奥拉帕尼作用机制 - Medchemexpress - MCE中国

1、Product Data SheetOlaparibCat. No.: HY-10162CAS No.: 763113-22-0分式: CHFNO分量: 434.46作靶点: PARP; Autophagy; Mitophagy作通路: Cell Cycle/DNA Damage; Epigenetics; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 33.33 mg/mL (76.72 mM)* means soluble, but satura

2、tion unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 2.3017 mL 11.5085 mL 23.0171 mL5 mM 0.4603 mL 2.3017 mL 4.6034 mL10 mM 0.2302 mL 1.1509 mL 2.3017 mL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；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据

3、您的实验动物和给药式选择适当的溶解案。以下溶解案都请先按照 In Vitro 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 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.75 mM); Clear solution此案可获得 2.5 mg/mL (5.75 mM，饱和度

4、未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (5.75 mM); Clear solutionPage 1 of 2 www.MedChemE此案可获得 2.5 mg/mL (5.75 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L

5、 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.75 mM); Clear solution此案可获得 2.5 mg/mL (5.75 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Olaparib (AZD2281; KU00

6、59436)种服有效的 PARP 抑制剂，抑制 PARP-1 和 PARP-2 的 IC50 分别为 5 和 1 nM。Olaparib 种噬 (autophagy) 和有丝分裂 (mitophagy) 激活剂。IC & Target PARP-2 PARP-1 tankyrase-1 Autophagy1 nM (IC50) 5 nM (IC50) 1.5 M (IC50)Mitophagy体外研究 Olaparib (AZD2281) is a single digit nanomolar inhibitor of both PARP-1 and PARP-2 that shows sta

7、ndalone activityagainst BRCA1-deficient breast cancer cell lines. Olaparib is applied to SW620 cell lysates, and identified the IC50 forPARP-1 inhibition to be around 6 nM and the total ablation of PARP-1 activity to be at concentrations of 30100 nM1.体内研究 Animals bearing SW620 xenografted tumors are

8、 treated with Olaparib (10 mg/kg, p.o.) in combination with NSC362856 (TMZ) (50 mg/kg, p.o.) once daily for 5 consecutive days, after which the tumors are left to grow out1.Olaparib increases vascular perfusion in Calu-6 tumors established in a DWC model. Administration of olaparib(50mg/kg, p.o.) as

9、 a single agent (top panel) or in combination with radiation (bottom panel) results in an increase influorescence intensity in the Calu-6 tumors2.PROTOCOLKinase Assay 1 This assay determined the ability of Olaparib to inhibit PARP-1 enzyme activity. PARP-2 activity inhibition is measuredby using a v

10、ariation of the PARP-1 assay in which PARP-2 protein (recombinant) is bound down by a PARP-2 specificantibody in a 96-well white-walled plate. PARP-2 activity is measured following 3H-NAD+ DNA additions. Afterwashing, scintillant is added to measure 3H-incorporated ribosylations. For tankyrase-1, an

11、 AlphaScreen assay isdeveloped in which HIS-tagged recombinant TANK-1 protein is incubated with biotinylated NAD+ in a 384-wellProxiPlate assay. Alpha beads are added to bind the HIS and biotin tags to create a proximity signal, whereas theinhibition of TANK-1 activity is directly proportional to th

12、e loss of this signal. All experiments are repeated at leastthree times1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 The PF50 value is the potentiation factor, which is calculated as the ratio of the IC50 of the control growth withalkyl

13、ating agent methylmethane sulfonate (MMS) divided by the IC50 of the MMS combined with the PARP inhibitor.HeLa B cells are used, and Olaparib is tested at a fixed 200 nM concentration for screening with MMS. For the testingof Olaparib on the SW620 colorectal cell line, the concentrations that are us

14、ed are 1, 3, 10, 100 and 300 nM. Cellgrowth is assessed by the use of the sulforhodamine B (SRB) assay1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemEAnimal Mice2Administration 2 Mice bearing 220-250 mm3 tumors are randomized i

15、nto 4 treatment groups (n=5): A; vehicle control (10% DMSO inPBS/10% 2-hydroxy-propyl-cyclodextrin daily for 5 days by oral gavage), B; Olaparib (50 mg/kg daily for 5 days byoral gavage), C; 10 Gy fractionated radiotherapy (2 Gy daily for 5 days), D; Olaparib and 10 Gy (52 Gy) fractionatedradiothera

16、py (with olaparib given 30 min prior to each daily 2 Gy dose of radiation). Tumor volume measurementsare determined daily until they reached 1000 mm3. The number of days for each individual tumor to quadruple insize from the start of the treatment (relative tumor volume4; RTV4) is calculated for the

17、 individual tumors in eachgroup.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Nature. 2018 Nov;563(7729):131-136. Cell. 2019 Jan 24;176(3):505-519.e22. Cancer Discov. 2018 Mar;8(3):354-369. Cancer Discov. 2017 Sep;7(9):984-998. Mol Cell. 2

18、019 Sep 19;75(6):1299-1314.e6.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Menear KA, et al. 4-3-(4-cyclopropanecarbonylpiperazine-1-carbonyl)-4-fluorobenzyl-2H-phthalazin-1-one: a novel bioavailable inhibitor of poly(ADP-ribose) polymerase-1. J Med Chem. 2008 Oct 23;51(20):6581-912. Senra JM, et al. Inhibition of PARP-1 by olaparib (AZD2281) increases the radiosensitivity of a lung tumor xenograft.