Ginseng peptide with alpha-glycosidase inhibitory activity, ginseng peptide analogue, composition and application
By screening and validating ginseng peptides, ginseng peptides IRPPGF, ACYPIF, and YPIW were identified, which solved the problem of insufficient research on α-glucosidase inhibitory activity in existing technologies and provided new possibilities for diabetes treatment and blood glucose regulation.
Patent Information
- Application Number
- CN202511161637.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-19
- Publication Date
- 2025-11-14
AI Technical Summary
There is limited research on ginseng peptides in the current technology, especially the lack of discovery and bioactivity identification of novel ginseng peptides with α-glucosidase inhibitory activity, which leads to insufficient application in the development of diabetes treatment drugs and nutritional foods.
Ginseng proteins were screened from protein databases, and peptides with α-glycosidase inhibitory activity were identified using in vitro digestion and PeptideRanker scoring. In vitro enzyme activity experiments were conducted to verify the activity of ginseng peptides with amino acid sequences of IRPPGF, ACYPIF, and YPIW. The inhibitory effect was further verified by in vitro enzyme activity inhibition experiments.
The α-glucosidase inhibitory activities of IRPPGF, ACYPIF, and YPIW ginseng peptides were successfully identified and verified, providing new possibilities for diabetes treatment and glycemic regulation, and have important application value.
Smart Images
Figure CN120943892A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, and mainly relates to α-glycosidase inhibitory peptides and their applications. Specifically, this invention relates to a peptide derived from ginseng with α-glycosidase inhibitory activity and its applications. Background Technology
[0002] Ginseng is the tuberous root and rhizome of a perennial herbaceous plant belonging to the Araliaceae family. It possesses rich nutritional value and significant biological activity. For example, ginseng exhibits various functional activities such as antioxidation, antitumor activity, antidepressant activity, anti-aging, blood sugar reduction, and liver damage protection. Therefore, the applications of ginseng and its products extend to food, health products, pharmaceuticals, and cosmetics.
[0003] The pharmacologically active substances in ginseng include saponins, polysaccharides, peptides, sterols, proteins, amino acids, flavonoids, nucleotides, organic acids, and vitamins. Ginseng peptides are polypeptides derived from ginseng and possess physiological activities such as anti-inflammatory, hypoglycemic, lipid-regulating, anti-cell proliferation, and antioxidant effects. Although ginseng peptides are important bioactive substances in ginseng, research on them is limited, particularly regarding the discovery of small molecule monomeric peptides and the identification of their bioactivity.
[0004] Alpha-glucosidase catalyzes the hydrolysis of α-1,4-glycosidic bonds, leading to the hydrolysis of oligosaccharides such as maltose and sucrose in the small intestine. Inhibiting α-glucosidase activity slows down glucose production and absorption, reduces postprandial blood glucose peaks, and regulates blood glucose levels. It also regulates fat production while modulating glucose metabolism. Therefore, α-glucosidase inhibitors could serve as a novel approach to treating diabetes. Ginseng peptides are diverse and exhibit varied activities; discovering and identifying novel ginseng peptides that can inhibit α-glucosidase activity is of significant value for developing drugs and nutritional foods for diabetes treatment. Summary of the Invention
[0005] The purpose of this invention is to provide a ginseng peptide, which has a certain inhibitory activity against α-glucosidase.
[0006] To achieve the above objectives, the present invention provides the following technical solution: In a first aspect, the present invention provides a ginseng peptide with α-glycosidase inhibitory activity, wherein the amino acid sequence of the ginseng peptide is IRPPGF, ACYPIF or YPIW.
[0007] In a second aspect, the present invention provides a method for screening the ginseng peptide, comprising the following steps: All ginseng proteins were screened from the protein database; The screened ginseng protein was digested into peptides using in vitro digestion. The obtained peptides were scored for bioactivity potential using the PeptideRanker tool, and peptides with scores greater than 0.9 were selected. The α-glycosidase inhibitory activity of peptides with a prediction score greater than 0.9 was used to obtain peptides; The obtained peptides were subjected to α-glycosidase inhibitory activity assays and verifications to obtain ginseng peptides with α-glycosidase inhibitory activity.
