1、Product Data SheetPAC-1Cat. No.: HY-13523CAS No.: 315183-21-2分式: CHNO分量: 392.49作靶點(diǎn): Caspase; Autophagy; Apoptosis作通路: Apoptosis; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 50 mg/mL (127.39 mM; Need ultrasonic)H2O : 0.1 mg/mL (insoluble)SolventMass

2、1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 2.5478 mL 12.7392 mL 25.4784 mL5 mM 0.5096 mL 2.5478 mL 5.0957 mL10 mM 0.2548 mL 1.2739 mL 2.5478 mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲(chǔ)備液；旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲(chǔ)存時(shí)，請(qǐng)?jiān)?6 個(gè)內(nèi)使，-20C 儲(chǔ)存時(shí)，請(qǐng)?jiān)?1 個(gè)內(nèi)使。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?。以下溶解?/p>

3、都請(qǐng)先按照 In Vitro 式配制澄清的儲(chǔ)備液，再依次添加助溶劑：為保證實(shí)驗(yàn)結(jié)果的可靠性，澄 的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (6.37 mM); Clear solution此案可獲得 2.5 mg/mL (6.37 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取

4、100 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (6.37 mM); Suspended solution; Need ultrasonicPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (6.37 mM) 的均勻懸濁液，懸濁液可于服和腹腔注射。以 1 mL 作液為例，取 1

5、00 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (6.37 mM); Clear solution此案可獲得 2.5 mg/mL (6.37 mM，飽和度未知) 的澄 溶液，此案不適于實(shí)驗(yàn)周 期在半個(gè)以上的實(shí)驗(yàn)。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 PAC-1種 procaspase-3

6、 激活劑，誘導(dǎo)癌細(xì)胞凋亡，EC50 為 2.08 M。IC & Target Procaspase-32.08 M (EC50)體外研究 PAC-1 activates procaspase-3 with an EC50 of 2.08 M. PAC-1 exhibits an enhanced zinc chelating ability (EC50= 7.08M). PAC-1 induces leukemia cell death with IC50 of 4.03 M, which is consistent with the values reported by otherinves

7、tigators. PAC-1 treatment also results in death of other malignant cells in a concentration-dependent mannerwith IC50s ranging from 4.03 to 53.44M. The overall mean IC50 in the fifteen malignant cell lines is 0.88 mM for WF-210 and 19.40 M for PAC-1. In contrast, the sensitivity of the normal human

8、cells (PBL, L-02, HUVEC and MCF 10A)to WF-210 is 2.6-fold lower (mean IC50=412.34 M) than PAC-1 (mean IC50=158.29 M)1. Procaspase-activatingcompound-1 (PAC-1) is the first direct caspase-activating compound discovered. PAC-1 treatment upregulates Ero1in multiple cell lines, whereas silencing of Ero1

9、 significantly inhibits calcium release from ER and cell death2.體內(nèi)研究 To evaluate the in vivo effect of WF-210 on the growth of malignant tumors, we examined the ability of WF-210 tosuppress tumor growth in mouse Hep3B and MDA-MB-435 xenograft models. These two cell lines expressprocaspase-3 at relat

10、ively high levels. Tumors induced by xenografts of the liver cancer cell Hep3B are allowed todevelop and grow to a size of 100 mm3, after which WF-210 (2.5 mg/kg) or PAC-1 (5.0 mg/kg) is given daily for twoweeks by intravenous (i.v.) administration. As shown in both PAC-1 and WF-210 significantly in

11、hibits the growth ofHep3B tumor xenografts1.PROTOCOLKinase Assay 1 Various concentrations of WF-210 or PAC-1 are added to procaspase-3 in buffer containing 50 mM HEPES, 0.1%CHAPS, 10% glycerol, 100 mM NaCl, 0.1 mM EDTA, 10 mM DTT pH 7.4,and incubated for 12 h at 37C. The finalvolume is 10 mL and the

12、 final concentration of procaspase-3 is 1 mM. Then 40 mL of the substrate Ac-DEVD-pNA(final concentration 0.4 mM) in buffer containing 50 mM HEPES pH 7.4, 100 mM NaCl, 10 mM DTT, 0.1 mM EDTAdisodium salt, 0.10% CHAPS, 10% glycerol is added and the absorbance of the plate is read at 405 nm for a tota

13、l of 1h. The slope of the linear portion for each well is determined as the enzyme activity1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Cell viability is measured using the MTT method or the Cell Titer-Glo luminescent assay. For the MT

14、T assay, the cells(1105 cells/mL) are seeded into 96- well culture plates. After overnight incubation, cells are treated with variousconcentrations of agents (PAC-1, WF-210 or other agents) for 24 or 72 h. Then 10 mL MTT solution (2.5 mg/mL inPBS) is added to each well, and the plates are incubated

15、for an additional 4 h at 37C. After centrifugation (2500 rpm,10min), the medium containing MTT is aspirated, and100mL DMSO is added. The optical density of each well ismeasured at 570 nm with a Biotek Synergy HT Reader. The Cell Titer-Glo kit is used to determine the relative levels ofPage 2 of 3 ww

16、w.MedChemEintracellular ATP as a biomarker for live cells1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice1Administration 1 To determine the in vivo anti-tumor activity of WF-210, viable human gallbladder cancer GBC-SD cells (5106/100mL PBS

17、per mouse), human breast cancer MDA-MB-435 cells (1107/100 mL PBS per mouse), human liver cancerHep3B cells (5106/100 mL PBS per mouse) and human breast cancer MCF-7 cells (1107/100 mL PBS per mouse)are subcutaneously (s.c.) injected into the right flank of 7- to 8-week old male SCID mice or Balb/c

18、nude mice. Cellnumbers are confirmed by trypan blue staining prior to injection. Specially, MCF-7 xenograft mice are alsoadministered with the hormone 17-beta-estradiol (3 mg/kg) on alternate days. When the average s.c. tumor volumereached 100 mm3, mice are randomly divided into various treatment an

19、d control groups (eight mice per group).Tumor size is measured once every two days with a caliper (calculated volume=shortest diameter2longestdiameter/2). Body weight, diet consumption and tumor size are recorded once every two days. After two or fourweeks, mice are sacrificed and tumors are excised

20、 and stored at -80C until further analysis.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Poult Sci. 2019 Aug 9. pii: pez465. Oncotarget. 2017 Feb 14;8(7):12311-12322.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Wang F, et al. A novel small-molecule activator of procaspase-3 induces apoptosis in cancer cells and reduces tumor growth in human breas