GSTO1-IN-1 is a potent glutathione S-transferase omega 1 (GSTO1) inhibitor with an IC₅₀ of 31 nM.

IC50: 31 nM (GSTO1)^[1]

GSTO1-IN-1 (C1-27) potently inhibits GSTO1 enzyme activity with an IC₅₀ value of 31 nM. GSTO1-IN-1 also potently competes with 5-chloromethylfluorescein diacetate (CMFDA) for binding to recombinant protein, as well as endogenous GSTO1 in the milieu of a soluble proteome. HCT116 cells treated with GSTO1-IN-1 also show a decrease in cell viability in a dose-dependent manner. GSTO1-IN-1 inhibits the clonogenic survival of HCT116 cells at sub-micromolar concentrations^[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 test whether GSTO1-IN-1 had in vivo efficacy, its effects are evaluated in a human colon cancer cell line xenograft model. GSTO1-IN-1 (20-45 mg/kg) is administered as a single agent to nude mice bearing HCT116 xenografts. After 5 weeks of treatment, tumor growth is significantly inhibited in GSTO1-IN-1-treated mice compared with the vehicle-treated group (P<0.05). GSTO1-IN-1 treatment is generally well tolerated by mice up to 45 mg/kg, with no overt signs of toxicity and no significant variations in average body weights throughout the duration of the study^[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.

311.18

C₁₀H₁₂Cl₂N₂O₃S

568544-03-6

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 : ≥ 300 mg/mL (964.07 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 (8.03 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (8.03 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% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.5 mg/mL (8.03 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (8.03 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 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.5 mg/mL (8.03 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (8.03 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]. Ramkumar K, et al. Mechanistic evaluation and transcriptional signature of a glutathione S-transferase omega 1 inhibitor. Nat Commun. 2016 Oct 5;7:13084.

Cell proliferation is assessed by a MTT assay. Cancer cells H630, HT29 and HCT116 are seeded in 96-well microtitre plates and, after overnight attachment, treated with GSTO1 inhibitors (e.g., GSTO1-IN-1; 0.1, 1, 10 and 100 μM). After 72 h, MTT solution (3 mg/mL; 20mL) is added to each well and cells are incubated for 3 h at 37°C. After incubation, media from each well is removed and the dark blue formazan crystals formed by live cells are dissolved in DMSO (150 mL per well). The absorbance intensity is measured at 570 nm on a microplate reader. Cell viability after 24 h treatment is assessed using ApoTox-Glo triplex assay. At least three independent dose-response experiments with each concentration tested in triplicate are performed for each cell line^[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]
In the pilot study, HCT116 cells (1×10⁶) in exponential phase are injected subcutaneously into the left flank of 8- to 10-week-old female nude mice (25-30 g) . The perpendicular diameters of the tumors are measured three times weekly using standard calipers and tumour volumes are calculated. Tumors are allowed to grow to a volume of 50 mm³ and mice are randomized into control (n=5) and GSTO1-IN-1 (n=3) treatment groups. GSTO1-IN-1 is administered intraperitoneally (20 mg/kg per day) for the first 2 weeks on a 5 days on/2 days off schedule. The dose is then increased to 25 mg/kg per day for the next 23 days and further escalated by 5 mg/kg per day to a final dose of 45 mg/kg for the remaining duration of treatment. Tumor volumes and body weights are measured three times weekly to monitor tumor burden and weight loss during treatment. At the end of the experiment, animals are killed and tumor, kidney and liver are collected, fixed and paraffin embedded for histology^[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]. Ramkumar K, et al. Mechanistic evaluation and transcriptional signature of a glutathione S-transferase omega 1 inhibitor. Nat Commun. 2016 Oct 5;7:13084.