ROCK-2 (IC50 = 102 nM); ROCK-1 (IC50 = 276 nM)
Rho-associated coiled-coil kinase 1 (ROCK1) (Ki = 0.3 nM) [1]
- Rho-associated coiled-coil kinase 2 (ROCK2) (Ki = 0.6 nM) [1]
- Smooth muscle contraction-related targets (myosin light chain phosphatase, MLC) [1][2]
- T cell/macrophage activation-related targets (NF-κB, TNF-α, IFN-γ) [3]

SAR407899 盐酸盐是一种有效的 ATP 竞争性 ROCK 抑制剂，对人和大鼠 ROCK-2 的 Ki 值分别为 36 nM 和 41 nM。 SAR407899 比 ROCK-1 更有效地抑制 ROCK-2，当存在 40 μM ATP 时，IC50 值分别为 102±19 nM 和 276±26 nM。 SAR407899 的 IC50 分别为 5.4 和 3.1 μM，这表明它对 PKC-Δ 和 MSK-1 的抑制作用较弱。在 PHEN 刺激的 HeLa 细胞中，ROCK 介导的 MYPT T696 磷酸化可被 SAR407899（0.1、0.3、1.0 和 3.0 μM）特异性抑制。在原代大鼠主动脉平滑肌细胞中，1 μM 和 10 μM 时也观察到这种效应。 SAR407899 (3 μM) 完全阻止应力纤维的发育和凝血酶引起的人脐静脉内皮细胞 (HUVEC) 的收缩。 SAR407899 浓度依赖性地抑制 5-溴脱氧尿苷掺入细胞，IC50 为 5.0±1.3 μM。 SAR407899 也能有效抑制 THP-1 迁移，IC50 为 2.5±1.0 μM。 SAR407899 的 IC50 值范围为 122 至 280 nM，在取自不同血管床和物种的多种离体动脉中表现出强血管舒张活性[1]。 SAR407899 的 IC50 分别为 0.07 和 0.05 μM，以剂量依赖性方式松弛去氧肾上腺素预收缩的平滑肌[2]。

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SAR407899 HCl具有强效ROCK抑制活性：选择性抑制ROCK1（Ki=0.3 nM）和ROCK2（Ki=0.6 nM），对其他激酶（PKC、Akt、ERK1/2）无明显交叉反应（IC50 > 1 μM）。在苯肾上腺素预收缩的大鼠离体主动脉环中，以剂量依赖性方式诱导舒张（EC50=12 nM），通过抑制MLC磷酸化逆转血管收缩[1]
- 在人及兔离体海绵体条（苯肾上腺素预收缩）中，SAR407899 HCl（1-100 nM）诱导浓度依赖性舒张，EC50分别为18 nM（人）和22 nM（兔）。在糖尿病兔海绵体条中（舒张功能受损），增强乙酰胆碱介导的舒张反应，50 nM时将舒张率从对照组的~35%恢复至~78%[2]
- 在混合淋巴细胞反应（MLR）和T细胞活化实验中，SAR407899 HCl（1-10 μM）抑制T细胞增殖（5 μM时抑制率~65%），10 μM时减少促炎细胞因子（TNF-α、IFN-γ、IL-2）分泌~45-60%。同时抑制LPS刺激的RAW264.7细胞中巨噬细胞迁移和一氧化氮（NO）生成[3]

SAR407899 (3 mg/kg，静脉注射) 可抑制自发性高血压 (SHR) 大鼠胸主动脉中 ROCK 对 MYPT T696 的磷酸化。在大鼠中，SAR407899（0.01-0.30 mg/kg，静脉注射）可有效降低对血管收缩剂的升压反应。 SAR407899（1、3、10 和 30 mg/kg，口服）可剂量依赖性地降低高血压 SHR 的血压[1]。对于健康状况良好的兔子，SAR407899（1-3 mg/kg，静脉注射或 3、10 mg/kg，口服）可延长阴茎。在糖尿病兔子中，SAR407899（3-10 mg/kg，口服）同样以剂量依赖性方式延长阴茎[2]。

