ProTAME

别名: pro-Tosyl-L-Arginine Methyl Ester ProTAME
目录号: V13285 纯度: ≥98%
ProTAME 是一种新型 APC/CFzr 和 APC/CCdc20 抑制剂,具有抗癌活性。
ProTAME CAS号: 1362911-19-0
产品类别: New1
产品仅用于科学研究,不针对患者销售
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产品描述
ProTAME 是一种新型 APC/CFzr 和 APC/CCdc20 抑制剂,具有抗癌活性。它作为 TAME 的细胞渗透性前药,TAME 是一种后期促进复合物/环体 (APC/C) 抑制剂,与 APC 结合优先抑制 APC/C(Cdc20)。它使细胞停滞在中期而不干扰纺锤体,但依赖于纺锤体装配检查点 (SAC)。 proTAME 与拓扑异构酶抑制剂、依托泊苷和阿霉素的组合可显着增加多发性骨髓瘤 (MM) 细胞系和原代细胞的细胞死亡,特别是如果首先通过 ProTAME 预处理增加 TOPIIα 水平。
生物活性&实验参考方法
体外研究 (In Vitro)
ProTAME 可阻止小鼠和牛卵母细胞以及小鼠 2 细胞胚胎进入后期。用 proTAME (0-100 μM) 处理的哺乳动物卵母细胞表现出剂量依赖性中期停滞和早期卵裂胚胎。此外,纺锤体组装检查点 (SAC) 活性对于药物诱导的中期停滞不是必需的 [1]。由于 ProTAME 抑制 APC/C,因此可以阻止小鼠卵母细胞减数分裂 I。与体细胞相反,卵母细胞和胚胎的停滞是不可逆的 [1]。在卵母细胞和胚胎中,proTAME (0–20 μM) 剂量依赖性地改变纺锤体形态特征 [1]。 ProTAME 有效克服了卵巢癌细胞中基于 Polo 样激酶 1 (PLK1) 的耐药性、胶质母细胞瘤细胞中基于 CDH1 过度磷酸化的耐药性以及弥漫性大 B 细胞淋巴瘤细胞中基于 CDC20 的耐药性所产生的耐药性。治疗品质[1]。 proTAME 抑制 OVCAR-3 细胞生长,IC50 为 12.5 μM[2]。
参考文献

[1]. ProTAME Arrest in Mammalian Oocytes and Embryos Does Not Require Spindle Assembly Checkpoint Activity. Int J Mol Sci. 2019 Sep 13;20(18):4537.

[2]. Blocking Mitotic Exit of Ovarian Cancer Cells by Pharmaceutical Inhibition of the Anaphase-Promoting Complex Reduces Chromosomal Instability. Neoplasia. 2019 Apr;21(4):363-375.

*注: 文献方法仅供参考, InvivoChem并未独立验证这些方法的准确性
化学信息 & 存储运输条件
分子式
C34H38N4O12S
分子量
726.75
精确质量
726.22
元素分析
C, 56.19; H, 5.27; N, 7.71; O, 26.42; S, 4.41
CAS号
1362911-19-0
PubChem CID
56924780
外观&性状
White to off-white solid powder
密度
1.3±0.1 g/cm3
折射率
1.587
LogP
6.05
tPSA
223
氢键供体(HBD)数目
3
氢键受体(HBA)数目
14
可旋转键数目(RBC)
23
重原子数目
51
分子复杂度/Complexity
1210
定义原子立体中心数目
1
SMILES
C(OC)(=O)[C@@H](NS(C1=CC=C(C)C=C1)(=O)=O)CCC/N=C(\NC(OCOC(CC1=CC=CC=C1)=O)=O)/NC(=O)OCOC(=O)CC1=CC=CC=C1
InChi Key
MHYOVHULCQSDRZ-NDEPHWFRSA-N
InChi Code
InChI=1S/C34H38N4O12S/c1-24-15-17-27(18-16-24)51(44,45)38-28(31(41)46-2)14-9-19-35-32(36-33(42)49-22-47-29(39)20-25-10-5-3-6-11-25)37-34(43)50-23-48-30(40)21-26-12-7-4-8-13-26/h3-8,10-13,15-18,28,38H,9,14,19-23H2,1-2H3,(H2,35,36,37,42,43)/t28-/m0/s1
化学名
methyl (2S)-5-[bis[(2-phenylacetyl)oxymethoxycarbonylamino]methylideneamino]-2-[(4-methylphenyl)sulfonylamino]pentanoate
别名
pro-Tosyl-L-Arginine Methyl Ester ProTAME
HS Tariff Code
2934.99.9001
存储方式

