Icodextrin-containing anti-adhesion liquid for abdominal surgery or laparoscopic surgery and preparation method of icodextrin-containing anti-adhesion liquid

Abstract

The invention belongs to the technical field of surgical anti-adhesion liquid, and particularly relates to icodextrin-containing anti-adhesion liquid for abdominal surgery or laparoscopic surgery and a preparation method of the icodextrin-containing anti-adhesion liquid. The anti-adhesion liquid is composed of a component A and a component B, wherein the component A and the component B are stored separately; the component A is a solution taking water as a solvent and comprises 20-100g / L of icodextrin, 0-20g / L of sodium lactate, 0.1-0.6 g / L of calcium chloride, 0.03-0.15 g / L of magnesium chloride and 0-60g / L of sodium chloride; the component B is a solution taking water as a solvent and comprises 0-54 g / L of sodium chloride, 10-20 g / L of sodium bicarbonate and 0-19 g / L of sodium lactate; the volume ratio of the component A to the component B is 1: (0.1-0.9); the pH value of the component A is 3.0-6.5, and the pH value of the component B is 4.5-9.0. The anti-adhesion liquid disclosed by the invention can play a role in preventing adhesion of tissues and organs while cleaning a wound surface.

Description

Technical Field

[0001] The invention belongs to the technical field of anti-adhesion liquid for surgery, and in particular relates to an anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin and a preparation method thereof. Background Art

[0002] Postoperative adhesions are a common and distressing complication after surgery, especially after gynecological abdominal surgery. These adhesions may not only lead to female infertility, but also cause complications such as intestinal adhesions, intestinal obstruction, and chronic abdominal and pelvic pain. These complications increase the difficulty of pelvic and abdominal reoperations. Furthermore, the large number of complications associated with postoperative adhesions places an increasing burden and considerable costs on surgeons and the healthcare system. Therefore, it is necessary to fully assess the risk of adhesions before surgery and implement effective anti-adhesion measures during surgery to reduce the incidence of adhesion complications.

[0003] Among the traditional measures to prevent abdominal adhesion, there are anti-adhesion isolation films made of various materials placed at the adhesion-prone areas, such as A hyaluronic acid-carboxymethyl cellulose membrane; and an anti-adhesion gel barrier used in areas prone to adhesion, such as clinically used hyaluronic acid, chitosan and other macromolecular materials and polyethylene glycol. These methods can effectively prevent and reduce adhesions. However, with the recent popularity of laparoscopic technology and its significant advantages, traditional open surgery has been increasingly replaced by laparoscopic surgery. In these surgeries, anti-adhesion procedures using membrane and gel barriers have shown certain limitations when combined with laparoscopic surgery. In comparison, liquid flotation anti-adhesion products are more suitable for postoperative anti-adhesion procedures in conjunction with laparoscopic surgery.

[0004] Due to its unique colloidal permeability and large molecular size, icodextrin is unable to diffuse directly through the peritoneal wall into the intraperitoneal microvasculature and be absorbed by the body. This allows it to maintain its clearance efficacy within the peritoneal cavity for a long time. The glycosidic bond of icodextrin is typically metabolized by amylase, which is not present in the human peritoneal cavity. Therefore, it can persist in the peritoneal cavity for several days and be slowly absorbed into the systemic circulation by the lymphatic system, where it is broken down by amylase and metabolized into glucose. During this long period, the icodextrin solution present in the peritoneal cavity can separate damaged surfaces and minimize contact between organs during the critical period of adhesion formation. The FDA has approved Adept 4%, an anti-adhesion product containing icodextrin, for postoperative use. However, this product currently has some significant drawbacks. First, compared with icodextrin peritoneal dialysis solution, there are two significant differences in the clinical application of icodextrin as the main component of anti-adhesion fluid: (1) icodextrin peritoneal dialysis solution is placed on the outside of the peritoneum during dialysis and exerts its clinical effect by colloidal osmotic pressure. Except for a small amount of absorption, most of it will not come into contact with the tissues in the abdominal cavity. When used as an anti-adhesion product, icodextrin needs to be infused into the abdominal cavity and directly come into contact with various organs and tissues in the abdominal cavity; (2) The residence time of icodextrin peritoneal dialysis solution during use is 8 to 12 hours, and no longer than 16 hours, and then it is excreted from the body. As an anti-adhesion product, the residence time of icodextrin anti-adhesion fluid must be at least 4 days until it is completely absorbed into the systemic circulation through the lymphatic system. The above clinical usage methods put forward higher biocompatibility requirements for icodextrin anti-adhesion products: (1) the composition and concentration level should be closer to the physiological fluids of the human body; (2) higher biocompatibility, especially pH compatibility, so as to reduce its irritation to abdominal tissues and organs.

