来氟米特作用机制 - Medchemexpress - MCE中国

1、Product Data SheetLeflunomideCat. No.: HY-B0083CAS No.: 75706-12-6分式: CHFNO分量: 270.21作靶点: Others作通路: Others储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 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制备储备液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请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据您的

3、实验动物和给药式选择适当的溶解案。以下溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (9.25 mM); Suspended solution; Need ultrasonic and warm

4、ing此案可获得 2.5 mg/mL (9.25 mM) 的均匀悬浊液，悬浊液可于服和腹腔注射。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (9.25 mM); Clear solutionPage 1 of 2 www.MedChemE此案可获得 2.5 mg/mL (9.

5、25 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (9.25 mM); Clear solution此案可获得 2.5 mg/mL (9.25 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGIC

6、AL ACTIVITY物活性 Leflunomide湿的作。种嘧啶合成抑制剂，通过抑制氢乳 酸脱氢酶 (dihydroorotate dehydrogenase) 起作，具有抗风体外研究 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户使本产品发表的科研献 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