TW202600162A - Anti-intestinal adhesion composition and its preparation method - Google Patents

Abstract

This invention discloses an anti-intestinal adhesion composition and its preparation method. The anti-intestinal adhesion composition comprises an arginine aspartic peptide and a hydrogel matrix, wherein the hydrogel matrix is composed of a chitosan and a β-glycerophosphate through a crosslinking reaction. By applying the anti-intestinal adhesion composition disclosed in this invention to the damaged area of an organ or tissue in an organism, a physical barrier can be formed between the damaged area and its adjacent tissues, thereby reducing adhesion between the damaged area and adjacent tissues, achieving the effects of promoting wound healing and improving prognosis.

Description

Anti-intestinal adhesion composition and its preparation method

This invention relates to a medical material, and more particularly to an anti-intestinal adhesion composition and a method for preparing the same.

According to statistics, the adhesion rate during abdominal and gastrointestinal surgeries is as high as 90%. If the adhesion is severe, it can cause abdominal pain, intestinal necrosis, and even death. Therefore, various methods are used during surgery to prevent organ adhesion. These methods include reducing surgery time, maintaining tissue moisture during surgery, minimizing damage during surgery, and using anti-adhesion products.

There are various types of anti-adhesion products on the market, which can be divided into patches, gels, and sprays based on their formulation. The advantage of patches is that they can be cut to size and applied directly to the wound, but the disadvantage is that they are more likely to cause a foreign body reaction. Gel products can be used on uneven wounds, but some gels are absorbed by the body too quickly, making it impossible to avoid adhesion. Spray products can be used on large, uneven wounds, but there may be problems with uneven spraying.

The main objective of this invention is to provide an anti-adhesion compound and its manufacturing method. Because the anti-adhesion compound has high biocompatibility, no cytotoxicity, anti-inflammatory, anti-adhesion and cell growth promotion properties, it can be used in surgery, on organ and/or tissue wounds to effectively prevent adhesion and promote healing.

Therefore, in order to achieve the above objectives, the present invention discloses an anti-intestinal adhesion composition, the main components of which include an arginine aspartic peptide and a hydrogel matrix, wherein the hydrogel matrix is formed by a chitosan and a β-phosphoglycerate through a crosslinking reaction.

The sequence of the arginine aspartic peptide is Arg-Gly-Asp, and the concentration is 0.01~0.2 μg/ml.

The concentration of β-glycerophosphate is 12-16% (w/v).

The concentration of chitosan is 2.0-2.5% (w/v).

Another embodiment of the present invention discloses a method for manufacturing the wound protection composition, which mainly includes the following steps:

Step a: Prepare a shellac/acetic acid solution with a concentration of 2.0-2.5% (w/v).

Step b: The chitosan/acetic acid solution, β-glycerol phosphate, and arginine aspartic peptide were mixed in a low-temperature environment to obtain a mixed solution, wherein:

The concentration of this arginine aspartic peptide is 0.01~0.2 μg/ml.

The concentration of this β-glycerophosphate is 12-16% (w/v);

The low-temperature environment refers to an environment with a temperature of approximately 4°C.

Step c: The mixed solution is subjected to a gelation reaction at a reaction temperature to obtain an RGD/CS/β-GP hydrogel, wherein the reaction temperature is 36-65°C.

This invention discloses an anti-enteric adhesion composition comprising an arginine aspartic peptide and a hydrogel matrix, wherein the hydrogel matrix is composed of a chitosan and a β-glycerophosphate. The anti-enteric adhesion composition disclosed in this invention has the characteristic of being liquid at low temperatures and solid at temperatures above room temperature. Therefore, it can be directly applied to damaged organs or tissues within the body, forming a physical barrier between the damaged area and adjacent tissues, thereby reducing adhesion between the damaged area and adjacent tissues, and achieving the effects of promoting wound healing and improving prognosis.

In one embodiment of the present invention, the anti-gut adhesion composition is composed of the arginine aspartic peptide and the hydrogel matrix.

The concentration of arginine aspartic peptide is 0.01~0.2 μg/ml, such as 0.01, 0.05, 0.1, 0.15, 0.2 μg/ml.

The gelation temperature and gelation time of the anti-enterotoluene composition can be changed by adjusting the concentrations of β-glycerol phosphate and chitosan. Specifically, the higher the concentration of chitosan and/or β-glycerol phosphate, the lower the gelation temperature and the shorter the gelation time of the anti-enterotoluene composition. Therefore, the anti-enterotoluene composition disclosed in this invention can be applied to different species and their wounds by adjusting the concentrations of its components.