[0005] Secondly, in liquid formulations, icodextrin is more stable under slightly acidic conditions. After high-temperature sterilization, in an environment with a pH above 5.5, icodextrin produces a large number of degradation products, including 5-hydroxymethylfurfural, formaldehyde, acetaldehyde, methylglyoxal, glyoxal, and 3-deoxyglucosidone. These glucose degradation products (GDPs) have varying degrees of cytotoxicity, and prolonged exposure can cause apoptosis of various epithelial cells within the peritoneum. Therefore, to ensure the stability of icodextrin, the pH of the product needs to be lowered. The stable pH of icodextrin in liquid form is between 3.5 and 5.0, but too low a pH can directly irritate organs and tissues within the abdominal cavity. Therefore, to balance the stability and compatibility of icodextrin, the pH of the product is usually adjusted between 5.0 and 5.5. However, studies have shown that this pH range still has a negative impact on the survival of human peritoneal mesothelial cells, and prolonged exposure can accelerate their apoptosis, with the rate of apoptosis far exceeding that of a solution with a neutral pH.

[0006] The marketed product Adept 4% uses single-chamber packaging, which is affected by the pH range and cannot solve the stability problem after adding sodium bicarbonate to the formula. The formula cannot achieve a more compatible level of human body fluid components, nor can it solve the stability problem of icodextrin in the formula under neutral pH conditions. In addition, the glucose degradation products in it are significantly higher than the physiological tolerance level.

[0007] Based on this, the present invention provides an anti-adhesion liquid with a mixed component and pH closer to the physiological state of body fluids through a new prescription composition and packaging compatibility form, prolongs the retention time of the anti-adhesion liquid in the abdominal cavity, improves its effectiveness and safety, and optimizes the production process to improve the stability of icodextrin under high-temperature sterilization conditions. Summary of the Invention

[0008] In order to overcome the shortcomings of the existing technology, the surgical anti-adhesion liquid containing icodextrin of the present invention is clinically applicable to various types of abdominal surgeries. It can be used to clean the surgical wound during abdominal surgery and can also be used for abdominal perfusion after various types of abdominal surgeries to prevent postoperative adhesion of organs and tissues in the abdominal cavity. According to the characteristics of the liquid preparation of this product, it is particularly suitable for preventing adhesion in the abdominal cavity after various types of laparoscopic surgeries.

[0009] One of the purposes of the present invention is to provide an anti-adhesion solution for surgery containing icodextrin.

[0010] One of the purposes of the present invention is achieved by the following technical solution:

[0011] An anti-adhesion solution for surgery containing icodextrin, comprising a component A and a component B, wherein the components A and B are stored separately;

[0012] The component A is a solution with water as solvent, including 20-100 g / L icodextrin, 0-20 g / L sodium lactate, 0.1-0.6 g / L calcium chloride, 0.03-0.15 g / L magnesium chloride, and 0-60 g / L sodium chloride; the component B is a solution with water as solvent, including 0-54 g / L sodium chloride, 10-20 g / L sodium bicarbonate, and 0-19 g / L sodium lactate;

[0013] The volume ratio of the component A to the component B is 1:0.1-0.9; the pH of the component A is 3.0-6.5, and the pH of the component B is 4.5-9.0.

[0014] Furthermore, the component A is a solution with water as solvent, comprising 20-75 g / L icodextrin, 1-3 g / L sodium lactate, 0.1-0.5 g / L calcium chloride, and 0.05-0.08 g / L magnesium chloride; the component B is a solution with water as solvent, comprising 25-40 g / L sodium chloride and 10-20 g / L sodium bicarbonate;

[0015] The volume ratio of the component A to the component B is 1:0.1-0.8; the pH of the component A is 3.0-5.0, and the pH of the component B is 6.0-9.0.

