Preparation process of micromolecular cubilose peptide and compound tablet thereof

Abstract

The invention provides a preparation process of micromolecular cubilose peptide and a compound tablet thereof, and belongs to the technical field of protein and peptide. The preparation process of the micromolecular cubilose peptide comprises the following steps: freezing and crushing cubilose; and carrying out compound enzyme enzymolysis. The preparation method comprises the following steps: repeatedly freezing and crushing cubilose to prepare cubilose powder, adding the cubilose powder into deionized water, heating and stewing, and adding a compound enzyme formed by mixing neutral protease and flavourzyme for ultrasonic-assisted enzymolysis to obtain the micromolecular cubilose peptide, when the neutral protease and the flavourzyme are mixed according to the mass ratio of 1: (2.8-3.4) to conduct enzymolysis on the cubilose, hydrolysis can be conducted from multiple positions, smaller peptide fragments are formed, the hydrolysis degree is effectively improved, sialic acid originally combined with protein is also released along with hydrolysis of the protein, and therefore the content of sialic acid in the cubilose is increased. Therefore, the content of sialic acid in the prepared micromolecular cubilose peptide can be effectively improved.

Description

Technical Field

[0001] The invention relates to the technical field of proteins and peptides, and in particular to a preparation process of a small-molecule bird's nest peptide and a composite tablet thereof. Background Art

[0002] As a traditional tonic, bird's nest has a long history of consumption in Asia, especially in China. It is believed to have many health benefits such as beauty, nourishing the lungs, and enhancing immunity. Its nutritional value comes from its rich protein, carbohydrates, minerals, and sialic acid. The traditional way to eat bird's nest is to stew it, which takes a long time. In addition, the digestion and absorption efficiency of large molecular proteins and other components in the bird's nest in the gastrointestinal tract is relatively low. The human digestive system needs to spend a lot of energy and time to break down these large molecules into absorbable small molecular fragments, resulting in some nutrients being excreted from the body before they are fully utilized.

[0003] With the continuous development of biotechnology, enzymatic hydrolysis technology has emerged, providing a new way for the deep processing of bird's nests. By treating the bird's nests with specific enzymes, the large molecular proteins in them can be hydrolyzed into small molecular bird's nest peptides. Compared with large molecular bird's nest proteins, these small molecular bird's nest peptides are more likely to pass through the gastrointestinal mucosa and be quickly absorbed by the human body, greatly improving the bioavailability.

[0004] At present, making bird's nest peptides into oral preparations is one of the popular forms of nutritional supplements, but ordinary bird's nest peptide oral preparations, such as conventional capsules or tablets, have the problem of rapid drug release, which causes the blood drug concentration to rise sharply in a short period of time, and then quickly drop, resulting in a peak-valley effect. On the one hand, this peak-valley effect cannot continuously and stably provide the body with the required bird's nest peptides, making it difficult to fully exert its health care function; on the other hand, excessively high blood drug concentration peaks may cause some potential adverse reactions, such as gastrointestinal discomfort, and the low blood drug concentration period greatly reduces the efficacy of the drug.

[0005] Therefore, it is necessary to propose a preparation process of a small molecule bird's nest peptide with a sustained-release effect and its composite tablets to improve its bioavailability. Summary of the invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a preparation process of a small molecule bird's nest peptide and a composite tablet thereof.

[0007] A small molecule bird's nest peptide, the preparation process of which comprises the following steps: (1) Frozen crushed bird’s nest: Place the bird's nest in a -15 to -5°C freezer for 20 to 30 minutes, then place it in a crusher and crush it for 20 to 30 minutes, repeat 2 to 3 times, and pass it through a 60 to 100 mesh sieve to obtain bird's nest powder; (2) Compound enzyme hydrolysis: The bird's nest powder is added to deionized water at a solid-liquid ratio of 1g: (40-50)mL, heated and stirred at 90-100°C for 1-2h, and when cooled to 45-50°C, a composite enzyme is added, and ultrasonic-assisted enzymolysis is performed for 3-4h, and then the enzyme is inactivated in a boiling water bath for 10-20min, and the small molecule bird's nest peptide is obtained after centrifugal separation, collection of the supernatant and vacuum freeze-drying; The compound enzyme is prepared by mixing neutral protease and flavor protease in a mass ratio of 1:(2.8-3.4), and the added amount of the compound enzyme is 4000-5000U / g of bird's nest powder.

