帕唑帕尼作用機制 - Medchemexpress - MCE中國

1、Product Data SheetPazopanibCat. No.: HY-10208CAS No.: 444731-52-6分式: CHNOS分量: 437.52作靶點: VEGFR; c-Kit; PDGFR; Autophagy; FGFR作通路: Protein Tyrosine Kinase/RTK; Autophagy儲存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據體外實驗 DMSO : 43 mg/mL (98.28 mM)* means soluble, but satur

2、ation unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲備液1 mM 2.2856 mL 11.4280 mL 22.8561 mL5 mM 0.4571 mL 2.2856 mL 4.5712 mL10 mM 0.2286 mL 1.1428 mL 2.2856 mL請根據產品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分裝保存，避免反復凍融造成的產品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲存時，請在 6 個內使，-20C 儲存時，請在 1 個內使。體內實驗請根

3、據您的實驗動物和給藥式選擇適當?shù)娜芙獍?。以下溶解案都請先按?In Vitro 式配制澄清的儲備液，再依次添加助溶劑：為保證實驗結果的可靠性，澄 的儲備液可以根據儲存條件，適當保存；體內實驗的作液，建議您現(xiàn)現(xiàn)配，當天使； 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. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (5.71 mM); Clear solution此案可獲得 2.5 mg/mL (5.71 mM，飽和度未知) 的澄

4、清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。2. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.71 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (5.71 mM，飽和度未知) 的澄 溶液，此案不適于實驗周 期在半個以上的實驗。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻

5、。BIOLOGICAL ACTIVITY物活性 Pazopanib (GW786034)是多靶點抑制劑，抑制 VEGFR1，VEGFR2，VEGFR3，PDGFR，c-Kit，F(xiàn)GFR1，c-Fms的IC50 分別為10，30，47，84，74，140，146 nM。IC & Target VEGFR1 VEGFR2 VEGFR3 PDGFR10 nM (IC50) 30 nM (IC50) 47 nM (IC50) 84 nM (IC50)FGFR1 c-Kit c-Fms140 nM (IC50) 74 nM (IC50) 146 nM (IC50)體外研究 Pazopanib shows

6、 good potency against all the human VEGFR receptors with an IC50 of 10, 30, and 47 nM for VEGFR-1, -2, and -3, respectively. Significant activity is also seen against the closely related tyrosine receptor kinases PDGFR,c-Kit, FGF-R1, and c-fms with IC50s of 84, 74, 140, and 146 nM, respectively. In

7、cellular assays, in addition to inhibitingthe VEGF-induced proliferation of HUVECs, Pazopanib potently inhibits VEGF-induced phosphorylation of VEGFR-2 inHUVEC cells with an IC50 of 8 nM. Pazopanib possesses good pharmacokinetics in rat, dog, and monkey with lowclearances (1.4-1.7 mL/min/kg) and goo

8、d oral bioavailabilities (72, 47, 65%) dosed at 10, 1, and 5 mg/kg, respectively.The cytochrome P450 profile is also improved with inhibition 10 M against the isozymes tested, with the exceptionof 2C9 (7.9 M)1.體內研究 Treatment of mice with 100 mg/kg of Pazopanib twice daily for five days results in si

9、gnificant inhibition in the degreeof vascularization. The antiangiogenic activity of Pazopanib is examined in mice bearing established humanxenografts (200250 mm3) using HT29 (colon carcinoma), A375P (melanoma), and HN5 (head and neck carcinoma)tumors following a standard three-week course of therap

10、y. The HN5 and HT29 xenografts responded better at alldoses compared to the A375P model, which is historically more resistant to VEGFR-2 inhibitors. As support that theobserved inhibition of xenograft growth is working through an antiangiogenic rather than antitumor mechanism, noantiproliferative ac

11、tivity is observed below 10 M for Pazopanib against these human tumor lines (HT29, HN5, A375P)growing in serum-containing media. No significant effect on the body weight of mice is observed, and the animalsappeared healthy and active throughout the study duration1. The quantity of adherent leukocyte

12、s in the Pazopanibeye drops group is less than untreated diabetic animals and more than the healthy animals. Average leukocytesadhered to the retinal vasculature in healthy animals is 37.27.8, whereas diabetic animals have an average value of10215.6, approximately 3-fold higher than healthy animals.

