Small molecule protein peptide and application thereof

Abstract

The invention belongs to the technical field of bioactive peptides, and relates to a small molecular protein peptide and application thereof, the amino acid sequence of the small molecular protein peptide is shown as SEQ ID NO.1, and the molecular weight is 547.6 Da. The invention provides a small molecule protein peptide which can effectively inhibit generation of fructose-induced bovine serum albumin glycosylation intermediates and can reduce modification of carbonyl and sulfydryl, oxidation products and cross-linked structures, and application of the small molecule protein peptide.

Description

Technical Field

[0001] This invention belongs to the field of bioactive peptide technology, and relates to a small molecule protein peptide and its application, particularly a small molecule protein peptide with non-enzymatic glycosylation inhibitory activity and its application. Background Technology

[0002] Non-enzymatic glycosylation and the accumulation of its end products (AGEs) are key pathological mechanisms leading to diabetic complications, aging, and various chronic diseases such as inflammation, Alzheimer's disease, and atherosclerosis. Non-enzymatic glycosylation and AGEs continuously damage the body's health by altering protein structure and function and inducing oxidative stress. Therefore, effectively inhibiting non-enzymatic glycosylation and blocking AGE formation is of great significance for the prevention and treatment of related diseases.

[0003] Currently, although some synthetic non-enzymatic glycosylation inhibitors (such as metformin and aminoguanidine) are used clinically, their long-term use is often accompanied by side effects such as gastrointestinal reactions and potential damage to liver and kidney function, which limits their safety for widespread and long-term use. Therefore, developing novel active ingredients with high safety and significant non-enzymatic glycosylation inhibition effects is of great practical significance.

[0004] Bioactive peptides, as a class of small-molecule, highly safe, easily absorbed, and diverse bioactivities, have become a hot topic in the development of novel functional factors. Some endogenous or animal-derived bioactive peptides (such as carnosine and glutathione) have been shown to effectively inhibit AGEs formation through multiple mechanisms, including acting as competitive targets for glycosylation and scavenging reactive carbonyl groups and free radicals. However, the types of peptides currently discovered with glycosylation-inhibiting activity are still relatively limited, making it difficult to meet practical application needs. There is an urgent need to further explore and develop novel small-molecule protein peptides with non-enzymatic glycosylation-inhibiting activity. Summary of the Invention

[0005] To address the aforementioned technical problems in the background art, this invention provides a small molecule protein peptide that can effectively inhibit the formation of fructose-induced bovine serum albumin glycosylation intermediates, reduce carbonyl and thiol group modifications, oxidation products, and cross-linked structures, as well as its applications.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: A small molecule protein peptide, characterized in that: the amino acid sequence of the small molecule protein peptide is shown in SEQ ID NO.1.

[0007] Preferably, the molecular weight of the small molecule protein peptide provided by the present invention is 547.6 Da.

[0008] The application of small molecule protein peptides in inhibiting non-enzymatic glycosylation activity, as previously described.

[0009] The previously described applications of small molecule protein peptides in scavenging superoxide anion radicals, scavenging hydroxyl radicals, capturing methylglyoxal, or causing fluorescence quenching of BSA.

[0010] A non-enzymatic glycosylation inhibitor, characterized in that: the non-enzymatic glycosylation inhibitor uses small molecule protein peptides as described above as its active ingredient.

[0011] Preferably, the non-enzymatic glycosylation inhibitor provided by the present invention is an inhibitor A for scavenging superoxide anion free radicals or scavenging hydroxyl free radicals, an inhibitor B for capturing methylglyoxal, or an inhibitor C for causing fluorescence quenching of BSA.

[0012] Preferably, the concentration of the small molecule protein peptide in inhibitor A provided by the present invention is 10-50 µg / mL, but is not limited to this concentration range; the concentration of the small molecule protein peptide in inhibitor B is 100-1500 µg / mL, but is not limited to this concentration range; and the concentration of the small molecule protein peptide in inhibitor C is 2.738-27.38 µg / mL, but is not limited to this concentration range.

