A protein peptide preparation for enhancing immunity and its preparation method

Abstract

The present invention relates to the technical field of protein peptide preparation, and specifically to an immunity-enhancing protein peptide preparation and a preparation method thereof. The protein peptide preparation comprises the following components by weight: 45-60 parts of bovine whey protein peptide, 8-15 parts of onion seed protein peptide, 5-8 parts of 7-hydroxychromone derivative, 0.01-0.03 parts of emulsifier, and 0.02-0.05 parts of stabilizer; the structural formula of the 7-hydroxychromone derivative is: By testing relevant biochemical indicators of an immunosuppressive mouse model, it was found that the combination of onion seed protein peptide, 7-hydroxychromone derivative, and bovine whey protein peptide can enhance the body's immune function by reducing damage to the immune organs of immunosuppressive mice and promoting the division and proliferation of spleen cells.

Description

Technical Field

[0001] The present invention relates to the technical field of protein peptide preparation, and in particular to an immunity-enhancing protein peptide preparation and a preparation method thereof. Background Art

[0002] Immunity is the body's own defense mechanism that can identify and eliminate foreign invading substances (such as viruses, bacteria, etc.), deal with aging, damaged, dead or degenerated self-cells, and identify and deal with mutated cells and virus-infected cells in the body. Strong immunity can effectively resist the invasion of pathogens such as bacteria and viruses, and the immune system can identify and eliminate mutated cells in the body, reducing the risk of cancer. In addition, maintaining immune balance can prevent a variety of diseases, including infectious diseases, autoimmune diseases and tumors. It can be seen that immunity plays a vital role in maintaining human health.

[0003] Current treatments for boosting immunity primarily include modern medicine and traditional Chinese medicine. Modern medicine includes immunomodulators to regulate immune system function and increase resistance, and oral vitamins and minerals to enhance immunity. Traditional Chinese medicine includes internal treatment with Chinese herbal medicine and external treatment with Chinese herbal medicine. Chinese herbal medicine treatment involves enhancing immune cell activity through the use of traditional Chinese medicine, Chinese patent medicines, and medicated diets, while external treatments regulate immune function through moxibustion, acupoint massage, acupuncture, cupping, and Chinese herbal foot baths. Protein peptide preparations are typically small molecule peptides prepared from proteins through enzymatic hydrolysis, chemical synthesis, or bioengineering techniques. These peptides possess specific biological activities and can be used in medical treatment, nutritional supplements, or functional foods. However, existing protein peptide preparations suffer from poor efficacy. Therefore, developing a protein peptide preparation that can be used long-term and enhance immunity is a technical problem that urgently needs to be addressed by those skilled in the art. Summary of the Invention

[0004] The first purpose of the present invention is to provide a protein peptide preparation for enhancing immunity, which can significantly improve the body's immunity.

[0005] The second object of the present invention is to provide a method for preparing a protein peptide preparation for enhancing immunity, which has simple steps and is easy to practice.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] A protein peptide preparation for enhancing immunity, comprising the following components by weight: 45-60 parts of bovine whey protein peptide, 8-15 parts of onion protein peptide, 5-8 parts of 7-hydroxychromone derivative, 0.01-0.03 parts of emulsifier, 0.02-0.05 parts of stabilizer, and 60-70 parts of water; the structural formula of the 7-hydroxychromone derivative is:

[0008]

[0009] Preferably, the preparation method of the 7-hydroxychromone derivative comprises the following steps:

[0010]

[0011] (1) 7-Hydroxychromone, pyridine, and 4-dimethylaminopyridine were added to ethyl acetate, cooled, and glycine was added to react. After purification, intermediate 1 was obtained;

[0012] (2) Triethylamine and D-(+)-glyceraldehyde are added to a methanol solution of the intermediate 1 to react, and a 7-hydroxychromone derivative is obtained after purification.

[0013] Preferably, the molar ratio of 7-hydroxychromone, pyridine, 4-dimethylaminopyridine and glycine in step (1) is 10:(12-15):(10-12):(10-12).

[0014] Preferably, the temperature after cooling in step (1) is 0-5°C; and the reaction time is 5-8h.

[0015] Preferably, the molar ratio of triethylamine, D-(+)-glyceraldehyde and intermediate 1 in step (2) is (20-25):(12-15):10; and the molar concentration of intermediate 1 in the methanol solution of intermediate 1 is 0.2-0.5 mmol / mL.