13、 Animals treated with 0.5 % w/v Pazopanib suspensiondemonstrate 69.59.5 leukocytes adhered in their retinal vasculature, which is found to be significantly lower thandiabetic animals2.PROTOCOLKinase Assay 1 VEGFR enzyme assays for VEGGR1, VEGFR2, and VEGFR3 are run in homogeneous time-resolved fluor

14、escence (HTRF)format in 384-well microtiter plates using a purified, baculovirus-expressed glutathione-S-transferase (GST) fusionprotein encoding the catalytic c-terminus of human VEGFR receptor kinases 1, 2, or 3. Reactions are initiated by theaddition of 10 L of activated VEGFR2 kinase solution fi

15、nal concentration, 1 nM enzyme in 0.1 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.5, containing 0.1 mg/mL bovine serum albumin (BSA), 300 Mdithiothreitol (DTT) to 10 L substrate solution final concentration, 360 nM peptide, (biotin-aminohexyl-EEEEYFELVAKKKK-NH2), 75 M ATP, 10

16、M MgCl2, and 1 L of titrated compound in DMSO. Plates are incubated atroom temperature for 60 min, and then the reaction is quenched by the addition of 20 L of 100 mM ethylenePage 2 of 3 www.MedChemEdiamine tetraacetic acid (EDTA). After quenching, 20 L HTRF reagents (final concentration, 15 nM Stre

17、ptavidin-linkedallophycocyanin, 1 nM Europium-labeled antiphosphotyrosine antibody diluted in 0.1 mg/mL BSA, 0.1 M HEPES, pH7.5) is added and the plates incubated for a minimum of 10 min. The fluorescence at 665 nM is measured with aWallac Victor plate reader using a time delay of 50 s1.MCE has not

18、independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 The effect of Pazopanib on cell proliferation is measured using 5-bromo-2-deoxyuridine (BrdU) incorporationmethod using commercially available kits. HUVEC is seeded in medium containing 5% fetal bovine se

19、rum (FBS) intype 1 collagen coated 96-well plates and incubated overnight at 37C, 5% CO2. The medium is aspirated from thecells, and various concentrations of Pazopanib in serum-free medium are added to each well. After 30 min, eitherVEGF (10 ng/mL) or bFGF (0.3 ng/mL) is added to the wells. Cells a

20、re incubated for an additional 72 h and BrdU (10 M) is added during the last 18 to 24 h of incubation. At the end of incubation, BrdU incorporation in cells is measuredby ELISA. Data are fitted with a curve described by the equation, y=Vmax(1(x/(K+x), where K is equal to the IC501.MCE has not indepe

21、ndently confirmed the accuracy of these methods. They are for reference only.Animal Mice1Administration 12 Tumors are initiated by injection of tumor cell suspension in 812 week old nude mice. When tumors reach a volumeof 100200 mm3, mice are randomized and divided into groups of eight. Pazopanib is

22、 administered once or twicedaily at 10, 30, or 100 mg/kg. Animals are euthanized by inhalation of CO2 at the completion of the study. Tumorvolume is measured twice weekly by calipers, using the equation: tumor volume (mm3)=(lengthwidth2)/2. Resultsare routinely reported as % inhibition=1(average gro

23、wth of the drug treated population/average growth of vehicletreated control population).Rats2Male Brown-Norway (BN; pigmented) rats weighing 200 to 250 g are acclimatized for at least two days prior to anyexperimental procedure. After overnight fasting for 12-16 h, an intraperitoneal injection of 30

24、 mg/mL solution ofStreptozotocin in 10 mM citrate buffer (pH 4.5) is administered (60 mg/kg body weight) to induce diabetes. After 3-4h of Streptozotocin injection, animals are put on a regular diet and 24 h after Streptozotocin injection, blood sample(5-10 L) is collected via tail vein. The blood g

25、lucose levels in the animals are determined with a glucose monitor.Animals with blood glucose levels greater than 250 mg/dL are considered diabetic. The animals are divided into threegroups. Group 1: Healthy (n=12), Group 2: Diabetic (n=12) and Group 3: Diabetic+Treatment (n=12). Treatment isstarted

26、 immediately after diabetes induction. Both eyes are dosed twice daily for 30 days with 0.5 % w/v Pazopanibsuspension (10 L volume in each eye) and animals in all groups are sacrificed on day 31, 16-17 h after last dose onday 30.MCE has not independently confirmed the accuracy of these methods. They are for reference only.戶使本產品發(fā)表的科研獻 Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Cell Syst. 2018 Apr 25;6(4):424-443.e