4-Methylumbelliferyl-beta-D-glucuronide(4-MUG) yog cov organic compound nrog cov qauv molecular C16H13NO7 thiab molar mass ntawm 333.27 g / mol rau CAS 6160-80-1. Nws feem ntau tshwm li dawb mus rau lub teeb daj hmoov los yog siv lead ua. Nws muaj dej solubility zoo thiab tuaj yeem yaj hauv dej. Cov qauv molecular ntawm cov tshuaj no muaj ib pawg glucuronic acid (GlcA) thiab ib pawg 4-methylumbelliferone (4-MeUmb). Cov pab pawg glucuronic acid txuas nrog 4- pawg methylumbelliferone los ntawm glycosidic bonds. Cov yam ntxwv spectral. Piv txwv li, nyob rau hauv lub teeb ultraviolet, lub compound nthuav tawm muaj zog fluorescence thiab tuaj yeem siv rau nws qhov kev soj ntsuam ntau. Nws qhov siab tshaj plaws excitation wavelength thiab siab tshaj plaws emission wavelength yog 365nm thiab 448nm, feem. Nws yog cov kua qaub tsis muaj zog nrog qis pKa tus nqi hauv dej. Raws li lub substrate nrog cov khoom fluorescent, nws muaj peev xwm siv dav hauv kev tshawb nrhiav thiab kev tshuaj ntsuam enzyme. Nws lub hauv paus ntsiab lus yooj yim, kev ua haujlwm yooj yim, siab rhiab heev, thiab quantifiability ua rau nws yog ib qho cuab yeej tseem ceeb hauv ntau yam xws li biomedical, tshuaj, thiab biological sciences.
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Tshuaj Formula |
C16H16O9 |
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Exact Mass |
352 |
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Molecular Luj |
352 |
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m/z |
352 (100.0%), 353 (17.3%), 354 (1.8%), 354 (1.4%) |
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Elemental Analysis |
C, 54.55; H, 4.58; O, 40.87 |
4-Methylumbelliferyl-beta-D-glucuronide(4-MUG) yog cov fluorescent substrate dav siv rau kev tshawb nrhiav thiab kev tshuaj ntsuam enzyme.
1. Kev kuaj xyuas cov enzymes
Lub hauv paus ntsiab lus: Kev tshawb nrhiav enzyme kev ua haujlwm ntawm 4-MUG yog raws li nws cov yam ntxwv ntawm kev ua hydrolyzed los ntawm cov enzymes tshwj xeeb ua cov substrate. Thaum lub enzyme ua rau 4-MUG, nws cleaves nws glycosidic bonds, tso tawm 4-methylumbelliferone (4-MU) nrog fluorescent zog. Los ntawm kev txheeb xyuas qhov kev siv thiab kev hloov pauv ntawm cov teeb liab fluorescence, kev ua haujlwm ntawm cov enzymes tuaj yeem soj ntsuam.
Daim ntawv thov teb:
(1) Kev soj ntsuam lom neeg lom neeg: Siv 4-MUG ua lub substrate, kev ua haujlwm ntawm cov enzymes tshwj xeeb hauv cov qauv lom (xws li ntshav, zis, cov ntaub so ntswg homogenate, thiab lwm yam) tuaj yeem kuaj pom. Qhov no yog qhov tseem ceeb heev rau kev kuaj kab mob, kev tshawb fawb tshuaj metabolism, thiab kev tshawb pom biomarker.
(2) Enzyme inhibitor kev tshuaj ntsuam: Los ntawm kev sib koom ua ke 4-MUG nrog enzyme inhibitors, cov nyhuv ntawm enzyme inhibitors ntawm kev ua enzyme tuaj yeem pom. Txoj kev no tuaj yeem siv los tshuaj xyuas thiab ntsuas qhov ua tau zoo ntawm enzyme inhibitors, muab kev taw qhia rau kev tsim tshuaj thiab kev tsim kho.