[0013] Preferably, the non-enzymatic glycosylation inhibitor provided by the present invention further includes pharmaceutically acceptable excipients, said excipients being one or more combinations of fillers, binders, disintegrants, and lubricants.

[0014] Based on the use of the non-enzymatic glycosylation inhibitors as described above in the preparation of drugs, functional foods and / or cosmetics that inhibit non-enzymatic glycosylation, said dosage forms being tablets, capsules, granules, oral liquids or injections.

[0015] Compared with the prior art, the present invention has the following beneficial effects: This invention discloses a small molecule protein peptide with the amino acid sequence shown in SEQ ID NO.1, namely FDPLG (phenylalanine-aspartic acid-proline-leucine-glycine). Experimental determination shows that the molecular weight of this small molecule protein peptide is 547.6 Da. Its specific amino acid composition and sequence structure endow it with excellent inhibitory activity. The small molecule protein peptide provided by this invention exhibits significant non-enzymatic glycosylation inhibitory activity, effectively inhibiting the formation of fructose-induced bovine serum albumin (BSA) glycosylation intermediates and reducing carbonyl and thiol modifications, oxidation products, and cross-linking structures. A concentration range of 100-2400 µg / mL of the small molecule protein peptide significantly inhibits the formation of fluorescent AGEs (P<0.05) and shows a significant dose-response effect (P<0.05). Its IC50 value for inhibiting glycosylation is 1.897 mM, significantly superior to aminoguanidine (IC50 of 5.083 mM) (P<0.05). The small molecule protein peptides provided by this invention can be obtained through artificial synthesis or biological extraction, and the preparation process is relatively simple. They can be used as active ingredients in the preparation of drugs that inhibit non-enzymatic glycosylation, and can also be applied to functional foods (such as foods with high blood sugar regulation function), cosmetics (such as skin care products with anti-aging effects), etc. This provides a new effective ingredient for solving the side effects of current glycosylation inhibitors, and has broad application prospects and significant practical value. The small molecule protein peptides provided by this invention can capture large amounts of the intermediate MGO generated during glycosylation, exhibiting a significant free radical scavenging effect, and effectively targeting the large amount of superoxide anion free radicals generated during glycosylation (…). Hydroxyl radicals (·OH) have a good scavenging effect and spontaneously and stably bind to protein BSA to form a complex with a stoichiometric ratio of about 1:1. The complex is very stable after simulated gastrointestinal digestion, providing a new and effective substance for solving the glycosylation problem. Attached Figure Description

[0016] Figure 1 This is the mass spectrum of the small molecule protein peptide FDPLG provided by this invention; Figure 2 This is the curve of the total fluorescence intensity of the small molecule protein peptide FDPLG provided by this invention on the non-enzymatic glycosylation system; Figure 3 The inhibition rate of the total fluorescence intensity of the non-enzymatic glycosylation system by the small molecule protein peptide FDPLG provided by this invention; Figure 4 This is a graph showing the effect of the small molecule protein peptide FDPLG provided by this invention on the IC50 of the non-enzymatic glycosylation system. Figure 5 The different concentrations of small molecule protein peptide FDPLG provided by this invention have a susceptibility to superoxide anion free radicals (FOPG). The absorbance value to be removed; Figure 6 The different concentrations of small molecule protein peptide FDPLG provided by this invention have a susceptibility to superoxide anion free radicals (FOPG). The removal rate of the cleanup; Figure 7 These are the absorbance values ​​of different concentrations of small molecule protein peptide FDPLG provided by this invention for scavenging hydroxyl radicals (·OH); Figure 8 The scavenging rate of hydroxyl radicals (·OH) by different concentrations of small molecule protein peptide FDPLG provided by this invention; Figure 9 This is a graph showing the capture capacity of different concentrations of small molecule protein peptide FDPLG for MGO provided by the present invention. Figure 10 The effect of different concentrations of small molecule protein peptide FDPLG on the fluorescence spectrum of BSA at 298 K was investigated. Figure 11 The Stern-Volmer curves of BSA at different concentrations of small molecule protein peptide FDPLG at 298 K are shown. Figure 12 The effect of different concentrations of small molecule protein peptide FDPLG on the fluorescence spectrum of BSA at 310K; Figure 13 The Stern-Volmer curves of BSA at different concentrations of small molecule protein peptide FDPLG at 310 K are shown. Figure 14 The effect of different concentrations of small molecule protein peptide FDPLG on the fluorescence spectrum of BSA at 304K; Figure 15 The Stern-Volmer curves of BSA at different concentrations of small molecule protein peptide FDPLG at 304K are shown. Figure 16 This is a chromatogram of the elution times of the small molecule protein peptide FDPLG provided by this invention after treatment with different proteases. Figure 17 This is a diagram showing the relative peak areas of the small molecule protein peptide FDPLG provided by this invention after treatment with different proteases. Detailed Implementation

