A plant-derived small molecule peptide composition for promoting the production of natural bezoar

Abstract

The present invention belongs to the field of biomedicine, and specifically relates to a plant-based small molecule peptide composition for promoting the production of natural bezoar. Small molecule peptides are extracted from Centella asiatica, and five small molecule peptide sequences F, G, H, I, and J are determined through raw material collection and processing, low-temperature drying and crushing, multi-condition extraction, activity screening, peptide component separation, and sequencing analysis. Experiments show that these peptide sequences and their compositions can significantly promote the production of natural bezoar, increase the formation rate and yield of bezoar, and reduce gallbladder inflammation. Compared with the prior art, the present invention has the advantages of high efficiency, safety, strong specificity, and high cost-effectiveness.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a technology and application of extracting a small molecule peptide sequence composition from Centella asiatica for promoting the production of natural bezoar. Background Art

[0002] As a treasure among traditional Chinese medicines, natural bezoar has extremely important medicinal value. It has many functions such as clearing the heart, removing phlegm, opening the orifices, cooling the liver, calming the wind, and detoxifying, and performs well in treating a variety of difficult diseases. For example, in Angong Niuhuang Pills, natural bezoar plays a key role and has significant therapeutic effects on severe diseases such as fever and coma; Niuhuang Jiedu Tablets use it as the main ingredient and have a good effect in relieving symptoms such as sore throat and mouth sores.

[0003] With the widespread spread of traditional Chinese medicine around the world and people's increasing attention to health, the market demand for natural bezoar has shown a spurt of growth. However, the probability of natural bezoar being formed naturally is extremely low. According to statistics, only about 1-2 cows out of every 100,000 cows can produce natural bezoar, and the output is extremely rare, which has caused its price to continue to soar, and the market has been in a state of tension with supply and demand for a long time. The high price has greatly increased the production cost of Chinese patent medicines containing natural bezoar, seriously restricting the clinical application and market promotion of these drugs.

[0004] At present, the production technology of natural bezoar has obvious deficiencies. Although the gallbladder implantation foreign body method attempts to stimulate the formation of bezoar by implanting foreign bodies into the gallbladder of cattle, this method is very likely to cause serious complications such as gallbladder infection and inflammation in cattle, which will not only lead to a sharp deterioration in the health of cattle, but may even cause the death of cattle. Moreover, the quality of bezoar produced by this method is uneven, and the content of effective ingredients varies greatly, making it difficult to meet the strict medicinal standards of natural bezoar. Although the in vitro cultivation method can simulate the internal environment of cattle to a certain extent to cultivate bezoar, and has a certain control ability over the quality of bezoar, this technology has extremely high requirements for equipment and technology, huge initial equipment investment, expensive production costs, and a long cultivation cycle, which cannot meet the growing demand for large-scale production.

[0005] Therefore, it is urgent to develop an efficient, safe and low-cost technology to promote the production of natural bezoar, which has important practical significance for meeting market demand and promoting the development of traditional Chinese medicine industry. Summary of the invention The present invention provides a small molecule peptide composition for promoting the production of natural bezoar, comprising the following small molecule peptide sequence:

[0006] Peptide sequence F: Ala - Thr - Glu - Lys - Pro - Arg - Ser - Tyr - Asn - Ile -Val - Leu - Phe - Met - Gly; Peptide sequence G: Gly - Glu - Asp - His - Pro - Ser - Thr - Val - Ile - Leu -Phe - Tyr - Trp - Lys - Arg; Peptide sequence H: Thr - Lys - Arg - Glu - Asp - Pro - Gly - Ser - Ile - Val -Leu - Phe - Tyr - Trp - His - Ala; Peptide sequence I: Pro - Arg - Lys - Glu - Asp - Gly - Ser - Thr - Val - Ile -Leu - Phe - Tyr - Trp - Asn - Gln; Peptide sequence J: Ser - Thr - Glu - Lys - Arg - Pro - Gly - Asp - Ile - Val - Leu - Phe - Tyr - Trp - His - Glu, The small molecule peptide sequence is derived from the extract of Centella asiatica.

