Protein peptide preparation for enhancing immunity and preparation method thereof

Abstract

The invention relates to the technical field of protein peptide preparation, in particular to a protein peptide preparation for enhancing immunity and a preparation method thereof. The protein peptide preparation is prepared from the following components in parts by weight: 45 to 60 parts of bovine whey protein peptide, 8 to 15 parts of fistular onion seed protein peptide, 5 to 8 parts of 7-hydroxychromone derivative, 0.01 to 0.03 part of emulsifier and 0.02 to 0.05 part of stabilizer, the structural formula of the 7-hydroxychromone derivative is # imgabs0. Related biochemical index detection on a mouse immunocompromised model finds that through cooperative use of the fistular onion seed protein peptide, the 7-hydroxychromone derivative and the bovine whey protein peptide, the damage of immune organs of the immunocompromised mouse can be reduced; the division and proliferation of splenocytes are promoted to enhance the immune function of the body.

Description

Technical Field

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

[0002] Immunity is the body's own defense mechanism, which 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] The current treatment methods for improving immunity mainly include modern medical methods and traditional Chinese medicine methods. Modern medical methods include immunomodulators to regulate the function of the immune system, increase the body's resistance, and oral vitamins and minerals to enhance immunity. Traditional Chinese medicine methods include internal conditioning with Chinese medicine and external treatment with Chinese medicine. Among them, Chinese medicine conditioning is to enhance the activity of immune cells by taking Chinese medicine, Chinese patent medicine, and medicinal diet, and external treatment with Chinese medicine is to regulate the body's immune function through moxibustion, acupoint massage, acupuncture, cupping, and Chinese medicine foot bath. Protein peptide preparations are usually small molecule peptides prepared by enzymatic hydrolysis, chemical synthesis or bioengineering technology of proteins. These peptides have specific biological activities and can be used in medical treatment, nutritional supplements or functional foods. However, existing protein peptide preparations have problems such as poor effects. Therefore, the development of a protein peptide preparation that can be used for a long time and enhance immunity is a technical problem that urgently needs to be solved 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-hydroxy chromone derivative comprises the following steps:

[0010]

[0011] (1) Add 7-hydroxy chromone, pyridine, and 4-dimethylaminopyridine to ethyl acetate. After cooling, add glycine for reaction. After purification, intermediate 1 is obtained.

[0012] (2) Add triethylamine and D-(+)-glyceraldehyde to the methanol solution of intermediate 1 for reaction. After purification, the 7-hydroxy chromone derivative is obtained.

[0013] Preferably, the molar ratio of 7-hydroxy chromone, 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; the reaction time is 5 - 8 h.

[0015] Preferably, 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.

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

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

[0018] Add scallion seed powder to petroleum ether for defatting to obtain defatted scallion seed powder; add the defatted scallion seed powder to phosphate buffer for extraction, filter to obtain the supernatant; add ammonium sulfate to the supernatant and stir for 1 - 2 h, centrifuge to obtain the precipitate; dissolve the precipitate with phosphate buffer, then dialyze for 36 - 48 h and lyophilize to obtain freeze-dried scallion seed powder; add pepsin enzymatic hydrolysate to the freeze-dried scallion seed powder, adjust the pH to 2 and enzymatically hydrolyze for 1 - 2 h, then add trypsin enzymatic hydrolysate, adjust the pH to 7.5 and enzymatically hydrolyze for 2 - 3 h, terminate the enzymatic hydrolysis, dialyze for 36 - 48 h, and lyophilize to obtain it.

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

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

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

[0022] Mix the raw materials evenly according to the weight ratio, and then sterilize them.