[0017] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0018] This invention provides a small molecule protein peptide, the mass spectrum of which is shown below. Figure 1 As shown, the information related to the small molecule protein peptide is illustrated. Its amino acid sequence, as shown in SEQ ID NO.1 of the sequence listing, is FDPLG (phenylalanine-aspartic acid-proline-leucine-glycine). The molecular weight of this small molecule protein peptide is 547.6 Da, and its specific amino acid composition and sequence structure endow it with excellent inhibitory activity. The small molecule protein peptide provided by this invention can effectively inhibit non-enzymatic glycosylation by scavenging free radicals, interacting with BSA, and capturing methylglyoxal (MGO). The small molecule protein peptide contains glycine and aspartic acid, which can efficiently capture MGO; it contains aromatic amino acids such as phenylalanine, hydrophobic amino acids such as phenylalanine, proline, and leucine, and acidic amino acids such as aspartic acid, and has a hydrophobic amino acid at the N-terminus, thus exhibiting good free radical scavenging ability. These amino acids and their sequences have a positive impact on free radical scavenging activity, thereby inhibiting glycosylation. The small molecule protein peptide FDPLG forms a ground-state complex with a binding site of BSA through a spontaneous, entropy-driven process via static quenching. Hydrophobic interactions are the primary force, thereby inhibiting glycosylation. Its IC50 value for inhibiting non-enzymatic glycosylation is 1.897 mM, significantly superior to aminoguanidine (IC50 5.083 mM) (P < 0.05). Based on these characteristics, this small molecule protein peptide can be used as an active ingredient in the preparation of compound drugs for non-enzymatic glycosylation and diabetic complications. The small molecule protein peptide provided by this invention has the advantages of expanding its application range in medicine, food, cosmetics, and other fields, and solves the problems of significant side effects of existing non-enzymatic glycosylation inhibitors and the lack of diverse bioactive peptides with glycosylation-inhibiting effects.

[0019] The technical solution provided by this invention will be described in detail below with reference to experiments and accompanying drawings: Determination of the inhibitory activity of small molecule protein peptide FDPLG against non-enzymatic glycosylation 1. Preparation of non-enzymatic glycosylation system BSA solution (20 mg / mL) was mixed with 0.5 M fructose in phosphate-buffered saline (PBS, 0.01 M, pH 7.4). Subsequently, small molecule protein peptides of varying concentrations (100–2400 μg / mL) were added and mixed with the prepared solution. The mixture was then incubated in a constant-temperature shaker at 37°C for 7 days with continuous shaking at 100 rpm. Aminoguanidine (AG) was used as a positive control. After incubation, the reaction was terminated by cooling the mixture to an ice bath, followed by further analysis.

[0020] 2. Experimental Methods 2.1 Determination of Glycosylation Inhibition Rate Glycosylation inhibition rate was expressed as the change in fluorescence of AGEs in the product. The excitation wavelength (λex) was set to 370 nm, and the emission range (λem) was 400–600 nm. The fluorescence intensity of the glycosylated AGEs was measured using a fluorescence spectrophotometer with a bandwidth of 10 nm. The inhibitory effect of total AGEs was evaluated at λex = 370 nm and λem = 440 nm. The inhibition rate (I%) was calculated using the following formula: Note: F0 and F represent the fluorescence values ​​in the absence of small molecule peptides and in the presence of small molecule peptides, respectively.