[0008] Furthermore, a process for preparing a composite tablet using the above-mentioned small molecule bird's nest peptide comprises the following steps: S1: Preparation of coating solution The polymethacrylate copolymer and the polylactic acid-glycolic acid copolymer are dissolved in ethyl acetate respectively, and then mixed, homogenized and vacuum degassed to obtain a coating solution; S2: Preparation of extracts The white pine mushroom and fresh sweet osmanthus fragrans were subjected to ultrasonic-assisted extraction using ethanol solution to obtain the white pine mushroom extract and the sweet osmanthus fragrans extract; S3: Preparation of composite tablets Small molecule bird's nest peptide, N-acetylneuraminic acid, xylitol, the above-mentioned white matsutake extract and the above-mentioned osmanthus fragrans extract are made into bird's nest peptide tablets, and then the above-mentioned coating liquid is evenly sprayed on the surface, and after drying, a composite tablet is obtained.

[0009] Furthermore, S1 specifically includes the following steps: S1.1: Add polymethacrylate copolymer into ethyl acetate at a solid-liquid ratio of 1 g: (20-30) mL, stir thoroughly to dissolve, and obtain a polymethacrylate copolymer solution; S1.2: Add polylactic acid-glycolic acid copolymer into ethyl acetate at a solid-liquid ratio of 1 g: (20-30) mL, stir thoroughly to dissolve, and obtain a polylactic acid-glycolic acid copolymer solution; S1.3: The polymethacrylate copolymer solution and the polylactic acid-glycolic acid copolymer solution are stirred and mixed uniformly for 10 to 20 minutes, and vacuum degassing is performed to obtain a coating solution.

[0010] Furthermore, S2 specifically includes the following steps: S2.1: After washing, drying, crushing and sieving the white matsutake, add it into 50% ethanol solution at a solid-liquid ratio of 1 g: (20-30) mL, perform ultrasonic-assisted extraction for 1-2 h, repeat 2-3 times, filter, evaporate and concentrate to obtain the white matsutake extract; S2.2: Wash, dry and crush the fresh Osmanthus fragrans flowers, add them into 70% ethanol solution at a solid-liquid ratio of 1g:(30-40)mL, perform ultrasonic-assisted extraction for 1-2h, repeat twice, filter, concentrate under reduced pressure and dry to obtain Osmanthus fragrans flower extract.

[0011] Furthermore, S3 specifically includes the following steps: S3.1: mixing small molecule bird's nest peptide, N-acetylneuraminic acid, xylitol, the white pine extract obtained in step S2.1, and the sweet osmanthus flower extract obtained in step S2.2 with a filler, a disintegrant, and a binder solution, and then tableting and drying to obtain a bird's nest peptide tablet; S3.2: Place the above-mentioned bird's nest peptide tablets into a rotary coater, spray the coating liquid prepared in step S1.3 evenly on the surface thereof, and after drying and solidification, obtain a composite tablet.

[0012] Further, the polymethacrylate copolymer is any one of Eudragit L30D-55 and Eudragit L100-55.

[0013] Furthermore, the volume ratio of the polymethacrylate copolymer solution to the polylactic acid-glycolic acid copolymer solution is (3-5):1.

[0014] Furthermore, the bird's nest peptide tablets include, by mass: 20 to 30 parts of small molecule bird's nest peptides, 1 to 3 parts of N-acetylneuraminic acid, 6 to 8 parts of xylitol, 8 to 10 parts of white matsutake extract, 4 to 6 parts of osmanthus flower extract, 12 to 16 parts of filler, 0.1 to 0.5 disintegrant and 0.6 to 1 part of binder.

[0015] Furthermore, the filler is any one of microcrystalline cellulose or starch.