For example, if the anti-gut adhesion composition disclosed in this invention is intended for human use, the concentration of the chitosan is 2.0-2.5 (w/v) and the concentration of the β-glycerophosphate is 16% (w/v).

The method for manufacturing the wound protection composition disclosed in this invention includes the following steps:

Step a: Prepare a chitosan/acetic acid solution, wherein the chitosan/acetic acid solution is prepared by dissolving chitosan in 0.1M acetic acid, and the concentration of the chitosan/acetic acid solution is 2.0-2.5% (w/v).

Step b: Mix the chitosan/acetic acid solution, β-glycerol phosphate and arginine aspartic peptide in a low temperature environment to obtain a mixed solution, wherein the sequence of the arginine aspartic peptide is Arg-Gly-Asp; and the concentration of the β-glycerol phosphate is 12-16% (w/v).

Step c: Perform a gelation reaction on the mixed solution to obtain an RGD/CS/β-GP hydrogel.

Specifically, in step b, the hydrogel matrix formed by chitosan and β-glycerol phosphate is crosslinked through ionic gels. That is, through the mixing of cationic polymer chitosan and anionic crosslinker β-glycerol phosphate, the negatively charged phosphate groups in β-glycerol phosphate interact with the positively charged amino groups in chitosan to form a hydrogel network.

In step b, the low-temperature environment refers to a temperature of approximately 4°C.

In step c, the gelation reaction is carried out by placing the mixed solution in an environment with a temperature of approximately 36-65°C.

The term "chitosan" refers to a biopolymer extracted from chitin, which can be obtained from the exoskeleton of crustaceans such as shrimp, crab, and lobster, or from the cell walls of fungi. Chitosan is biodegradable and biocompatible, and is therefore often used as a wound dressing or a drug delivery carrier.

The term "arginine aspartic peptide," also known as RGD, RGD peptide, or RGD peptide, refers to a peptide composed of arginine, glycine, and aspartic acid. Its amino acid sequence is represented as Arg-Gly-Asp, and its structure is shown in the following formula (I):

….Formula (I)

The term "β-glycerol phosphate" refers to an organic compound composed of a glycerol molecule with a phosphate group linked to a β-carbon atom (the second carbon atom from the end). Its chemical formula is _C3H7O6P , and it is called _{2,3 -} dihydroxypropyl dihydrogen phosphate.

The following are some experimental examples and illustrations to illustrate the technical features and effects of this invention.

The cells used in the following examples are readily available to those with ordinary knowledge of the art to which this invention pertains, and therefore do not require patent registration.

The animal experiments in the following examples all comply with the relevant ethical norms for animal testing.

Example 1: Preparation of CS/β-GP hydrogel

Chitosan was dissolved in 0.1M acetic acid and stirred at room temperature until homogeneous, typically for about 12 hours, to obtain a chitosan solution (CS solution) with a concentration of 2.0% or 2.5% (w/v). At 4°C, different concentrations of β-glycerophosphate (β-GP) were added to 30 ml of 2.0% (w/v) or 2.5% (w/v) CS solution, respectively. After mixing, a gelation reaction was carried out in a constant temperature water bath at 37°C to obtain CS/β-GP hydrogel.

The thermosensitivity temperature, gelation time, and degradation gelation time of CS/β-GP hydrogels prepared with different ratios of chitosan and β-glycerol phosphate were tested. The results are shown in Table 1 below.

Table 1: Analysis of the properties of each CS/β-GP hydrogel Chitosan (%) β-glycerol phosphate (%) Temperature (°C) Gel time (seconds) Biodegradation (%, 7 days) Aperture (μm) Porosity (%) 2 12 65 102 37 56.64±8.82 81±3 14 60 61 43 53.37±5.54 79±2 16 45 42 46 51.69±4.83 80±2 2.5 12 60 74 39 59.64±7.52 80±2 14 45 46 45 56.33±6.78 78±3 16 36 twenty two 48% 54.61±4.72 77±2

As shown in Table 1, the CS/β-GP hydrogels prepared from different ratios of chitosan and β-glycerol phosphate all have similar porosity and pore size. The concentration of β-glycerol phosphate added is negatively correlated with the thermosensitive temperature of the hydrogel, meaning that the higher the concentration of β-glycerol phosphate added, the lower the thermosensitive temperature of the hydrogel. As shown in Table 1, when the concentration of β-glycerol phosphate is 16% (w/v) and the concentration of chitosan solution is 2.5% (w/v), the gelation temperature of the hydrogel obtained is closest to the physiological body temperature of 37°C. Therefore, a concentration of 16% (w/v) of β-glycerol phosphate and a concentration of 2.5% (w/v) of chitosan solution is the optimal ratio for preparing the gel matrix in the anti-gut adhesion composition disclosed in this invention.