[0021] 2.2 Determination of the free radical scavenging ability of small molecule protein peptide FDPLG 2.2.1 Superoxide anion radical ( In the MGO–lysine model, O2⁻· levels were detected using the NBT assay. 100 µL of lysine (0.73 mg / mL) and 90 µL of MGO (0.36 mg / mL) were mixed with different concentrations of small molecule protein peptides (0–50 µg / mL), and 1 mL of PBS buffer (10 mM, pH 7.4) was added. The mixture was incubated at 37°C for 3 hours. Then, 200 μL of NBT (200.0 µM) was added, and the reaction was continued for 30 minutes. UV spectra were measured from 500 to 800 nm. The O2⁻· scavenging rate was calculated using the following formula: .

[0022] Note: A0 and A represent the absorbance in the absence of small molecule peptides and in the presence of small molecule peptides, respectively.

[0023] 2.2.2 Hydroxyl radical (·OH): 180 µL of MgO (0.36 mg / mL), 200 µL of lysine (0.73 mg / mL), 50 µM ferric chloride, and small molecule peptides of varying concentrations (0-50 µg / mL) were mixed in 1 mL of PBS buffer (10 mM, pH 7.4) and incubated at 37°C for 3 hours. Then, 200 µL of 2-deoxy-D-ribose (20.0 mM) was added, and incubation continued for another 3 hours. The reaction mixture was then added to 200 µL of 1% TBA and 200 µL of 2.8% TCA, boiled at 100°C for 10 minutes, and then rapidly cooled. The UV spectra from 510 to 600 nm were measured, and the absorbance at 524 nm was used to assess the ·OH scavenging rate.

[0024] .

[0025] Note: A0 and A represent the absorbance in the absence of small molecule peptides and in the presence of small molecule peptides, respectively.

[0026] 2.3 Determination of the ability of small molecule protein peptides FDPLG to capture MGO After incubation with small molecule protein peptides and methylglyoxal (MGO), residual MGO was detected using an Ultimate 3000 high-performance liquid chromatograph. Separation was performed using a ChromCore C18 column (4.6 × 150 mm, 5 μm, NanoChrom, China). Mobile phase A: 0.2% aqueous acetic acid; Mobile phase B: acetonitrile; Gradient program: 0–5 min: phase B 5% → 45%, 5–20 min: phase B 45% → 60%, 20–25 min: phase B 60% → 5%; Detection wavelength: 315 nm; Injection volume: 20 μL; Flow rate: 1.0 mL / min. MGO capture rate (I%) was calculated by the peak area ratio of derivative 2-MQ and internal standard 5-methylquinoline (5-MQ), corresponding to retention times of 7.50 and 8.57 min, respectively.

[0027] Note: C0 and C represent the MGO content in the absence and presence of small molecule peptides, respectively.

[0028] 2.4 Determination of the interaction between small molecule protein peptide FDPLG and BSA Different concentrations of small molecule protein peptides were incubated with BSA solution at 298K, 310K, and 304K for 20 minutes, with a peptide-free control group included. Fluorescence intensity was measured under the following conditions: excitation wavelength set to 280 nm, emission spectrum range of 300–500 nm, excitation and emission slit widths both set to 5.0 nm, and scan rate maintained at 1000 nm / min. All fluorescence data were corrected for internal filtering effects, and the fluorescence spectra were further refined by subtracting appropriate blank fluorescence. The blank group consisted of peptide solution and PBS buffer of the same concentration. The following formula was used to exclude interference from internal structural factors: Fco = （ ） / 2 In the formula, Fco is the calibrated fluorescence value; Fob is the actual measured fluorescence value; AEx is the ultraviolet absorbance value of the excitation spectrum; and AEm is the ultraviolet absorbance value of the emission wavelength.

[0029] The fluorescence quenching mechanism was analyzed using the Stern-Volmer formula at three temperatures: 298 K, 310 K, and 304 K. = = In the formula, F0 represents the fluorescence intensity without the small molecule peptide; F represents the fluorescence intensity with the small molecule peptide; and K represents the fluorescence intensity with the small molecule peptide. sv : Quenching constant; K q : Quenching rate constant; τ0 (2.9 × 10 −9 s): mean protein lifetime; [Q]: sample concentration.