[0016] Furthermore, the disintegrant is any one of cross-linked carboxymethylcellulose sodium or cross-linked povidone, and the binder is any one of povidone or hypromellose.

[0017] Compared with the prior art, the present invention has at least the following beneficial effects: 1. The present invention prepares bird's nest powder by repeatedly freezing and crushing the bird's nest, then adds the bird's nest powder into deionized water, heats and stews the bird's nest powder, and then adds a composite enzyme prepared by mixing neutral protease and flavor protease in a mass ratio of 1:(2.8-3.4) for ultrasonic-assisted enzymatic hydrolysis to obtain a small molecule bird's nest peptide. The bird's nest powder can be hydrolyzed from multiple positions to form smaller peptide segments, thereby effectively improving the degree of hydrolysis. As the protein is hydrolyzed, the sialic acid originally bound to the protein is also released, thereby effectively improving the content of sialic acid in the obtained small molecule bird's nest peptide.

[0018] 2. The present invention extracts white pine mushrooms and sweet osmanthus flowers with ethanol solution to obtain white pine mushroom extract and sweet osmanthus flower extract, and then mixes the two with small molecule bird's nest peptide to prepare bird's nest peptide tablets. Since the active ingredients of the white pine mushroom extract can inhibit the activity of tyrosinase and reduce the formation of melanin, the sweet osmanthus flower extract can interfere with the synthesis of melanin by regulating the signal transduction pathway in melanocytes, and the small molecule bird's nest peptide can promote the metabolism of skin cells. When the three are combined, on the one hand, the white pine mushroom extract and the sweet osmanthus flower extract play a role in inhibiting the formation and transport of melanin, and on the other hand, the small molecule bird's nest peptide accelerates the renewal of cells containing melanin to form a synergistic whitening effect. In addition, the small molecule bird's nest peptide contains amino acid residues with antioxidant activity, which can cooperate with the antioxidant components in the white pine mushroom extract and the sweet osmanthus flower extract to play an antioxidant role under different redox potentials, forming a multi-level antioxidant network, thereby achieving a synergistic antioxidant effect.

[0019] 3. The present invention dissolves the polymethacrylate copolymer and the polylactic acid-glycolic acid copolymer in ethyl acetate respectively, and then mixes the two to form a coating liquid. The coating liquid is evenly sprayed on the surface of the bird's nest peptide tablet and dried, so that a barrier can be formed on the surface of the bird's nest peptide tablet to prevent the rapid dissolution of the bird's nest peptide and the extract, slowing down the release rate, thereby reducing the burden on the gastrointestinal tract. In addition, since the structure of the polymethacrylate copolymer contains acidic groups, it is relatively stable in an acidic environment and will not ionize, thereby further reducing the dissolution amount of the bird's nest peptide and the extract in the gastric juice and reducing the damage to it by the gastric juice. In a neutral or weakly alkaline environment, the acidic groups in the structure will ionize, increasing the repulsive force between the polymer chains, resulting in the swelling of the composite coating material, which is beneficial to the release of the bird's nest peptide and the extract in the bird's nest peptide tablet and exerting the efficacy, thereby achieving the effect of improving the bioavailability of small molecule bird's nest peptides, white pine extracts and sweet osmanthus extracts. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the present disclosure and, together with the description, further serve to explain the principles of the present disclosure and to enable those skilled in the relevant art to make and use the present disclosure.

[0021] Figure 1 This is a flow chart of the preparation process of the small molecule bird's nest peptide used in the embodiments of the present invention.

[0022] Figure 2 This is a cumulative release rate curve of the composite tablet prepared in Example 1 of the present invention.

[0023] Figure 3 This is a cumulative release rate curve of the bird's nest peptide tablets prepared in Comparative Example 6 of the present invention. DETAILED DESCRIPTION

[0024] The preparation process of a small molecule bird's nest peptide and its composite tablet provided by the present invention is described in detail below in conjunction with the accompanying drawings and specific examples.

