Ang-(1-7) receptor ( IC50 = 21±35 nM )

AVE 0991 是一种非肽化合物，可对内皮细胞产生类似于 Ang-(1-7) 的作用。 AVE 0991 和未标记的 Ang-(1-7) 竞争 [125I]-Ang-(1-7) 与牛主动脉内皮细胞膜的高亲和力结合，IC50 分别为 21±35 和 220±280 nM。 AVE 0991 钠盐和 Ang-(1-7)（均为 10 μM）释放的 NO 和 O2- 峰值浓度没有显着差异（NO：295±20 和 270±25 nM；O2-：18±2 和 20 ±4nM）。然而，与 Ang-(1-7) 相比，AVE 0991 释放的生物活性 NO 量大约高出 5 倍[1]。

与载体治疗的动物相比，AVE 0991 (0.58 nmol/g) 使 WT 小鼠的水利尿作用显着减少（0.06±0.03 mL 对比 0.27±0.05；每组 n=9；P<0.01）。 AVE 0991 (AVE) 的抗利尿作用与尿液渗透压的增加相关（媒介物治疗小鼠中为 1669±231.0 mOsm/KgH2O 对比 681.1±165.8 mOsm/KgH2O；P<0.01）。 Mas 的基因缺失消除了 AVE 0991 在水负荷期间的抗利尿作用（AVE 0991 处理的小鼠为 0.37±0.10 mL [n=9] 对比 0.27±0.03 mL [n=11]）。正如在 C57BL/6 小鼠中所观察到的，与媒介物处理的动物相比，在水负载的 Swiss 小鼠中施用 AVE 0991 (0.58 nmol/g) 也会导致尿量显着减少（0.13±0.05 mL [n=16] 0.51±0.04 mL [n=40]；P<0.01)[2]。用 AVE-0991 治疗一周后，灌注压显着降低（媒介物处理大鼠为 56.55±0.86 对比 68.73±0.69 mmHg），收缩压增加（媒介物处理大鼠为 11.40±0.05 对比 9.84±0.15 g）。张力上升率（+dT/dt；184.30±0.50 vs. 155.20±1.97 g/s（媒介物处理大鼠））、张力下降率（?dT/dt；179.60±1.39 vs. 150.80±2.42） g/s（在媒介物处理的大鼠中）。还观察到心率 (HR) 略有增加（媒介物处理的大鼠为 220.40±0.71 次/分钟 vs. 214.20±0.74 次/分钟[3]。

使用巨细胞病毒启动子将大鼠 Mas cDNA 稳定转染至中国仓鼠卵巢 (CHO) 细胞和猴肾 (COS) 细胞中。新霉素用于选择转染的细胞。有或没有 AVE 0991（AVE，10^-10 至 ^-5 mol/L）、¹²⁵I-Ang-(1-7 ) (0.5×10^-9 mol/L) 在 24 孔板中于 0.3 mL 补充有 0.2% BSA、0.005 的无血清培养基 (DMEM) 中于 4°C 培养 60 分钟％杆菌肽、0.1 mol/L PMSF 和 0.5 mol/L 邻菲咯啉与 Mas 转染的 COS 细胞。两次冷的无血清 DMEM 洗涤后，使用 0.1% Triton X-100 搅动细胞。伽马计数器用于测量结合放射性。在存在或不存在 AVE (10^-6 mol/L) 的情况下使用 2×10^-9 mol/L 罗丹明标记的 Ang-(1-7)，CV11974 (10^-6 mol/L) 或 PD123319 (10^-6 mol/L)，Mas 转染的 CHO 中罗丹明-Ang-(1-7) 的结合细胞是在同等条件下进行的。计算 NSB 需要浓度为 10^-6 mol/L 的 Ang-(1-7) [1]。

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.

[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.

C29H32N4O5S2

580.71818

580.18

C, 59.98; H, 5.55; N, 9.65; O, 13.78; S, 11.04

304462-19-9

AVE 0991 sodium salt; 306288-04-0

Solid powder

CCNC(=O)NS(=O)(=O)C1=C(C=C(S1)CC(C)C)C2=CC=C(C=C2)CN3C(=C(N=C3C4=CC=CC=C4)OC)C=O

QTOZBSNPDCWHPV-UHFFFAOYSA-N

InChI=1S/C29H32N4O5S2/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)

1-ethyl-3-[3-[4-[(5-formyl-4-methoxy-2-phenylimidazol-1-yl)methyl]phenyl]-5-(2-methylpropyl)thiophen-2-yl]sulfonylurea

AVE0991; AVE0991; AVE-0991

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO: ~41.7 mg/mL (~71.8 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.31 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 25.0 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.

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Solubility in Formulation 2: 2.5 mg/mL (4.31 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (4.31 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.

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。