[0016] Preferably, the reaction time in step (2) is 18 to 24 hours.

[0017] Preferably, the preparation method of the onion protein peptide is as follows:

[0018] The method comprises adding scallion powder to petroleum ether for defatting to obtain defatted scallion powder; adding the defatted scallion powder to phosphate buffer for extraction, filtering to obtain a supernatant; adding ammonium sulfate to the supernatant, stirring for 1 to 2 hours, and obtaining a precipitate by centrifugation; dissolving the precipitate with phosphate buffer, dialyzing for 36 to 48 hours, and freeze-drying to obtain scallion freeze-dried powder; adding pepsin hydrolyzate to the scallion freeze-dried powder, adjusting the pH to 2, and then hydrolyzing for 1 to 2 hours, then adding trypsin hydrolyzate, adjusting the pH to 7.5, and then hydrolyzing for 2 to 3 hours, stopping the hydrolysis, dialyzing for 36 to 48 hours, and freeze-drying to obtain the product.

[0019] Preferably, the ratio of the onion seed powder to petroleum ether is 1 g: (5-10) mL; the defatting temperature is 35-50° C., and the extraction time is 2-3 h; the pH of the phosphate buffer is 8, and the amount of ammonium sulfate added is such that the saturation of ammonium sulfate in the supernatant reaches 80-85%.

[0020] Preferably, the stabilizer is selected from one of xanthan gum, apple pectin, and gum arabic, and the emulsifier is soybean lecithin or lecithin.

[0021] The preparation method of the above-mentioned protein peptide preparation for enhancing immunity comprises the following steps:

[0022] According to the above weight ratio, mix the raw materials evenly and sterilize them.

[0023] The beneficial technical effects of the present invention are:

[0024] The immune-enhancing protein peptide preparation of the present invention contains ingredients such as onion seed protein peptide, 7-hydroxychromone derivatives, and bovine whey protein peptide. Testing of relevant biochemical indicators in immunocompromised mice revealed that the combined use of onion seed protein peptide, 7-hydroxychromone derivatives, and bovine whey protein peptide can enhance the immune function of immunocompromised mice by reducing damage to their immune organs and promoting the division and proliferation of spleen cells. DETAILED DESCRIPTION

[0025] The following is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it is not intended that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art of the present invention, without departing from the inventive concept, several simple deductions or replacements can also be made, all of which should be considered to belong to the scope of protection of the present invention. The specific conditions not indicated in the examples are carried out according to conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, unless otherwise specified, are conventional products obtained through commercial channels.

[0026] (1) Embodiment

[0027] Example 1

[0028] Example 1 provides a protein peptide preparation for enhancing immunity. The protein peptide preparation is composed of the following components in parts by weight: 51 parts of bovine whey protein peptide, 12 parts of onion protein peptide, 6 parts of 7-hydroxychromone derivative, 0.02 parts of soybean lecithin, and 0.03 parts of xanthan gum; the structural formula of the 7-hydroxychromone derivative is:

[0029]

[0030] The preparation method of the above-mentioned 7-hydroxychromone derivative is as follows:

[0031]

[0032] (1) 7-Hydroxychromone (CAS: 261-973-7), pyridine, 4-dimethylaminopyridine, glycine, and ethyl acetate were added to ethyl acetate in a ratio of 10 mmol: 14 mmol: 11 mmol: 11 mmol: 20 mL. The mixture was cooled to 4°C, and glycine was added under stirring for 6 h. The reaction solution was poured into ice water, adjusted to pH 7 with 0.1 mol / L hydrochloric acid, allowed to stand, and the organic layer was separated. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined and washed three times with saturated brine, dried over anhydrous sodium sulfate, and distilled under reduced pressure. Recrystallization with ethanol gave Intermediate 1 with a yield of 62.53%.

[0033] The NMR and mass spectrometry results of intermediate 1 are as follows:

[0034] 1 HNMR (C 11 H9NO4,400MHz,d6-DMSO)δ8.70(s,2H),8.03(d,1H),7.73(d,1H),7.42(dd,1H),7.20(d,1H),6.35(d,1H),4.24(s,2H); MS(ESI)m / z=220.05[M+H] + , the above results confirmed that the target product was obtained.