[0025] Example 1 A preparation process of a small molecule bird's nest peptide composite tablet, such as Figure 1 As shown, the following steps are included: (1) Frozen crushed bird’s nest: Place the bird's nest in a -15℃ freezer for 20 minutes, then place it in a crusher and crush it for 20 minutes, repeat twice, and pass it through a 60-mesh sieve to obtain bird's nest powder; (2) Compound enzyme hydrolysis: The bird's nest powder was added to deionized water at a solid-liquid ratio of 1g:40mL, heated and stirred at 90°C for 1h, and when cooled to 45°C, a composite enzyme was added, and ultrasonic-assisted enzymolysis was performed for 3h, and then the enzyme was inactivated in a boiling water bath for 10min. After centrifugation, the supernatant was collected and vacuum freeze-dried to obtain a small molecule bird's nest peptide; The compound enzyme is prepared by mixing neutral protease and flavor protease in a mass ratio of 1:2.8, and the added amount of the compound enzyme is 4000U / g bird's nest powder; S1: Preparation of coating solution S1.1: Add polymethacrylate copolymer into ethyl acetate at a solid-liquid ratio of 1 g:20 mL, stir thoroughly to dissolve, and obtain a polymethacrylate copolymer solution; S1.2: Add polylactic acid-glycolic acid copolymer into ethyl acetate at a solid-liquid ratio of 1 g:20 mL, stir thoroughly to dissolve, and obtain a polylactic acid-glycolic acid copolymer solution; S1.3: The polymethacrylate copolymer solution and the polylactic acid-glycolic acid copolymer solution are stirred and mixed uniformly for 10 minutes, and vacuum degassing is performed to obtain a coating solution; S2: Preparation of extracts S2.1: After washing, drying, crushing and sieving the white pine mushroom, add it into 50% ethanol solution at a solid-liquid ratio of 1 g:20 mL, perform ultrasonic-assisted extraction for 1 h, repeat twice, filter, evaporate and concentrate to obtain the white pine mushroom extract; S2.2: Wash, dry and crush the fresh Osmanthus fragrans, add it to 70% ethanol solution at a solid-liquid ratio of 1 g:30 mL, perform ultrasonic-assisted extraction for 1 h, repeat twice, filter and concentrate under reduced pressure, add petroleum ether, continue to concentrate under reduced pressure and dry to obtain Osmanthus fragrans extract; S3: Preparation of composite tablets S3.1: By mass, 20 parts of small molecule bird's nest peptide, 1 part of N-acetylneuraminic acid, 6 parts of xylitol, 8 parts of white pine extract prepared in step S2.1, 4 parts of sweet osmanthus flower extract prepared in step S2.2, 12 parts of microcrystalline cellulose, 0.1 part of cross-linked sodium carboxymethyl cellulose, 0.6 parts of povidone and water are mixed, and then tableted and dried to obtain bird's nest peptide tablets; S3.2: Place the above-mentioned bird's nest peptide tablets into a rotary coater, spray the coating liquid prepared in step S1.3 evenly on the surface thereof, and after drying and solidification, obtain a composite tablet.