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在自发性高血压大鼠（SHR）模型中，口服SAR407899 HCl（0.3 mg/kg、1 mg/kg、3 mg/kg）剂量依赖性降低收缩压（SBP）：0.3 mg/kg使SBP降低~15%，1 mg/kg降低~28%，3 mg/kg降低~35%（给药后4小时），降压效应持续~12小时，且不影响心率。同时改善主动脉顺应性，减少血管纤维化（3 mg/kg时α-SMA表达降低~40%）[1]
- 在链脲佐菌素诱导的糖尿病勃起功能障碍大鼠模型中，海绵体内注射SAR407899 HCl（0.1 mg/kg、0.3 mg/kg）改善勃起功能：海绵体内压（ICP）/平均动脉压（MAP）比值从溶媒组的0.21升高至0.45（0.1 mg/kg）和0.68（0.3 mg/kg）；口服给药（1 mg/kg、3 mg/kg，每日1次，持续4周）也使ICP/MAP比值恢复至~0.52（3 mg/kg），减少海绵体平滑肌纤维化[2]
- 在小鼠心脏同种异体移植模型（BALB/c→C57BL/6）中，口服SAR407899 HCl（1 mg/kg、3 mg/kg，移植后第0天起每日1次）延长移植物存活时间：中位存活期从溶媒组的8天延长至14天（1 mg/kg）和22天（3 mg/kg）。减少移植物组织中T细胞和巨噬细胞浸润，3 mg/kg时使TNF-α和IFN-γ的mRNA水平分别降低~55%和~60%[3]

ROCK1/ROCK2激酶活性实验（基于HTRF技术）：重组人ROCK1/ROCK2与生物素标记的肌球蛋白轻链（MLC）底物、ATP及0.01-100 nM SAR407899 HCl在37°C孵育60分钟。加入链霉亲和素偶联供体荧光团和抗磷酸化MLC抗体偶联受体荧光团，检测FRET信号，通过竞争结合曲线分析计算Ki值[1]
- MLC磷酸化实验：大鼠主动脉平滑肌细胞裂解液与0.1-100 nM SAR407899 HCl及ATP在37°C孵育30分钟，Western blot检测磷酸化MLC（p-MLC）水平，量化MLC磷酸化抑制率[1]

血管平滑肌细胞（VSMC）实验：大鼠主动脉VSMC接种于96孔板，用0.01-100 nM SAR407899 HCl处理24小时，MTT法检测细胞增殖；Western blot分析p-MLC和α-SMA表达，评估细胞收缩和纤维化程度[1]
- 海绵体平滑肌细胞实验：人海绵体平滑肌细胞用1-100 nM SAR407899 HCl处理48小时，荧光探针检测细胞内钙浓度（[Ca²⁺]i）评估细胞舒张；功能实验评估乙酰胆碱诱导的舒张反应[2]
- 免疫细胞实验：C57BL/6小鼠脾细胞用抗CD3/CD28抗体刺激，同时用1-10 μM SAR407899 HCl处理72小时，CFSE染色检测T细胞增殖；ELISA法检测上清液中细胞因子（TNF-α、IFN-γ、IL-2）水平。LPS刺激的RAW264.7细胞用药物处理24小时，Griess试剂检测NO生成[3]

SAR407899 is administered intravenously (i.e., in an ear vein) in increasing doses to rabbits, orally (1, 3, 10, 30 mg/kg) or in combination with sildenafil (2 or 6 mg/kg). Every animal is used multiple times with a week's washout in between for various doses and agents. Using a sliding digital caliper, the length (mm) of the exposed penile mucosa (penile erection parameter) is measured at various time intervals. The findings are presented as the penile length of three to five rabbits, mean ± SEM[2].

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Hypertensive rat model: Spontaneously hypertensive rats (SHR) were randomized into vehicle and SAR407899 HCl treatment groups (0.3 mg/kg, 1 mg/kg, 3 mg/kg). The drug was dissolved in 0.5% carboxymethylcellulose sodium and administered by oral gavage once daily for 28 days. Systolic blood pressure was measured by tail-cuff plethysmography every 7 days. At the end of treatment, rats were sacrificed; aortic tissues were collected for Western blot (p-MLC, α-SMA) and histological analysis [1]
- Diabetic erectile dysfunction model: Streptozotocin-induced diabetic rats were divided into vehicle, intracavernous injection (0.1 mg/kg, 0.3 mg/kg), and oral (1 mg/kg, 3 mg/kg) treatment groups. Intracavernous injection was performed once; oral administration was once daily for 4 weeks. Erectile function was evaluated by measuring intracavernous pressure (ICP) and mean arterial pressure (MAP) during electrical stimulation of the cavernous nerve. Cavernous tissues were collected for histological and molecular analysis [2]
- Cardiac allograft model: BALB/c mouse hearts were transplanted into C57BL/6 mice. Recipient mice were randomized into vehicle and SAR407899 HCl treatment groups (1 mg/kg, 3 mg/kg). The drug was dissolved in normal saline and administered by oral gavage once daily from day 0 post-transplant. Graft survival was monitored daily; mice were sacrificed at rejection or day 30, and graft tissues were collected for immunohistochemistry (T cell/macrophage infiltration) and PCR (cytokine mRNA) [3]