[0035] (2) Triethylamine and D-(+)-glyceraldehyde were added to a methanol solution of intermediate 1 at a molar ratio of 22:14:10. The molar concentration of intermediate 1 in the methanol solution of intermediate 1 was 0.3 mmol / mL. The reaction was carried out at room temperature for 20 h. The reaction solution was concentrated and chromatographed on a silica column (V 二氯甲烷 :V 乙醇 =3:1) analysis, and 7-hydroxychromone derivative was obtained with a yield of 60.41%.

[0036] The NMR and mass spectrometry results of 7-hydroxychromone derivatives are as follows:

[0037] 1 HNMR (C 14 H 13 NO6,400MHz,d6-DMSO)δ8.03(d,1H),7.73(d,1H),7.42(dd,1H),7.20(d,1H),6.53(d,1H),6.35(d,1H), 6.18(s,1H),4.95(s,1H),4.10-3.85(m,2H),3.65-3.63(q,1H),2.51(s,2H),MS(ESI)m / z=291.07[M+H] +, the above results confirmed that the target product was obtained.

[0038] The preparation method of the onion protein peptide is as follows:

[0039] The onion powder was added to petroleum ether at a ratio of 1 g:8 mL, and the mixture was defatted at 45°C to obtain defatted onion powder. The defatted onion powder was added to phosphate buffer at a pH of 8 at a ratio of 1 g:20 mL, and the mixture was extracted at room temperature for 2.5 h. The filtered phase was obtained by filtration, and the filtered phase was centrifuged at 4°C (5000 r / min, 30 min) to obtain a supernatant. Ammonium sulfate (saturation of 82%) was added to the supernatant, stirred for 1.5 h, and centrifuged at 4°C (5000 r / min, 30 min). ) was centrifuged to obtain a precipitate; the precipitate was dissolved in a phosphate buffer solution with a pH of 8 at a ratio of 1 g to 20 mL of precipitate and phosphate buffer, and the precipitate was dialyzed with a 3500Da dialysis bag for 40 hours and freeze-dried to obtain scallion freeze-dried powder; 4% pepsin hydrolyzate was added to the scallion freeze-dried powder, the pH was adjusted to 2, and the powder was hydrolyzed in a 36°C water bath with shaking for 1.5 hours, and then 4% trypsin hydrolyzate was added, the pH was adjusted to 7.5, and the powder was hydrolyzed in a 36°C water bath with shaking for 2.5 hours. After the hydrolysis was completed, the temperature was raised to 95°C for reaction for 6 minutes to terminate the hydrolysis, and the powder was dialyzed with a 2500Da dialysis bag for 40 hours and freeze-dried to obtain the powder.

[0040] Example 1 also provides a method for preparing the above-mentioned protein peptide preparation for enhancing immunity, which is as follows:

[0041] According to the above weight ratio, mix the raw materials evenly and sterilize them.

[0042] Example 2

[0043] Example 2 provides a protein peptide preparation for enhancing immunity. The protein peptide preparation is composed of the following components in parts by weight: 45 parts of bovine whey protein peptide, 8 parts of onion protein peptide, 5 parts of 7-hydroxychromone derivative, 0.01 parts of lecithin, and 0.02 parts of apple pectin; the structural formula of the 7-hydroxychromone derivative is the same as that of Example 1.

[0044] The preparation method of the above-mentioned 7-hydroxychromone derivative is as follows:

[0045] (1) 7-Hydroxychromone (CAS: 261-973-7), pyridine, 4-dimethylaminopyridine, glycine, and ethyl acetate were added to ethyl acetate in a molar ratio of 10 mmol: 12 mmol: 10 mmol: 10 mmol: 20 mL. The mixture was cooled to 0°C, and glycine was added under stirring for 8 h. The reaction solution was poured into ice water, adjusted to pH 7 with 0.1 mol / L hydrochloric acid, allowed to stand, and the organic layer was separated. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined and washed three times with saturated brine, dried over anhydrous sodium sulfate, and distilled under reduced pressure, followed by recrystallization with ethanol to obtain intermediate 1 with a yield of 57.26%. The results of nuclear magnetic resonance and mass spectrometry were the same as those in Example 1.

