NADPH → Tetrahydrofolic acid

HRID 742297Notes

Reaction equation

HRID 742297 reaction equation
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]

Participants

5

Reactant 4

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

Product 1

HCID 135444742Tetrahydrofolic acid

Conditions

Temperature
37 °C

Procedure

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 mi. 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.

Workup

  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

Sources & evidence

ORD record
ord-0f75588ec29b41278fbda6d9d8f8d378
Source dataset
uspto-grants-1996_09
Patent number
US05554615

Reaction notes

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