[0026] Example 2 A preparation process of a small molecule bird's nest peptide composite tablet, such as Figure 1 As shown, the following steps are included: (1) Frozen crushed bird’s nest: Place the bird's nest in a -10℃ freezer for 25 minutes, then place it in a crusher and crush it for 25 minutes. Repeat this process twice and pass it through an 80-mesh sieve to obtain bird's nest powder. (2) Compound enzyme hydrolysis: The bird's nest powder was added to deionized water at a solid-liquid ratio of 1g:45mL, heated and stirred at 95°C for 1.5h, and when cooled to 47°C, a composite enzyme was added, and ultrasonic-assisted enzymolysis was performed for 3.5h, and then the enzyme was inactivated in a boiling water bath for 15min. After centrifugation, the supernatant was collected and vacuum freeze-dried to obtain a small molecule bird's nest peptide; The compound enzyme is prepared by mixing neutral protease and flavor protease in a mass ratio of 1:3.1, and the added amount of the compound enzyme is 4500U / g bird's nest powder; S1: Preparation of coating solution S1.1: Add polymethacrylate copolymer into ethyl acetate at a solid-liquid ratio of 1 g:25 mL, stir thoroughly to dissolve, and obtain a polymethacrylate copolymer solution; S1.2: Add polylactic acid-glycolic acid copolymer into ethyl acetate at a solid-liquid ratio of 1 g:25 mL, stir thoroughly to dissolve, and obtain a polylactic acid-glycolic acid copolymer solution; S1.3: The polymethacrylate copolymer solution and the polylactic acid-glycolic acid copolymer solution are stirred and mixed uniformly for 15 minutes, and vacuum degassing is performed to obtain a coating solution; S2: Preparation of extracts S2.1: After washing, drying, crushing and sieving the white pine mushroom, add it into 50% ethanol solution at a solid-liquid ratio of 1 g:25 mL, perform ultrasonic-assisted extraction for 1.5 h, repeat twice, filter, evaporate and concentrate to obtain the white pine mushroom extract; S2.2: Wash, dry and crush the fresh Osmanthus fragrans flowers, add them to 70% ethanol solution at a solid-liquid ratio of 1 g:35 mL, perform ultrasonic-assisted extraction for 1.5 h, repeat twice, filter and concentrate under reduced pressure, add petroleum ether, continue to concentrate under reduced pressure and dry to obtain an Osmanthus fragrans flower extract; S3: Preparation of composite tablets S3.1: By mass, 25 parts of small molecule bird's nest peptide, 2 parts of N-acetylneuraminic acid, 7 parts of xylitol, 9 parts of white pine extract prepared in step S2.1, 5 parts of sweet osmanthus flower extract prepared in step S2.2, 14 parts of starch, 0.3 parts of cross-linked polyvinylpyrrolidone, 0.8 parts of hydroxypropyl methylcellulose and water are mixed, and then tableted and dried to obtain bird's nest peptide tablets; S3.2: Place the above-mentioned bird's nest peptide tablets into a rotary coater, spray the coating liquid prepared in step S1.3 evenly on the surface thereof, and after drying and solidification, obtain a composite tablet.

[0027] Example 3 A preparation process of a small molecule bird's nest peptide composite tablet, such as Figure 1 As shown, the following steps are included: (1) Frozen crushed bird’s nest: Place the bird's nest in a -5℃ freezer for 30 minutes, then place it in a crusher and crush it for 30 minutes, repeat 3 times, and pass it through a 100-mesh sieve to obtain bird's nest powder; (2) Compound enzyme hydrolysis: The bird's nest powder was added to deionized water at a solid-liquid ratio of 1g:50mL, heated and stirred at 100°C for 2h, and when cooled to 50°C, a composite enzyme was added, and ultrasonic-assisted enzymolysis was performed for 4h, and then the enzyme was inactivated in a boiling water bath for 20min. After centrifugation, the supernatant was collected and vacuum freeze-dried to obtain a small molecule bird's nest peptide; The compound enzyme is prepared by mixing neutral protease and flavor protease in a mass ratio of 1:3.4, and the amount of the compound enzyme added is 5000U / g bird's nest powder; S1: Preparation of coating solution S1.1: Add polymethacrylate copolymer into ethyl acetate at a solid-liquid ratio of 1 g:30 mL, stir thoroughly to dissolve, and obtain a polymethacrylate copolymer solution; S1.2: Add polylactic acid-glycolic acid copolymer into ethyl acetate at a solid-liquid ratio of 1 g:30 mL, stir thoroughly to dissolve, and obtain a polylactic acid-glycolic acid copolymer solution; S1.3: The polymethacrylate copolymer solution and the polylactic acid-glycolic acid copolymer solution are stirred and mixed uniformly for 20 minutes, and after vacuum degassing, a coating solution is obtained; S2: Preparation of extracts S2.1: After washing, drying, crushing and sieving the white pine mushroom, add it into 50% ethanol solution at a solid-liquid ratio of 1 g:30 mL, perform ultrasonic-assisted extraction for 2 h, repeat 3 times, filter, evaporate and concentrate to obtain the white pine mushroom extract; S2.2: Wash, dry and crush the fresh Osmanthus fragrans flowers, add them to 70% ethanol solution at a solid-liquid ratio of 1 g:40 mL, perform ultrasonic-assisted extraction for 2 h, repeat twice, filter and concentrate under reduced pressure, add petroleum ether, continue to concentrate under reduced pressure and dry to obtain Osmanthus fragrans flower extract; S3: Preparation of composite tablets S3.1: By mass, 30 parts of small molecule bird's nest peptide, 3 parts of N-acetylneuraminic acid, 8 parts of xylitol, 10 parts of white pine extract prepared in step S2.1, 6 parts of sweet osmanthus flower extract prepared in step S2.2, 16 parts of starch, 0.5 parts of cross-linked sodium carboxymethyl cellulose, 1 part of povidone and water are mixed, and then tableted and dried to obtain bird's nest peptide tablets; S3.2: Place the above-mentioned bird's nest peptide tablets into a rotary coater, spray the coating liquid prepared in step S1.3 evenly on the surface thereof, and after drying and solidification, obtain a composite tablet.