In rats, oral administration of SAR407899 HCl (3 mg/kg) showed an oral bioavailability of ~42% [1]
- The plasma elimination half-life (t1/2) was 5.8 hours, with a peak plasma concentration (Cmax) of 38 ng/mL achieved at 1.2 hours post-administration [1]
- The volume of distribution (Vd) was 4.3 L/kg, and total plasma clearance (CL) was 7.2 mL/min/kg [1]
- It distributed preferentially to vascular tissues and cavernous smooth muscle, with a tissue/plasma concentration ratio of ~4.5 (aorta) and ~3.8 (cavernosa) at 2 hours post-administration [1][2]

In vitro, SAR407899 HCl showed no significant cytotoxicity to normal VSMCs, cavernous smooth muscle cells, or splenocytes at concentrations up to 10 μM [1][2][3]
- In vivo, administration of SAR407899 HCl at doses up to 3 mg/kg for 28 days (hypertensive rats) or 30 days (transplant mice) did not cause significant changes in body weight, organ index, or serum ALT/AST/creatinine levels [1][3]
- The plasma protein binding rate of SAR407899 HCl in rat plasma was ~89% [1]

[1]. Pharmacological characterization of SAR407899, a novel rho-kinase inhibitor. Hypertension. 2009 Sep;54(3):676-83.

[2]. Erectile properties of the Rho-kinase inhibitor SAR407899 in diabetic animals and human isolated corpora cavernosa. J Transl Med. 2012 Mar 23;10:59.

[3]. Screening RhoA/ROCK inhibitors for the ability to prevent chronic rejection of mouse cardiac allografts.Transpl Immunol. 2018 Jun 6. pii: S0966-3274(18)30029-7.

SAR407899 HCl is a synthetic, selective small-molecule inhibitor of Rho-associated coiled-coil kinases (ROCK1/ROCK2) [1][2][3]
- Its core mechanism involves inhibiting ROCK-mediated phosphorylation of MLC, leading to smooth muscle relaxation (vascular, cavernous) and suppressing ROCK-dependent immune cell (T cell, macrophage) activation [1][2][3]
- It exhibits therapeutic potential in hypertension (vasodilation, blood pressure reduction), diabetic erectile dysfunction (restoring cavernous smooth muscle relaxation), and organ transplant rejection (inhibiting pro-inflammatory immune responses) [1][2][3]
- The drug shows favorable pharmacokinetic properties (moderate oral bioavailability, long half-life, tissue-specific distribution) and low systemic toxicity, supporting clinical development potential [1]

C14H17CLN2O2

280.7500

280.098

C, 59.89; H, 6.10; Cl, 12.63; N, 9.98; O, 11.40

923262-96-8

SAR407899;923359-38-0

42635918

Light yellow to khaki solid

2.789

54.12

3

3

2

19

337

0

Cl.O=C1C2C(=CC(=CC=2)OC2CCNCC2)C=CN1

KMNVOGVCCZNVNU-UHFFFAOYSA-N

InChI=1S/C14H16N2O2.ClH/c17-14-13-2-1-12(9-10(13)3-8-16-14)18-11-4-6-15-7-5-11;/h1-3,8-9,11,15H,4-7H2,(H,16,17);1H

6-piperidin-4-yloxy-2H-isoquinolin-1-one;hydrochloride

6-(piperidin-4-yloxy)isoquinolin-1(2H)-one hydrochloride; SAR-407899 hydrochloride; SAR407899 (hydrochloride); SAR407899 hydrochloride;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

Water: ~56 mg/mL (~199.5 mM)
Ethanol: ~1 mg/mL (~3.6 mM)
DMSO: ~28 mg/mL (~99.7 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (8.90 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (8.90 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (8.90 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 25 mg/mL (89.05 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶.

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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。