Ace-inhibitory peptide yvll derivatives and uses thereof
By acetylation or formylation of the ACE inhibitory peptide YVLL, ACE inhibitory peptide YVLL derivatives are formed, which significantly improves the ACE inhibitory activity and blood pressure lowering effect, solving the problem of the insignificant effect of the ACE inhibitory peptide YVLL in the prior art.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- FISHERIES RESEARCH INSTITURE OF FUJIAN
- Filing Date
- 2023-08-03
- Publication Date
- 2026-05-26
AI Technical Summary
The existing ACE inhibitory peptide YVLL has insignificant ACE inhibitory activity and blood pressure lowering effect, and there is a need to develop more efficient ACE inhibitory peptide derivatives.
By acetylation or formylation of the ACE inhibitory peptide YVLL, derivatives of the ACE inhibitory peptide YVLL are formed, preferably with the chemical formulas C28H44N4O7 and C32H51N5O8S, which improve the IC50 value of ACE inhibition rate by 12.1 μmol/L and 16.0 μmol/L, respectively.
The ACE inhibitory activity of the ACE inhibitory peptide YVLL derivative was significantly enhanced, and the antihypertensive effect was significantly improved, increasing by 394.7% and 373.8% respectively, with the maximum antihypertensive value increasing by 145.2% and 82.4%.
Smart Images
Figure CN117003816B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of peptide derivative technology, and particularly relates to an ACE inhibitory peptide YVLL derivative and its applications. Background Technology
[0002] Angiotensin-converting enzyme (ACE) inhibitory peptides are small polypeptides formed through proteolysis. They have a significant blood pressure-lowering effect, and more importantly, compared with other common antihypertensive drugs, ACE inhibitory peptides have no toxic side effects and do not affect normal blood pressure. ACE plays an important role in blood pressure regulation. By removing two amino acids (His-Leu) from its C-terminus, it can convert the originally inactive angiotensin I into the active angiotensin II, causing vasoconstriction and raising blood pressure. ACE also inactivates bradykinin, which has a vasodilatory function, similarly causing a rise in blood pressure. ACE inhibitory peptides can block these two biochemical reaction processes caused by ACE, thus lowering blood pressure.
[0003] A bioactive tetrapeptide with publication number CN113480598A has an amino acid sequence of YVLL and exhibits certain ACE inhibitory activity and antihypertensive effects, but the effects are not significant.
[0004] Therefore, those skilled in the art are dedicated to developing new ACE inhibitory peptide derivatives with higher ACE inhibitory activity and better blood pressure-lowering effects. Summary of the Invention
[0005] The purpose of this invention is to propose an ACE inhibitory peptide YVLL derivative and its application, so as to overcome at least one of the above-mentioned defects in the prior art.
[0006] To achieve this objective, the present invention adopts the following technical solution:
[0007] The ACE inhibitory peptide YVLL derivative provided by this invention has the structural formula shown in Formula 1 or Formula 2:
[0008]
[0009] Preferably, the ACE inhibitory peptide YVLL derivative is obtained by acetylation or formylation of the ACE inhibitory peptide YVLL.
[0010] Preferably, the chemical formula of the acetylated ACE inhibitory peptide YVLL derivative is C 28 H44N4O7, with a molecular weight of 548.68, has an IC50 of ACE inhibition rate. 50 The value is 12.1 μmol / L.
[0011] Preferably, the chemical formula of the formylated ACE inhibitory peptide YVLL derivative is C32 H51N5O8S, with a molecular weight of 665.85, has an IC50 of ACE inhibition rate of [missing value]. 50 The value was 16.0 μmol / L.
[0012] This invention also provides the application of the above-mentioned ACE inhibitory peptide YVLL derivative in the preparation of antihypertensive drugs.
[0013] Preferably, the drug is an injection, powder, granule, pill, oral preparation, tablet, capsule, suppository, spray, or ointment.
[0014] The present invention also provides an antihypertensive pharmaceutical composition comprising the above-mentioned ACE inhibitory peptide YVLL derivative and a pharmaceutically acceptable carrier.