[0008] In a third aspect, the present invention provides a ginseng peptide analog, wherein the ginseng peptide analog is a repeating sequence of the amino acid sequence IRPPGF, ACYPIF or YPIW.
[0009] In a fourth aspect, the present invention provides a composition comprising the ginseng peptide analogue and the ginseng peptide.
[0010] In a fifth aspect, the present invention provides the use of the ginseng peptide or the ginseng peptide analog or the composition in the preparation of an α-glucosidase inhibitor.
[0011] In a sixth aspect, the present invention provides an α-glycosidase inhibitor comprising any one or a combination of ginseng peptides having the amino acid sequences IRPPGF, ACYPIF, and YPIW; or an analogue of the ginseng peptide or the composition thereof.
[0012] In a preferred embodiment of the present invention, the α-glycosidase inhibitor further includes acceptable excipients.
[0013] In a seventh aspect, the present invention provides the use of the ginseng peptide, or the ginseng peptide analog, or the composition, or the α-glucosidase inhibitor, in the preparation of a product for regulating glucose and lipid metabolism.
[0014] In a preferred embodiment of the present invention, the product is a drug or health product.
[0015] More preferably, the product includes excipients acceptable in the pharmaceutical or health product fields.
[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention utilizes in vitro simulated digestion technology of ginseng protein to obtain a ginseng peptide fragment library, and further identifies three ginseng peptide monomers with α-glycosidase activity inhibitory effects through online virtual screening and in vitro enzyme activity inhibition experiments.
[0017] This invention further validated the ginseng peptides with potential α-glycosidase inhibitory activity obtained from the initial screening using in vitro enzyme activity inhibition experiments. It was found that three ginseng peptide monomers with amino acid sequences of IRPPGF, ACYPIF, and YPIW had certain α-glycosidase activity inhibitory effects. Attached Figure Description
[0018] Figure 1 This is a process for discovering and identifying ginseng peptides with α-glycosidase inhibitory activity.
[0019] Figure 2 This is the chemical structure diagram of ginseng peptide IRPPGF monomer.
[0020] Figure 3 This is the chemical structure diagram of the ginseng peptide ACYPIF monomer.
[0021] Figure 4 This is the chemical structure diagram of the ginseng peptide YPIW monomer.
[0022] Figure 5 It is the inhibitory effect of ginseng peptide IRPPGF monomer on 1 unit / mL α-glucosidase activity.
[0023] Figure 6 It is the inhibitory effect of ginseng peptide ACYPIF monomer on 1 unit / mL α-glucosidase activity.
[0024] Figure 7 It is the inhibitory effect of ginseng peptide YPIW monomer on 1 unit / mL α-glucosidase activity.
[0025] Figure 8 It is the inhibitory effect of ginseng peptide IRPPGF monomer on α-glucosidase activity at 0.5 unit / mL.
[0026] Figure 9 It is the inhibitory effect of ginseng peptide ACYPIF monomer on α-glucosidase activity at 0.5 unit / mL.
[0027] Figure 10 It is the inhibitory effect of ginseng peptide YPIW monomer on α-glucosidase activity at 0.5 unit / mL. Detailed Implementation
[0028] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.
[0029] Ginseng possesses a variety of functional activities, including antioxidant, antitumor, antidepressant, anti-aging, hypoglycemic, and liver damage protection. The pharmacologically active substances in ginseng include saponins, polysaccharides, peptides, sterols, proteins, amino acids, flavonoids, nucleotides, organic acids, and vitamins. Ginseng peptides are polypeptides derived from ginseng and are also active substances in ginseng, possessing physiological activities such as anti-inflammatory, hypoglycemic, lipid-regulating, anti-cell proliferation, and antioxidant effects.
