Capzimin is a potent and moderately specific proteasome isopeptidase Rpn11 inhibitor.

Target: Rpn11^[1]

Capzimin (3027) shows 80-fold selectivity for Rpn11 over Csn5, 10-fold over AMSH and 6-fold over BRCC36 (IC₅₀=30 μM, 4.5 μM and 2.3 μM respectively. Capzimin is screened against the NCI panel of 60 cancer cell lines. The median GI₅₀ is 3.3 μM. Capzimin exhibits promising activity in leukemia cells including the SR and K562 cell lines (GI₅₀ values of 0.67 μM and 1 μM respectively), as well as several solid tumor cell lines including NCI-H460 (non-small cell lung cancer; GI₅₀= 0.7 μM) and MCF7 (breast cancer; GI₅₀=1.0 μM). Immunoblotting for the processed form of caspase 3 and caspase-cleaved poly ADP-ribose polymerase in HCT116 cells confirm that Capzimin not only blocks cell growth, but also induces apoptosis^[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.

628.81

C₃₀H₂₄N₆O₂S₄

2084868-04-0

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 : ≥ 55.67 mg/mL (88.53 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 (3.98 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (3.98 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 (3.98 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (3.98 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]. Li J, et al. Capzimin is a potent and specific inhibitor of proteasome isopeptidase Rpn11. Nat Chem Biol. 2017 May;13(5):486-493.

Fluorescence polarization assays are performed in low-volume 384 well solid black plates in quadruplicate. The assays are performed in buffer containing 50 mM Tris-HCl pH7.5, 1mM MgCl₂, 50 μM ATP, 1 mM DTT and 0.01% NP-40. The components for the assay are added in the follow sequence: 1) 5 μL compound (Capzimin, et al.) (in 3% DMSO) at different concentrations, 2) 5 μL of diluted human 26S proteasome, and 3) 5μL of substrate (3 nM Ub4- peptideOG). 100 μM Zn(cyclen)²⁺ is present in the titration reaction for the experiments performed with Zn(cyclen)²⁺. Fluorescence polarization is measured at 30°C with excitation at 480 nm and emission at 520 nm. Collected data is normalized to DMSO control and fitted to a dose-response curve to determine the IC₅₀ value^[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.

HCT116 cells are treated with different concentrations of 3021 or Capzimin for 72 hours in normal or low serum medium and then mixed with CellTiter-Glo reagent to estimate cell proliferation. Measured luminescence values are normalized to DMSO control and data are fitted to a dose-response equation to determine the GI₅₀^[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]. Li J, et al. Capzimin is a potent and specific inhibitor of proteasome isopeptidase Rpn11. Nat Chem Biol. 2017 May;13(5):486-493.