[0015] The beneficial effects of this invention are as follows:
[0016] 1. The acetylated and formylated derivatives of the ACE inhibitory peptide YVLL showed significantly enhanced ACE inhibitory activity, increasing by 394.7% and 373.8% respectively compared to the ACE inhibitory peptide YVLL. The antihypertensive effect was also significantly improved, with the maximum antihypertensive value increasing by 145.2% and 82.4% respectively compared to the ACE inhibitory peptide YVLL.
[0017] 2. The acetylated derivative of the ACE inhibitory peptide YVLL can interact with amino acid residues at the ACE active site via hydrogen bonds, hydrophobic bonds, and van der Waals forces. The acetylated derivative of the ACE inhibitory peptide YVLL can form a 7-hydrogen bond network (His383, His353, Tyr523, His513, Asn277, Glu376) with ACE; it can also form strong hydrophobic interactions and van der Waals potentials with 13 amino acid residues, including the active site Ala354, Gln281, and Glu162. Furthermore, the peptide Leu5 binds to Zn at its binding site. 2+ The strong polar interactions formed disrupt the intended function of Zn ions and can also lead to the inhibition of ACE activity. Compared with the unmodified form, the hydrogen bond network formed between the acetylated ACE inhibitory peptide YVLL and ACE was slightly reduced, but the hydrophobic interactions and van der Waals potential interaction sites were significantly increased, and both could interact with Zn. 2+ Ions react.
[0018] 3. The total binding free energy ΔGbinding of the acetylated derivative of the ACE inhibitory peptide YVLL was -66.8087 kJ / mol, which was 40.5% higher than that before modification (-47.55 kJ / mol).
[0019] 4. YVLL formylated derivatives can function via hydrogen bonds, hydrophobic bonds, van der Waals forces, and amino acid residues at the ACE active site. Figure 7 ).like Figure 7 As shown, YVLL formylated derivatives can form a 6-hydrogen bond network (Arg125, Ser630, Tyr547, Glu206) with ACE; YVLL formylated derivatives can also form strong hydrophobic interactions and van der Waals potentials with amino acid residues such as Glu206, Ser630, and Arg125. Attached Figure Description
[0020] Figure 1 This is a graph showing the ACE inhibitory activity of different YVLL derivatives of this invention.
[0021] Figure 2 This is a graph showing the ACE inhibitory activity of different concentrations of YVLL derivatives of this invention.
[0022] Figure 3 This is a graph showing the hypotensive effect of different YVLL derivatives of the present invention on SHR.
[0023] Figure 4 This is a schematic diagram of the binding pattern of the YVLL acetylated derivative of the present invention with ACE.
[0024] Figure 5 yes Figure 4 A magnified view of a portion of the image.
[0025] Figure 6 This invention describes the inhibition mode and mechanism of action of the YVLL acetylated derivative with ACE.
[0026] Figure 7 This is the rmsd curve of the kinetic process of the YVLL acetylated derivative of this invention.
[0027] Figure 8 This invention describes the inhibition mode and mechanism of action of the YVLL formylated derivative with ACE.
[0028] Figure 9 This is the rmsd curve of the kinetic process of the YVLL formylation derivative of this invention. Detailed Implementation
[0029] The present invention will now be further described in conjunction with the accompanying drawings and specific embodiments.
[0030] The ACE inhibitory peptide YVLL (Tyr-Val-Leu-Leu) derivative provided in this embodiment has the structural formula shown in Formula 1 (acetylated ACE inhibitory peptide YVLL derivative) or Formula 2 (formylated ACE inhibitory peptide YVLL derivative):
[0031]
[0032] Among them, the ACE inhibitory peptide YVLL derivative is obtained by acetylation or formylation of the ACE inhibitory peptide YVLL. The chemical formula of the acetylated ACE inhibitory peptide YVLL derivative is C 28 H44N4O7, with a molecular weight of 548.68, has an IC50 of ACE inhibition rate. 50 The value is 12.1 μmol / L. The chemical formula of the formylated ACE inhibitory peptide YVLL derivative is C 32 H51N5O8S, with a molecular weight of 665.85, has an IC50 of ACE inhibition rate of [missing value]. 50 The value was 16.0 μmol / L.
