 beats/min as opposed to 214.20±0.74 beats/min[3].
Animal Protocol	Mice: The mice used are male Swiss mice, male Mas-KO (Mas-/-) mice on the pure genetic background C57BL/6, and male WT C57BL/6 control mice (Mas+/+). In conscious mice, an intraperitoneal water injection (0.05 mL/g of body weight [BW]) causes water diuresis. Predetermined volumes of water load (0.01 mL/g BW) are used to administer drugs in the same injection. In the first set of experiments, either vehicle for AVE 0991 (10 μM KOH, 0.01 mL/g; n = 9 control; n = 9 Mas-KO) or 0.58 nmol/g AVE 0991 (n = 9 control; n = 11 Mas-KO mice) is administered to WT (C57BL/6, control group) or Mas-KO mice. Swiss mice in the second set receive the following treatments: (1) vehicle (n = 36); (2) 0.58 nmol/g AVE 0991 (n = 16); (3) 46 pmol/g Ang-(1-7) antagonist A-779 (n = 4); (4) 2 nmol/g DuP-753 or CGP 48933 (n = 5); (5) 2 nmol/g AT2 receptor antagonists PD123319 or PD123177 (n = 9); (6) AVE 0991 combined with A-779; (7) AVE 0991 combined with DuP-753 or CGP 48933 (n = 4 for each); (8) or AVE 0991 combined with PD123319 (n = 5) or PD123177 (n = 4). The dosage of AVE 0991 is determined by means of pilot studies conducted on Swiss mice. Rats: We use male Wistar rats weighing between 250 and 300 grams. Rats are given AVE-0991 (1 mg/kg, n = 9) or vehicle (0.9% NaCl, n = 11) orally via gavage.
References	[1]. AVE 0991, a nonpeptide mimic of the effects of angiotensin-(1-7) on the endothelium. Hypertension. 2002 Dec;40(6):847-52. [2]. Nonpeptide AVE 0991 is an angiotensin-(1-7) receptor Mas agonist in the mouse kidney. Hypertension. 2004 Oct;44(4):490-6. [3]. The nonpeptide angiotensin-(1-7) receptor Mas agonist AVE-0991 attenuates heart failure induced by myocardial infarction. Am J Physiol Heart Circ Physiol. 2007 Feb;292(2):H1113-9. [4]. AVE 0991 attenuates oxidative stress and neuronal apoptosis via Mas/PKA/CREB/UCP-2 pathway after subarachnoid hemorrhage in rats. Redox Biol. 2018 Sep 28;20:75-86.

Solubility Data

Solubility (In Vitro)

DMSO: ≥ 55 mg/mL (~91.3 mM) H2O: ~50 mg/mL (~83 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.45 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (3.45 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (3.45 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.6592 mL	8.2960 mL	16.5920 mL
	5 mM	0.3318 mL	1.6592 mL	3.3184 mL
	10 mM	0.1659 mL	0.8296 mL	1.6592 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.