在 ATRA 敏感的 APL NB4 细胞中，bestatin 促进 ATRA 诱导的分化并防止 ATRA 驱动的 p38 MAPK 激活。 ATRA 抗性 APL MR2 细胞具有贝他汀不可逆的分化阻断作用。当CD13与抗CD13抗体WM-15连接时，p38 MAPK被磷酸化，Bestatin对p38 MAPK磷酸化的抑制作用减弱，并且Bestatin对ATRA诱导NB4细胞分化的增强作用被完全消除[2]。用贝他汀 (600 μM) 处理的细胞会经历细胞周期进程延迟，因为它们的分裂频率和生长速率降低。 Bestatin 在 0-600 μM 浓度范围内对盘状 D. discoideum 细胞无害，但它确实会抑制这些细胞的有丝分裂频率和固有的多核性。在 PsaA-GFP 和 GFP 表达细胞的裂解物中，bestatin 使氨肽酶活性分别降低对照的 69.39% ± 10.5% 和 39.93% ± 18.7%[4]。

与糖尿病载体治疗的小鼠相比，bestatin (20 μM) 显着抑制 MMP-9 特异性溶胶带密度，并显着降低糖尿病小鼠中的 CD13 表达。在糖尿病小鼠中，用甜菜素治疗可显着降低乙酰肝素酶和 VEGF 的表达。在糖尿病小鼠的视网膜中，玻璃体内注射贝他汀疗法可显着降低 VEGF 和 HIF-1α 的表达。此外，玻璃体内注射贝他汀治疗可有效抑制糖尿病小鼠视网膜中乙酰肝素酶的表达增加[1]。在对 SRBC 产生抗原增强体液反应之前，给予贝他汀（10、1 和 0.1 mg/kg，腹腔注射）会增加产生溶血性抗 SRBC 抗体 (PFC) 的脾细胞数量和 2-ME 抗性血清血凝素滴度（剂量为0.1毫克/千克）。当小鼠注射环磷酰胺后不同天给予贝他汀（1和0.1 mg/kg）5次时，药物对PFC数量的抑制作用保持不变。事实上，在抗原刺激后第七天，贝他汀（1 mg/kg）会导致总抗 SRBC 血凝素进一步下降。

收获细胞、洗涤并在 NP-40 裂解缓冲液（50 mM Tris-HCl [pH 7.5]、150 mM NaCl、0.5% NP-40）中裂解。使用 Bradford 测定法对总细胞蛋白进行定量，并制备 1 mg/mL 的蛋白等分试样。将 10 微升总细胞蛋白与 290 μL 底物溶液（0.1 mg/mL 二硫苏糖醇 [DTT]、0.1 mg/mL 白蛋白和 1 mM 丙氨酸-β-萘酰胺）混合。 15 分钟和 30 分钟后进行荧光测量（340 nm 激发，400 nm 发射）。 15 分钟和 30 分钟测量值之间的直线斜率用于代表氨肽酶活性。在进行荧光氨肽酶测定之前，将总细胞蛋白与 bestatin、amastatin、嘌呤霉素、EDTA 和/或 ZnCl2 预孵育 20 分钟。

将生长中的细胞（1×106 至 2×106 细胞/mL）稀释至 1.0×103 细胞/mL，并转移（3 mL）至 12 孔多孔板（2.5 厘米直径/孔）的孔中。细胞用 0、10、50、100、300 或 600 μM Bestatin 处理，并在 21°C 下以 180 rpm 振荡生长 48 小时。血细胞计数器用于测量 0、24 和 48 小时后的细胞密度。

[1]. Hossain A, et al. Protective effects of bestatin in the retina of streptozotocin-induced diabetic mice. Exp Eye Res. 2016 Aug;149:100-6
[2]. Qian X, et al. Inhibition of p38 MAPK Phosphorylation Is Critical for Bestatin to Enhance ATRA-Induced Cell Differentiation in Acute Promyelocytic Leukemia NB4 Cells. Am J Ther. 2016 May-Jun;23(3):e680-9.
[3]. Lis M, et al. The effects of bestatin on humoral response to sheep erythrocytes in non-treated and cyclophosphamide-immunocompromised mice. Immunopharmacol Immunotoxicol. 2013 Feb;35(1):133-8
[4]. Poloz Y, et al. Bestatin inhibits cell growth, cell division, and spore cell differentiation in Dictyostelium discoideum. Eukaryot Cell. 2012 Apr;11(4):545-57

C18H25F3N2O6

422.3961

223763-80-2

Bestatin;58970-76-6;Bestatin hydrochloride;65391-42-6

FC(C(=O)O[H])(F)F.O([H])[C@]([H])(C(N([H])[C@]([H])(C(=O)O[H])C([H])([H])C([H])(C([H])([H])[H])C([H])([H])[H])=O)[C@@]([H])(C([H])([H])C1C([H])=C([H])C([H])=C([H])C=1[H])N([H])[H]

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)

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 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).

View More

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 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).

View More

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.

---

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。