[0033] This embodiment also provides the application of the above-mentioned ACE inhibitory peptide YVLL derivative in the preparation of antihypertensive drugs. The drugs are prepared as injections, powders, granules, pills, oral preparations, tablets, capsules, suppositories, sprays, or ointments.
[0034] This embodiment also provides an antihypertensive drug composition comprising the above-mentioned ACE inhibitory peptide YVLL derivative and a pharmaceutically acceptable carrier.
[0035] I. Materials and Methods
[0036] 1.1 Main Materials and Reagents
[0037] A summary table of experimental drugs and reagents is shown in Table 1:
[0038] Table 1 Summary of Experimental Drugs and Reagents
[0039]
[0040]
[0041] 1.2 Main Instruments
[0042] The summary table of experimental instruments is shown in Table 2:
[0043] Table 2 Summary of Experimental Instruments
[0044]
[0045] 1.3 Experimental Methods
[0046] 1.3.1 Structural Modification and Solid-Phase Synthesis of Peptides
[0047] The polypeptide sequence YVLL (Jinruis Biotechnology Co., Ltd.) was synthesized and identified using a solid-phase synthesis method. It was then modified by acetylation, amidation, myristylation, palmitoylation, formylation, and phosphorylation. The purity of the polypeptide was determined by RP-HPLC, and the molecular weight of the polypeptide was verified by LC-MS.
[0048] 1.3.2 Analysis of ACE inhibitory activity of peptide derivatives
[0049] The peptide derivatives were formulated to a concentration of 1 mg / mL for ACE inhibition activity assay, and peptide derivatives with high ACE inhibition activity were screened for further study.
[0050] 1.3.3 Analysis of the antihypertensive effect of peptide derivatives
[0051] The selected peptide derivatives were subjected to antihypertensive activity assays using essential hypertensive rats (SHR) as a model, and peptide derivatives with good antihypertensive activity were screened out.
[0052] 1.3.4 Molecular docking of peptide derivatives
[0053] The selected peptide derivatives were subjected to molecular docking studies to compare the differences in intermolecular interactions before and after modification.
[0054] 1.3.5 Molecular Dynamics of Peptide Derivatives
[0055] The selected peptide derivatives were subjected to molecular dynamics analysis to study the effect of the modified groups on the total binding free energy.
[0056] 1.4 Data Analysis
[0057] Data is The obtained data were subjected to multiple regression analysis in Design-Expert 8.0, and SPSS 22.0 software was used for data statistics and Origin 2022 for plot design.
[0058] II. Experimental Results
[0059] 2.1 Activity analysis of different YVLL derivatives
[0060] 2.1.1 ACE inhibitory activity analysis of YVLL derivatives
[0061] YVLL was modified by acetylation, amidation, myristylation, palmitoylation, formylation, and phosphorylation. The effects of each derivative on the ACE inhibition rate were then determined after chemical synthesis. Results are as follows: Figure 1As shown, acetylation and formylation of YVLL significantly enhanced ACE inhibitory activity, increasing it by 394.7% and 373.8%, respectively. Amidation, myristylation, palmitoylation, and phosphorylation of YVLL all showed a decreasing trend in activity, decreasing by 88.0%, 48.7%, 14.3%, and 26.9%, respectively. Therefore, acetylated and formylated derivatives of YVLL were selected for further research.
[0062] 2.1.2 IC of YVLL derivatives 50 Measurement
[0063] The chemical formulas of YVLL acetylated and formylated derivatives are C0 and Cforylated, respectively. 28 H44N4O7 (molecular weight 548.68) and C 32 H51N5O8S (molecular weight 665.85). The ACE inhibition rate of different concentrations of YVLL acetylated and formylated derivatives was determined, and the results are as follows: Figure 2 As shown, with increasing concentration, the ACE inhibition rate of YVLL acetylated and formylated derivatives gradually increased. The IC50 values of YVLL acetylated and formylated derivatives were calculated. 50 The concentrations were 12.1 and 16.0 μmol / L, respectively.
[0064] 2.1.3 Analysis of the antihypertensive effect of YVLL derivatives
[0065] Based on the ACE activity assay results, YVLL and its acetylated and formylated derivatives were selected for a study on their antihypertensive effects. The results are as follows: Figure 3 As shown, the antihypertensive effect of YVLL was significantly enhanced after acetylation and formylation modification, with the maximum antihypertensive value increasing by 145.2% and 82.4% respectively compared with YVLL. Due to the high antihypertensive effect of the acetylated YVLL derivative, it was selected for further research.
