Tripeptides with angiotensin converting enzyme inhibition activity and their use and composition
An angiotensin and activity-inhibiting technology, which is applied in the field of functional foods and pharmaceutical compositions, can solve problems such as poor patient compliance, and achieve the effect of inhibiting ACE activity
Image
Examples
preparation example Construction
[0017] The preparation method of the tripeptide provided by the present invention includes but not limited to: separating the polypeptide from the hydrolyzate by hydrolyzing the milk protein; or adopting a chemical synthesis method. The chemical synthesis method has unique advantages in the synthesis of small molecular polypeptides, and can quickly synthesize a high-purity single sequence in a short period of time, so the chemical synthesis method is preferred.
[0018] The chemical synthesis method can be tBoc solid-phase synthesis method or Fmoc solid-phase synthesis method, preferably Fmoc solid-phase synthesis method. The present invention can utilize the principle, steps and operating conditions of the Fmoc solid-phase synthesis method described in detail in Merrifield B. Solid phase synthesis [J]. Science, 1986, 232 (4748): 341-347.
[0019] The present invention also provides compositions containing at least one tripeptide described in the present invention as an active...
Embodiment 1
[0029] This example is used to illustrate the preparation process of the tripeptide of the present invention.
[0030] (1) Fmoc solid-phase synthesis process
[0031] Referring to the principle, steps and operating conditions of the Fmoc solid-phase synthesis method described in detail in Merrifield B. Solid phase synthesis [J]. Science, 1986, 232 (4748): 341-347, synthesize TDY, FKI, LSK, and LKY in sequence , IKF, IPA, IKY, LSF, LKP, WYY, RVY, IKQ or MMA.
[0032] (2) Purification of tripeptide
[0033] The tripeptide synthesized in step (1) was purified by semi-preparative reversed phase high performance liquid chromatography (RP-HPLC) under the following conditions. A Shimadzu (LC-10A) liquid chromatograph was used with BECKMAN C18 preparative column, mobile phase A: 0.1% trifluoroacetic acid (TFA) / ultrapure water; mobile phase B: 0.1% TFA / acetonitrile. Gradient program: 5%-50% B for 30min, flow rate 5mL / min, sample volume 3mL, use 230nm wavelength to detect and collect...
Embodiment 2
[0040] This example is used to determine the ACE inhibitory activity of the tripeptide prepared in Example 1.
[0041]Referring to the assay method of ACE inhibitory activity established by Wu et al. in 2002 (Wu J P, Aluko R E, Muir A D. Improved method for direct high-performance liquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions[J].Journal of Chromatography A, 2002, 950: 125-130.). Take 130 μL of 5 mmol / L HHL solution, add 20 μL of 20 mmol / L tripeptide, mix well, and keep warm in a constant temperature water bath at 37°C for 10 min. Then 10 μL of ACE enzyme solution was added, reacted in a constant temperature water bath at 37° C. for 30 min, and then 150 μL of 1 mol / L HCl was added to terminate the reaction to obtain a reaction solution. The results were analyzed by HPLC system. At the same time, 20 μL of 0.1mol / L phosphate buffer was used instead of tripeptide as a control group. The formula for calculating ACE inhibitory activity is as fol...
PUM
| Property | Measurement | Unit |
|---|---|---|
| control rate | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- IPC
- C07K5/083; C07K5/087; C07K5/097; C07K5/09; A23L1/305; A61K38/06; A61P9/12
- Inventors
- 姜鹭; 任发政