[0007] Furthermore, the content ratio of each peptide sequence in the small molecule peptide composition is: peptide sequence F accounts for 10%-30%, peptide sequence G accounts for 10%-30%, peptide sequence H accounts for 10%-30%, peptide sequence I accounts for 10%-30%, and peptide sequence J accounts for 10%-30%.

[0008] Furthermore, the method of preparing the small molecule peptide composition is characterized by comprising the following steps: Collect the aboveground part of the Centella asiatica that is growing vigorously and free of pests and diseases, rinse it thoroughly with clean water to remove the surface soil, impurities and microorganisms, and cut it into small sections of 2-3 cm; Using vacuum freeze drying technology, the small pieces of Centella asiatica are dried at -40°C to -20°C to a moisture content of less than 8%, and then crushed into coarse powder that can pass through a 80-100 mesh sieve; Using organic solvents such as ethanol, methanol, ethyl acetate and mixed solvents in different proportions, the crude powder was subjected to extraction tests by changing the extraction temperature (35°C - 55°C), time (15 - 30 hours), and solid-liquid ratio (1:6 (g / mL) - 1:10 (g / mL)), and the extract was collected by centrifugation or filtration, and concentrated under reduced pressure to obtain an extract concentrate; The extract concentrate is initially separated by a macroporous adsorption resin column, and a suitable macroporous adsorption resin is selected according to the polarity and molecular size of the peptide components. The extract is gradient eluted with ethanol solutions of different concentrations, and the eluate that may contain peptide components is collected; The above eluate was further separated by gel filtration chromatography, using gel media such as Sephadex G-75 and SephacrylS-150, and the peptides were separated according to their molecular size differences. The eluate at different time periods was collected for preliminary grouping. The grouped peptide components are separated again by ion exchange chromatography. According to the charge properties and quantity of peptide molecules, cation exchange resin SP-Sepharose or anion exchange resin Q-Sepharose is selected, the pH value and ionic strength of the eluent are adjusted, and different elution peak solutions are collected to obtain relatively purified peptide components. The relatively purified peptide components were sequenced by Edman degradation or mass spectrometry sequencing, and the peptide sequences F, G, H, I, and J were determined by bioinformatics analysis, and then mixed to obtain a small molecule peptide composition.

[0009] Furthermore, in the preliminary separation step using a macroporous adsorption resin column, a polar macroporous adsorption resin is selected for peptide components with a relatively high polarity, and a non-polar macroporous adsorption resin is selected for non-polar or weakly polar peptide components.

[0010] Furthermore, in mass spectrometry sequencing, peptide samples are first pre-treated by desalting and concentration, and then injected into the mass spectrometer. Electrospray ionization (ESI) or matrix-assisted laser desorption ionization (MALDI) is selected for ionization, and professional software is used to analyze the mass spectrometry data.

[0011] Furthermore, the small molecule peptide composition is used to promote the production of natural bezoar.

[0012] Furthermore, the small molecule peptide composition of claim 1 is formulated into a solution and administered to cattle by intravenous injection at a dose of 3 mg / kg body weight, twice a week for 10 consecutive weeks.

[0013] A method for promoting the production of natural bezoar, characterized in that chitosan with good biocompatibility and biodegradability is selected to prepare a microsphere carrier with a diameter of 1-2 mm, the small molecule peptide sequence described in claim 1 is loaded separately or mixed onto the microsphere carrier, the microsphere carrier loaded with the small molecule peptide sequence or the combination is surgically implanted near the gallbladder of the cattle, the experimental cattle are carefully cared for after the operation, and if necessary, the small molecule peptide obtained by extraction and purification is formulated into a solution, and the drug is administered by intravenous injection at a dose of 3 mg / kg body weight twice a week for 10 consecutive weeks.

[0014] Furthermore, the small molecule peptide sequence is loaded onto the microsphere carrier by physical adsorption or chemical coupling. The physical adsorption method mixes the microsphere carrier with the small molecule peptide solution for a certain period of time under appropriate temperature and stirring conditions. The chemical coupling method utilizes the chemical reaction between the surface active groups of the carrier and the specific functional groups of the small molecule peptide to achieve covalent connection.

[0015] Furthermore, the pharmaceutical preparation also contains a pharmaceutically acceptable carrier, excipient or diluent, and the dosage form is an injection, capsule, tablet or granule.