[0066] 2.2 Molecular docking study of YVLL acetylated derivatives with ACE
[0067] YVLL acetylated derivatives can act via hydrogen bonds, hydrophobic bonds, van der Waals forces, and amino acid residues at the ACE active site. Figures 4-5 ).like Figure 6 As shown, YVLL acetylated derivatives can form a 7-hydrogen bond network (His383, His353, Tyr523, His513, Asn277, Glu376) with ACE; YVLL acetylated derivatives can also form strong hydrophobic interactions and van der Waals potentials with 13 amino acid residues, including the active sites Ala354, Gln281, and Glu162. Furthermore, the peptide Leu5 binds to Zn at its binding site.2+ The formation of strong polar interactions disrupts the intended function of Zn ions and can also inhibit ACE activity. Compared to the unmodified form, the hydrogen bond network between the YVLL acetylated derivative and ACE is slightly reduced, but the hydrophobic interactions and van der Waals potential interaction sites are significantly increased, and both can interact with Zn. 2+ Ions react.
[0068] 2.3 Molecular dynamics study of YVLL acetylated derivatives and ACE
[0069] To examine the interaction between YVLL acetylated derivatives and ACE from a dynamic perspective and to quantitatively analyze the contribution of interacting residues to peptide binding, molecular dynamics simulations were performed on the YVLL acetylated derivative complex. For example... Figure 7 As shown, in the initial stage of the simulation (0-20 ns), there were some conformational changes, and the RMSD fluctuations were slightly large. However, after 20 ns, the RMSD curve tended to stabilize, and the protein conformation gradually stabilized, indicating that the YVLL acetylated derivative bound to the ACE protein and formed a stable complex. The total binding free energy ΔGbinding of the ACE inhibitory peptide YVLL acetylated derivative was -66.8087 kJ / mol, of which the electrostatic potential energy was -150.5873 kJ / mol and the van der Waals potential energy was -45.0012 J / mol. The electrostatic potential energy played a greater role in the complex binding process. Compared with the unmodified state (-47.55 kJ / mol), the total binding free energy of the YVLL acetylated derivative increased by 40.5%, which is one of the reasons for the significant increase in the ACE inhibition rate of the YVLL acetylated derivative.
[0070] Molecular docking study of 2,4YVLL formylated derivatives with ACE
[0071] YVLL formylated derivatives can act via hydrogen bonds, hydrophobic bonds, van der Waals forces, and amino acid residues at the ACE active site. Figure 8 ).like Figure 8 As shown, YVLL formylated derivatives can form a 6-hydrogen bond network (Arg125, Ser630, Tyr547, Glu206) with ACE; YVLL formylated derivatives can also form strong hydrophobic interactions and van der Waals potentials with amino acid residues such as Glu206, Ser630, and Arg125.
[0072] Molecular dynamics study of 2,5YVLL formylated derivatives and ACE
[0073] To examine the dynamic interaction between YVLL formylated derivatives and ACE, and to quantitatively analyze the contribution of interacting residues to peptide binding, molecular dynamics simulations were performed on the YVLL formylated derivative complex. Figure 9 As shown, in the initial stage of the simulation (0-20 ns), there were some conformational changes and the RMSD fluctuated slightly. However, after 20 ns, the RMSD curve tended to stabilize, and the protein conformation gradually stabilized, indicating that the YVLL formylated derivative bound to the ACE protein and formed a stable complex.
[0074] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims
1. An ACE-inhibitory peptide YVLL derivative, characterized in that, The structural formula is as follows: 。 2. The application of the ACE inhibitory peptide YVLL derivative according to claim 1 in the preparation of antihypertensive drugs.
3. The application according to claim 2, characterized in that, The drug is an injection, powder, granule, pill, tablet, capsule, suppository, spray, or ointment.
4. A blood pressure-lowering drug composition, characterized in that, This includes the ACE-inhibiting peptide YVLL derivative as described in claim 1, and pharmaceutically acceptable carriers.