[0046] (2) Triethylamine and D-(+)-glyceraldehyde were added to a methanol solution of intermediate 1 at a molar ratio of 20:12:10. The molar concentration of intermediate 1 in the methanol solution of intermediate 1 was 0.2 mmol / mL. The reaction was carried out at room temperature for 18 h. The reaction solution was concentrated and chromatographed on a silica column (V 二氯甲烷 :V 乙醇 =3:1) analysis, 7-hydroxychromone derivative was obtained with a yield of 54.83%. The results of nuclear magnetic resonance and mass spectrometry were the same as those in Example 1.

[0047] The preparation method of the onion protein peptide is as follows:

[0048] The onion powder was added to petroleum ether at a ratio of 1 g:5 mL, and the mixture was defatted at 35°C to obtain defatted onion powder. The defatted onion powder was added to phosphate buffer at a pH of 8 at a ratio of 1 g:20 mL, and the mixture was extracted at room temperature for 2 h. The filtered phase was obtained by filtration, and the filtered phase was centrifuged at 4°C (5000 r / min, 30 min) to obtain a supernatant. Ammonium sulfate (saturation of 80%) was added to the supernatant, stirred for 1 h, and centrifuged at 4°C (5000 r / min, 30 min). ) was centrifuged to obtain a precipitate; the precipitate was dissolved in a phosphate buffer solution with a pH of 8 at a ratio of 1 g to 20 mL of precipitate and phosphate buffer, and the precipitate was dialyzed with a 3500Da dialysis bag for 36 hours and freeze-dried to obtain scallion freeze-dried powder; 4% pepsin hydrolyzate was added to the scallion freeze-dried powder, the pH was adjusted to 2, and the powder was hydrolyzed in a 35°C water bath with shaking for 2 hours, and then 4% trypsin hydrolyzate was added, the pH was adjusted to 7.5, and the powder was hydrolyzed in a 35°C water bath with shaking for 3 hours. After the hydrolysis was completed, the temperature was raised to 92°C for reaction for 8 minutes to terminate the hydrolysis, and the powder was dialyzed with a 2500Da dialysis bag for 36 hours and freeze-dried to obtain the powder.

[0049] Example 2 also provides a method for preparing the above-mentioned protein peptide preparation for enhancing immunity, which is as follows:

[0050] According to the above weight ratio, mix the raw materials evenly and sterilize them.

[0051] Example 3

[0052] Example 3 provides a protein peptide preparation for enhancing immunity. The protein peptide preparation is composed of the following components in parts by weight: 60 parts of bovine whey protein peptide, 15 parts of onion protein peptide, 8 parts of 7-hydroxychromone derivative, 0.03 parts of soybean lecithin, and 0.05 parts of gum arabic; the structural formula of the 7-hydroxychromone derivative is the same as that of Example 1.

[0053] The preparation method of the above-mentioned 7-hydroxychromone derivative is as follows:

[0054] (1) 7-Hydroxychromone (CAS: 261-973-7), pyridine, and 4-dimethylaminopyridine were added to ethyl acetate in a molar ratio of 10 mmol: 15 mmol: 12 mmol: 12 mmol: 20 mL. The mixture was cooled to 5°C, and glycine was added under stirring for 5 h. The reaction solution was poured into ice water, adjusted to pH 7 with 0.1 mol / L hydrochloric acid, allowed to stand, and the organic layer was separated. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined and washed three times with saturated brine, dried over anhydrous sodium sulfate, and distilled under reduced pressure, followed by recrystallization with ethanol to obtain intermediate 1 with a yield of 56.22%. The results of nuclear magnetic resonance and mass spectrometry were the same as those in Example 1.

[0055] (2) Triethylamine and D-(+)-glyceraldehyde were added to a methanol solution of intermediate 1 at a molar ratio of 25:15:10. The molar concentration of intermediate 1 in the methanol solution of intermediate 1 was 0.5 mmol / mL. The reaction was carried out at room temperature for 24 h. The reaction solution was concentrated and chromatographed on a silica column (V 二氯甲烷 :V 乙醇 =3:1) analysis, 7-hydroxychromone derivative was obtained with a yield of 51.97%. The NMR results were the same as those in Example 1.

