來氟米特作用機(jī)制 - Medchemexpress - MCE中國

1、Product Data SheetLeflunomideCat. No.: HY-B0083CAS No.: 75706-12-6分式: CHFNO分量: 270.21作靶點(diǎn): Others作通路: Others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 : 50 mg/mL (185.04 mM)Methanol : 2 mg/mL (7.40 mM; Need ultrasonic and warming)* means soluble, but saturati

2、on unknown.SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 3.7008 mL 18.5041 mL 37.0083 mL5 mM 0.7402 mL 3.7008 mL 7.4017 mL10 mM 0.3701 mL 1.8504 mL 3.7008 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)根據(jù)您的

3、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: 2.5 mg/mL (9.25 mM); Suspended solution; Need ultrasonic and warm

4、ing此案可獲得 2.5 mg/mL (9.25 mM) 的均勻懸濁液，懸濁液可于服和腹腔注射。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(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (9.25 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (9.

5、25 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。3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (9.25 mM); Clear solution此案可獲得 2.5 mg/mL (9.25 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 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 油中，混合均勻。BIOLOGIC

6、AL ACTIVITY物活性 Leflunomide濕的作。種嘧啶合成抑制劑，通過抑制氫乳 酸脫氫酶 (dihydroorotate dehydrogenase) 起作，具有抗風(fēng)體外研究 Leflunomide is actually a prodrug that has been shown to inhibit proliferation of mononuclear and T-cells.Leflunomide is an inhibitor of several protein tyrosine kinases, with IC50 values between 30 mM and

7、100 mM in vitrocellular and enzymatic assays1. Leflunomide is capable of inhibiting anti-CD3- and interleukin-2 (IL-2)-stimulated Tcell proliferation. Leflunomide is able to inhibit p59fyn and p56lck activity in in vitro tyrosine kinase assays.Leflunomide also inhibits Ca2+ mobilization in Jurkat ce

8、lls stimulated by anti-CD3 antibody but not in thosestimulated by ionomycin. Leflunomide also inhibits distal events of anti-CD3 monoclonal antibody stimulation,namely, IL-2 production and IL-2 receptor expression on human T lymphocytes. Leflunomide also inhibits tyrosinephosphorylation in CTLL-4 ce

9、lls stimulated by IL-22. Leflunomide is an immunomodulatory drug that may exert itseffects by inhibiting the mitochondrial enzyme dihydroorotate dehydrogenase (DHODH), which plays a key role in thede novo synthesis of the pyrimidine ribonucleotide uridine monophosphate (rUMP). Leflunomide prevents t

10、heexpansion of activated and autoimmune lymphocytes by interfering with the cell cycle progression due to inadequateproduction of rUMP and utilizing mechanisms involving p533.PROTOCOLKinase Assay 1 DHODase activity is measured by the DCIP colorimetric assay. This is a coupled assay in which oxidatio

11、n of DHO andsubsequent reduction of ubiquinone are stoichiometrically equivalent to the reduction of DCIP. Reduction of DCIP isaccompanied by a loss of absorbance at 610 nm (=21500 M/cm). The assay is performed in a 96-well microtiter plateat ambient temperature (ca. 25C). Stock solutions of 10 mM l

12、eflunomide and A771726 are prepared in dimethylsulfoxide (DMSO) and these are diluted with reaction buffer (100 mM Tris and 0.1 % Triton X-100, pH 8.0) to prepareworking stocks of the inhibitors at varying concentrations. For each reaction, the well contained 10 nM DHODase, 68M DCIP, 0.16 mg/mL gela

13、tin, the stated concentration of ubiquinone, 10 L of an inhibitor working stock to give thestated final concentration, and reaction buffer. After a 5-min equilibration period, the reaction is initiated by additionof DHO to the stated final concentrations. The total volume of reaction mixture for eac

14、h assay is 150 L, and the finalDMSO concentration is 0.01% (v/v). The reaction progress is followed by recording the loss of absorbance at 610nm over a 10-min period (during which the velocity remained linear). Velocities are reported as the change inabsorbance at 610 nm per minute, and each reporte

15、d value is the average of three replicates. In experiments wherethe DHO or ubiquinone concentration is varied, the other substrate is held constant at 200 M. To determine theinhibitor potency of leflunomide and A771726, the effects of varying concentrations of the two compounds on theinitial velocit

16、y of the DHODase reaction is measured over a concentration range of 0.011.0 M. In these experimentsthe DHO and ubiquinone concentrations are held constant at 200 and 100 M, respectively.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Page 2 of 3 www.Med

17、ChemE戶使本產(chǎn)品發(fā)表的科研獻(xiàn) Haematologica. 2018 Sep;103(9):1472-1483. Cancer Lett. 2018 Mar 28;417:21-34. Harvard Medical School LINCS LIBRARYSee more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Davis JP, et al. The immunosuppressive metabolite of leflunomide is a potent inhibitor o

18、f human dihydroorotate dehydrogenase. Biochemistry. 1996 Jan30;35(4):1270-3.2. Xu X, et al. Inhibition of protein tyrosine phosphorylation in T cells by a novel immunosuppressive agent, leflunomide. J Biol Chem. 1995 May26;270(21):12398-403.3. Fox RI, et al. Mechanism of action for leflunomide in rheumato