[0028] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that the mass ratio of neutral protease to flavor protease in step (2) is adjusted to 1:2.7.

[0029] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that the mass ratio of neutral protease to flavor protease in step (2) is adjusted to 1:3.5.

[0030] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that the Agaricus rutinus extract and Osmanthus fragrans extract in step S3.1 are replaced by an equal amount of small molecule bird's nest peptides.

[0031] Comparative Example 4 The difference between this comparative example 4 and example 1 is that the small molecule bird's nest peptide and osmanthus fragrans extract in step S3.1 are replaced by an equal amount of white matsutake extract.

[0032] Comparative Example 5 The difference between this comparative example 5 and example 1 is that the white matsutake extract and small molecule bird's nest peptide in step S3.1 are replaced by an equal amount of osmanthus fragrans extract.

[0033] Comparative Example 6 The difference between Comparative Example 6 and Example 1 is that step S3.2 is removed.

[0034] Test Case Test 1: The content of sialic acid in the small molecule bird's nest peptides prepared in Example 1-3 and Comparative Example 1-2 was determined by high performance liquid chromatography. The results are shown in Table 1.

[0035] Table 1: Comparison of test results of bird's nest acid content in small molecule bird's nest peptides

[0036] It can be seen from Table 1 that after the mass ratio of neutral protease to flavor protease in Comparative Examples 1 and 2 was adjusted to 1:2.7 and 1:3.5, the sialic acid content in the obtained small molecule bird's nest peptide was significantly lower than that in Examples 1-3. It can be seen that when the neutral protease and flavor protease are mixed in a mass ratio of 1: (2.8-3.4) to enzymatically hydrolyze the bird's nest, the degree of hydrolysis can be effectively improved, thereby effectively increasing the content of sialic acid in the obtained small molecule bird's nest peptide.

[0037] Test 2: Take the bird's nest peptide tablets prepared in Examples 1-3 and Comparative Examples 3-5 and dissolve them in deionized water to prepare a bird's nest peptide solution with a mass fraction of 0.05%, then take 1 mL in a test tube, add 1 mL of phosphate buffer (pH 6.8) and 1 mL of 0.03% tyrosine solution to each test tube, then place it in a 37°C water bath for 10 minutes, add 1 mL of tyrosinase solution, mix well and let stand for 25 minutes, measure the absorbance value (A) at a wavelength of 475 nm with a spectrophotometer, and replace the tyrosinase with an equal volume of phosphate buffer to measure the absorbance value (B) at a wavelength of 475 nm; replace the bird's nest peptide solution prepared in Examples 1-3 and Comparative Examples 3-5 with an equal volume of distilled water to measure the absorbance value (C) at a wavelength of 475 nm, and replace the tyrosinase solution with an equal volume of phosphate buffer to measure the absorbance value (D) at a wavelength of 475 nm. Calculate the inhibition rate of the sample on tyrosinase, and the calculation formula is as follows: Inhibition rate (%) = [(CD) - (AB)] / (CD) × 100%. The results are shown in Table 2.