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

[0024] The protein peptide preparation for enhancing immunity of the present invention contains components such as scallion seed protein peptide, 7-hydroxy chromone derivative, and milk whey protein peptide. Through the detection of relevant biochemical indexes on the immune deficiency mouse model, it is found that the combined use of scallion seed protein peptide, 7-hydroxy chromone derivative, and milk whey protein peptide can enhance the immune function of the body by reducing the damage of immune organs in immune deficiency mice and promoting the division and proliferation of spleen cells. Specific Embodiments

[0025] The following content is a further detailed description of the present invention in combination with specific preferred embodiments. It cannot be determined that the specific implementation of the present invention is only limited to these descriptions. For those of ordinary skill in the technical field to which the present invention belongs, without departing from the concept of the present invention, several simple deductions or substitutions can be made, which should all be regarded as belonging to the protection scope of the present invention. The specific conditions not specified in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, unless otherwise specified, are all conventional products obtained through commercial channels.

[0026] (I) Embodiment

[0027] Embodiment 1

[0028] Embodiment 1 provides a protein peptide preparation for enhancing immunity. By weight, the protein peptide preparation consists of the following components: 51 parts of milk whey protein peptide, 12 parts of scallion seed protein peptide, 6 parts of 7-hydroxy chromone derivative, 0.02 parts of soy phospholipid, and 0.03 parts of xanthan gum; the structural formula of the 7-hydroxy chromone derivative is:

[0029]

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

[0031]

[0032] (1) According to the dosage ratio of 7-hydroxy chromone, pyridine, 4-dimethylaminopyridine, glycine, and ethyl acetate of 10 mmol: 14 mmol: 11 mmol: 11 mmol: 20 mL, add 7-hydroxy chromone (CAS: 261-973-7), pyridine, and 4-dimethylaminopyridine to ethyl acetate, cool to 4 °C, and then add glycine under stirring conditions to react for 6 h; pour the reaction solution into ice water, adjust the pH to 7 with 0.1 mol / L hydrochloric acid, let it stand, separate the organic layer, extract the aqueous phase with ethyl acetate, combine the organic phases, wash with saturated brine 3 times, dry with anhydrous sodium sulfate, and recrystallize with ethanol after vacuum distillation to obtain Intermediate 1 with a yield of 62.53%.

[0033] The NMR and MS results of Intermediate 1 are as follows:

[0034] 1 HNMR(C 11 H9NO4, 400 MHz, d6-DMSO) δ 8.70 (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 confirm the obtained target product.

[0035] (2) According to the molar ratio of triethylamine, D-(+)-glyceraldehyde, and Intermediate 1 of 22:14:10, add triethylamine and D-(+)-glyceraldehyde to the methanol solution of Intermediate 1. The molar concentration of Intermediate 1 in the methanol solution of Intermediate 1 is 0.3 mmol / mL, and react at room temperature for 20 h; concentrate the reaction solution and perform chromatography (V 二氯甲烷 : V 乙醇 = 3:1) analysis on a silica column to obtain the 7-hydroxy chromone derivative with a yield of 60.41%.

[0036] The NMR and MS results of the 7-hydroxy chromone derivative are as follows:

[0037] 1 HNMR(C 14 H 13 NO6, 400 MHz, d6-DMSO) δ 8.03 (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 confirm the obtaining of the target product.

[0038] The preparation method of the above-mentioned scallion seed protein peptide is as follows:

