Pemetrexed disodium hemipenta hydrate
作者：齊岳
發(fā)布時(shí)間：2023-09-11
09:24:14
點(diǎn)擊數(shù)：
產(chǎn)品名稱：Pemetrexed disodium hemipenta hydrate
產(chǎn)品描述：
| 產(chǎn)品描述 | Pemetrexed Disodium Hydrate is a new-type antifolate and antimetabolite for TS, DHFR, and GARFT. The Ki of Pemetrexed Disodium Hydrate for TS, DHFR and GARFT is 1.3 nM, 7.2 nM and 65 nM, respectively. |
| 靶點(diǎn)活性 | TS:1.3 nM(Ki), DHFR:7.2 nM(Ki), GARFT:65 nM(Ki) |
| 體外活性 | 在人H460非小細(xì)胞肺*異種移植模型中,Pemetrexed disodium能夠抑制生長. |
| 體內(nèi)活性 | \在CCRF-CEM白血病,GC3/C1結(jié)腸*,和HCT-8回盲部結(jié)腸*細(xì)胞中,Pemetrexed disodiumIC50分別為25 nM,34 nM和220 nM,表現(xiàn)出抗活性。 |
| 激酶實(shí)驗(yàn) | Enzyme Assays and Methods.: TS activity is assayed using a spectrophotometric method,which involved monitoring the increase in absorbance at 340 nm resulting from formation of the product,7,8-dihydrofolate.The assay buffer contains 50 mM N-tris[hydroxymethyljmethyl-2-aminoethanesulfonic acid,25 mM MgC12,6.5 mM formaldehyde,1 mM EDTA,and 75 mM 2-mercaptoethanol,pH 7.4.The concentrations of deoxyuridylate monophosphate,6R-MTHF,and hIS are 100 μM,30 μM and 30 nM (1.7 milliunits/mL),respectively.At the 6R-MTHF concentration,an uninhibited reaction and six concentrations of inhibitor are assayed.Ki app values are determined by fitting the data to the Morrison equation using nonlinear regression analysis with the aid of the program ENZFITTER.Ki values are calculated using the equation: Ki app= Ki(1 + [S]/Km),where [S] is equal to 30 μM and Km is equal to 3 μM.DHFR activity is assayed spectrophotometrically by monitoring the dis appearance of the substrates NADPH and 7,8-dihydrofolate at 340 nm.The reaction takes place at 25°C in 0.5 mL of 50 mM potassium phosphate buffer,which contains 150 mM KC1 and 10 nM 2-mercaptoethanol,pH 7.5,and 14 nM (0.34 milliunitlmL) DHFR.The NADPH concentration is 10 μM and 7,8-dihydrofolate is varied at 5,10,or 15 μM.At each 7,8-dihydrofolate concentration,an uninhibited reaction and seven concentrations of inhibitor are assayed.The ENZFITI'ER microcomputer program is used to obtain Ki app values by fitting the data to the Morrison equation by nonlinear regression analysis.Ki app= Ki(1 + [S]/Km),where [S] is equal to the concentration of 7,8-dihydrofolate used and Km of 7,8-dihydrofolate is equal to 0.15 μM.GARFT activity is assayed spectrophotometrically by monitoring the increase of absorbance resulting from formation of the product 5,8-dideazafolate at 295 nm.The reaction solvent contains 75 mM HEPES,20% glycerol,and 50 mM a-thioglygerol,pH 7.5,at 25°C. |
| 細(xì)胞實(shí)驗(yàn) | Dose-response curves are generated to determine the concentration required for 50% inhibition of growth (IC50). Pemetrexed disodium is dissolved initially in DMSO at a concentration of 4 mg/mL and further diluted with cell culture medium to the desired concentration. CCRF-CEM leukemia cells in complete medium are added to 24-well Cluster plates in a total volume of 2.0 mL. Pemetrexed disodium at various concentrations are added to duplicate wells so that the final volume of DMSO is 0.5%. The plates are incubated for 72 hour at 37 °C in an atmosphere of 5% CO2 in air. At the end of the incubation, cell numbers are determined on a ZBI Coulter counter. For several studies, IC50s are determined for each compound in the presence of either 300 μM AICA, 5 μM thymidine, 100 μM hypoxanthine, or combination of 5 μM hymidine plus 100 μM hypoxanthine. For adherent tumor cells, a modification of the original MTT colorimetric assay is used to measure cell cytotoxicity. The human tumor cells are seeded in 100 μL assay medium/well in 96-well flat-bottomed tissue culture plates. The assay medium contains folic acid-free RPMI 1640 supplemented with 10% FCS and either 2 nM folinic acid or 2.3 μM folic acid as the sole folate source. Well 1A is left blank. Stock solutions of antifolates are prepared in Dulbecco's PBS at 1 mg/mL, and a series of 2-fold dilutions are subsequently made in PBS. Ten-μL aliquots of each concentration are added to triplicate wells. Plates are incubated for 72 hours at 37 °C in a humidified atmosphere of 5% CO2-in-air. MTT is dissolved in PBS at 5 mg/mL, 10 μL of stock MTF solution are added to each well of an assay, and the plates are incubated at 37 °C for 2 additional hours. Following incubation, 100 μL of DMSO are added to each well. After thorough formazan solubilization, the plates are read on a Dynatech MR600 reader, using a test wavelength of 570 nm and a reference wavelength of 630 nm.The IC50 is determined as the concentration of drug required to inhibit cell growth by 50% compared to an untreated controls.(Only for Reference) |
| 別名 | 培美曲塞二鈉一水合物, Pemetrexed Disodium Hydrate, Pemetrexed sodium hydrate, LY-231514 Disodium Hydrate, 培美曲塞二鈉水合物 |
| 分子量 | 518.43 |
| 分子式 | C20H21N5O6·2Na·5/2H2O |
| CAS No. | 357166-30-4 |
存儲(chǔ)
Powder: -20°C for 3 years | In solvent: -80°C for 2 years
溶解度
DMSO: insoluble
( < 1 mg/mL refers to the product slightly soluble or insoluble )
聯(lián)系我們：
郵箱：2519696869@qq.com
QQ: 2519696869
電話：18066853083
微信：18066853083
公司介紹：
西安齊岳生物科技有限公司是集化學(xué)科研和定制與一體的高科技化學(xué)公司。業(yè)務(wù)范圍包括化學(xué)試劑和產(chǎn)品的研發(fā)、生產(chǎn)、銷售等。涉及產(chǎn)品為通用試劑的分銷、非通用試劑的定制與研發(fā)，涵蓋生物科技、化學(xué)品、中間體和化工材料等領(lǐng)域。
主營產(chǎn)品：COF、MOF單體系列：三蝶烯衍生物、金剛烷衍生物、四苯甲烷衍生物、peg、上轉(zhuǎn)換、石墨烯、光電材料、點(diǎn)擊化學(xué)、凝集素、載玻片、蛋白質(zhì)交聯(lián)劑、脂質(zhì)體、蛋白、多肽、氨基酸、糖化學(xué)等。