[0056] The preparation method of the onion protein peptide is as follows:

[0057] The onion powder was added to petroleum ether at a ratio of 1 g:10 mL, and the mixture was defatted at 50°C to obtain defatted onion powder. The defatted onion powder was added to phosphate buffer at a pH of 8 at a ratio of 1 g:20 mL, and the mixture was extracted at room temperature for 3 h. The filtered phase was obtained by filtration, and the filtered phase was centrifuged at 4°C (5000 r / min, 30 min) to obtain a supernatant. Ammonium sulfate (saturation of 85%) was added to the supernatant, stirred for 2 h, and centrifuged at 4°C (5000 r / min, 30 min). n) centrifuging to obtain a precipitate; dissolving the precipitate in a phosphate buffer solution having a pH of 8 at a ratio of 1 g to 20 mL of precipitate and phosphate buffer, dialyzing the solution with a 3500 Da dialysis bag for 48 hours, and lyophilizing to obtain scallion seed lyophilized powder; adding a 4% pepsin hydrolyzate to the scallion seed lyophilized powder, adjusting the pH to 2, and performing enzymolysis in a 37° C. water bath with shaking for 1 hour; then adding a 4% trypsin hydrolyzate, adjusting the pH to 7.5, and continuing the enzymolysis in a 37° C. water bath with shaking for 2 hours; after the enzymolysis is completed, heating the solution to 96° C. for 5 minutes to terminate the enzymolysis, dialyzing the solution with a 2500 Da dialysis bag for 48 hours, and lyophilizing to obtain the product.

[0058] Example 3 also provides a method for preparing the above-mentioned protein peptide preparation for enhancing immunity, which is as follows:

[0059] According to the above weight ratio, mix the raw materials evenly and sterilize them.

[0060] (2) Comparative Example

[0061] Comparative Example 1

[0062] Comparative Example 1 provides a preparation, which consists of the following components in parts by weight: 69 parts of bovine whey protein peptide, 0.02 parts of soybean lecithin, and 0.03 parts of xanthan gum.

[0063] Comparative Example 1 also provides a preparation method of the above-mentioned preparation, which is specifically the same as Example 1.

[0064] Comparative Example 2

[0065] Comparative Example 2 provides a preparation, which consists of the following components in parts by weight: 51 parts of bovine whey protein peptide, 6 parts of 7-hydroxychromone derivative, 0.02 parts of soybean lecithin, and 0.03 parts of xanthan gum.

[0066] Comparative Example 2 also provides a preparation method for the above-mentioned preparation, which is specifically the same as Example 1.

[0067] Comparative Example 3

[0068] Comparative Example 3 provides a preparation, which consists of the following components in parts by weight: 51 parts of bovine whey protein peptide, 12 parts of onion protein peptide, 6 parts of 7-hydroxychromone, 0.02 parts of soybean lecithin, and 0.03 parts of xanthan gum.

[0069] Comparative Example 3 also provides a preparation method for the above-mentioned preparation, which is specifically the same as Example 1.

[0070] Comparative Example 4

[0071] Comparative Example 4 provides a preparation, which consists of the following components in parts by weight: 51 parts of bovine whey protein peptide, 18 parts of vitamin C, 0.02 parts of soybean lecithin, and 0.03 parts of xanthan gum.

[0072] Comparative Example 4 also provides a preparation method for the above-mentioned preparation, which is specifically the same as Example 1.

[0073] (3) Test examples

[0074] 1. Test subjects

[0075] A total of 90 male BABL / c mice weighing 18-22 g were adaptively fed for 5 days and randomly divided into a normal group, a model group, Example groups 1-3, and Comparative Example groups 1-4, with 10 mice in each group.

[0076] 2. Establishment of Immunocompromised Model

[0077] The normal group was intraperitoneally injected with normal saline (0.2 mL / 10 g). Except for the normal group, the mice in the other groups were intraperitoneally injected with cyclophosphamide solution (100 mg / kg each time) once a day for 3 consecutive days to establish the immunodeficiency model in mice.

[0078] 3. Dosage

[0079] Normal group: gavage with an equal amount of normal saline once a day for 28 days;

[0080] Model group: gavage with an equal amount of normal saline once a day for 28 days;

[0081] Groups 1 to 3 of Examples were given the protein peptide preparation of Groups 1 to 3 by oral gavage at a dose of 2 g / kg, once a day for 28 consecutive days;

[0082] Comparative Example 1-4 Groups: The preparation of Comparative Example 1-4 Groups was administered orally at a dose of 2 g / kg, once a day for 28 consecutive days;