[0016] Beneficial effects: High efficiency: It can significantly improve the formation rate and yield of natural bezoar. The experimental results show that after using the small molecule peptide sequence and composition of the present invention, the formation rate of bezoar increased from 20% of the control group to 65% - 70%, and the average weight also increased significantly, especially the small molecule peptide combination group had a more prominent effect.

[0017] Safety: The small molecule peptide extracted from plants is combined with a biodegradable carrier with good biocompatibility, which has no obvious adverse effects on the health of cattle and avoids serious complications such as infection and inflammation that may be caused by traditional methods such as gallbladder implantation. The carrier gradually degrades in the body and will not cause long-term potential harm to animals.

[0018] Specificity: Through target design, small molecule peptides can specifically act on key targets related to the formation of bezoar, such as enzymes and receptors, improving the pertinence and effectiveness of the action. Different small molecule peptide sequences play a role in promoting bile acid synthesis, bilirubin uptake, gallbladder contraction and other different links, and work together to comprehensively promote the formation of bezoar.

[0019] Cost-effectiveness: The extraction process is relatively simple, the raw materials of Centella asiatica are widely available, and the production cost is low. The price of the carrier material is relatively low, the preparation process is not complicated, and it has good economic benefits and market application prospects. Compared with the in vitro cultivation method, it does not require high equipment investment and complex technical operations, which reduces the production threshold. DETAILED DESCRIPTION Example 1

[0020] Preparation of Centella asiatica extract Raw material collection and processing: Carefully select Centella asiatica plants that are growing vigorously and free of pests and diseases. Collect the above-ground parts, and ensure the integrity of the plants during the collection process. Rinse the collected Centella asiatica quickly and thoroughly with clean water to remove the dirt, impurities and various microorganisms attached to the surface. After washing, cut it into small pieces about 2-3 cm in length for subsequent processing.

[0021] Drying and crushing: Using vacuum freeze drying technology, small segments of Centella asiatica are dried at a low temperature of -40℃ to -20℃ to reduce the water content to less than 8%. This drying method can retain the biologically active ingredients in Centella asiatica to the greatest extent. The dried Centella asiatica is processed by crushing equipment to make coarse powder, and its particle size is controlled to be able to pass through an 80-100 mesh screen to ensure that the coarse powder can fully contact with the extraction solvent during the subsequent extraction process, thereby improving the extraction efficiency.

[0022] Extraction process optimization: Extraction experiments were conducted on the crude powder of Centella asiatica using a variety of organic solvents (such as ethanol, methanol, ethyl acetate, etc.) and mixed solvents in different proportions. Multiple sets of parallel experiments were designed by changing key conditions such as extraction temperature, time, and solid-liquid ratio. For example, the ethanol concentration was set to 30%, 40%, 50%, 60%, and 70%, the solid-liquid ratio was from 1:6 (g / mL) to 1:10 (g / mL), the extraction time was from 15 hours to 30 hours, and the extraction temperature was from 35°C to 55°C. After the extraction was completed, the extract was collected by centrifugation or filtration, and the extract was concentrated under reduced pressure using a rotary evaporator or other equipment to obtain the Centella asiatica extract concentrate under different conditions.

[0023] Activity screening Establishment of in vitro cell model: Based on hepatocytes, a cell culture system was constructed that can accurately simulate the bile acid synthesis, bilirubin uptake and other processes closely related to the formation of bezoar. The concentrated extracts of Centella asiatica obtained under different conditions were applied to the cell model. When studying the activity related to bile acid synthesis, the activity changes of intracellular cholesterol 7α-hydroxylase (CYP7A1) were focused on; when studying the activity related to bilirubin uptake, key indicators such as the changes in the cell's uptake of bilirubin were carefully observed. By comparing the differences in cell-related indicators after the action of different extracts, extracts with potential activity to promote the formation of bezoar were screened out.

[0024] Animal pre-experiment: Select a small number of healthy experimental animals (such as mice or rats) and administer the selected Centella asiatica extracts with potential activity to the animals through appropriate administration routes (such as intraperitoneal injection, oral gavage, etc.). During the administration period, closely observe the physiological state and behavioral changes of the animals to ensure that the extracts have no obvious toxicity. At the same time, regularly collect blood, bile and other samples from the animals to test biochemical indicators related to the formation of bezoars, such as bile acid content and bilirubin level. Based on the results of the animal pre-experiment, further determine the Centella asiatica extracts with significant activity in promoting the formation of bezoars.