[0030] 2.5 Determination of the digestive stability of small molecule protein peptide FDPLG A small molecule protein peptide solution (0.1 mg / ml) was prepared, and after adjusting the pH to 2.0, pepsin (1:30000) was added. The solution was incubated at 37 ℃ for 2 h, and then the sample was removed. Subsequently, the pH of the peptide solution after pepsin hydrolysis was adjusted to 7.0, and trypsin (1:250) was added. The solution was then incubated at 37 ℃ for 2 h and stored at 4 ℃ for later use. Finally, the extracted peptide sample solution was filtered through a 0.22 µm pore size membrane and injected into an HPLC apparatus for analysis. Specific conditions are shown in Table 1. The peak area and elution time of the HPLC chromatogram were used to determine whether the peptide had degraded after pepsin and trypsin hydrolysis.

[0031] Table 1. HPLC conditions for determining the digestibility of small molecule protein peptides. Name, Conditions, Instrument: Ultimate 3000 High Performance Liquid Chromatograph (Thermo Fisher Scientific, USA); Column: ChromCore C18 column (4.6 × 150 mm, 5 μm, NanoChrom, China); Column Temperature: 25℃; Detection Wavelength: 214 nm; Injection Volume: 20 μl; Flow Rate: 0.7 ml / min mobile phase Mobile Phase A: 2% acetonitrile + 98% water + 0.05% TFA Mobile Phase B: 90% acetonitrile + 10% water + 0.05% TFA Testing procedures 0.2 - 30 min: Phase B 12% -> 42% 30 - 40 min: Phase B 42% -> 95% 40 - 40.1 min: Phase B 95% -> 12% 40.1 - 50 min: Phase B 12% -> 12% 3 Results 3.1 Inhibitory activity of small molecule protein peptide FDPLG against non-enzymatic glycosylation See Figure 2 , Figure 3 as well as Figure 4 (Note: Different letters indicate significant differences (P<0.05)). The fluorescence intensity of total AGEs in BSA was lower, while the fluorescence intensity in Gly-BSA increased significantly, indicating widespread AGE formation. However, with increasing peptide concentration, the fluorescence intensity of total AGEs gradually decreased, and the inhibition rate of AGE formation increased significantly in a concentration-dependent manner, reaching 61.92% at 1000 μg / mL. The IC50 value determined by regression fitting in SPSS was 1.897 mM, significantly better than that of aminoguanidine (5.083 mM) (P<0.05).

[0032] 3.2 Free radical scavenging ability of small molecule protein peptide FDPLG See Figure 5 , Figure 6 , Figure 7 as well as Figure 8 (Note: Different letters indicate significant differences (P<0.05)). The scavenging effect of the small molecule protein peptide FDPLG on superoxide anion radicals (O2⁻) and hydroxyl radicals (·OH) was evaluated by ultraviolet spectroscopy. The experimental results showed that with the increase of the concentration of the small molecule protein peptide FDPLG, its scavenging ability against these two free radicals gradually increased, while the ultraviolet intensity decreased significantly, indicating that the small molecule protein peptide FDPLG can effectively scavenge superoxide anion radicals and hydroxyl radicals.

[0033] Regarding the scavenging effect on superoxide anion radicals, the UV intensity gradually decreased as the concentration of the small molecule protein peptide FDPLG increased from 10 µg / mL to 50 µg / mL, indicating that FDPLG can effectively scavenge superoxide anion radicals. The scavenging effect was most significant at a concentration of 50 µg / mL, with a scavenging rate approaching 60%. This trend was positively correlated with the increase in FDPLG concentration. Regarding the scavenging effect on hydroxyl radicals, the UV intensity gradually decreased with increasing FDPLG concentration, showing that FDPLG can also scavenge hydroxyl radicals. At a concentration of 50 µg / mL, the scavenging rate of FDPLG for hydroxyl radicals reached approximately 30%. This result further confirms the broad activity of FDPLG in scavenging free radicals, and its scavenging effect is significantly enhanced with increasing concentration. The scavenging effects of different concentrations of FDPLG on free radicals varied significantly, and the concentration dependence was obvious. The differences in scavenging rates among different concentration groups were statistically significant (P < 0.05). Small molecule protein peptides contain aromatic amino acids such as phenylalanine, hydrophobic amino acids such as phenylalanine, proline, and leucine, and acidic amino acids such as aspartic acid. Furthermore, they contain hydrophobic amino acids at their N-terminus, thus exhibiting good free radical scavenging ability. These amino acids and their sequences positively influence free radical scavenging activity, thereby inhibiting non-enzymatic glycosylation.