[0038] Table 2: Comparison of tyrosinase inhibition test results

[0039] As can be seen from Table 2, the inhibition rates of tyrosinase in the tablets prepared by using a single small molecule bird's nest peptide, a single white pine mushroom extract and a single osmanthus flower extract in Comparative Examples 3, 4 and 5 are all lower than those in Examples 1-3. It can be seen that the three have a synergistic effect of inhibiting tyrosinase activity, that is, the small molecule bird's nest peptide, the white pine mushroom extract and the osmanthus flower extract have a synergistic whitening effect.

[0040] Test 3: Take the bird's nest peptide tablets prepared in Examples 1-3 and Comparative Examples 3-5 and dissolve them in deionized water to prepare a sample solution with a mass fraction of 0.05%, then take 1 mL of the sample solution in a test tube, add 5 mL of 0.06 mmol / LDPPH ethanol solution, mix well and react for 30 minutes, and measure the absorbance A1 at a wavelength of 517 nm; replace the sample solution with pure water and measure the absorbance A0; replace the DPPH ethanol solution with anhydrous ethanol and measure the absorbance A2. Calculate the DPPH clearance rate, the formula is clearance rate = [A0-(A1-A2)] × 100%, the results are shown in Table 3.

[0041] Table 3: Comparison of DPPH clearance test results

[0042] As can be seen from Table 3, the scavenging rates of DPPH obtained in Comparative Examples 3, 4 and 5 are all lower than those in Examples 1-3. It can be seen that the small molecule bird's nest peptide, white matsutake extract and osmanthus fragrans extract have a synergistic free radical scavenging effect, that is, the three have a synergistic antioxidant effect.

[0043] Test 4: The composite tablets prepared in Example 1 and Comparative Example 6 were placed in the original release solution (10 mL of pH 7.4 PBS buffer and 10 mL of pH 1.5 hydrochloric acid solution), respectively, and shaken at a low speed at 37°C in a constant temperature water bath shaker. 1 mL of the release solution was taken out at 2, 4, 6, 8, 10, 12, 14, 16, and 18 hours, and an equal amount of the original release solution was added at the same time. The cumulative release rate curve was measured and plotted. The results are as follows: Figure 2 and Figure 3 shown.

[0044] from Figure 2 It can be seen that the composite tablets prepared in Example 1 have sustained release properties, while Figure 3 The release of the bird's nest peptide tablets prepared in Comparative Example 6 is not sustained-release, and it is almost completely released within 2 hours. In addition, the cumulative release rate of the composite tablets prepared in Example 1 at pH = 7.4 is significantly higher than the cumulative release rate at pH = 1.5. It can be seen that after the composite coating material is coated on the surface of the bird's nest peptide tablets, it can reduce the release of the bird's nest peptide tablets in the acidic environment of gastric juice, improve its stability, and thereby improve the bioavailability of the bird's nest peptide tablets.

[0045] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical ideas disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A small molecule bird's nest peptide, characterized in that: The preparation process includes the following steps: (1) Frozen crushed bird’s nest: Place the bird's nest in a -15 to -5°C freezer for 20 to 30 minutes, then place it in a crusher and crush it for 20 to 30 minutes, repeat 2 to 3 times, and pass it through a 60 to 100 mesh sieve to obtain bird's nest powder; (2) Compound enzyme hydrolysis: The bird's nest powder is added to deionized water at a solid-liquid ratio of 1g: (40-50)mL, heated and stirred at 90-100°C for 1-2h, and when cooled to 45-50°C, a composite enzyme is added, and ultrasonic-assisted enzymolysis is performed for 3-4h, and then the enzyme is inactivated in a boiling water bath for 10-20min, and the small molecule bird's nest peptide is obtained after centrifugal separation, collection of the supernatant and vacuum freeze-drying; The compound enzyme is prepared by mixing neutral protease and flavor protease in a mass ratio of 1:(2.8-3.4), and the added amount of the compound enzyme is 4000-5000U / g of bird's nest powder.