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

运输条件
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
溶解度数据
溶解度 (体外实验)
DMSO : ~100 mg/mL (~137.60 mM)
溶解度 (体内实验)
配方 1 中的溶解度: ≥ 3.75 mg/mL (5.16 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 37.5 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中,混合均匀。

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。
制备储备液 1 mg 5 mg 10 mg
1 mM 1.3760 mL 6.8799 mL 13.7599 mL
5 mM 0.2752 mL 1.3760 mL 2.7520 mL
10 mM 0.1376 mL 0.6880 mL 1.3760 mL

1、根据实验需要选择合适的溶剂配制储备液 (母液):对于大多数产品,InvivoChem推荐用DMSO配置母液 (比如:5、10、20mM或者10、20、50 mg/mL浓度),个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;

2、如果您找不到您想要的溶解度信息,或者很难将产品溶解在溶液中,请联系我们;

3、建议使用下列计算器进行相关计算(摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等);

4、母液配好之后,将其分装到常规用量,并储存在-20°C或-80°C,尽量减少反复冻融循环。

计算器

摩尔浓度计算器可计算特定溶液所需的质量、体积/浓度,具体如下:

  • 计算制备已知体积和浓度的溶液所需的化合物的质量
  • 计算将已知质量的化合物溶解到所需浓度所需的溶液体积
  • 计算特定体积中已知质量的化合物产生的溶液的浓度
使用摩尔浓度计算器计算摩尔浓度的示例如下所示:
假如化合物的分子量为350.26 g/mol,在5mL DMSO中制备10mM储备液所需的化合物的质量是多少?
  • 在分子量(MW)框中输入350.26
  • 在“浓度”框中输入10,然后选择正确的单位(mM)
  • 在“体积”框中输入5,然后选择正确的单位(mL)
  • 单击“计算”按钮
  • 答案17.513 mg出现在“质量”框中。以类似的方式,您可以计算体积和浓度。

稀释计算器可计算如何稀释已知浓度的储备液。例如,可以输入C1、C2和V2来计算V1,具体如下:

制备25毫升25μM溶液需要多少体积的10 mM储备溶液?
使用方程式C1V1=C2V2,其中C1=10mM,C2=25μM,V2=25 ml,V1未知:
  • 在C1框中输入10,然后选择正确的单位(mM)
  • 在C2框中输入25,然后选择正确的单位(μM)
  • 在V2框中输入25,然后选择正确的单位(mL)
  • 单击“计算”按钮
  • 答案62.5μL(0.1 ml)出现在V1框中
g/mol

分子量计算器可计算化合物的分子量 (摩尔质量)和元素组成,具体如下:

注:化学分子式大小写敏感:C12H18N3O4  c12h18n3o4
计算化合物摩尔质量(分子量)的说明:
  • 要计算化合物的分子量 (摩尔质量),请输入化学/分子式,然后单击“计算”按钮。
分子质量、分子量、摩尔质量和摩尔量的定义:
  • 分子质量(或分子量)是一种物质的一个分子的质量,用统一的原子质量单位(u)表示。(1u等于碳-12中一个原子质量的1/12)
  • 摩尔质量(摩尔重量)是一摩尔物质的质量,以g/mol表示。
/

配液计算器可计算将特定质量的产品配成特定浓度所需的溶剂体积 (配液体积)

  • 输入试剂的质量、所需的配液浓度以及正确的单位
  • 单击“计算”按钮
  • 答案显示在体积框中
动物体内实验配方计算器(澄清溶液)
第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量)
第二步:请输入动物体内配方组成(配方适用于不溶/难溶于水的化合物),不同的产品和批次配方组成不同,如对配方有疑问,可先联系我们提供正确的体内实验配方。此外,请注意这只是一个配方计算器,而不是特定产品的确切配方。
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+
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计算结果:

工作液浓度 mg/mL;

DMSO母液配制方法 mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度,请首先与我们联系。

体内配方配制方法μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL ddH2O,混匀澄清。

(1) 请确保溶液澄清之后,再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶;
            (2) 一定要按顺序加入溶剂 (助溶剂) 。

