鹽酸帕唑帕尼作用機(jī)制 - Medchemexpress - MCE中國(guó)

1、Product Data SheetPazopanib HydrochlorideCat. No.: HY-12009CAS No.: 635702-64-6分式: CHClNOS分量: 473.98作靶點(diǎn): PDGFR; VEGFR; FGFR; c-Kit; c-Fms; Autophagy作通路: Protein Tyrosine Kinase/RTK; 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 : 10 mg/mL (21.10 mM; Ne

2、ed ultrasonic)SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 2.1098 mL 10.5490 mL 21.0979 mL5 mM 0.4220 mL 2.1098 mL 4.2196 mL10 mM 0.2110 mL 1.0549 mL 2.1098 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

3、據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?。以下溶解案都?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: 1 mg/mL (2.11 mM); Clear solution此案可獲得 1 mg/mL (2.11 mM，飽和度未知)

4、 的澄清溶液。以 1 mL 作液為例，取 100 L 10.0 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: 1 mg/mL (2.11 mM); Clear solution此案可獲得 1 mg/mL (2.11 mM，飽和度未知) 的澄清溶液。Page 1 of 2 www.MedChemE以 1 mL 作液為例，取 100 L 10.0 m

5、g/mL 的澄均勻。DMSO 儲(chǔ)備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 1 mg/mL (2.11 mM); Clear solution此案可獲得 1 mg/mL (2.11 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 10.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Pazopanib Hydrochloride (GW786034 H

6、ydrochloride)是多靶點(diǎn)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 good potency against all the h

7、uman 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 cellular assays, in addition to

8、 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 good oral bioavailabilities (72, 4

9、7, 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(nèi)研究 Treatment of mice with 100 mg/kg of Pazopanib twice daily for five days results in significant inhibition in the deg

10、reeof 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 therapy. The HN5 and HT29 xenografts

11、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 activity is observed below 10 M f

12、or 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 leukocytes in the Pazopanibeye drops gro

13、up 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. Animals treated with 0.5 % w/v

14、 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 are initiated by the addition of 10 L of activated VEGFR2 kinase solution final concentration,1 nM enzym

15、e in 0.1 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.5, containing 0.1 mg/mLbovine serum albumin (BSA), 300 M dithiothreitol (DTT) to 10 L substrate solution final concentration, 360 nMPage 2 of 3 www.MedChemEpeptide, (biotin-aminohexyl-EEEEYFELVAKKKK-NH2), 75 M ATP, 10 M MgCl

16、2, and 1 L of titrated compound(Pazopanib) in DMSO. Plates are incubated at room temperature for 60 min, and then the reaction is quenched bythe addition of 20 L of 100 mM ethylene diamine tetraacetic acid (EDTA). After quenching, 20 L HTRF reagents(final concentration, 15 nM Streptavidin-linked all

17、ophycocyanin, 1 nM Europium-labeled antiphosphotyrosineantibody diluted in 0.1 mg/mL BSA, 0.1 M HEPES, pH 7.5) is added and the plates incubated for a minimum of 10min. The fluorescence at 665 nM is measured with a plate reader1.MCE has not independently confirmed the accuracy of these methods. They

18、 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 serum (FBS) intype 1 collagen coated 96-well plates and incub

19、ated 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 are incubated for an additional 72 h and BrdU (10 M) is adde

20、d 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 independently confirmed the accuracy of these methods. They are f

21、or 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 administered once or twicedaily at 10, 30, or 100 mg/kg. A

22、nimals 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 growth of the drug treated population/average growth of vehicl

23、etreated 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 mg/mL solution ofStreptozotocin in 10 mM citrate buffer (p

24、H 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 glucose levels in the animals are determined with a glucose

25、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 immediately after diabetes induction. Both eyes are dosed

26、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(chǎn)品發(fā)表的科研獻(xiàn) Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093. Cell Syst. 2018 Apr 25;6(4):424-443.e7. Cel