[0083] 4. Biochemical index detection method

[0084] 4.1. Serum hemolysin and IL-6 levels

[0085] On the 24th day after oral gavage, 5 mice were randomly selected from each group and sensitized with 5% SRBC (0.2 mL / mouse) via intraperitoneal injection. The mice were fasted for 12 hours that evening after the last oral gavage. The next day, blood was collected from the mice's orbits, and the serum was separated and diluted 100-fold with saline. 1 mL of the diluted serum was mixed with 0.5 mL of 10% SRBC and 0.5 mL of 10% complement serum. A blank control tube (with equal amounts of complement, SRBC, and 1 mL of saline) was also set up. The samples were incubated in a 37°C incubator for 0.5 h, then the reaction was terminated in a 4°C refrigerator. Finally, the samples were centrifuged at 2000 r / min for 10 min. 1 mL of the supernatant was collected and allowed to stand for 10 min. The control tube was used as a blank to adjust the zero value. The OD value of each tube was measured at 540 nm and the results were recorded. The results are shown in Table 1.

[0086] The remaining 5 mice in each group were fasted for 12 hours that night after the last gavage. Blood was collected from the eye sockets the next day, and the serum was separated. The IL-6 content of the mice in each group was detected according to the kit instructions. The results are shown in Table 1.

[0087] 4.2 Body weight and organ index testing

[0088] After blood collection, the fasting body weight of each group of mice was weighed and recorded as the final body weight. The mice were then sacrificed by cervical dislocation, and the spleen and thymus were removed. The blood was washed away with saline, and the excess saline was absorbed with filter paper before weighing. The organ index was calculated using the following formula:

[0089] Organ index (mg / g) = organ mass (mg) / final body weight (g). The results are shown in Table 1.

[0090] 4.3 Spleen Cell Cycle

[0091] After organ weighing, the spleens of mice in each group were ground on a glass slide and a cell suspension was prepared in physiological saline for later use. The spleen cell suspension was filtered, washed, fixed with 70% cold ethanol, and stored at 4°C until ready for use. Before loading, propidium iodide stain containing RNase was added, and the cells were incubated at 4°C for 30 minutes. After incubation, the cells were filtered and analyzed on a flow cytometer for cell cycle analysis. 30,000 cells were obtained for each sample, and cell cycle analysis was performed using MULTICYCLE software. The results are shown in Table 2.

[0092] 5. Results Analysis

[0093] Table 1

[0094] Grouping Spleen index (mg / g) Thymus index (mg / g) <![CDATA[Serum hemolysin (OD 540 )]]> IL-6 (pg / mL) Normal group 6.76 1.91 0.96 274.15 Model Group 2.53 1.35 0.21 171.60 Example 1 4.60 1.69 0.65 250.23 Example 2 4.51 1.65 0.62 248.17 Example 3 4.47 1.63 0.60 246.85 Comparative Example 1 3.42 1.38 0.36 196.41 Comparative Example 2 4.06 1.50 0.45 224.38 Comparative Example 3 4.29 1.59 0.50 239.97 Comparative Example 4 3.58 1.41 0.41 208.23

[0095] Table 2

[0096]

[0097]

[0098] As can be seen from Table 1, compared with the normal group, the spleen index and thymus index of the model group were significantly reduced, indicating that the immunosuppression model of mice was successfully constructed. Compared with the model group, after gavage with the protein peptide preparations of Examples 1 to 3, the spleen index and thymus index of each mouse increased significantly, indicating that the protein peptide preparation of the present invention can reduce the damage to the immune organs of immunosuppressed mice. Compared with the model group, after gavage with the protein peptide preparations of Examples 1 to 3, the serum hemolysin and IL-6 levels in the serum of each mouse increased significantly, indicating that the protein peptide preparation of the present invention can promote the recovery of immune function. In summary, the protein peptide preparation of the present invention can not only reduce the damage to the immune organs of immunosuppressed mice, but also increase the serum hemolysin and IL-6 levels of mouse spleen cells, thereby improving the body's immune function.

[0099] As shown in Table 2, compared with the normal group, the model group had a significantly increased number of splenocytes in the G0+G1 phase, and a significantly decreased number of splenocytes in the S+G2+M phase. Compared with the model group, after oral administration of the protein peptide preparations of Examples 1 to 3, the number of splenocytes in the S+G2+M phase increased significantly. In summary, the protein peptide preparations of the present invention can promote the division and proliferation of splenocytes in immunocompromised mice.