Example 2: Preparation of RGD/CS/β-GP hydrogel

Chitosan was dissolved in 0.1M acetic acid and stirred at room temperature until homogeneous, typically for about 12 hours, to obtain a chitosan solution with a concentration of 2.0% or 2.5% (w/v). At 4°C, 16% (w/v) β-glycerophosphate and arginine aspartate peptide (RGD, concentration 0.1 μg/ml) were added to 2.0% (w/v) and 2.5% (w/v) CS solutions (30 ml), respectively. After mixing, a gelation reaction was carried out in a constant temperature water bath at 37°C to obtain RGD/CS/β-GP hydrogel.

Example 3: FTIR Spectroscopic Analysis

FTIR analysis was performed on RGD peptides, chitosan, β-glycerol phosphate, the various CS/β-GP hydrogels prepared in Example 1, and the RGD/CS/β-GP hydrogel prepared in Example 2. The RGD/CS/β-GP hydrogel was prepared from 2.5% (w/v) chitosan solution, 16% (w/v) β-glycerol phosphate, and 0.1 μg/ml RGD peptide. The FTIR analysis results are shown in Figures 1 to 4. The results in Figures 1 to 4 show that the hydrogels prepared from chitosan and β-glycerol phosphate have stable hydrogel structures.

Example 4: Solubility Analysis

The hydrogels prepared in Example 1 were immersed in phosphate buffer (PBS) and their swelling rate was observed. The results are shown in Figures 5 and 6.

As shown in Figures 5 and 6, the CS/β-GP hydrogels prepared with different ratios of chitosan and β-glycerol phosphate exhibited the lowest solubility and swelling rates after 24 hours of soaking. Among them, the hydrogels prepared with a β-glycerol phosphate concentration of 16% (w/v) and a chitosan solution concentration of 2% or 2.5% (w/v) showed the lowest solubility and swelling rates. These results indicate that higher concentrations of chitosan and β-glycerol phosphate in the CS/β-GP hydrogel result in a denser network structure, reduced water absorption, and thus better stability and lower swelling.

Example 5: Hydrogel Structure Analysis

The hydrogels prepared in Example 1 were analyzed by electron microscopy, and the results are shown in Figure 7.

As shown in Figure 7, the hydrogel prepared with 2.5% (w/v) chitosan and 16% (w/v) β-glycerol phosphate exhibits a clear and uniform three-dimensional structure, with a pore structure of uniform size evenly distributed throughout the hydrogel matrix. This demonstrates that the hydrogel prepared with 2.5% (w/v) chitosan and 16% (w/v) β-glycerol phosphate has the most stable structure.

Example 6: NMR Analysis

Hydrogels containing RGD and hydrogels without RGD were analyzed by nuclear magnetic resonance spectroscopy. The matrix of the hydrogel consisted of 2.5% (w/v) chitosan and 16% (w/v) β-glycerophosphate. The analytical results are shown in Figure 8.

As shown in Figure 8, the hydrogel containing arginine aspartic peptide has an aromatic peak, while the hydrogel without arginine aspartic peptide does not have an aromatic peak. This result shows that the method described in Example 2 can indeed successfully integrate RGD peptide into the hydrogel matrix prepared by 2.5% (w/v) chitosan and 16% (w/v) β-glycerophosphate.

Example 7: In vitro degradation experiment

Each hydrogel prepared in Example 1 was freeze-dried and weighed (W0); then each hydrogel sample was divided into two groups and placed in 20 mL of phosphate buffer (pH≥7.4), one group without lysozyme and the other group with lysozyme (3% (w/v)); then the degradation reaction was carried out at 37°C for 7 days, during which the stirring speed was 50 rpm; the weight (Wt) of each group of hydrogels was recorded during the degradation period, and the degradation rate was calculated using the following formula. The results are shown in Figures 9 and 10.