[0016] Furthermore, the anti-adhesion liquid consists of two chambers of component A and component B, which are separated by a virtual weld. Before use, they are squeezed through and shaken to mix evenly before use.

[0017] Furthermore, the volume ratio of component A to component B is 1:0.1-0.5; the pH of component A is 3.5-4.5, and the pH of component B is 7.0-9.0; after components A and B are mixed, the pH of the mixture is 6.5-7.5.

[0018] Furthermore, the pH range is adjusted by at least one of hydrochloric acid solution, lactic acid solution, sodium hydroxide solution, sodium bicarbonate solution, citrate solution, carbon dioxide gas, succinate solution, fumarate solution, malate solution, and oxaloacetate solution.

[0019] The present invention contains the prescription quantity of icodextrin for the surgical anti-adhesion liquid icodextrin. Although its solubility is higher, in order to reduce its viscosity in the preparation process so as to be easier to fill, it is placed in a large chamber. Magnesium chloride and calcium chloride may form calcium carbonate and magnesium carbonate precipitation in long-term placement with the carbonate in the sodium bicarbonate aqueous solution, and therefore need to be placed in different chambers respectively. And icodextrin is because it needs to be stored under weakly acidic conditions, and sodium bicarbonate can not stably exist under acidic conditions, therefore selection is icodextrin, magnesium chloride, calcium chloride is placed in a large chamber, and is placed separately with sodium bicarbonate.

[0020] Aqueous sodium lactate and sodium bicarbonate solutions are both alkaline and can theoretically be placed in the same chamber. However, studies have shown that sterilization of these solutions in the same chamber results in poor stability. In particular, after mixing, sodium bicarbonate decomposes into carbon dioxide, which significantly overflows, significantly increasing the pH of the mixed solution. Sodium lactate can tolerate lower pH levels, so the present invention further prefers placing sodium lactate and icodextrin in the same chamber, allowing them to coexist under acidic conditions.

[0021] In the present invention, sodium bicarbonate is incorporated into the formulation as a component, rather than a pH regulator, based on the composition of electrolytes in human body fluids. To ensure that the fluid infused into the peritoneal cavity does not interfere with the acid-base balance of human body fluids, sodium bicarbonate is added, and its concentration is maintained to be substantially consistent with the bicarbonate levels in the blood and body fluids. The ratio of the two in the mixed solution maintains a more stable pH level and buffering capacity for the anti-adhesion fluid. Furthermore, a certain concentration of sodium lactate is retained in the formulation. This concentration avoids the use of excessive sodium bicarbonate for pH control, which could result in an elevated carbon dioxide partial pressure in body fluids.

[0022] Regarding the volume ratio of component A to component B, first determine that component A is a large chamber to facilitate the dissolution of a larger amount of icodextrin. Component A is an acidic liquid and component B is an alkaline liquid. After the two are mixed, the pH is close to neutral. Under the premise of using less pH regulator, the stability of product quality can be met. Therefore, the optimal volume ratio of components A and B is subject to the limitations of the above conditions.

[0023] Furthermore, the A component further comprises 2-5 g / L modified yeast glucan. The modified yeast glucan is prepared as follows: yeast glucan is added to a solvent, mixed evenly, and then a dehydroabietic acid derivative, triethylamine, and 4-dimethylaminopyridine are added to react, and the reaction solution is post-treated to obtain the modified yeast glucan.

[0024] Furthermore, the post-treatment process is to pour ice water into the reaction solution; then add anhydrous ethanol, let it stand and centrifuge, dialyze the precipitate, and freeze-dry the dialyzate.

[0025] Furthermore, the mass ratio of the yeast glucan, dehydroabietic acid derivative, triethylamine, and 4-dimethylaminopyridine is 1:(2.5-3):(2.6-3.5):(2.6-3.5), the concentration of the yeast glucan in the solvent is 0.075-0.1 g / mL; the solvent is N,N-dimethylformamide; the reaction temperature is 40-60°C, and the reaction time is 2-4 hours.

[0026] Furthermore, when the yeast glucan is added to N,N-dimethylformamide, the temperature of the reaction system is 10-15°C.