[0034] 3.3 Capacity of small molecule protein peptides FDPLG to capture MGO See Figure 9 (Note: Different letters indicate significant differences (P<0.05)) This shows the effect of different concentrations (100 µg / mL, 200 µg / mL, 500 µg / mL, 800 µg / mL, 1000 µg / mL, 1500 µg / mL) of the small molecule protein peptide FDPLG on MGO clearance. Figure 9 The clearance rate of MGO in the small molecule protein peptide FDPLG increased significantly with increasing concentration, and the higher the concentration, the more significant the clearance effect. At 100 µg / mL, the clearance rate was approximately 10%. The clearance rate gradually increased with increasing concentration. At a concentration of 1500 µg / mL, the clearance rate of MGO reached approximately 50%, and the difference in clearance rate between different concentrations was significant (P < 0.05). Because the small molecule protein peptide FDPLG contains glycine and aspartic acid, it can efficiently capture MGO; therefore, FDPLG can inhibit non-enzymatic glycosylation by clearing MGO.

[0035] 3.4 Interaction between small molecule protein peptide FDPLG and BSA The quenching constant and related thermodynamic parameters of the interaction between the small molecule protein peptide FDPLG and BSA at different temperatures are shown in Table 2.

[0036] Table 2 Quenching constants and related thermodynamic parameters of the interaction between small molecule protein peptides FDPLG and BSA at different temperatures. T(K) Ksv (10 4 L / mol) Kq (10 12 L / mol) R a n Ka (10 4 L / mol) R b ΔH (kJ / mol) ΔS (J / mol / K) AG (kJ / mol) F52980.48601.73580.99531.00340.49150.995810.127104.65-21.0593040.45911.6359 0.99461.07180.87960.9901-21.6873100.44501.58930.9951.14321.83150.9919-22.315 See Figure 10 , Figure 11 , Figure 12 , Figure 13 , Figure 14 as well as Figure 15 As the concentration of the small molecule protein peptide FDPLG increased, the fluorescence intensity of BSA gradually decreased, indicating that FDPLG may have interacted with BSA, leading to fluorescence quenching. Higher concentrations resulted in lower fluorescence intensity, showing a negative correlation between concentration and fluorescence intensity. Furthermore, the fluorescence intensity decreased with increasing temperature. At 298 K, Ksv was 0.4860 × 10⁻⁶. 4 Kq is 1.7358 × 10 12 The binding is relatively strong; however, as the temperature increases, Ksv and Kq decrease slightly, but Kq is significantly greater than the maximum scattering collision quenching constant of 2.0 × 10⁻⁶. 10 L / (mol s). The above results indicate that the fluorescence quenching of BSA by the small molecule protein peptide FDPLG is a static quenching process involving the formation of a ground-state complex. Thermodynamic parameters show that the enthalpy change ΔH > 0 (10.127 kJ / mol), entropy change ΔS > 0 (104.65 J / mol·K), and Gibbs free energy change ΔG are all negative (-21.059 kJ / mol, -21.687 kJ / mol, -22.315 kJ / mol), and change with increasing temperature. When ΔH > 0 and ΔS > 0, hydrophobic interaction is the dominant force. The number of binding sites is close to 1, and the linear correlation coefficient R0 is high. b The value ranges from 0.9901 to 0.9958. This indicates that the binding of the small molecule protein peptide FDPLG to BSA is a spontaneous, entropy-driven process with a single binding site.

