Hydrochloric Acid → Coenzyme A

HRID 1694334

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HRID 1694334 반응식
C[C@@](O)(CC([S:10][CH2:11][CH2:12][NH:13][C:14]([CH2:16][CH2:17][NH:18][C:19]([C@H:21]([OH:57])[C:22]([CH2:25][O:26][P:27]([O:30][P:31]([O:34][CH2:35][C@H:36]1[O:40][C@@H:39]([N:41]2[C:45]3[N:46]=[CH:47][N:48]=[C:49]([NH2:50])[C:44]=3[N:43]=[CH:42]2)[C@H:38]([OH:51])[C@@H:37]1[O:52][P:53]([OH:56])([OH:55])=[O:54])([OH:33])=[O:32])([OH:29])=[O:28])([CH3:24])[CH3:23])=[O:20])=[O:15])=O)CC(O)=O.C(O)C(N)(CO)CO.Cl.[Mg+2].[Cl-].[Cl-].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.SC[C@H]([C@@H](CS)O)O.C(SCCNC(=O)CCNC(=O)[C@H](O)C(C)(C)COP(O)(=O)OP(O)(=O)OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1OP(O)(O)=O)(=O)CC(C)=O.Cl>CS(C)=O>[CH3:24][C:22]([C@@H:21]([OH:57])[C:19]([NH:18][CH2:17][CH2:16][C:14]([NH:13][CH2:12][CH2:11][SH:10])=[O:15])=[O:20])([CH2:25][O:26][P:27]([O:30][P:31]([O:34][CH2:35][C@H:36]1[O:40][C@@H:39]([N:41]2[C:45]3[N:46]=[CH:47][N:48]=[C:49]([NH2:50])[C:44]=3[N:43]=[CH:42]2)[C@H:38]([OH:51])[C@@H:37]1[O:52][P:53]([OH:56])([OH:55])=[O:54])([OH:33])=[O:32])([OH:29])=[O:28])[CH3:23]

참가 물질

9

반응물 7

HCID 313Hydrochloric Acid2회HCID 6049Ethylenediaminetetraacetic AcidHCID 92153acetoacetyl-CoAHCID 93573Tromethamine hydrochlorideHCID 445127HMG-CoAHCID 446094D-1,4-DithiothreitolHCID 5360315Magnesium Chloride

생성물 1

HCID 87642Coenzyme A

시약·촉매·용매 1

HCID 679Dimethyl Sulfoxide용매

실험 절차

Intrinsic activity of HMG-CoA synthase is measured in the following in vitro assay. Enzyme protein (ca. 12.2 μg) is added to a solution containing 117 mM Tris-HCl (pH 8.0), 11.7 mM MgCl2, 1.17 mM Ethylenediaminetetraacetic acid (EDTA), 0.58 mM dithiothreitol, in the presence or absence of the test compound (added as, e.g., a 2 μg/ml solution in dimethylsulfoxide). The incubation is in a volume of 0.085 ml at 30° C. in a shaking water bath. After 5 minutes, 15 μl of a solution containing acetoacetyl-CoA and 0.1 μCi of 1[14C]-acetyl-CoA is added to give final concentrations of 0.1 and 0.4 mM, respectively. The incubation is continued for 2 more minutes and the reaction stopped by the addition of 50 μl of the assay mixture to 0.2 ml of 6N HCl in a glass scintillation vial. The vial is heated for 1 hour at 110° C. after which time 0.2 ml more of 6N HCl is again added to each vial and the heating continued for another hour. Following this, 1.0 ml of 0.9% saline is added to each vial and finally 10 ml of scintillation liquid. Radioactivity is determined in a Packard Tri-Carb liquid scintillation counter. Percent inhibition is calculated a standard formula. IC50 values are determined by plotting the log of the concentration of the test compound verses the percentage inhibition and fitting a straight line to the resulting data by using the least squares method.

후처리

  1. additionEnzyme protein (ca. 12.2 μg) is added to a solution
  2. additionin the presence or absence of the test compound (added as, e.g.
  3. customThe incubation is in a volume of 0.085 ml at 30° C. in a shaking water bath
  4. additionis added
  5. customto give final concentrations of 0.1 and 0.4 mM
  6. additionis again added to each vial and the heating
  7. waitcontinued for another hour
  8. additionis added to each vial and finally 10 ml of scintillation liquid

출처 및 근거

ORD 기록
ord-b2f0cae6b7df438cb6552561aa3dc895
출처 데이터셋
uspto-grants-2012_04
특허 번호
US08158677B2

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