Shevchenko K V, Nagaev I Yu, Andreeva L A, Shevchenko V P, Myasoedov N F
Institute of Molecular Genetics of the Russian Academy of Sciences, Moscow, Russia.
Biomed Khim. 2019 Apr;65(3):180-201. doi: 10.18097/PBMC20196503180.
New data on peptide drugs have been summarized; their high stability is due to both the introduction of Pro-Gly-Pro in various amino acid sequences and the modification of the glyproline fragment itself. Pro-Gly-Pro-Leu, ACTH(6-9)Pro-Gly-Pro, 5-oxo-Pro-Arg-Pro and 5-oxo-Pro-His-Pro-NH2 were used as proline-containing peptides. Tritiated peptides were obtained: Pro-Gly-Pro-Leu with specific radioactivity of 135 Ci/mmol, ACTH(6-9)Pro-Gly-Pro - 26 Ci/mmol, 5-oxo-Pro-Arg-Pro - 60 Ci/mmol and 5-oxo-Pro-His-Pro-NH2 - 75 Ci/mmol. The concentration of Pro-Gly-Pro-Leu, ACTH(6-9)Pro-Gly-Pro, 5-oxo-Pro-Arg-Pro and 5-oxo-Pro-His-Pro-NH2 in the blood was found to be about 200 times more than in the brain for intranasal administration, and in average 600 times more for intravenous administration. The stability of proline-containing peptides in vitro experiments was determined using different commercially available peptidases (leucine aminopeptidases, dipeptidases, carboxypeptidases B and Y), and using nasal mucus, microsomal fraction of the rat brain (IMPC) and rat blood plasma. During peptidase hydrolysis of Pro-Gly-Pro-Leu, the main metabolites were Gly-Pro-Leu, Pro-Gly-Pro, Gly-Pro and Pro-Gly. For ACTH(6-9)Pro-Gly-Pro, the main metabolites were Phe-Arg-Trp-Pro-Gly-Pro and Trp-Pro-Gly-Pro. In peptidase hydrolysis of 5-oxo-Pro-His-Pro-NH2, the major metabolite was 5-oxo-Pro-His-Pro. It was shown that with different methods of peptides administration the composition of the metabolites formed is different. Based on the data obtained, resistance to enzymatic cleavage of peptides and their metabolic pathways were evaluated. Thus, these new data have shown that the above approaches can be used to prolong the action of glyprolines in living objects. In this case, the degradation of proline-containing peptides occurs mainly not due to the action of proteases, but due to other ways of degradation. In general, the data presented in the review indicate the promise of intranasal way of introducing biologically active peptides into the brain of living organisms.
已总结了肽类药物的新数据；它们的高稳定性归因于在各种氨基酸序列中引入了脯氨酸 - 甘氨酸 - 脯氨酸（Pro - Gly - Pro）以及对甘脯氨酸片段本身的修饰。脯氨酸 - 甘氨酸 - 脯氨酸 - 亮氨酸（Pro - Gly - Pro - Leu）、促肾上腺皮质激素（6 - 9）脯氨酸 - 甘氨酸 - 脯氨酸（ACTH(6 - 9)Pro - Gly - Pro）、5 - 氧代脯氨酸 - 精氨酸 - 脯氨酸（5 - oxo - Pro - Arg - Pro）和5 - 氧代脯氨酸 - 组氨酸 - 脯氨酸 - 氨基（5 - oxo - Pro - His - Pro - NH2）被用作含脯氨酸的肽。获得了氚标记的肽：脯氨酸 - 甘氨酸 - 脯氨酸 - 亮氨酸的比放射性为135居里/毫摩尔，促肾上腺皮质激素（6 - 9）脯氨酸 - 甘氨酸 - 脯氨酸为26居里/毫摩尔，5 - 氧代脯氨酸 - 精氨酸 - 脯氨酸为60居里/毫摩尔，5 - 氧代脯氨酸 - 组氨酸 - 脯氨酸 - 氨基为75居里/毫摩尔。发现经鼻给药时，脯氨酸 - 甘氨酸 - 脯氨酸 - 亮氨酸、促肾上腺皮质激素（6 - 9）脯氨酸 - 甘氨酸 - 脯氨酸、5 - 氧代脯氨酸 - 精氨酸 - 脯氨酸和5 - 氧代脯氨酸 - 组氨酸 - 脯氨酸 - 氨基在血液中的浓度比在脑中高约200倍，静脉给药时平均高600倍。在体外实验中，使用不同的市售肽酶（亮氨酸氨肽酶、二肽酶、羧肽酶B和Y）以及鼻黏液、大鼠脑微粒体部分（IMPC）和大鼠血浆来测定含脯氨酸肽的稳定性。在脯氨酸 - 甘氨酸 - 脯氨酸 - 亮氨酸的肽酶水解过程中，主要代谢产物是甘氨酸 - 脯氨酸 - 亮氨酸、脯氨酸 - 甘氨酸 - 脯氨酸、甘氨酸 - 脯氨酸和脯氨酸 - 甘氨酸。对于促肾上腺皮质激素（6 - 9）脯氨酸 - 甘氨酸 - 脯氨酸，主要代谢产物是苯丙氨酸 - 精氨酸 - 色氨酸 - 脯氨酸 - 甘氨酸 - 脯氨酸和色氨酸 - 脯氨酸 - 甘氨酸 - 脯氨酸。在5 - 氧代脯氨酸 - 组氨酸 - 脯氨酸 - 氨基的肽酶水解过程中，主要代谢产物是5 - 氧代脯氨酸 - 组氨酸 - 脯氨酸。结果表明，采用不同的肽给药方法时，形成的代谢产物组成不同。基于所获得的数据，评估了肽对酶解的抗性及其代谢途径。因此，这些新数据表明上述方法可用于延长甘脯氨酸在生物体内的作用。在这种情况下，含脯氨酸肽的降解主要不是由于蛋白酶的作用，而是由于其他降解方式。总体而言，综述中呈现的数据表明经鼻将生物活性肽引入生物体脑内具有前景。