[0027] Furthermore, the structural formula of the dehydroabietic acid derivative is

[0028] The preparation process of the dehydroabietic acid derivative comprises the following steps:

[0029] (1) adding lithium aluminum tetrahydride to tetrahydrofuran, and then adding dehydroabietic acid to react to obtain dehydroabietic acid reduced alcohol;

[0030] (2) Adding dehydroabietic acid reducing alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate into N,N-dimethylformamide for reaction, and purifying the reaction solution after the reaction is completed to obtain a dehydroabietic acid derivative.

[0031] Furthermore, in step (1), the molar ratio of dehydroabietic acid to lithium aluminum tetrahydride is 1:4-4.5, and the amount ratio of dehydroabietic acid to tetrahydrofuran is 1 mmol:5-5.5 mL; in step (1), when dehydroabietic acid is added, the temperature of the reaction system is -2-3°C, and the reaction time is 4-6 hours.

[0032] Furthermore, in step (2), the molar ratio of the dehydroabietic acid reducing alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate is 1:(1-1.1):(2.5-3); and the concentration of the dehydroabietic acid reducing alcohol in N,N-dimethylformamide is 0.3-0.5 mol / L.

[0033] Furthermore, in step (2), the inorganic carbonate is potassium carbonate or sodium carbonate, the reaction temperature is 60-80° C., and the reaction time is 12-18 h.

[0034] The second object of the present invention is to provide a method for preparing an anti-adhesion liquid for surgery containing icodextrin. The method is simple and feasible, and provides a new preparation method for the anti-adhesion liquid for surgery.

[0035] The method for preparing the above-mentioned anti-adhesion liquid for surgery containing icodextrin comprises the following steps:

[0036] a. Add the raw material components to water according to the ratio, stir evenly, adjust the pH value to the range, and then filter and fill to obtain component A;

[0037] b. Add the raw material components to water according to the ratio, stir evenly, adjust the pH to the range, filter and fill to obtain component B;

[0038] c. Fill component A and component B separately and sterilize them. Mix component A and component B when using.

[0039] Furthermore, during packaging, components A and B are packaged in the same PVC or non-PVC soft bag, which is then divided into a large chamber and a small chamber using a cold welding process. Component A is packaged in the large chamber, and component B is packaged in the small chamber. The cold weld strength between components A and B is 3 to 20 N / 15 mm, and the cold weld opening force is 300 to 3000 N. Before use, the cold weld is opened by squeezing to allow the solutions in the large and small chambers to flow through, and the mixture is then shaken for uniform mixing before use.

[0040] Furthermore, the soft bag is made of non-PVC material and has a filling volume of 500-5000 ml, with preferred sizes being 1000 ml, 1500 ml, 2000 ml, and 3000 ml. Furthermore, the osmotic pressure of the liquid in the large chamber is 50-70 mOsm / L, and the osmotic pressure of the liquid in the small chamber is 900-1200 mOsm / L; after the large and small chambers are combined, the osmotic pressure of the liquid is 260-310 mOsm / L.

[0041] Furthermore, the sterilization temperature is 115-121° C., and the time is 10-30 minutes; and the packaging and filling are carried out under Class C + Class A clean conditions.

[0042] Furthermore, the surgical anti-adhesion fluid containing icodextrin is suitable for various clinical abdominal surgeries. It can be used to clean surgical wounds during abdominal surgery and can also be used for peritoneal perfusion after various abdominal surgeries to prevent postoperative adhesion of intra-abdominal organs and tissues. According to the characteristics of the surgical anti-adhesion fluid of the present invention, it is particularly suitable for peritoneal perfusion after various laparoscopic surgeries to achieve an anti-adhesion effect.

[0043] Compared with the prior art, the present invention has the following beneficial effects:

[0044] 1. The present invention provides an icodextrin-containing surgical anti-adhesion fluid, which is primarily used to prevent adhesions after various laparoscopic surgeries. The anti-adhesion fluid contains ingredients such as icodextrin and modified yeast glucan, which, together with the other components of the anti-adhesion fluid, not only cleanses wound surfaces but also prevents postoperative adhesions. Specifically, the modified yeast glucan introduced in the present invention is primarily modified by using a dehydroabietic acid derivative to modify yeast glucan, where the sulfonic acid groups of the dehydroabietic acid derivative react with the hydroxyl groups of the yeast glucan to form a sulfonate ester. On the one hand, the water solubility of yeast glucan is improved, so that its hydrophilic groups, sulfonate groups and hydroxyl groups, are cross-linked with icodextrin to form a stable cross-linking system, thereby improving the stability of icodextrin. On the other hand, the unique hydrophobic tricyclic skeleton and rigid structure of dehydroabietic acid can effectively increase the retention time of yeast glucan in the body, thereby increasing the duration of its synergistic effect with icodextrin, continuously isolating the damaged tissue surface, reducing direct contact between the uterus and tissue after surgery, inhibiting fibrin deposition and adhesion formation, and achieving an effective and safe postoperative anti-adhesion effect.