生物数据图片
  • The impact of prodrug tosyl-l-arginine methyl ester (proTAME) on the meiotic maturation of mouse and bovine oocytes. (A) The frequency of polar body extrusion (PBE) in mouse untreated oocytes (n = 43) and oocytes treated with 5 μM (n = 42) and 20 μM (n = 44) proTAME was scored. Oocyte maturation was monitored by live cell microscopy and 88% of cells in the control group and 0% in 5 μM and 20 μM proTAME underwent PBE. Data were obtained in two independent experiments. The right side panel shows representative examples of oocytes with and without PB. Scale bar: 20 μm. The difference between the control group and both 5 μM and 20 μM proTAME is statistically significant (α < 0.05; *** p < 0.0001). (B) The frequency of PBE in bovine untreated oocytes (n = 97) and oocytes treated with 50 μM (n = 92) and 100 μM (n = 83) proTAME was scored. PBE was scored after 20 h of maturation. A total of 97% of control cells, 86% of cells in 50 μM and 0% of cells in 100 μM proTAME underwent PBE. Data were obtained in two independent experiments. The right side panel shows representative examples of oocytes with and without PB. Scale bar: 20 μm. The difference between the control and 50 μM proTAME is statistically significant (α < 0.05; ** p = 0.0080); the difference between the control and 100 μM proTAME is also statistically significant (α < 0.05; *** p < 0.0001). [1].ProTAME Arrest in Mammalian Oocytes and Embryos Does Not Require Spindle Assembly Checkpoint Activity. Int J Mol Sci. 2019 Sep 13;20(18):4537.
  • The impact of proTAME on the mitotic division of mouse two-cell embryos. (A) Frames from a time lapse microscopy experiment showing the cleavage of the untreated mouse embryo and embryos treated with 5 μM, 10 μM and 20 μM proTAME. Scale bar: 20 μm. (B) The frequency of cleaving, morphologically abnormal and not cleaving blastomeres was scored in control embryos (n = 60), embryos treated with 5 μM (n = 58), 10 μM (n = 60) and 20 μM (n = 60) proTAME. In the control group, 92% of blastomeres were cleaving with no morphological abnormalities, and 8% of blastomeres were not dividing. In 5 μM proTAME, 15% of blastomeres were cleaving with no morphological abnormalities, 45% of blastomeres showed morphological abnormalities and 40% of blastomeres were arrested. In 10 μM proTAME, 3% of blastomeres were cleaving, 25% of blastomeres showed morphological abnormalities and 72% of blastomeres were arrested. In 20 μM proTAME, 100% of blastomeres were arrested. Data were collected in two independent experiments. The difference between the control group and 5 μM, 10 μM, and 20 μM proTAME is statistically significant (α < 0.05; *** p < 0.0001).[1].ProTAME Arrest in Mammalian Oocytes and Embryos Does Not Require Spindle Assembly Checkpoint Activity. Int J Mol Sci. 2019 Sep 13;20(18):4537.
  • Oocytes arrested by proTAME show a low level of anaphase promoting complex/cyclosome (APC/C) activity. (A) Time frames from live cell microscopy showing the expression levels of microinjected SECURIN (grey) in various stages of meiotic maturation. Upper panels show the control cell, lower panels show the oocyte exposed to 5 μM proTAME. Scale bar: 20 μm. (B) Average SECURIN curves of control cells (blue, n = 5) and proTAME-treated cells (red, n = 11) with error bars. Time is relative to GVBD (a movie frame with GVBD or the first frame after GVBD represents time 0). Presented are representative results from a single experiment, which was repeated three times. (C) Time frames from live cell microscopy showing expression levels of microinjected SECURIN (grey) after the removal of proTAME from the culture media. Upper panels show the control cell, lower panels show cells previously exposed to 5 μM proTAME. Scale bar: 20 μm. (D) The left panel shows the average SECURIN curve with error bars of control cells (blue, n = 5) after proTAME removal; the right panel shows the average SECURIN curve with error bars of proTAME-treated cells (red, n = 11) after the removal of proTAME. The time is relative to the first frame after the removal of the inhibitor. Presented are representative results from a single experiment, which was repeated three times. [1].ProTAME Arrest in Mammalian Oocytes and Embryos Does Not Require Spindle Assembly Checkpoint Activity. Int J Mol Sci. 2019 Sep 13;20(18):4537.
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