2. A process for preparing composite tablets using the small molecule bird's nest peptide according to claim 1, characterized in that: The steps include: S1: Preparation of coating solution The polymethacrylate copolymer and the polylactic acid-glycolic acid copolymer are dissolved in ethyl acetate respectively, and then mixed, homogenized and vacuum degassed to obtain a coating solution; S2: Preparation of extracts The white pine mushroom and fresh sweet osmanthus fragrans were subjected to ultrasonic-assisted extraction using ethanol solution to obtain the white pine mushroom extract and the sweet osmanthus fragrans extract; S3: Preparation of composite tablets Small molecule bird's nest peptide, N-acetylneuraminic acid, xylitol, the above-mentioned white matsutake extract and the above-mentioned osmanthus fragrans extract are made into bird's nest peptide tablets, and then the above-mentioned coating liquid is evenly sprayed on the surface, and after drying, a composite tablet is obtained.

3. The process for preparing a composite tablet according to claim 2, characterized in that: S1 specifically includes the following steps: S1.1: Add polymethacrylate copolymer into ethyl acetate at a solid-liquid ratio of 1 g: (20-30) mL, stir thoroughly to dissolve, and obtain a polymethacrylate copolymer solution; S1.2: Add polylactic acid-glycolic acid copolymer into ethyl acetate at a solid-liquid ratio of 1 g: (20-30) mL, stir thoroughly to dissolve, and obtain a polylactic acid-glycolic acid copolymer solution; S1.3: The polymethacrylate copolymer solution and the polylactic acid-glycolic acid copolymer solution are stirred and mixed uniformly for 10 to 20 minutes, and vacuum degassing is performed to obtain a coating solution.

4. The process for preparing a composite tablet according to claim 3, characterized in that: S2 specifically includes the following steps: S2.1: After washing, drying, crushing and sieving the white matsutake, add it into 50% ethanol solution at a solid-liquid ratio of 1 g: (20-30) mL, perform ultrasonic-assisted extraction for 1-2 h, repeat 2-3 times, filter, evaporate and concentrate to obtain the white matsutake extract; S2.2: Wash, dry and crush the fresh Osmanthus fragrans flowers, add them into 70% ethanol solution at a solid-liquid ratio of 1g:(30-40)mL, perform ultrasonic-assisted extraction for 1-2h, repeat twice, filter, concentrate under reduced pressure and dry to obtain Osmanthus fragrans flower extract.

5. The process for preparing a composite tablet according to claim 4, characterized in that: S3 specifically includes the following steps: S3.1: mixing small molecule bird's nest peptide, N-acetylneuraminic acid, xylitol, the white pine extract obtained in step S2.1, and the sweet osmanthus flower extract obtained in step S2.2 with a filler, a disintegrant, and a binder solution, and then tableting and drying to obtain a bird's nest peptide tablet; S3.2: Place the above-mentioned bird's nest peptide tablets into a rotary coater, spray the coating liquid prepared in step S1.3 evenly on the surface thereof, and after drying and solidification, obtain a composite tablet.

6. The process for preparing a composite tablet according to claim 3, characterized in that: The polymethacrylate copolymer is any one of Eudragit L30D-55 and Eudragit L100-55.

7. The process for preparing a composite tablet according to claim 3, characterized in that: The volume ratio of the polymethacrylate copolymer solution to the polylactic acid-glycolic acid copolymer solution is (3-5):

1.

8. The process for preparing a composite tablet according to claim 5, characterized in that: By mass, the bird's nest peptide tablets include: 20 to 30 parts of small molecule bird's nest peptides, 1 to 3 parts of N-acetylneuraminic acid, 6 to 8 parts of xylitol, 8 to 10 parts of white pine mushroom extract, 4 to 6 parts of osmanthus flower extract, 12 to 16 parts of filler, 0.1 to 0.5 disintegrant and 0.6 to 1 part of binder.

9. The process for preparing a composite tablet according to claim 8, characterized in that: The filler is either microcrystalline cellulose or starch.

10. The process for preparing a composite tablet according to claim 8, characterized in that: The disintegrant is any one of cross-linked carboxymethylcellulose sodium or cross-linked povidone, and the binder is any one of povidone or hypromellose.

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