Acetonitrile → Methotrexate

HRID 1032025

Reaction equation

HRID 1032025 reaction equation
N[C:2]1NC(=O)C2C(=NC=CN=2)N=1.[NH2:13][C@H:14]([C:20]([O-:22])=[O:21])[CH2:15][CH2:16][C:17]([O-:19])=[O:18].CC#N.[CH:26]1[C:31]([C:32]([OH:34])=O)=[CH:30][CH:29]=[C:28]([NH:35][CH2:36][C:37]2[N:42]=[C:41]3[C:43]([NH2:48])=[N:44][C:45]([NH2:47])=[N:46][C:40]3=[N:39][CH:38]=2)[CH:27]=1>CC([O-])=O.[Na+].[Cl-].[Cl-].[Zn+2]>[CH3:2][N:35]([C:28]1[CH:29]=[CH:30][C:31]([C:32]([NH:13][C@H:14]([C:20]([OH:22])=[O:21])[CH2:15][CH2:16][C:17]([OH:19])=[O:18])=[O:34])=[CH:26][CH:27]=1)[CH2:36][C:37]1[CH:38]=[N:39][C:40]2[N:46]=[C:45]([NH2:47])[N:44]=[C:43]([NH2:48])[C:41]=2[N:42]=1

Participants

7

Reactant 4

HCID 6342AcetonitrileHCID 1697484-(N-((2,4-diamino-6-pteridinyl)methyl)amino)benzoic acidHCID 5460544L-glutamate(2-)HCID 135398660Pterin

Product 1

HCID 126941Methotrexate

Reagents, catalysts & solvents 2

HCID 3007855ZincchlorideCatalystHCID 517045Sodium AcetateSolvent

Procedure

Small samples (1 mg) of the end products of Examples 1 and 2 were treated at room temperature in 1M NaOAc (10 mL) containing ZnCl2 (10 mg) with freshly thawed carboxypeptidase G1 enzyme solution (0.3 μL, 4500 U/mL). In less than 15 min, glutamate hydrolysis was nearly complete according to HPLC analysis (20% MeCN 0.1M NH4OAc, pH 7.5), which showed the disappearance of >95% of the starting material (Example 1, 2.5 min; Example 2, 2.89 min) and the appearance of new peaks (3.0 and 3.3 min) assumed to be the 2-desamino and 2-desamino-2-methyl derivates of 4-amino-4-deoxypteroic acid, respectively. Under identical conditions, clinical grade methotrexate (mainly the L-form, 3.0 min) was cleaved to 4-amino-4-deoxy-N10 -methylpteroic acid (4.0 min), whereas D MTX (4.0 min, pre-formed from clinical grade MTX by carboxypeptidase G1 treatment) was resistant tc further treatment with the enzyme. Since the enzyme carboxypeptidase G1 is well known to cleave L-MTX to 4-amino-4-deoxy-N10 -methylpteroic acid under conditions that leave D-MTX unaffected, these results show that my desamino analogues are essentially pure L-enantiomers.

Workup

  1. waitIn less than 15 min
  2. customthe appearance of new peaks (3.0 and 3.3 min)

Sources & evidence

ORD record
ord-7cd562cd343f4b4f99ff414b94e9ceeb
Source dataset
uspto-grants-1990_09
Patent number
US04956461

Linked notes

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