[0039] According to the dosage ratio of scallion seed powder to petroleum ether of 1 g: 8 mL, add the scallion seed powder to petroleum ether, degrease at 45 °C to obtain defatted scallion seed powder; according to the dosage ratio of defatted scallion seed powder to phosphate buffer of 1 g: 20 mL, add the defatted scallion seed powder to phosphate buffer with a pH of 8, extract at room temperature for 2.5 h, filter to obtain the filtered phase, and centrifuge the filtered liquid phase at 4 °C (5000 r / min, 30 min) to obtain the supernatant; add ammonium sulfate (saturation degree of 82%) to the supernatant and stir for 1.5 h, centrifuge at 4 °C (5000 r / min, 30 min) to obtain a precipitate; according to the dosage ratio of the precipitate to phosphate buffer of 1 g: 20 mL, dissolve the precipitate with phosphate buffer with a pH of 8, dialyze with a 3500 Da dialysis bag for 40 h, and lyophilize to obtain freeze-dried scallion seed powder; add 4% pepsin enzymatic hydrolysate to the freeze-dried scallion seed powder, adjust the pH to 2, enzymatically hydrolyze in a water bath at 36 °C with shaking for 1.5 h, then add 4% trypsin enzymatic hydrolysate, adjust the pH to 7.5, continue to enzymatically hydrolyze in a water bath at 36 °C with shaking for 2.5 h, after the enzymatic hydrolysis is completed, raise the temperature to 95 °C and react for 6 min to terminate the enzymatic hydrolysis, dialyze with a 2500 Da dialysis bag for 40 h, and lyophilize to obtain the product.

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

[0041] According to the above weight ratio, mix each raw material evenly, and sterilize it to obtain the product.

[0042] Example 2

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

[0044] The preparation method of the above 7-hydroxy chromone derivative is specifically as follows:

[0045] (1) According to the molar ratio of 7-hydroxy chromone, pyridine, 4-dimethylaminopyridine, glycine, and ethyl acetate of 10 mmol: 12 mmol: 10 mmol: 10 mmol: 20 mL, add 7-hydroxy chromone (CAS: 261-973-7), pyridine, and 4-dimethylaminopyridine to ethyl acetate, cool to 0 °C, and then add glycine under stirring conditions and react for 8 h; pour the reaction solution into ice water, adjust the pH to 7 with 0.1 mol / L hydrochloric acid, let it stand, separate the organic layer, extract the aqueous phase with ethyl acetate, combine the organic phases, wash them 3 times with saturated brine, dry with anhydrous sodium sulfate, and recrystallize with ethanol after vacuum distillation to obtain Intermediate 1, with a yield of 57.26%. The NMR and mass spectrometry results are the same as those in Example 1.

[0046] (2) According to the molar ratio of triethylamine, D-(+)-glyceraldehyde, and Intermediate 1 of 20:12:10, add triethylamine and D-(+)-glyceraldehyde to the methanol solution of Intermediate 1. The molar concentration of Intermediate 1 in the methanol solution of Intermediate 1 is 0.2 mmol / mL, and react at room temperature for 18 h; concentrate the reaction solution and perform chromatography (V 二氯甲烷 :V 乙醇 = 3:1) analysis on a silica gel column to obtain the 7-hydroxy chromone derivative, with a yield of 54.83%. The NMR and mass spectrometry results are the same as those in Example 1.

[0047] The preparation method of the above-mentioned scallion protein peptide is as follows:

[0048] According to the dosage ratio of scallion powder to petroleum ether of 1 g: 5 mL, add scallion powder to petroleum ether and degrease at 35 °C to obtain defatted scallion powder; according to the dosage ratio of defatted scallion powder to phosphate buffer of 1 g: 20 mL, add defatted scallion powder to the phosphate buffer with a pH of 8 and extract at room temperature for 2 h. After filtration, obtain the filtered phase, and centrifuge the filtered liquid phase at 4 °C (5000 r / min, 30 min) to obtain the supernatant; add ammonium sulfate (saturation degree of 80%) to the supernatant and stir for 1 h, and centrifuge at 4 °C (5000 r / min, 30 min) to obtain a precipitate; according to the dosage ratio of the precipitate to the phosphate buffer of 1 g: 20 mL, dissolve the precipitate with the phosphate buffer with a pH of 8, dialyze with a 3500 Da dialysis bag for 36 h, and freeze-dry to obtain freeze-dried scallion powder; add 4% pepsin enzyme solution to the freeze-dried scallion powder, adjust the pH to 2, and enzymatically hydrolyze in a water bath at 35 °C with shaking for 2 h. Then add 4% trypsin enzyme solution, adjust the pH to 7.5, and continue to enzymatically hydrolyze in a water bath at 35 °C with shaking for 3 h. After the enzymatic hydrolysis is completed, raise the temperature to 92 °C and react for 8 min to terminate the enzymatic hydrolysis, dialyze with a 2500 Da dialysis bag for 36 h, and freeze-dry to obtain the product.