[0030] Alpha-glucosidase catalyzes the hydrolysis of α-1,4-glycosidic bonds, leading to the hydrolysis of oligosaccharides such as maltose and sucrose in the small intestine. Inhibiting α-glucosidase activity slows down glucose production and absorption, reduces postprandial blood glucose peaks, and regulates blood glucose levels. It also regulates fat production while modulating glucose metabolism. Therefore, α-glucosidase inhibitors could serve as a novel approach to treating diabetes. Ginseng peptides are diverse and exhibit varied activities; discovering and identifying novel ginseng peptides that can inhibit α-glucosidase activity is of significant value for developing drugs and nutritional foods for diabetes treatment.
[0031] Based on this, the present invention provides a ginseng peptide with α-glycosidase inhibitory activity, wherein the amino acid sequence of the ginseng peptide is IRPPGF, ACYPIF or YPIW.
[0032] The present invention also provides a ginseng peptide analog, wherein the ginseng peptide analog has an amino acid sequence that is a repeating sequence of IRPPGF, ACYPIF or YPIW.
[0033] This invention further identified three ginseng peptide monomers with α-glycosidase activity inhibitory effects through online virtual screening and in vitro enzyme activity inhibition experiments.
[0034] Example 1 Using computer virtual technology to screen for a novel ginseng peptide with α-glycosidase inhibitory activity, such as... Figure 1 As shown, it includes the following steps: (1) Search the Uniprot protein database and screen out all verified ginseng proteins; (2) Using the bioinformatics tool Peptide Cutter, the digestive enzymes selected are pepsin, trypsin and chymotrypsin. The ginseng protein retrieved in step (1) is digested into peptides through in vitro digestion to obtain a ginseng polypeptide fragment library. (3) The peptides obtained in step (2) were scored for bioactivity potential using the PeptideRanker tool, and peptides with scores greater than 0.9 were selected. (4) Submit the peptides selected in step (3) to the BIOPEP-UWM database and use the database’s bioactivity prediction module to predict the α-glycosidase inhibitory activity of the peptides. (5) The ginseng peptides with α-glycosidase inhibitory activity screened in step (4) are sorted and synthesized to obtain ginseng peptides with α-glycosidase inhibitory activity.
[0035] The amino acid sequences of the obtained ginseng peptides are IRPPGF, ACYPIF, YPIW, YPFPDF, QPPF, HFPPTF, ALPPF, LYPW, DGAGPPPF, and ILPPF.
[0036] Further in vitro enzyme activity inhibition experiments confirmed that the ginseng peptides with amino acid sequences of IRPPGF, ACYPIF, and YPIW all exhibited good α-glycosidase inhibitory activity. Their monomeric chemical structures are as follows: Figures 2-4 As shown.
[0037] Experimental Example 1 Test of the inhibitory activity of ginseng peptides against 1 unit / mL α-glucosidase Ginseng peptides were prepared into polypeptide solutions of different concentrations: 0, 31.25, 62.5, 125, 250, 500, and 1000 μg / mL. 50 μL of peptide solutions of different concentrations and 100 μL of α-glucosidase solution with an enzyme activity of 1 unit / mL were mixed and reacted in a water bath at 37.0 °C for 10 min. The reaction solution was then mixed with 50 μL of 5 mmol / L p-nitrophenyl-α-D-glucopyranoside solution and reacted in a water bath at 37.0 °C for 20 min. The reaction was terminated by adding 200 μL of 0.2 mol / L sodium carbonate solution to the resulting reaction solution. The final reaction solution was analyzed to determine the release of p-nitrophenol at a wavelength of 405 nm using a microplate reader. The inhibition rate of α-glucosidase activity was calculated using the following formula:
[0038] Concentration-inhibition rate curves were plotted based on different concentrations of polypeptide solutions and the corresponding α-glucosidase inhibition rates.
[0039] Figure 5 The figure shows the peptide concentration-inhibition rate curves plotted for IRPPGF peptide at different concentrations and corresponding α-glucosidase inhibition rates. When the peptide concentration reaches 1000 μg / ml, the α-glucosidase inhibition effect is the best, reaching 48.00%.