Degradation rate (%) = (Wt)/W0 × 100%

As shown in Figures 9 and 10, the degradation rate of each hydrogel in an environment without lysozyme was only about 10% after 7 days. In contrast, under conditions without lysozyme, the degradation rate of each hydrogel in an environment containing lysozyme reached 50% or more after 7 days, indicating that lysozyme promotes the degradation of hydrogels and destroys their structure.

Example 8: Cell Experiment

(i) After sterilizing the CS/β-GP hydrogel solutions prepared in Example 1, each CS/β-GP hydrogel (100 μL) was used to cover a 24-well plate. After gelation at 37°C for 30 minutes, 500 μL of a cell suspension of 3T3 cells or Hs68 cells (cell density of 1 × ^10⁴ cells/mL) was added to each well. The 24-well plate was then incubated at 37°C in a 5% CO₂ incubator for 24 hours. The cell viability on each hydrogel was then measured using a CCK-8 assay kit. The results are shown in Figures 11 and 12.

(ii) Hs68 cells were treated with different concentrations (0, 0.4, 2, 10) of arginine aspartic peptide for 24 hours and their survival rate was observed. The results are shown in Figure 13.

(iii) After sterilizing the RGD/CS/β-GP hydrogel solution prepared in Example 2, Hs68 cells were cultured in each RGD/CS/β-GP hydrogel solution for 24 hours according to the procedure in (i) above; the cell viability of Hs68 was then analyzed, and the results are shown in Figure 14.

As shown in Figures 11 to 14, the RGD/CS/β-GP hydrogel, CS/β-GP hydrogel, and arginine aspartate peptide disclosed in this invention do not affect cell survival rate, meaning they are not cytotoxic. However, Figures 11 and 12 show that cell survival rate decreases slightly with increasing β-glycerophosphate concentration in the hydrogel (approximately...). While the percentage of CS/β-GP hydrogel is 5%, overall, regardless of the ratio used, cell survival rates can still be maintained above 90%, indicating that the CS/β-GP hydrogel system is not cytotoxic. Furthermore, cell survival rate is related to the hydrogel structure, meaning that the higher the degree of crosslinking in the hydrogel, the better the cell survival rate. Therefore, when the hydrogel matrix is composed of 2.5% (w/v) chitosan solution, 16% (w/v) β-glycerol phosphate and arginine aspartate peptide, it not only does not reduce cell survival rate but also enhances cell growth and proliferation.

Example 9: Animal Experiments

Multiple C57BL/6 mice were randomly divided into 5 groups. Except for the control group, abdominal adhesion patterns were established in the abdomen of each group of mice during week 1 by abrading the cecum and removing adjacent abdominal walls. The mice were then treated with the following different conditions for 3 weeks:

Group 1: This is the control group; the mice did not receive any treatment.

Group 2: The wound was sutured without any medication;

Group 3: The wound was sutured after injecting 0.2 ml of hydrogel, which was prepared from 2.5% (w/v) CS, 16% (w/v) β-GP and 0.1 μg/ml RGD;

Group 4: The wound was sutured after injecting 0.2 ml of hydrogel, which was prepared from 2.0% (w/v) CS, 16% (w/v) β-GP and 0.1 μg/ml RGD peptide;

Group 5: The wound was sutured after injecting commercially available hydrogel (PROTAHERE absorbable adhesive barrier).

After the experiment, the abdominal adhesions of each group of mice were confirmed, as shown in Figure 15. In Group 1, no treatment was given, so adhesions appeared between the injured cecum and the abdominal wall, indicating a successful establishment of the abdominal adhesion animal model. In Groups 3 and 4, because the RGD/CS/β-GP hydrogel described in this invention was administered, no adhesions appeared at the injured organs in the abdomen. This shows that after the RGD/CS/β-GP hydrogel is injected into the organism, the increased body temperature causes the hydrogel to form a protective film between the injected organs or tissues. In other words, the RGD/CS/β-GP hydrogel described in this invention can act as a barrier between the injured cecum and surrounding tissues.

Furthermore, although no adhesions were observed in the injured cecum and abdominal wall of mice in group 5, the commercially available hydrogel showed signs of detachment. In contrast, the hydrogel applied to the abdomens of mice in groups 3 and 4 was successfully absorbed by the human body. This demonstrates that the RGD/CS/β-GP hydrogel disclosed in this invention has high biocompatibility and exhibits superior adhesion and high safety compared to commercially available hydrogels.

