TMS ((E)-2,3′,4,5′-tetramethoxystilbene) is a selective and competitive CYP1B1 inhibitor with an IC₅₀ of 6 nM and a K_i value of 3 nM. TMS shows a lesser extent inhibitory effect on CYP1A1 (IC₅₀50[1]^[2][3].

TMS, an analogue of resveratrol, is considered to be a potential cancer preventive agent since it is a potent inhibitor of CYP1B1. To assess survival of MCF-7 cells exposed to 1 μM benzo[a]pyrene (BP), 1 μM BP+1 μM TMS and 1 μM BP+4 μM TMS, cells ae incubated for up to 72 h without a media change. Luminescence units from exposed cells, expressed as a percentage of luminescence units from solvent (DMSO)-treated cells at the same time intervals. In all exposure groups, cell viability remains >90% for the first 24 h, but by 72 h, cell survival drops to 60-70%^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

To determine the contribution of CYP1B1 in development of hypertension in spontaneously hypertensive rats (SHR), the effect of TMS is examined on in SHR and WKY rats. Systolic BP steadily increases in SHR from 4 weeks of age. Starting from 8 weeks of age, daily injections of TMS reduce systolic BP in SHR to levels observed at the beginning of the experiment (207±7 vs. 129±2 mmHg). Systolic BP is not altered in WKY injected with TMS or its vehicle (129±7 vs. 127±4 mmHg) ^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

300.35

C₁₈H₂₀O₄

24144-92-1

Room temperature in continental US; may vary elsewhere.

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	6 months
	-20°C	1 month

In Vitro: 

DMSO : ≥ 50 mg/mL (166.47 mM)

* “≥” means soluble, but saturation unknown.

*

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.5 mg/mL (8.32 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (8.32 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (8.32 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (8.32 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Sanghee Kim, et al. Design, synthesis, and discovery of novel trans-stilbene analogues as potent and selective human cytochrome P450 1B1 inhibitors. J Med Chem. 2002 Jan 3;45(1):160-4.

  [2]. Einem Lindeman T, et al. The resveratrol analogue, 2,3′,4,5′-tetramethoxystilbene, does not inhibit CYP gene expression, enzyme activity and benzo[a]pyrene-DNA adduct formation in MCF-7 cells exposed to benzo[a]pyrene. Mutagenesis. 2011 Sep;26(5):629-35.

  [3]. Jennings BL, et al. Cytochrome P450 1B1 contributes to increased blood pressure and cardiovascular and renal dysfunction in spontaneously hypertensive rats. Cardiovasc Drugs Ther. 2014 Apr;28(2):145-61.

Cell viability is determined using the Cell Titer Glo Luminescent Cell Viability Assay. In brief, MCF-7 cells are seeded in 12-well plates (75 000 cells per well) in triplicate. Attached cells are exposed to 1 μM BP in the presence of 0, 1 or 4 μM TMS. At 4, 12, 24 or 72 h, RIPA Lysis Buffer (1x) is added to lyse the cells. A diluted sample of homogeneous cell lysate is transferred in duplicate to a 96-well plate and combined with an equivalent volume of Cell Titer Glo Luminescence. Luminescence measure using the Tropix 717 Microplate Luminometer. To examine viability of cells exposed to BP alone, the 72-h treatment is repeated twice, and for each experiment cells are assayed in triplicate. The values are expressed as % of the DMSO-alone solvent control^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
36 three-weeks-old male spontaneously hypertensive rats (SHR) and 36 age-matched WKY are used throughout this research. Each strain of rats is split into two groups; one group is injected daily with TMS (600 μg/kg) and the other with vehicle (DMSO; 100 μL) beginning at 8 weeks of age. Systolic BP and mean arterial pressure (MAP) are measured twice a week from 4 weeks of age; a noninvasive tail cuff method is used^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Sanghee Kim, et al. Design, synthesis, and discovery of novel trans-stilbene analogues as potent and selective human cytochrome P450 1B1 inhibitors. J Med Chem. 2002 Jan 3;45(1):160-4.

  [2]. Einem Lindeman T, et al. The resveratrol analogue, 2,3′,4,5′-tetramethoxystilbene, does not inhibit CYP gene expression, enzyme activity and benzo[a]pyrene-DNA adduct formation in MCF-7 cells exposed to benzo[a]pyrene. Mutagenesis. 2011 Sep;26(5):629-35.

  [3]. Jennings BL, et al. Cytochrome P450 1B1 contributes to increased blood pressure and cardiovascular and renal dysfunction in spontaneously hypertensive rats. Cardiovasc Drugs Ther. 2014 Apr;28(2):145-61.