[0100] Compared with Examples 1 to 3, Comparative Example 1 replaces onion protein peptide and 7-hydroxychromone derivatives with bovine whey protein peptide to prepare the preparation, Comparative Example 2 omits the preparation prepared by onion protein peptide, Comparative Example 3 replaces 7-hydroxychromone derivatives with 7-hydroxychromone preparation, and Comparative Example 4 replaces onion protein peptide and 7-hydroxychromone derivatives with vitamin C preparation. Although they can also enhance the body's immunity, the effect is poor. In summary, the combined use of onion protein peptide, 7-hydroxychromone derivatives, and bovine whey protein peptide can significantly enhance the body's immunity.

[0101] Finally, it should be noted that the above embodiments are intended only to illustrate the technical solutions of the present invention and are not intended to limit the same. The basic principles and main features of the present invention have been described above using specific implementation schemes. Modifications or substitutions may be made based on the present invention, but such modifications or substitutions do not detract from the essence of the corresponding technical solutions from the scope of protection claimed by the present invention.

Claims

1. A protein peptide preparation for enhancing immunity, characterized in that: The invention is composed of the following components by weight: 45-60 parts of bovine whey protein peptide, 8-15 parts of onion protein peptide, 5-8 parts of 7-hydroxychromone derivative, 0.01-0.03 parts of emulsifier, and 0.02-0.05 parts of stabilizer; the structural formula of the 7-hydroxychromone derivative is: ； The preparation method of the onion protein peptide is as follows: The onion powder is added to petroleum ether for defatting to obtain defatted onion powder; the defatted onion powder is added to a phosphate buffer solution with a pH of 8 for extraction, and filtered to obtain a supernatant; ammonium sulfate is added to the supernatant, stirred for 1-2 hours, and centrifuged to obtain a precipitate; the precipitate is dissolved in a phosphate buffer solution with a pH of 8, dialyzed through a 3500Da dialysis bag for 36-48 hours, and freeze-dried to obtain onion freeze-dried powder; 4% pepsin hydrolyzate is added to the onion freeze-dried powder, the pH is adjusted to 2, and then enzymolyzed for 1-2 hours, and then 4% trypsin hydrolyzate is added, the pH is adjusted to 7.5, and then enzymolyzed for 2-3 hours, the enzymolysis is terminated, and the powder is dialyzed through a 2500Da dialysis bag for 36-48 hours, and freeze-dried to obtain the powder; The stabilizer is selected from one of xanthan gum, apple pectin and gum arabic, and the emulsifier is soybean lecithin or lecithin.

2. The protein peptide preparation for enhancing immunity according to claim 1, characterized in that The preparation method of the 7-hydroxychromone derivative comprises the following steps: (1) 7-Hydroxychromone, pyridine, and 4-dimethylaminopyridine were added to ethyl acetate, cooled, and glycine was added to react. After purification, intermediate 1 was obtained; (2) Triethylamine and D-(+)-glyceraldehyde are added to a methanol solution of intermediate 1 to react, and a 7-hydroxychromone derivative is obtained after purification.

3. The protein peptide preparation for enhancing immunity according to claim 2, characterized in that The molar ratio of 7-hydroxychromone, pyridine, 4-dimethylaminopyridine and glycine in step (1) is 10: (12-15): (10-12): (10-12).

4. The protein peptide preparation for enhancing immunity according to claim 2, characterized in that The temperature after cooling in step (1) is 0-5°C; the reaction time is 5-8h.

5. The protein peptide preparation for enhancing immunity according to claim 2, characterized in that The molar ratio of triethylamine, D-(+)-glyceraldehyde, and intermediate 1 in step (2) is (20-25): (12-15): 10; the molar concentration of intermediate 1 in the methanol solution of intermediate 1 is 0.2-0.5 mmol / mL.

6. The protein peptide preparation for enhancing immunity according to claim 2, characterized in that The reaction time in step (2) is 18 to 24 hours.

7. The protein peptide preparation for enhancing immunity according to claim 1, characterized in that The ratio of the onion seed powder to petroleum ether is 1 g: (5-10) mL; the degreasing temperature is 35-50° C., and the extraction time is 2-3 hours; and the amount of ammonium sulfate added is such that the saturation of ammonium sulfate in the supernatant reaches 80-85%.

8. The method for preparing the immunity-enhancing protein peptide preparation according to any one of claims 1 to 7, characterized in that: The following steps are involved: According to the above weight ratio, mix the raw materials evenly and sterilize them.

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