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

[0050] Mix the raw materials evenly according to the above weight parts ratio, and then sterilize them to obtain the product.

[0051] Example 3

[0052] Example 3 provides a protein peptide preparation for enhancing immunity. By weight, the protein peptide preparation consists of the following components: 60 parts of bovine whey protein peptide, 15 parts of scallion seed protein peptide, 8 parts of 7-hydroxy chromone derivative, 0.03 part of soybean phospholipid, and 0.05 part of arabic gum; the structural formula of the 7-hydroxy chromone derivative is the same as that in Example 1.

[0053] The preparation method of the above 7-hydroxy chromone derivative is specifically as follows:

[0054] (1) According to the molar ratio of 7-hydroxy chromone:pyridine:4-dimethylaminopyridine:glycine:ethyl acetate of 10 mmol:15 mmol:12 mmol:12 mmol:20 mL, add 7-hydroxy chromone (CAS: 261-973-7), pyridine, and 4-dimethylaminopyridine to ethyl acetate, cool to 5 °C, and then add glycine under stirring conditions and react for 5 h; pour the reaction solution into ice water, adjust the pH to 7 with 0.1 mol / L hydrochloric acid, let it stand, separate the organic layer, extract the aqueous phase with ethyl acetate, combine the organic phases, wash them 3 times with saturated brine, dry them with anhydrous sodium sulfate, distill under reduced pressure, and then recrystallize with ethanol to obtain Intermediate 1, with a yield of 56.22%, and the NMR and mass spectrometry results are the same as those in Example 1.

[0055] (2) According to the molar ratio of triethylamine:D-(+)-glyceraldehyde:Intermediate 1 of 25:15:10, add triethylamine and D-(+)-glyceraldehyde to the methanol solution of Intermediate 1. The molar concentration of Intermediate 1 in the methanol solution of Intermediate 1 is 0.5 mmol / mL, and react at room temperature for 24 h; concentrate the reaction solution and perform chromatography (V 二氯甲烷 :V 乙醇 = 3:1) analysis to obtain the 7-hydroxy chromone derivative, with a yield of 51.97%, and the NMR results are the same as those in Example 1.

[0056] The preparation method of the above scallion seed protein peptide is as follows:

[0057] According to the dosage ratio of scallion powder to petroleum ether of 1 g:10 mL, add the scallion powder to petroleum ether, degrease at 50 °C to obtain defatted scallion powder; according to the dosage ratio of defatted scallion powder to phosphate buffer of 1 g:20 mL, add the defatted scallion powder to a phosphate buffer with a pH of 8, extract at room temperature for 3 h, filter to obtain a filtrate phase, and centrifuge the filtrate phase at 4 °C (5000 r / min, 30 min) to obtain a supernatant; add ammonium sulfate (saturation 85%) to the supernatant, stir for 2 h, and centrifuge at 4 °C (5000 r / min, 30 min) to obtain a precipitate; according to the dosage ratio of the precipitate to phosphate buffer of 1 g:20 mL, dissolve the precipitate with a phosphate buffer with a pH of 8, dialyze with a 3500 Da dialysis bag for 48 h, and lyophilize to obtain freeze-dried scallion powder; add a 4% pepsin digestion solution to the freeze-dried scallion powder, adjust the pH to 2, enzymatically hydrolyze in a water bath at 37 °C with shaking for 1 h, then add a 4% trypsin digestion solution, adjust the pH to 7.5, continue to enzymatically hydrolyze in a water bath at 37 °C with shaking for 2 h, after the enzymatic hydrolysis is completed, raise the temperature to 96 °C and react for 5 min to terminate the enzymatic hydrolysis, dialyze with a 2500 Da dialysis bag for 48 h, and lyophilize to obtain the product.