(3) Kev tshawb fawb Enzyme kinetics: Los ntawm kev siv ntau qhov sib txawv ntawm 4-MUG li substrates, cov kinetic tsis ntawm enzyme, xws li Michaelis tas li (Km) thiab qhov siab tshaj plaws cov tshuaj tiv thaiv (Vmax), tuaj yeem kawm. Cov ntaub ntawv no pab kom nkag siab txog cov txheej txheem thiab kev ua haujlwm ntawm enzyme catalyzed cov tshuaj tiv thaiv.
2. Kev tshuaj ntsuam xyuas enzyme
(1) Kev tshuaj ntsuam xyuas dhau los: 4-MUG yog qhov tsim nyog rau kev tshuaj ntsuam xyuas cov enzyme siab tshaj plaws vim nws muaj cov khoom siv fluorescence thiab tuaj yeem ua tiav sai thiab tshawb pom. Los ntawm kev sib koom ua ke ntau ntawm cov neeg sib tw sib txuas nrog 4-MUG, cov khoom sib txuas nrog cov kev cai enzyme tuaj yeem kuaj xyuas.
(2) Enzyme inhibitor kev tshuaj ntsuam xyuas: Siv 4-MUG ua lub substrate, cov tebchaw nrog cov enzyme inhibitory tuaj yeem tshuaj xyuas. Txoj kev no tuaj yeem siv los tshawb pom cov enzyme inhibitors tshiab thiab muab cov neeg sib tw molecules rau kev tsim tshuaj thiab kev loj hlob.
(3) Enzyme activator screening: Ntxiv nrog rau cov enzyme inhibitors, 4-MUG tuaj yeem siv los tshuaj xyuas cov tebchaw nrog cov teebmeem enzyme. Los ntawm kev txheeb xyuas qhov kev txhim kho ntawm cov teeb liab fluorescence, cov tebchaw uas tuaj yeem txhim kho cov haujlwm enzyme tuaj yeem txheeb xyuas, muab txoj hauv kev tshiab rau kev tswj hwm kev ua haujlwm enzyme.
3. Biofluorescence labeling: 4-Methyllumbelliferol-beta-D-glucuronide muaj cov khoom fluorescence thiab feem ntau siv los ua biofluorescence marker. Nws muaj peev xwm emit xiav xiav fluorescence nyob rau hauv lub excitation ntawm ultraviolet lub teeb, thiab lub emission spectrum overlaps ho nrog lub excitation spectrum, uas yog zoo rau fluorescence nrhiav thiab tsom xam. Kev sojntsuam fluorescent tuaj yeem ua tau los ntawm kev khi 4-Methyllumbelliferol-beta-D-glucuronide nrog cov tshuaj tiv thaiv kab mob, cov protein, lossis lwm yam biomolecules, uas tuaj yeem siv rau kev kuaj pom thiab thaj chaw ntawm lub hom phiaj molecules hauv cov qauv tshuaj lom neeg.
4. Kev tshawb fawb txog tshuaj metabolism:4-Methylumbelliferyl-beta-D-glucuronidefeem ntau yog siv los ua tus qauv sib xyaw rau kev tshawb fawb tshuaj metabolism. Hauv kev tshawb fawb txog tshuaj metabolism, qhov sib xyaw no tuaj yeem simulate cov qauv ntawm cov metabolites ntuj thiab ua tus qauv rau cov khoom nruab nrab lossis cov khoom kawg uas yuav raug tsim tawm thaum cov tshuaj metabolism. Los ntawm kev kawm thiab qhia txog cov txheej txheem metabolic ntawm cov tshuaj hauv cov kab mob, peb tuaj yeem nkag siab tob txog lawv txoj hauv kev metabolic, kinetics, thiab bioavailability.
Laboratory synthesis txoj kev ntawm4-Methylumbelliferyl-beta-D-glucuronidesuav nrog cov kauj ruam hauv qab no:
(1) Kev taw qhia txog cov pab pawg tiv thaiv: Xaiv cov pab pawg tiv thaiv tsim nyog los tiv thaiv glucuronic acid thiab tsis txhob muaj kev cuam tshuam sab nraud hauv cov kev tshwm sim tom ntej. Cov pab pawg tiv thaiv muaj xws li ester pawg, ether pawg, thiab lwm yam. Xaiv methyl ester ua pab pawg tiv thaiv los hloov glucuronic acid rau methyl ester glucuronic acid. Cov tshuaj tiv thaiv no siv methanol thiab carboxylic anhydride ua reactants, nrog sulfuric acid ua catalyst.