[0040] Figure 6 The inhibitory effect of ginseng peptide ACYPIF monomer on α-glucosidase activity at 1 unit / mL was observed. The inhibitory effect on α-glucosidase was best when the peptide concentration reached 1000 μg / ml, reaching 15.33%.
[0041] Figure 7 The inhibitory effect of ginseng peptide YPIW monomer on α-glucosidase activity at 1 unit / mL was observed. The inhibitory effect on α-glucosidase was best when the peptide concentration reached 1000 μg / ml, reaching 22.89%.
[0042] Experimental Example 2 Assay for the inhibitory activity of ginseng peptides against 0.5 unit / mL α-glucosidase Ginseng peptides were prepared into polypeptide solutions of different concentrations: 0, 31.25, 62.5, 125, 250, 500, and 1000 μg / mL. 50 μL of peptide solutions of different concentrations and 100 μL of α-glucosidase solution with an enzyme activity of 0.5 units / mL were mixed and reacted in a water bath at 37.0 ℃ for 10 min. The reaction solution was then mixed with 50 μL of 5 mmol / L p-nitrophenyl-α-D-glucopyranoside solution and reacted in a water bath at 37.0 ℃ for 20 min. The reaction was terminated by adding 200 μL of 0.2 mol / L sodium carbonate solution to the resulting reaction solution.
[0043] The release of p-nitrophenol from the final reaction solution was measured at a wavelength of 405 nm using an ELISA reader. The inhibition rate of α-glucosidase activity was calculated using the following formula:
[0044] The inhibitory effects of ginseng peptide IRPPGF monomer, ACYPIF monomer, and YPIW monomer on 0.5 unit / mL α-glucosidase activity are shown in the following order: Figures 8-10 As shown, the α-glucosidase inhibition effect was best when the concentration of IRPPGF monomeric peptide reached 250 μg / ml, reaching 53.86%; the α-glucosidase inhibition effect was best when the concentration of ACYPIF monomeric peptide reached 125 μg / ml, reaching 21.15%; and the α-glucosidase inhibition effect was best when the concentration of YPIW monomeric peptide reached 1000 μg / ml, reaching 29.37%.
[0045] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.
Claims
1. A ginseng peptide with α-glycosidase inhibitory activity, characterized in that, The amino acid sequence of the ginseng peptide is IRPPGF, ACYPIF, or YPIW.
2. A method for screening ginseng peptides according to claim 1, characterized in that, Includes the following steps: All ginseng proteins were screened from the protein database; The screened ginseng protein was digested into peptides using in vitro digestion. The obtained peptides were scored for bioactivity potential using the PeptideRanker tool, and peptides with scores greater than 0.9 were selected. The α-glycosidase inhibitory activity of peptides with a prediction score greater than 0.9 was used to obtain peptides; The obtained peptides were subjected to α-glycosidase inhibitory activity assays and verifications to obtain ginseng peptides with α-glycosidase inhibitory activity.
3. A ginseng peptide analog, characterized in that, The ginseng peptide analogue is a repeating sequence of IRPPGF, ACYPIF, or YPIW as described in claim 1.
4. A composition, characterized in that, The composition is composed of the ginseng peptide analog of claim 3 and the ginseng peptide of claim 1.
5. The use of the ginseng peptide of claim 1, or the ginseng peptide analog of claim 3, or the composition of claim 4 in the preparation of an α-glucosidase inhibitor.
6. An α-glycosidase inhibitor, characterized in that, It includes any one or a combination of several of the ginseng peptides with the amino acid sequences of IRPPGF, ACYPIF, and YPIW as described in claim 1; or The ginseng peptide analogue of claim 3; or The composition of claim 4.
7. The α-glycosidase inhibitor according to claim 6, characterized in that, The α-glucosidase inhibitor also includes acceptable excipients.
8. The use of the ginseng peptide of claim 1, or the ginseng peptide analog of claim 3, or the composition of claim 4, or the α-glucosidase inhibitor of claim 6 in the preparation of a product for regulating glucose and lipid metabolism.