[0045] 2. The present invention adopts a dual-chamber packaging form, wherein the acidic component A and the alkaline component B containing icodextrin are stored separately by cold welding and mixed before clinical use to achieve a neutral pH value (6.5-7.5). This not only inhibits the degradation of traditional icodextrin (which is directly used with the pH controlled within the range of 5.0-5.5) to produce harmful GDP-like substances, but also reduces the irritation of low pH to blood vessels, tissues, etc., thereby improving the biocompatibility of the product.

[0046] 3. The present invention provides a method for preparing an anti-adhesion fluid for surgical use containing icodextrin, utilizing a common large-volume infusion production process and GMP management requirements. During the production process, key process control parameters, such as feeding temperature, feeding sequence, liquid precision filtration cycle time, and pH control range, are further refined based on product characteristics. The method also controls the opening force of finished product weld defects, the vacuum level of the high-barrier film packaging (affecting the heat penetration level and uniformity during sterilization), and sterilization conditions (ensuring the sterility level of the large-volume liquid). This comprehensive control of these process parameters ensures product quality and reliability.

[0047] 4. The anti-adhesion liquid of the present invention provides a new preparation idea for the preparation of anti-adhesion liquid for surgery. The anti-adhesion liquid can be used for intraoperative lavage and flushing of various abdominal surgeries (open / laparoscopic), as well as postoperative abdominal perfusion, to reduce and prevent postoperative adhesions. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 This is a physical picture of the anti-adhesion liquid for surgery of the present invention;

[0049] Figure 2 is the infrared spectrum of the modified yeast glucan of the present invention;

[0050] Figure 3 Graph showing the biocompatibility results of the anti-adhesion liquid of the present invention;

[0051] Figure 4 This is a diagram showing the detection of formaldehyde and acetaldehyde in the sterilized sample of Example 2 of the present invention;

[0052] Figure 5 This is a diagram showing the detection status of 2-glucosone (2-k-DG), 3-deoxyglucosone (3-DG), glyoxal (GO), methylglyoxal (MGO) and other impurities in the sample of Example 2 of the present invention. DETAILED DESCRIPTION

[0053] Below, in conjunction with the accompanying drawings and specific embodiments, the present invention is further described. It should be noted that, under the premise of no conflict, the various embodiments described below or the various technical features can be arbitrarily combined to form new embodiments. Specific conditions not specified in the embodiments are carried out according to conventional conditions or conditions recommended by the manufacturer. Reagents or instruments used, unless otherwise specified, are conventional products obtained through commercial channels.

[0054] The molecular weight of the icodextrin of the present invention is 23-40 kDa and is prepared by Qingdao Liteng Pharmaceutical Technology Co., Ltd.; the sodium lactate Ringer's injection is purchased from Shandong Weigao Pharmaceutical Co., Ltd.

[0055] Preparation Example 1

[0056] The preparation process of dehydroabietic acid derivatives is as follows:

[0057]