[0025] Peptide component separation Preliminary separation using macroporous adsorption resin column: The Centella asiatica extracts that have been screened for activity are initially separated using macroporous adsorption resin columns. According to different polarities and molecular sizes, the appropriate type of macroporous adsorption resin is accurately selected. For peptide components with greater polarity, polar macroporous adsorption resins are selected; for non-polar or weakly polar peptide components, non-polar macroporous adsorption resins are selected. Gradient elution is performed using ethanol solutions of different concentrations, and solutions of different elution peaks are collected to preliminarily separate the eluent that may contain peptide components.

[0026] Gel filtration chromatography separation: The eluate obtained from the preliminary separation that may contain peptide components is further separated by gel filtration chromatography. Select a suitable gel medium (such as Sephadex G-75, Sephacryl S-150, etc.) and separate according to the size difference of peptide molecules. Smaller molecular peptides will diffuse in the pores of gel particles and take a longer elution time; larger molecular peptides cannot enter the pores of gel particles and take a shorter elution time. By collecting eluates at different time periods, the peptide components are preliminarily grouped according to molecular size.

[0027] Further separation by ion exchange chromatography: The different groups of peptide components obtained by gel filtration chromatography are further separated by ion exchange chromatography. According to the nature and quantity of the charge carried by the peptide molecules, a suitable ion exchange medium (such as cation exchange resin SP-Sepharose, anion exchange resin Q-Sepharose, etc.) is selected. By adjusting the pH value and ionic strength of the eluent, peptide components with different charge properties and quantities are separated on the ion exchange column, and the solutions corresponding to different elution peaks are collected to obtain relatively purified peptide components.

[0028] Sequencing analysis Application of sequencing technology: Advanced sequencing technology is used to sequence the relatively purified peptide components obtained through the above separation steps. Currently, the commonly used peptide sequencing technologies include Edman degradation and mass spectrometry sequencing. Edman degradation is to determine the sequence of the peptide by gradually degrading and identifying the amino acid residues at the N-terminus of the peptide chain. Mass spectrometry sequencing is to use a mass spectrometer to ionize peptide molecules and infer the sequence of the peptide by analyzing information such as the mass-to-charge ratio of the ions.

[0029] Mass spectrometry sequencing process: When using mass spectrometry sequencing, the peptide samples are first pretreated to make them suitable for mass spectrometry analysis. For example, the purity and concentration of the sample are improved by desalting and concentration. The treated sample is then injected into the mass spectrometer and a suitable ionization method is selected (such as electrospray ionization ESI or matrix-assisted laser desorption ionization MALDI, etc.). After obtaining the mass spectrum, the mass spectrometry data is analyzed using professional data analysis software. By analyzing the parent ion mass, fragment ion mass and other information of the peptide segment, combined with the known amino acid mass database, the amino acid sequence of the peptide molecule can be inferred.