Example 10: Motor Coordination and Balance Test

During the experiment, mice in each group of Example 9 underwent a rotating rod test, which involved placing the mice on a rotating rod and accelerating the rod from 4 rpm to 20 rpm within 3 minutes. The time it took for the mice to fall off the rotating rod was recorded, and the results are shown in Figure 16.

As shown in Figure 16, the running time of the second group of mice on the rotating rod was significantly lower than that of the first group, by approximately 20-30%. Compared to the first group, the running time of the third group did not show a significant decrease, indicating that the motor coordination and balance disorders of the third group were not affected by the abdominal injury and its wound. Although the motor performance of the fourth and fifth groups of mice was slightly lower than that of the third group, with running times decreasing by approximately 5% respectively, their overall motor performance was still significantly improved compared to the second group.

The results in Figure 16 show that the RGD/CS/β-GP hydrogel system disclosed in this invention can promote wound healing, so that the injury will not affect the individual's athletic ability and performance. Moreover, the higher the concentration of RGD/CS/β-GP hydrogel used, the better it can not only prevent the adhesion of abdominal tissues, but also improve the wound healing ability, thus having a better effect on improving athletic performance.

Example 11: Staining of Tissue Sections

After the experiment in Example 9 was completed, cecal tissue was taken from each group of mice and stained with hematoxylin and eosin. The results are shown in Figure 17. In addition, the abdominal adhesion index of each group of mice was calculated, as shown in Figure 18 and Table 2.

Table 2: Histological examination scores of the cecum and adhesion sites in each group of mice. organ Organizational performance Group 1 Group 2 Group 3 Group 4 Group 5 Peritoneum Inflammation, peritoneal, multifocal. - 5 2 1 3 Necrosis, peritoneum, multifocal - 5 2 - 3 Foreign body, test article - 2 2 1 2 skin Granuloma, suture, multifocal - 2 2 1 2 Intestine Inflammation, serosa, multifocal - 3 2 1 2

As shown in Figures 17 and 18, the tissue sections of the second group of mice contained fibroblasts and inflammatory cells, indicating that the wound was in the early stage of healing. However, the smooth muscle fused with the abdominal wall muscle, indicating significant adhesion between the intestine and surrounding tissues. Compared with the second group of mice, the third and fourth groups of mice showed better wound healing, with a significant reduction in the size of the adhesion area. In particular, the third group of mice showed the least infection and the least tissue adhesion, and the injured cecum had completed regeneration by the fourth week of the experiment, showing an intact intestinal wall structure. These results show that the RGD/CS/β-GP hydrogel system disclosed in this invention can achieve both anti-adhesion and anti-infection effects.

The above results confirm that the RGD/CS/β-GP hydrogel disclosed in this invention does indeed possess biocompatibility, absorbability, anti-adhesion, and anti-inflammatory properties simultaneously. Therefore, the RGD/CS/β-GP hydrogel disclosed in this invention can be directly applied to damaged organs within the body and can effectively reduce wound infection and tissue adhesion. In other words, the RGD/CS/β-GP hydrogel disclosed in this invention can be applied in various surgical procedures and can effectively act as a barrier between tissues to improve wound recovery rates and patient prognosis.

Example 12: Protein Expression and mRNA Expression

The mRNA and protein expression of the TGF-β1/Smad3 pathway and key signaling molecules (including ERK1/2, PI3K and p38) in each group of mice were detected by qRT-PCR and Western ink dot assay, respectively. The results are shown in Figures 19 and 20.

As shown in Figures 19 and 20, the RGD/CS/β-GP hydrogel system disclosed in this invention can effectively regulate the TGFβ1/Smad3 pathway, significantly reducing the expression levels of TGF-β1 and phosphorylated Smad3. This means that the RGD/CS/β-GP hydrogel system disclosed in this invention can downregulate the TGFβ1/Smad3 pathway; and the RGD/CS/β-GP hydrogel system disclosed in this invention... /CS/β-GP hydrogel can regulate the expression of downstream signaling molecules of the TGFβ1/Smad3 pathway: ERK1/2, PI3K, and p38. This result confirms that the RGD/CS/β-GP hydrogel disclosed in this invention affects key cell signaling pathways involved in adhesion and wound healing by forming a physical barrier between injured organs and tissues.

