托法替尼作用機(jī)制 - Medchemexpress - MCE中國(guó)

1、Product Data SheetTofacitinibCat. No.: HY-40354CAS No.: 477600-75-2分式: CHNO分量: 312.37作靶點(diǎn): JAK; Apoptosis作通路: Epigenetics; JAK/STAT Signaling; Stem Cell/Wnt; 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 : 65 mg/mL (208.09 mM; Need ultrasonic)H2O : 0.15

2、 mg/mL (0.48 mM; Need ultrasonic and warming)SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 3.2013 mL 16.0067 mL 32.0133 mL5 mM 0.6403 mL 3.2013 mL 6.4027 mL10 mM 0.3201 mL 1.6007 mL 3.2013 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)?/p>

3、 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?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(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過(guò)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: 2.17 mg/mL (6.95 mM); Clear sol

4、ution此案可獲得 2.17 mg/mL (6.95 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 21.7 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: 2.17 mg/mL (6.95 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.17 mg/mL (6.95

5、 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 21.7 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: 2.17 mg/mL (6.95 mM); Clear solution此案可獲得 2.17 mg/mL (6.95 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 21.7 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 Tofacitinib是JAK3/2/1 抑制劑，IC50 分別為1，20 和 112 nM。IC & Target JAK3 JAK2 JAK1 Rock-II1 nM (IC50) 20 nM (IC50) 112 nM (IC50) 3400 nM (IC50)Lck3870 nM (IC50)體外研究 Tofacitinib (CP-690550) citrate binds potentially at JAK3 and JAK2 as 2.2 nM and 5 nM (Kd). The report includesadditional binding

7、 for Tofacitinib at Camk1 (Kd of 5,000 nM), DCamkL3 (Kd of 4.5 nM), Mst2 (Kd of 4,300 nM), Pkn1 (Kd of 200 nM), Rps6ka2 (Kin.Dom.2-C-terminal) (Kd of 1,400 nM), Rps6ka6 (Kin.Dom.2-C-terminal) (Kd of 1,200 nM),Snark (Kd of 420 nM), Tnk1 (Kd of 640 nM) and Tyk2 (Kd of 620 nM)1.體內(nèi)研究 Animals that are tr

8、eated with Tofacitinib show a significantly lower production of anti-drug antibodies (ADAs)compare with PEG-treated control mice (for five weeks after initial immunization, p0.01, n=8). Moreover ADAsbecome detectable earliest on day 28. A difference of 1000- to 200-fold in titers to SS1P is apparent

9、 from days 21through 35, respectively. Compare to SS1P, mice injected with keyhole limpet hemocyanin (KLH) generate a morerapid antibody response. Yet, the administration of Tofacitinib reduces anti-KLH titers compare to controls (p0.05 onday 21, p4 hours3.PROTOCOLKinase Assay 1 Kinase activity is r

10、ecorded via a competition binding assay of selected kinases that are fused to a proprietary tag.Measurements of the amount of kinase binds to an immobilized, active-site directed ligand in the presence andabsence of the test compound (e.g., Tofacitinib) provide a % of DMSO control for binding of lig

11、and. Activitiesbetween 0 and 10 are selected for Kd determinations. Dendrogram representations are generated by an in-housevisualization tool designated PhyloChem1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Human CD4+ positive cells ar

12、e enriched from peripheral blood mononuclear cells obtained from a healthy donor bymagnetic separation (CD4+ MACS beads). CD4+ cells are activated for 3 days with plate bound anti-CD3 and antiCD28 antibodies (5 ug/mL each), and then expanded for another 4 days in the presence of IL-2 (50 U/mL). Cell

13、s arerested overnight in 1% RPMI, and pre-incubated with Tofacitinib or DMSO control for 1 hour at indicatedconcentrations (5 nM, 50 nM, 500 nM; DMSO concentration is equal in all preparations) and then activated with IL-2Page 2 of 3 www.MedChemE(1000 u/mL) or IL-12 (100 ng/mL) for 15 minutes. Cells

14、 (10106/condition) are lysed in 1% Triton-x lysis buffer andequal amounts of cell lysate are run in NuPage Bis-Tris gel (4-12% gradient). Proteins are transferred ontonitrocellulose membrane. Detection is done with indicated antibodies using Odyssey western blotting system1.MCE has not independently

15、 confirmed the accuracy of these methods. They are for reference only.Animal Mice2Administration 23 Female BALB/c mice (6-8 weeks old) are used. Mice receive Tofacitinib in PEG300 (100 mg/mL) or vehicle alone(PEG300) by osmotic pump infusion (Alzet Model 2004, 0.25 L/hour, 28 days). Four days prior

16、to immunization, miceare anesthetized and their dorsal surface is shaved. A one cm incision is made on the back to create a subcutaneouspocket and insert the pump. The incision site is closed with wound clips. Mice are injected weekly (i.p.) with SS1Precombinant immunotoxin (RIT; 5 g/mouse) beginnin

17、g on day 0; control mice received injections of saline alone.Every week before SS1P or vehicle immunization, 50 L of blood is drawn to obtain serum samples. Sera are stored at80C until analyzed.Rats3Adjuvant-induced arthritis (AIA) is induced in female Lewis rats. Rats are randomized according to hi

18、nd paw volumeand assigned to Tofacitinib or vehicle treatment regimens. Groups of 7-8 rats per treatment group, and normal naiverats (n=4 per group), are euthanized either 4 hours, 4 days, or 7 days after beginning treatment (days 16, 20, and 23after immunization, respectively). Tofacitinib is suspe

19、nded in 0.5% methylcellulose/0.025% Tween 20 for in vivostudies. Once-daily oral administration of vehicle or Tofacitinib (6.2 mg/kg) is initiated on day 16 followingimmunization and continued through day 23. Paw volumes are reassessed 4 and 7 days after the beginning oftreatment (days 20 and 23 aft

20、er immunization, respectively). For micro-computed tomography (micro-CT) imaging, aswell as tartrate-resistant acid phosphatase (TRAP) staining in paw tissue, AIA is induced in a separate cohort of Lewisrats.MCE has not independently confirmed the accuracy of these methods. They are for reference on

21、ly.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. Free Radic Biol Med. 2019 May 9. pii: S0891-5849(19)30555-6. J Immunol. 2013 Oct 1;191(7):3568-77. Inflamm Bowel Dis. 2020 Feb 11;26(3):407-422.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Jiang JK, et al. Examining the chirality, conformation and selective kinase inhibition of 3-(3R,4R)-4-methyl-3-(methyl(7H-pyrrolo2,3-dpyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile (CP-690,550). J Med Chem. 2008 Dec 25;51(24):8012-