[0030] Bioinformatics analysis: Bioinformatics analysis was performed on the peptide sequences obtained by sequencing. By comparing with known protein and peptide sequence databases (such as NCBI's protein database, etc.), it was determined whether it was a new sequence. At the same time, the structural characteristics of the peptide sequence, such as amino acid composition, hydrophilicity, and secondary structure prediction, were analyzed to provide a basis for subsequent research on its function and mechanism of action. After a lot of screening and sequencing work, the following 5 unreported small molecule peptide sequences were finally determined: Peptide sequence F: Ala - Thr - Glu - Lys - Pro - Arg - Ser - Tyr - Asn - Ile -Val - Leu - Phe - Met - Gly Peptide sequence G: Gly - Glu - Asp - His - Pro - Ser - Thr - Val - Ile - Leu - Phe - Tyr - Trp - Lys - Arg Peptide sequence H: Thr - Lys - Arg - Glu - Asp - Pro - Gly - Ser - Ile - Val - Leu - Phe - Tyr - Trp - His - Ala Peptide sequence I: Pro - Arg - Lys - Glu - Asp - Gly - Ser - Thr - Val - Ile -Leu - Phe - Tyr - Trp - Asn - Gln Peptide sequence J: Ser - Thr - Glu - Lys - Arg - Pro - Gly - Asp - Ile - Val - Leu - Phe - Tyr - Trp - His - Glu Carrier implantation and small molecule peptide combination synergistic effect Carrier selection and preparation: Select biocompatible and degradable materials, such as chitosan, to prepare microsphere carriers with a diameter of about 1-2 mm. Load the above small molecule peptide sequences separately or mixed onto the microsphere carrier. The loading process uses physical adsorption or chemical coupling methods to ensure that the small molecule peptide can be stably bound to the surface or inside of the carrier. For example, for the physical adsorption method, the microsphere carrier and the small molecule peptide solution can be mixed for a certain period of time under appropriate temperature and stirring conditions to allow the small molecule peptide to adsorb to the carrier surface; for the chemical coupling method, the active groups on the carrier surface can be used to react chemically with the specific functional groups of the small molecule peptide to achieve covalent bonding.

[0031] Synergistic Experimental Design: Experimental animal grouping: 80 healthy Simmental cattle weighing between 400 and 500 kg were selected and randomly divided into 8 groups, with 10 in each group. They were control group, vehicle control group, peptide sequence F group, peptide sequence G group, peptide sequence H group, peptide sequence I group, peptide sequence J group and small molecule peptide combination (F + G + H + I + J) group.

[0032] Experimental methods: Carrier control group: The microsphere carrier without small molecule peptide loading was surgically implanted near the gallbladder of the cow, and the surgical procedure strictly followed the aseptic operation specifications.

[0033] Peptide sequence group and combination group: The microsphere carrier loaded with the corresponding small molecule peptide sequence or small molecule peptide combination was also implanted near the gallbladder of the cattle through surgery. After the surgery, all experimental cattle were carefully cared for to ensure that they recovered to normal physiological state.

[0034] Supplemental administration (if necessary): To further enhance the effect, for the peptide sequence group and the combination group, after the vector is implanted, the extracted and purified small molecule peptides can be formulated into a solution and administered by intravenous injection. The dose is 3 mg / kg body weight, twice a week, for 10 consecutive weeks. The control group and the vector control group were injected with an equal amount of normal saline.

[0035] Sample collection and testing: During the experiment, blood and bile samples were collected every 3 weeks to detect the activity of related enzymes in the blood and the content of bile acid, bilirubin and other components in the bile.

[0036] After the experiment, the experimental cows were slaughtered, the gallbladders were removed, the formation of bezoars was carefully observed, the weight and size of the bezoars were accurately measured, and a comprehensive chemical composition analysis was conducted.

[0037] Part of the gallbladder tissue was taken for pathological section examination to observe the inflammation of the gallbladder tissue and the compatibility of the carrier with the tissue.

[0038] Experimental Results Changes in blood and bile composition Peptide sequence F group: After 3 weeks of administration, the activity of CYP7A1 in the blood began to increase slowly, and increased by 35% compared with the control group at 10 weeks. The content of bile acid in bile began to increase significantly after 6 weeks of administration, and increased by 30% compared with the control group at 10 weeks.

[0039] Peptide sequence G group: After 3 weeks of administration, the liver cells' ability to absorb bilirubin began to increase, and the bilirubin content in the blood gradually decreased, and was 38% lower than the control group at 10 weeks. The bilirubin content in bile increased significantly after 6 weeks of administration, and was 35% higher than the control group at 10 weeks.

[0040] Peptide sequence H group: After 3 weeks of administration, the frequency of gallbladder contraction began to increase, and increased by 25% at 6 weeks and 30% at 10 weeks compared with the control group. Bile excretion increased significantly after 6 weeks of administration and increased by 35% at 10 weeks compared with the control group.

[0041] Peptide sequence I group: After 3 weeks of administration, the oxidation decomposition rate of bilirubin in bile began to decrease, and it decreased by 32% at 6 weeks and 40% at 10 weeks compared with the control group. The content of bilirubin in bile increased significantly after 6 weeks of administration, and increased by 32% at 10 weeks compared with the control group.

