NADPH → Tetrahydrofolic acid

HRID 2294088ノート

反応式

HRID 2294088 の反応式
P([O-])([O-])([O-])=O.[K+].[K+].[K+].SC(O)C.C1N=C(N)C2N=CN([C@@H]3O[C@H](COP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H](O)[C@H]3OP(O)(O)=O)C=2N=1.[C:61]([OH:92])(=[O:91])[CH2:62][CH2:63][C@H:64]([NH:68][C:69]([C:71]1[CH:90]=[CH:89][C:74]([NH:75][CH2:76][C:77]2[CH2:78][NH:79][C:80]3[N:81]=[C:82]([NH:84][C:85]([C:87]=3[N:88]=2)=[O:86])[NH2:83])=[CH:73][CH:72]=1)=[O:70])[C:65]([OH:67])=[O:66]>>[C:61]([OH:92])(=[O:91])[CH2:62][CH2:63][C@H:64]([NH:68][C:69]([C:71]1[CH:72]=[CH:73][C:74]([NH:75][CH2:76][CH:77]2[NH:88][C:87]3[C:85](=[O:86])[NH:84][C:82]([NH2:83])=[N:81][C:80]=3[NH:79][CH2:78]2)=[CH:89][CH:90]=1)=[O:70])[C:65]([OH:67])=[O:66]

参加物

5

反応物 4

HCID 5884NADPHHCID 62657Potassium Phosphate, TribasicHCID 4713212Mercapto-ethanolHCID 135398604Dihydrofolic Acid

生成物 1

HCID 135444742Tetrahydrofolic acid

反応条件

温度
37 °C

実験手順

A solution of purified DHFR was added to a solution comprising a potassium phosphate buffer solution (pH: 7.5, 75 mM), mercaptoethanol (7.5 mM) and NADPH (0.25 mM) to make up to a total amount of 0.57 ml. Each of the serial dilutions of the compound according to the present invention in an amount of 0.015 ml was added to the mixture prepared above. The mixtures thus prepared were preliminarily heated at 37° C. for 5 minutes, followed by the addition of 0.015 ml of a 25 μM solution of dihydrofolic acid. The obtained mixtures were kept at 37° C. for 5 minutes to conduct a reaction. The resulting mixtures were each examined for the decrease in the absorbency at 339 nm per minute. The amount of tetrahydrofolic acid formed was determined on the basis of the decrease and compared with that of a control solution (containing none of the compounds according to the present invention) to calculate the 50% DHFR-inhibitory concentration (IC50). The results are given in Tables 1-A and 1-B.

後処理

  1. additionA solution of purified DHFR was added to a solution
  2. additionwas added to the mixture
  3. customprepared above
  4. customThe mixtures thus prepared
  5. customa reaction

出典と根拠

ORDレコード
ord-dd241ba94c854b938c6de6733db9ef67
出典データセット
uspto-grants-1997_11
特許番号
US05688800

反応ノート

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