Qhov sib npaug ntawm cov tshuaj tiv thaiv yog raws li nram no:
(CH3)2C(OH)-CH2-COOH + H2YOG4→ (CH3)2C(O)-CH2- COJ + CH3OH
(2) Coupling cov tshuaj tiv thaiv: Coupling cov tshuaj tiv thaiv ntawm 4-methylumbelliferone thiab glucuronic acid nrog pab pawg tiv thaiv. Cov kauj ruam no feem ntau yuav tsum tau siv cov khoom sib txuas xws li DCC, EDC, thiab lwm yam los pab txhawb cov tshuaj tiv thaiv. Cov tshuaj tiv thaiv yuav tsum tau tswj kom tsim nyog los xyuas kom meej qhov purity thiab tawm los ntawm cov khoom. Cov tshuaj tiv thaiv no siv DCC ua tus neeg sawv cev sib txuas,
(3) Qhov sib npaug ntawm cov tshuaj tiv thaiv yog raws li nram no:
(CH3)2C(O)-CH2-COOH + NH2-CH=CH-COOH + DCC + C6H5N3O → (CH3)2C(O)-CH2-CO-NH-CH=CH-COOH + H2O + NH4OH
(CH3)2C(O)-CH2-CO-NH-CH=CH-COOH + NH2(CH3) → (CH3)2C(O)-CH2-CO-NH-CH=CH-CO-NH(CH3) + H2O
(CH3)2C(O)-CH2-CO-NH-CH=CH-CO-NH(CH3) + HBr → (CH3)2C(O)-CH2-CO-NH-CH=CH-CO-Br(CH3) + HBr
(CH3)2C(O)-CH2-CO-NH-CH=CH-CO-Br(CH3) → (CH3)2C(O)-CH2-CO[C16H8-(NH-)-6]-Br(CH3) + NaCl + NaBr
(3) Kev tshem tawm cov pab pawg tiv thaiv: Los ntawm kev siv cov sib tov ntawm sodium methanol thiab sodium ethanol ua cov catalyst, methyl glucuronic acid methyl ester raug tshem tawm ntawm 4-Methyllumbelliferyl beta D-glucuronide. Qhov sib npaug ntawm cov tshuaj tiv thaiv yog raws li nram no:
(CH3)2C(O)-CH2-CO[C16H8-(NH-)-6]-Br(CH3) + UA3ONa + C2H5UA → (CH3)2C(O)-CH2-CO[C16H8-(NH-)-6] + 2NaBr + 2CH3OH + C2H5OH
(4) Purification: Cov khoom nyoos yog purified los ntawm kem chromatography thiab recrystallization kom tau high-purity 4-Methyllumbelliferol-beta-D-glucuronide. Cov txheej txheem tshwj xeeb yog raws li hauv qab no:
Dissolve cov khoom crude nyob rau hauv ib tug tsim nyog tus nqi ntawm cov kuab tshuaj thiab cais nws los ntawm kem chromatography. Xaiv cov adsorbents tsim nyog thiab eluents, ntxuav tawm impurities thiab unreacted raw cov ntaub ntawv nyob rau hauv ib theem kom tau cov khoom ntshiab. Recrystallization yog cov txheej txheem ntawm kev ua kom cov khoom ntshiab hauv qhov tsim nyog ntawm cov kuab tshuaj thiab tau txais cov khoom ntshiab los ntawm kev ua kom txias thiab crystallization. Recrystallization tuaj yeem tshem tawm me me ntawm cov khoom siv raw thiab cov khoom siv los ntawm cov khoom tsis zoo, txhim kho qhov purity ntawm cov khoom.
Cim npe nrov: 4-methylumbelliferyl-beta-d-glucuronide cas 6160-80-1, lwm tus neeg, manufacturers, hoobkas, lag luam wholesale, yuav, nqe, tej, kev muag khoom