[0042] Peptide sequence J group: After 3 weeks of administration, the levels of TNF-α and IL-6 in gallbladder tissue began to decrease, and at 6 weeks they were 25% and 20% lower than those in the control group, respectively, and at 10 weeks they were 40% and 35% lower, respectively.

[0043] Small molecule peptide combination group: CYP7A1 activity in the blood increased rapidly after 3 weeks of administration, and increased by 50% compared with the control group at 10 weeks. The liver cell's ability to absorb bilirubin was significantly enhanced, and the bilirubin content in the blood was reduced by 50% compared with the control group at 10 weeks. The frequency of gallbladder contraction increased by 40% compared with the control group at 10 weeks, bile excretion increased by 45%, bilirubin oxidation decomposition rate decreased by 50%, and TNF-α and IL-6 contents in gallbladder tissue decreased by 50% and 45%, respectively.

[0044] Bezoar formation Control group: Only 2 cows formed bezoar, with an average weight of 2g.

[0045] Vehicle control group: 3 cows formed bezoars with an average weight of 2.5 g.

[0046] Peptide sequence group F: 6 cows formed bezoars with an average weight of 4.5 g and a maximum weight of 6 g.

[0047] Peptide sequence group G: 7 cows formed bezoars with an average weight of 4.8 g and a maximum weight of 7 g.

[0048] Peptide sequence group H: Six cows formed bezoars with an average weight of 4.6 g and a maximum weight of 6.5 g.

[0049] Peptide sequence group I: 7 cows formed bezoars with an average weight of 4.7 g and a maximum weight of 7.2 g.

[0050] Peptide sequence group J: 8 cows formed bezoars with an average weight of 5 g and a maximum weight of 8 g.

[0051] Small molecule peptide combination group: 9 cows formed bezoars, with an average weight of 6.5g and a maximum weight of 10g.

[0052] Gallbladder histopathological examination results Control group: There was obvious inflammatory cell infiltration in the gallbladder tissue and the gallbladder mucosa was severely damaged.

[0053] Vector control group: Gallbladder tissue inflammation was mild, and a small amount of fibrous tissue was wrapped around the microsphere carrier, which did not significantly affect the normal function of the gallbladder.

[0054] Peptide sequence groups F, G, H, I and J: Gallbladder tissue inflammation was mild, gallbladder mucosa was basically intact, and microsphere carriers had good compatibility with tissues.

[0055] Small molecule peptide combination group: gallbladder tissue inflammation was the mildest, gallbladder mucosa was smooth, tissue structure was normal, and tissue reaction around the microsphere carrier was minimal.

[0056] Experimental conclusion The five small molecule peptide sequences extracted from Centella asiatica and the compositions composed of them can significantly promote the production of natural bezoar and effectively improve the yield and quality of bezoar. Among them, the small molecule peptide composition has the most significant effect. It can not only continuously release small molecule peptides through implanted carriers, but also more effectively reduce gallbladder inflammation under the synergistic effect of auxiliary intravenous administration, and create a good internal environment for the formation of bezoar. At the same time, the composition can comprehensively promote the processes related to the formation of bezoar, such as bile acid synthesis, bilirubin uptake and transport, and gallbladder contraction, thereby greatly improving the formation rate and quality of bezoar.

Claims

1. A small molecule peptide composition for promoting the production of natural bezoar, characterized in that: Contains the following small molecule peptide sequences: Peptide sequence F: Ala - Thr - Glu - Lys - Pro - Arg - Ser - Tyr - Asn - Ile - Val- Leu - Phe - Met - Gly; Peptide sequence G: Gly - Glu - Asp - His - Pro - Ser - Thr - Val - Ile - Leu - Phe- Tyr - Trp - Lys - Arg; Peptide sequence H: Thr - Lys - Arg - Glu - Asp - Pro - Gly - Ser - Ile - Val - Leu- Phe - Tyr - Trp - His - Ala; Peptide sequence I: Pro - Arg - Lys - Glu - Asp - Gly - Ser - Thr - Val - Ile - Leu- Phe - Tyr - Trp - Asn - Gln; Peptide sequence J: Ser - Thr - Glu - Lys - Arg - Pro - Gly - Asp - Ile - Val - Leu - Phe - Tyr - Trp - His - Glu, The small molecule peptide sequence is derived from the extract of Centella asiatica.