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

[0059] Mix the raw materials evenly according to the above weight ratio, and sterilize to obtain the product.

[0060] (II) Comparative Examples

[0061] Comparative Example 1

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

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

[0064] Comparative Example 2

[0065] Comparative Example 2 provides a preparation. By weight, the preparation consists of the following components: 51 parts of bovine whey protein peptide, 6 parts of 7-hydroxy chromone derivative, 0.02 parts of soybean phospholipid, and 0.03 parts of xanthan gum.

[0066] Comparative Example 2 also provides a preparation method of the above preparation, which is the same as that of Example 1.

[0067] Comparative Example 3

[0068] Comparative Example 3 provides a preparation. By weight, the preparation consists of the following components: 51 parts of milk whey protein peptide, 12 parts of scallion seed protein peptide, 6 parts of 7-hydroxy chromone, 0.02 part of soy lecithin, and 0.03 part of xanthan gum.

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

[0070] Comparative Example 4

[0071] Comparative Example 4 provides a preparation. By weight, the preparation consists of the following components: 51 parts of milk whey protein peptide, 18 parts of vitamin C, 0.02 part of soy lecithin, and 0.03 part of xanthan gum.

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

[0073] (III) 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 then randomly divided into a normal group, a model group, Example 1 - 3 groups, and Comparative Example 1 - 4 groups, with 10 mice in each group.

[0076] 2. Establishment of Immunosuppressed Model

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

[0078] 3. Administration Method

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

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

[0081] Example 1 - 3 groups: gavaged with the protein peptide preparations of Example 1 - 3 groups at a dose of 2 g / kg, once a day for 28 days;

[0082] Comparative Example 1 - 4 groups: gavaged with the preparations of Comparative Example 1 - 4 groups at a dose of 2 g / kg, once a day for 28 days;

[0083] 4. Biochemical Index Detection Method

[0084] 4.1. Serum Hemolysin and IL-6 Contents

[0085] On the 24th day of gavage, 5 mice were randomly selected from each group and sensitized by intraperitoneal injection of 5% SRBC (0.2 mL / mouse). After the last gavage, the mice were fasted for 12 h overnight. The next day, blood was collected from the orbital cavity of the mice. After separating the serum, it was diluted 100 times with normal saline. 1 mL of the diluted serum was mixed with 0.5 mL of 10% SRBC and 0.5 mL of 10% complement serum. At the same time, a blank control tube was set up (adding an equal amount of complement, SRBC and 1 mL of normal saline). The above samples were placed in an incubator at 37 °C for 0.5 h and then the reaction was terminated in a refrigerator at 4 °C. Finally, centrifuged at 2000 r / min for 10 min, 1 mL of the supernatant was taken and placed for 10 min. Using the control tube as a blank to zero, the OD values of each tube were measured at 540 nm, and the results were recorded. The results are shown in Table 1.

[0086] On the night after the last gavage of the remaining 5 mice in each group, the mice were fasted for 12 h. The next day, blood was collected from the orbital cavity, the serum was separated, and 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 detection

[0088] After blood collection, the fasting body weight of the mice in each group was weighed and recorded as the final body weight. Then, the mice in each group were sacrificed by cervical dislocation, the spleen and thymus were removed, washed with normal saline to remove the blood, and dried with filter paper to remove the excess normal saline, and then weighed. The organ index was calculated. The calculation formula of the organ index is:

[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 weighing the organs, the spleens of the mice in each group were ground with a glass slide and made into a cell suspension with normal saline for standby. Take the spleen cell suspension, after filtration, washing, and fixation with 70% cold ethanol, store it at 4 °C for standby. Before loading the machine, add propidium iodide staining solution containing RNase and incubate at 4 °C for 30 min. After incubation, filter and then measure the cell cycle of each group on a flow cytometer. 30,000 cells were obtained for each sample, and the cell cycle was analyzed with MULTICYCLE software. The results are shown in Table 2.

