Yuan M M, Wang M, Liu J Y, Zhang Z L
Department of Chemical Biology,Peking University School of Pharmaceutical Sciences, Beijing 100191, China.
Department of Chemical Biology,Peking University School of Pharmaceutical Sciences, Beijing 100191, China; Peking University State Key Laboratory of Natural and Biomimetic Drugs, Beijing 100191, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2017 Aug 18;49(4):714-718.
To optimize and establish the best hydrolysis method of diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydrofolate through the optimization of simple compound of diethyl N-(4-aminobenzoyl)-L-glutamate.
To increase the low yield of hydrolysis reaction of diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydrofolate due to the by-products and difficult purification, we studied the effect of NaOH and KOH, two kinds of alkalis, three concentrations between 0.175-1 mol/L and five types of reaction time involved in 20, 30, 60, 120 and 180 min on the common side chain diethyl N-(4-aminobenzoyl)-L-glutamate. A high performance liquid chromatography was established for measuring the target product and the by-products in reaction liquid in different reaction conditions. Finally, on the basis of the best hydrolysis method of diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydrofolate, we completed the optimization of the hydrolysis reaction conditions of diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydrofolate.
We developed the best reaction condition for the hydrolysis of diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydrofolate, which could be carried out easily and efficiently. The results indicated that treated with the optimized condition of 0.3 mol/L KOH in 60 min at the room temperature, diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydrofolate was converted into its diacid derivative in 95.6 % yield, which turned to be a better reaction condition compared with the previous reaction condition. The structures of those compounds were identified to be correct by 1H nuclear magnetic resonance(1H NMR), 13C nuclear magnetic resonance(13C NMR) and electrospray ionization time of flight mass spectrometry (ESI-MS). The purity of the diacid derivative of the compound was determined to be 96% by high performance liquid chromatography(HPLC).The new hydrolysis reaction condition could not only avoid the formation of single ester hydrolysis product and amide bond hydrolysis product, but also improve the yield of the hydrolysis reaction.
We have developed an efficient reaction for the hydrolysis of diethyl ester 4-amino-N-formyl-N, N-dideazatetrahydro. Since the final step of the synthesis of classical folic acid antagonists is always the catalyzed hydrolysis of the side chain glutamate, the reaction also has great significance for anti-folic acid anti-tumor inhibitors synthesis.
通过对N-（4-氨基苯甲酰基）-L-谷氨酸二乙酯简单化合物的优化，优化并建立4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯的最佳水解方法。
针对4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯水解反应产率低、副产物多、纯化困难的问题，研究了0.175 - 1mol/L三种浓度的NaOH和KOH两种碱以及20、30、60、120和180分钟五种反应时间对常见侧链N-（4-氨基苯甲酰基）-L-谷氨酸二乙酯的影响。建立了高效液相色谱法用于测定不同反应条件下反应液中的目标产物和副产物。最后，在4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯最佳水解方法的基础上，完成了4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯水解反应条件的优化。
开发出了4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯水解的最佳反应条件，该条件易于操作且高效。结果表明，在室温下用0.3mol/L KOH处理60分钟的优化条件下，4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯转化为其二酸衍生物的产率为95.6%，与先前的反应条件相比，这是一个更好的反应条件。通过1H核磁共振（1H NMR）、13C核磁共振（13C NMR）和电喷雾电离飞行时间质谱（ESI-MS）确定这些化合物的结构正确。通过高效液相色谱（HPLC）测定该化合物二酸衍生物的纯度为96%。新的水解反应条件不仅避免了单酯水解产物和酰胺键水解产物的形成，还提高了水解反应的产率。
开发了一种高效的4-氨基-N-甲酰基-N，N-二去氮四氢叶酸二乙酯水解反应。由于经典叶酸拮抗剂合成的最后一步总是侧链谷氨酸的催化水解，该反应对抗叶酸抗肿瘤抑制剂的合成也具有重要意义。