without

Figure 1 shows the FTIR spectra of chitosan and β-glycerol phosphate. Figure 2 shows the FTIR spectra of CS/β-GP hydrogels prepared with different ratios of chitosan and β-glycerol phosphate. Figure 3 shows the FTIR spectra of arginine aspartic peptide. Figure 4 shows the FTIR spectra of RGD/CS/β-GP hydrogel. Figure 5 shows the results of analyzing the solubility and swelling rate of CS/β-GP hydrogels prepared with 2% (w/v) chitosan and different concentrations of β-glycerol phosphate. Figure 6 shows the results of analyzing the solubility and swelling rate of CS/β-GP hydrogels prepared with 2.5% (w/v) chitosan and different concentrations of β-glycerol phosphate. Figure 7 shows the electron microscope observation results of CS/β-GP hydrogels prepared with different ratios of chitosan and β-glycerol phosphate. Figure 8 shows the NMR analysis results of hydrogels containing and without arginine aspartate peptide. Figure 9 shows the in vitro degradation rate analysis results of each hydrogel in an environment containing lysozyme. Figure 10 shows the in vitro degradation rate analysis results of each hydrogel in an environment without lysozyme. Figure 11 shows the analysis results of the effect of each hydrogel on Hs68 cell survival. Figure 12 shows the analysis results of the effect of each hydrogel on 3T3 cell survival. Figure 13 shows the cell viability analysis results after treating Hs68 cells with different concentrations of arginine aspartate peptide. Figure 14 shows the cell viability analysis results after treating Hs68 cells with various hydrogels containing arginine aspartate peptide. Figure 15 shows the abdominal adhesions in each group of mice. Figure 16 shows the results of the rotation bar test in each group of mice. Figure 17 shows the results of skin tissue section staining in each group of mice. Figure 18 shows the adhesion index of the abdomen in each group of mice. Figure 19 shows the relative mRNA expression levels of TGF-β1, Smad3, MAPK1, MAPK14, and PI3Kr1 in the abdominal adhesion tissue of each group of mice. Figure 20 shows the relative protein expression levels of TGF-β1, Smad3, ErK1/2, P38, and PI3K in the abdominal adhesion tissues of mice in each group.

Claims (10)

An anti-intestinal adhesion compound for use between a damaged organ and adjacent tissues, comprising an arginine aspartic peptide and a hydrogel matrix, wherein: the hydrogel matrix is composed of a chitosan and a β-glycerophosphate; the sequence of the arginine aspartic peptide is Arg-Gly-Asp. The anti-intestinal adhesion composition as described in claim 1 is composed of the arginine aspartic peptide and the hydrogel matrix. The anti-intestinal adhesion composition as described in claim 1 or 2, wherein the concentration of β-glycerophosphate is 12-16% (w/v). The intestinal adhesion-resistant composition as described in claim 1 or 2, wherein the concentration of the chitosan is 2.0-2.5% (w/v). The intestinal adhesion prevention composition as described in claim 1 or 2, wherein the concentration of the arginine aspartic peptide is 0.01~0.2 μg/ml. A method for preparing an anti-intestinal adhesion composition includes the following steps: Step a: Preparing a chitosan/acetic acid solution; Step b: Mixing the chitosan/acetic acid solution, β-glycerol phosphate, and an arginine aspartic peptide in a low-temperature environment to obtain a mixed solution, wherein the arginine aspartic peptide has the sequence Arg-Gly-Asp and a concentration of 0.01~0.2 μg/ml; Step c: Performing a gelation reaction on the mixed solution to obtain an RGD/CS/β-GP hydrogel. The method for preparing the anti-gut adhesion composition as described in claim 5, wherein the chitosan/acetic acid solution is prepared by dissolving chitosan in 0.1M acetic acid, and the concentration of the chitosan/acetic acid solution is 2.0-2.5% (w/v). The method for preparing the anti-intestinal adhesion composition as described in claim 5, wherein the concentration of β-glycerophosphate is 12-16% (w/v). The method for manufacturing the anti-gut adhesion composition as described in claim 5, wherein the low-temperature environment in step b refers to a temperature of approximately 4°C. The method for manufacturing the anti-adhesion composition as described in claim 5, wherein the gelation reaction in step c is carried out by placing the mixed solution in an environment with a temperature of about 36-65°C.