[0092] 5. Result 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 decreased, indicating that an immune hypofunction model of mice was successfully constructed. Compared with the model group, after intragastric administration of the protein peptide preparations in Examples 1-3, the spleen index and thymus index of each mouse were significantly increased, indicating that the protein peptide preparation of the present invention can reduce the damage of immune organs in immune hypofunction mice. Compared with the model group, after intragastric administration of the protein peptide preparations in Examples 1-3, the contents of serum hemolysin and IL-6 in the serum of each mouse were significantly increased, 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 of immune organs in immune hypofunction mice, but also increase the contents of serum hemolysin and IL-6 in mouse splenocytes, thereby improving the body's immune function.

[0099] As can be seen from Table 2, compared with the normal group, the number of splenocytes in the G0+G1 phase of the model group was significantly increased, and the number of splenocytes in the S+G2+M phase was significantly decreased. Compared with the model group, after intragastric administration of the protein peptide preparations in Examples 1-3, the number of splenocytes in the S+G2+M phase was significantly increased. In summary, the protein peptide preparation of the present invention can promote the division and proliferation of splenocytes in immune hypofunction mice.

[0100] Compared with Examples 1-3, in Comparative Example 1, the preparation obtained by replacing scallion seed protein peptide and 7-hydroxy chromone derivative with bovine whey protein peptide, in Comparative Example 2, the preparation obtained by omitting scallion seed protein peptide, in Comparative Example 3, the preparation obtained by replacing 7-hydroxy chromone derivative with 7-hydroxy chromone, and in Comparative Example 4, the preparation obtained by replacing scallion seed protein peptide and 7-hydroxy chromone derivative with vitamin C can also improve the body's immunity, but the effect is not good. In summary, the combined use of scallion seed protein peptide, 7-hydroxy chromone derivative and bovine whey protein peptide can significantly improve the body's immunity.

[0101] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them. The basic principles and main features of the present invention have been described with specific implementation schemes above. On the basis of the present invention, some modifications or replacements can be made, but these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the present invention claimed.

Claims

1. A protein peptide preparation for enhancing immunity, characterized in that: The invention comprises 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:

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 are added to ethyl acetate, and after cooling, glycine is added to react, and intermediate 1 is obtained after purification; (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.

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; and 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 preparation method of the onion protein peptide is as follows: Adding onion powder to petroleum ether for defatting to obtain defatted onion powder; adding the defatted onion powder to phosphate buffer for extraction, filtering to obtain a supernatant; adding ammonium sulfate to the supernatant and stirring for 1 to 2 hours, and obtaining a precipitate by centrifugation; The precipitate is dissolved in phosphate buffer, dialyzed for 36 to 48 hours, and freeze-dried to obtain onion seed freeze-dried powder; pepsin hydrolyzate is added to the onion seed freeze-dried powder, the pH is adjusted to 2, and then enzymolysis is performed for 1 to 2 hours, and then trypsin hydrolyzate is added, the pH is adjusted to 7.5, and then enzymolysis is performed for 2 to 3 hours, the enzymolysis is terminated, the powder is dialyzed for 36 to 48 hours, and then freeze-dried to obtain the powder.

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

9. The protein peptide preparation for enhancing immunity according to claim 1, characterized in that: The stabilizer is selected from one of xanthan gum, apple pectin and gum arabic, and the emulsifier is soybean lecithin or lecithin.

10. The method for preparing the protein peptide preparation for enhancing immunity according to any one of claims 1 to 9, characterized in that: The following steps are involved: According to the above weight ratio, the raw materials are mixed evenly and sterilized.

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