[0037] 3.5 Digestive stability of small molecule protein peptide FDPLG See Figure 16 as well as Figure 17 (Note: Identical letters indicate no significant difference (P<0.05)). To assess the stability of FDPLG after gastrointestinal digestion, an in vitro simulation of the gastrointestinal digestive pathway was performed. HPLC was used to detect the absorption peak area and elution time after treatment with pepsin and trypsin. Figure 16 As shown, the FDPLG was first subjected to pepsin digestion for 2 h, and it was found that, compared with the blank group, the liquid chromatogram and peak area did not change significantly after pepsin digestion for 1 h and 2 h; then the FDPLG was subjected to trypsin digestion for 1 h and 2 h respectively, and the peak time was consistent and the peak area did not change significantly. The above results show that the FDPLG has good gastrointestinal digestion stability and can exert biological activity in the body in its integrity.

[0038] Obviously, the small molecule protein peptide FDPLG provided by the present application has good application prospects in inhibiting non-enzymatic glycosylation activity, especially in scavenging superoxide anion free radicals, scavenging hydroxyl free radicals, capturing methyl glyoxal or causing fluorescence quenching of BSA, and has good use effect. At the same time, the small molecule protein peptide provided by the present application can also form a non-enzymatic glycosylation inhibitor, which can be an inhibitor A for scavenging superoxide anion free radicals or scavenging hydroxyl free radicals, an inhibitor B for capturing methyl glyoxal or an inhibitor C for causing fluorescence quenching of BSA. In addition, the non-enzymatic glycosylation inhibitor also includes a pharmaceutically acceptable adjuvant, which is a combination of one or more of a filler, a binder, a disintegrant and a lubricant. Further, based on the non-enzymatic glycosylation inhibitor, the non-enzymatic glycosylation inhibitor can also be used in the preparation of non-enzymatic glycosylation inhibitors, functional foods and / or cosmetics, and the application dosage form is tablet, capsule, granule, oral liquid or injection.

[0039] Although the embodiments of the present application have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to the embodiments without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.

Claims

1. A small molecule protein peptide, characterized in that: The amino acid sequence of the small molecule protein peptide is shown in SEQ ID NO.

1.

2. The small molecule protein peptide according to claim 1, characterized in that: The molecular weight of the small molecule protein peptide is 547.6 Da.

3. The application of the small molecule protein peptide according to claim 1 or 2 in inhibiting non-enzymatic glycosylation activity.

4. The use of the small molecule protein peptide according to claim 1 or 2 in scavenging superoxide anion free radicals, scavenging hydroxyl free radicals, capturing methylglyoxal, or causing fluorescence quenching of BSA.

5. A non-enzymatic glycosylation inhibitor, characterized in that: The non-enzymatic glycosylation inhibitor uses the small molecule protein peptide described in claim 1 or 2 as its active ingredient.

6. The non-enzymatic glycosylation inhibitor according to claim 5, characterized in that: The non-enzymatic glycosylation inhibitor is an inhibitor A used to scavenge superoxide anion radicals or hydroxyl radicals, an inhibitor B used to capture methylglyoxal, or an inhibitor C used to cause fluorescence quenching of BSA.

7. The non-enzymatic glycosylation inhibitor according to claim 6, characterized in that: The concentration of small molecule protein peptide in inhibitor A is 10-50 µg / mL; the concentration of small molecule protein peptide in inhibitor B is 100-1500 µg / mL; and the concentration of small molecule protein peptide in inhibitor C is 2.738-27.38 µg / mL.

8. The non-enzymatic glycosylation inhibitor according to claim 5, 6, or 7, characterized in that: The non-enzymatic glycosylation inhibitor also includes pharmaceutically acceptable excipients, which are one or more combinations of fillers, binders, disintegrants, and lubricants.

9. The use of the non-enzymatic glycosylation inhibitor as described in any one of claims 5-8 in the preparation of drugs, functional foods and / or cosmetics that inhibit non-enzymatic glycosylation, wherein the dosage form of the application is tablets, capsules, granules, oral liquids or injections.

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