2. The small molecule peptide composition according to claim 1, characterized in that: The content ratio of each peptide sequence in the small molecule peptide composition is: peptide sequence F accounts for 10%-30%, peptide sequence G accounts for 10%-30%, peptide sequence H accounts for 10%-30%, peptide sequence I accounts for 10%-30%, and peptide sequence J accounts for 10%-30%.

3. A method for preparing the small molecule peptide composition according to claim 1, characterized in that: The following steps are involved: Collect the aboveground part of the Centella asiatica that is growing vigorously and free of pests and diseases, rinse it thoroughly with clean water to remove the surface soil, impurities and microorganisms, and cut it into small sections of 2-3 cm; Using vacuum freeze drying, the small pieces of Centella asiatica are dried at -40°C to -20°C to a moisture content of less than 8%, and then crushed into a coarse powder that can pass through a 80-100 mesh sieve; Using ethanol, methanol, and ethyl acetate as organic solvents, the crude powder is subjected to extraction tests by changing the extraction temperature, time, and material-liquid ratio, collecting the extract by centrifugation or filtration, and concentrating under reduced pressure to obtain an extract concentrate; The extract concentrate is initially separated by a macroporous adsorption resin column, and the macroporous adsorption resin is selected according to the polarity and molecular size of the peptide components. The elution is gradient eluted with an ethanol solution, and the eluate that may contain the peptide components is collected; The above eluate was further separated by gel filtration chromatography, using Sephadex G-75 and Sephacryl S-150 gel media to separate the peptides according to their molecular size, and the eluate at different time periods was collected for preliminary grouping; The grouped peptide components are separated again by ion exchange chromatography. According to the charge properties and quantity of peptide molecules, cation exchange resin SP-Sepharose or anion exchange resin Q-Sepharose is selected, the pH value and ionic strength of the eluent are adjusted, and different elution peak solutions are collected to obtain relatively purified peptide components. The relatively purified peptide components were sequenced by Edman degradation or mass spectrometry sequencing, and the peptide sequences F, G, H, I, and J were determined by bioinformatics analysis, and then mixed to obtain a small molecule peptide composition.

4. The preparation method according to claim 3, characterized in that: In the preliminary separation step of the macroporous adsorption resin column, polar macroporous adsorption resin is selected for peptide components with greater polarity, and non-polar macroporous adsorption resin is selected for non-polar or weakly polar peptide components.

5. The preparation method according to claim 3, characterized in that: In mass spectrometry sequencing, peptide samples are first desalted and concentrated before being injected into a mass spectrometer and then ionized using electrospray ionization (ESI) or matrix-assisted laser desorption ionization (MALDI). Professional software is then used to analyze the mass spectrometry data.

6. Use of the small molecule peptide composition according to claim 1 in promoting the production of natural bezoar.

7. A method for promoting the production of natural bezoar, characterized in that: The small molecule peptide composition according to claim 1 is formulated into a solution, and administered to cattle by intravenous injection at a dose of 3 mg / kg body weight, twice a week for 10 consecutive weeks.

8. A method for promoting the production of natural bezoar, characterized in that: Chitosan with good biocompatibility and biodegradability is selected to prepare a microsphere carrier with a diameter of 1-2 mm, and the small molecule peptide sequence described in claim 1 is loaded onto the microsphere carrier separately or mixed, and the small molecule peptide obtained by extraction and purification is formulated into a solution, which is administered by intravenous injection at a dose of 3 mg / kg body weight, twice a week, for 10 consecutive weeks.

9. The synergy enhancement method according to claim 8, characterized in that: The small molecule peptide sequence is loaded onto the microsphere carrier by physical adsorption or chemical coupling. The physical adsorption method is to mix the microsphere carrier with the small molecule peptide solution under appropriate temperature and stirring conditions.

10. A pharmaceutical preparation comprising the small molecule peptide composition according to claim 1, characterized in that: The pharmaceutical preparation further comprises a pharmaceutically acceptable carrier, excipient or diluent, and the dosage form is injection, capsule, tablet or granule.