1、Product Data SheetTopotecan HydrochlorideCat. No.: HY-13768ACAS No.: 119413-54-6分式: CHClNO分量: 457.91作靶點(diǎn): Topoisomerase; Autophagy; Apoptosis作通路: Cell Cycle/DNA Damage; Autophagy; Apoptosis儲(chǔ)存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性數(shù)據(jù)體外實(shí)驗(yàn) H2O : 50 m

2、g/mL (109.19 mM; Need ultrasonic)DMSO : 25 mg/mL (54.60 mM; Need ultrasonic)SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 2.1838 mL 10.9192 mL 21.8384 mL5 mM 0.4368 mL 2.1838 mL 4.3677 mL10 mM 0.2184 mL 1.0919 mL 2.1838 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 mo

3、nths; -20C, 1 month (protect from light)。-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)娜芙獍浮Ｒ韵氯芙獍付颊?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(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可 以通過(guò)加熱和/或超聲的式助溶1. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in sa

4、line)Solubility: 2.5 mg/mL (5.46 mM); Clear solution此案可獲得 2.5 mg/mL (5.46 mM，飽和度未知) 的澄清溶液。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 20% 的 SBE-CD 理鹽溶液中，混合均勻。2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.46 mM); Clear solution此案可獲得 2.5 mg/mL (5.46 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

5、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。Page 1 of 2 www.MedChemEBIOLOGICAL ACTIVITY物活性 Topotecan Hydrochloride (SKF 104864A Hydrochloride)是具有抗癌活性的 拓?fù)洚悩?gòu)酶I 抑制劑。IC & Target Topoisomerase I體外研究 Topotecan Hydrochloride (SKF 104864A Hydrochloride) obviously inhibits proliferation of

6、 not only human gliomacells but also glioma stem cells (GSCs) in a dose- and time-dependent manner. According to the IC50 values at 24 h, 3M of Topotecan Hydrochloride is selected as the optimal administration concentration. In addition, TopotecanHydrochloride induces cell cycle arrest in G0/G1 and

7、S phases and promoted apoptosis. Results show that the cellviability is inhibited by Topotecan Hydrochloride in a dose-dependent manner. 2, 20 and 40 M of TopotecanHydrochloride obviously inhibits the cell viability compared with the control groups. The IC50 values of TopotecanHydrochloride at 24 h

8、are 2.730.25 M of U251 cells, 2.950.23 M of U87 cells, 5.460.41 M of GSCs-U251 and5.950.24 M of GSCs-U87. Thus 3 M of Topotecan Hydrochloride is selected as the optimal administrationconcentration in the subsequent experiments1.體內(nèi)研究 NUB-7 metastatic model, the animals belonging to all the 4 groups a

9、re sacrificed after 14 days treatment. Comparedwith the control, Low dose metronomic (LDM) Topotecan Hydrochloride and TP+Pazopanib (PZ) liver weights aresignificantly lower in TP+PZ-treated animals, compared with PZ. Microscopic tumors are visible in the livers of micebelonging to all the groups ex

10、cept TP+PZ confirming the ability of Topotecan Hydrochloride+PZ to control livermetastasis. In a previous dose-response study, the daily dose of oral metronomic Topotecan Hydrochloride (0.5, 1.0,and 1.5 mg/kg) causes greater reduction in microvascular density compared with weekly maximum-tolerated d

11、oseregimen (7.5 and 15 mg/kg) in an ovarian cancer model, but the mice treated with 1.5 mg/kg daily, oral TopotecanHydrochloride show decreased food intake, and a lesser antitumor effect2.PROTOCOLCell Assay 1 The U251, U87, GSCs-U251 and GSCs-U87 cells are seeded at a density of 2104 cells per well

12、in 96-well platesseparately, and incubated for 24 h. Cells are administered with Shikonin and Topotecan Hydrochloride (0.02, 0.2, 2,20, 40 M). After the treatment, 10 L of cell counting kit-8 (CCK-8) is added into each well for additional 1-hourincubation at 37C. The optical density (OD) is read wit

13、h a microplate reader at 450 nm1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice2Administration 2 For subcutaneous xenograft studies, we used SK-N-BE, SH-SY5Y, KHOS, and RH30. 1106 cells are implantedsubcutaneously into the inguinal fat pad

14、of each of nonobese diabetic/severe combined immune deficient(NOD/SCID) mice. When tumors reached 0.5 cm in diameter, the animals are randomized into 4 groups and treateddaily by oral gavage. The animals are grouped as: Control group, LDM Topotecan group or LDM TP (1 mg/kgTopotecan), Pazopanib group

15、 or PZ (150 mg/kg Pazopanib) and combination group or TP + PZ (1 mg/kg TopotecanHydrochloride + 150 mg/kg Pazopanib). To compare pulse Topotecan with LDM TP in KHOS osteosarcoma model,PZ is replaced by weekly oral dose of pulse Topotecan or Pulse TP (15 mg/kg Topotecan). The criteria for endpointare

16、 tumor sizes exceeding 2.0 cm in diameter or animals showing signs of morbidity. The tumor sizes are measuredon a daily basis until the endpoint or sacrifice. The long (D) and short diameters (d) are measured with calipers.Tumor volume (cm3) is calculated as V=0.5Dd2. When the endpoint is reached or

17、 at the end of the treatment, theanimals are sacrificed by cervical dislocation.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.MedChemE戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Nat Commun. 2019 Aug 21;10(1):3761. J Exp Clin Cancer Res. 2018 Dec 20;37(1):321. FASEB J

18、. 2020 Feb. J Mol Med (Berl). 2019 Aug;97(8):1183-1193. J Virol. 2019 Mar 13. pii: JVI.02230-18.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Zhang FL, et al. PLoS One. 2013 Nov 26;8(11):e81815.Topoisomerase I inhibitors, Shikonin and Topotecan, inhibit growth and induce apoptosis of gliomacells andglioma stem cells.2. Kumar S, et al. Metronomic oral topotecan with pazopanib is an active antiangiogenic regimen in mouse models of aggressive pedia