二氢叶酸还原酶（DHFR）在氨基酸和叶酸的生物合成中扮演了重要角色。在烟酰胺二核苷酸磷酸辅助因子的存在下，它通过将二氢叶酸还原为四氢叶酸来参与其中，并已通过多种临床研究验证使用DHFR作为治疗癌症和各种细菌感染的靶点。在本综述中，我们揭示了2001年以来具有二氨基嘧啶和喹噁啉核的合成和天然DHFR抑制剂的专利。此外，本综述还强调了过去五年中许多DHFR抑制剂的临床进展。从2001年到2021年，许多活性化学支架已经被引入，并被暴露为进入临床试验的有力DHFR抑制剂的领先候选者。此外，研究人员也极为关注开发一类具有更高选择性和效力的DHFR抑制剂。这一发展包括合成具有效力的合成和天然化合物作为DHFR抑制剂。根据文献综述，我们可以预期未来会出现大量新颖的活性分子，具有更卓越的药理学特性来针对这个酶。

Dihydrofolate reductase (DHFR) plays an important role in the biosynthesis of amino acid and folic acid. It participates by reducing dihydrofolate to tetrahydrofolate, in the presence of nicotinamide dinucleotide phosphate cofactor, and has been verified by various clinical studies to use DHFR as a target for the treatment of cancer and various bacterial infections.In this review, we have disclosed patents of synthetics and natural DHFR inhibitors with diaminopyrimidine and quinazoline nucleus from 2001. Additionally, this review highlights the clinical progression of numerous DHFR inhibitors received from the last five years.From 2001 to 2021, numerous active chemical scaffolds have been introduced and are exposed as lead candidates that have entered clinical trials as potent DHFR inhibitors. Moreover, researchers have paid considerable attention to the development of a new class of DHFR inhibitors with higher selectivity and potency. This development includes synthesis of synthetic as well as natural compounds that are potent DHFR inhibitors. On the basis of literature review, we can anticipate that there are a huge number of novel active molecules available for the future that could possess superior abilities to target this enzyme with a profound pharmacological profile.