西地那非作用機(jī)制 - Medchemexpress - MCE中國

1、Product Data SheetSildenafilCat. No.: HY-15025CAS No.: 139755-83-2分式: CHNOS分量: 474.58作靶點(diǎn): Phosphodiesterase (PDE); Autophagy; Apoptosis作通路: Metabolic Enzyme/Protease; Autophagy; Apoptosis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 : 29 mg/mL (61.11 mM)H2O : 0

2、.1 mg/mL (insoluble)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 2.1071 mL 10.5356 mL 21.0713 mL5 mM 0.4214 mL 2.1071 mL 4.2143 mL10 mM 0.2107 mL 1.0536 mL 2.1071 mL請根據(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 mo

3、nth。-80C 儲(chǔ)存時(shí)，請?jiān)?6 個(gè)內(nèi)使，-20C 儲(chǔ)存時(shí)，請?jiān)?1 個(gè)內(nèi)使。體內(nèi)實(shí)驗(yàn)請根據(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韵氯芙獍付颊埾劝凑?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. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (5.27

4、mM); Clear solution此案可獲得 2.5 mg/mL (5.27 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 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. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (5.27 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (5.27 mM

5、，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 Sildenafil (UK-92480)種有效的磷酸酯酶 5 (PDE5)抑制劑，IC50 為 5.22 nM。IC & Target IC50: 5.22 nM (PDE 5)1體外研究 Pretreatment with 1 M Sildenafil potentiates the phosphorylation of ERK1/ERK2, an increas

6、e in the percentage ofcells in S phase and cell proliferation, compared with serotonin stimulation alone (P0.05). Pretreatment with 1 MSildenafil citrate followed by serotonin stimulation leads to dramatic increase in OD value to 0.33, significantlydifferent compared with serotonin stimulation alone

7、 (P0.05). 1 M Sildenafil obviously enhances the upregulation ofERK1/ERK2 phosphorylation induced by serotonin2.體內(nèi)研究 In the dog model of erection, Sildenafil citrate significantly increases ICP and ICP/BP but shows no significant effecton BP compared with vehicle1. Sildenafil treatment significantly

8、decreases the number of TL+-cells at 10 but not 0.5 mg/kg. At this time point, cells positive for the M1-like marker COX-2+ are found in the ischemic core in PBS-treatedanimals, whereas they are mostly observed in the penumbra in 10 mg/kg (but not 0.5 mg/kg) Sildenafil-treatedanimals. In contrast, 8

9、 days after pMCAo the number of microglia/macrophages stained by Iba-1 are significantlyreduced by Sildenafil treatment (0.5 and/or 10 mg/kg dose)3. Sildenafil citrate has been reported to decrease flapnecrosis in preclinical animal models by increasing the secretion of growth factors (FGF and VEGF)

10、, and histologicallyis shown to be effective in rat cavernous nerve architecture4.PROTOCOLCell Assay 2 Cells at approximately 90% confluence are harvested with 0.1% trypsin/0.01% ethylene diamine tetraacetic acid(EDTA) solution and seeded into a 96-well plate at a density of 2104 cells/well and grow

11、n in RPMI-1640 containing10% FBS for three days, followed by serum starvation for three days. Cells are then incubated for different time withvarious concentration of serotonin or 1 M Sildenafil followed by serotonin with or without U0126, as indicated.Control cells are treated in the same way excep

12、t sterile PBS replaced the drug. After treatment, medium is changed tofresh medium, and cells are incubated with 5 g/L of MTT for four hours. MTT is then dissolved with 150 L of 10%DMSO for 20 minutes. The optical densities (OD) in the 96-well plates are determined using a microplate reader at570 nm

13、2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice3Administration 34 Ischemia is induced in C57Bl/6 mice on postnatal (P) day 9 by permanent middle cerebral artery occlusion (pMCAo),and followed by either PBS or Sildenafil intraperitoneal (i.

14、p.) injections. In the first set of experiments, animals arerandomly divided into five groups and treated with either PBS or a single dose of Sildenafil citrate (0.5, 2.5, 10, and 15mg/kg), given intraperitoneally (i.p.) 5 min after pMCAo. In the second set of experiments, animals are randomlydivide

15、d into three groups and treated with either PBS or a single dose of Sildenafil citrate (0.5 and 10 mg/kg, i.p.) 5min after pMCAo.Rats4Thirty male Sprague-Dawley rats weighing between 210 and 240 g are used. Rats from all groups are anesthetizedwith xylazine + ketamine and then a crush injury is crea

16、ted by using a one-minute long vascular clamp to the rightsciatic nerve. One day before the procedure, rats from Group 1 are started on a 28-day treatment consisting of aPage 2 of 3 www.MedChemEdaily dose of 20 mg/kg body weight Sildenafil given orally via nasogastric tube, while the rats from Group

17、 2 arestarted on an every-other-day dose of 10 mg/kg body weight Sildenafil citrate. Rats from Group 3 did not receive anydrugs. Subjects in all 3 groups are fed ad libitum with normal rat chow and tap water. Forty-two days after the nervedamage is created, the rats underwent a static sciatic index

18、(SSI) test, sedation and motor coordination tests, andaccelerated rotarod tests. Rats are sacrificed under anesthesia and their sciatic nerves are removed surgically.Histopathologic analyses of the nerves and bone densitometry evaluation of the extremities are then performed.MCE has not independentl

19、y confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Wang Z, et al. The Selectivity and Potency of the New PDE5 Inhibitor TPN729MA. J Sex Med. 2013 Nov;10(11):2790-7.2. Li BB, et al. Sildenafil potentiates the proliferative effect of porcine pulmonary artery smooth muscle cells ind