[0058] (1) In a nitrogen atmosphere, lithium aluminum tetrahydride (450 mmol) was added to 500 mL of tetrahydrofuran, the temperature of the reaction system was controlled to 0°C, dehydroabietic acid (100 mmol) was added in 5 equal portions in batches, and the mixture was stirred for 10 min after the addition was completed. The mixture was gradually heated to room temperature and reacted at room temperature for 5 h. The reaction system was cooled to 0°C in an ice-water bath, and water was slowly added to quench the reaction (the temperature during the quenching process was controlled at 10°C) until no obvious gas was released. The ice-water bath was removed, and the mixture was heated to room temperature. After stirring for 10 min, 1 M hydrochloric acid solution was added to adjust the pH of the mixture to 7. The mixture was concentrated to remove most of the tetrahydrofuran. The aqueous phase after most of the tetrahydrofuran was removed was extracted with ethyl acetate three times. The organic phases were combined and washed with saturated brine, and then dried over anhydrous sodium sulfate. After vacuum concentration, the mixture was purified by distillation to obtain dehydroabietic acid reduced alcohol. The characterization results of dehydroabietic acid reduced alcohol are as follows: 1 HNMR (C 20 H 30 O, 300MHz, DMSO-d6): δ7.03 (s, 1H), 6.90-6.86 (m, 2H), 4.22 (s, 1H), 3.52-3.42 (m, 2H), 2.88-2.78 (m, 3H), 2.01-1.99(m, 1H), 1.73-1.29(m, 10H), 1.18(d, 6H), 1.09(t, 1H), 0.89(s, 3H); ESI-MS(m / z): 287.23[M+H] + The above results confirm that the target product is obtained;

[0059] (2) Dehydroabietic acid reducing alcohol (100 mmol), 2-bromo-1-ethanesulfonic acid (CAS: 26978-65-4, 105 mmol) and potassium carbonate (280 mmol) were added to 300 mL of N,N-dimethylformamide and reacted at 70°C for 16 h. The reaction solution was diluted with a mixed organic solution (prepared by mixing dichloromethane and isopropanol in a volume ratio of 4:1), and then a saturated aqueous ammonium chloride solution was added and stirred evenly. The mixture was separated using a separatory funnel, and the saturated ammonium chloride solution was extracted with the mixed organic solution. The mixed organic phase was washed with saturated brine, concentrated under reduced pressure, and purified by column chromatography (90% n-heptane: 9% ethyl acetate: 1% acetic acid) to obtain a dehydroabietic acid derivative. The characterization results of the dehydroabietic acid derivative are as follows: 1 HNMR (C 22 H 34O4S, 300MHz, DMSO-d6): δ8.6 (s, 1H), 7.03 (s, 1H), 6.90-6.86 (m, 2H), 5.90 (d, 1H), 5.66 (d, 1H), 3.80 (t, 2H), 2.88-2.78 (m, 3H), 2.62(t, 2H), 2.01-1.99(m, 1H), 1.73-1.29(m, 10H), 1.18(d, 6H), 1.09(t, 1H), 0.89(s, 3H); ESI-MS(m / z): 393.18[MH] - .The above results confirmed that the target product was obtained.

[0060] Preparation Example 2

[0061] The preparation process of dehydroabietic acid derivatives is as follows:

[0062] (1) In a nitrogen atmosphere, lithium aluminum tetrahydride (400 mmol) was added to 500 mL of tetrahydrofuran, the temperature of the reaction system was controlled to -2°C, dehydroabietic acid (100 mmol) was added in 5 equal portions in batches, stirred for 10 min after the addition was completed, and the temperature was gradually raised to room temperature. The reaction was reacted at room temperature for 4 h; the reaction system was cooled to 5°C in an ice-water bath, and water was slowly added to quench the reaction (the temperature during the quenching process was controlled at 10°C) until no obvious gas was released. The ice-water bath was removed, and the mixed system was heated to room temperature and stirred for 10 min. After that, 1 M hydrochloric acid solution was added to adjust the pH of the mixed system to 7. The mixed system was concentrated to remove most of the tetrahydrofuran, and the aqueous phase after most of the tetrahydrofuran was removed was extracted with ethyl acetate three times. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then purified by distillation to obtain dehydroabietic acid reduced alcohol. The characterization results of the dehydroabietic acid reduced alcohol were the same as those in Preparation Example 1.

[0063] (2) Dehydroabietic acid reducing alcohol (100 mmol), 2-bromo-1-ethanesulfonic acid (100 mmol) and sodium carbonate (250 mmol) were added to 300 mL of N,N-dimethylformamide and reacted at 60° C. for 18 h. The reaction solution was diluted with a mixed organic solution (prepared by mixing dichloromethane and isopropanol in a volume ratio of 4:1), and then a saturated aqueous ammonium chloride solution was added and stirred evenly. The mixture was separated using a separatory funnel, and the saturated ammonium chloride solution was extracted with the mixed organic solution. The mixed organic phase was washed with saturated brine, concentrated under reduced pressure, and purified by column chromatography (90% n-heptane: 9% ethyl acetate: 1% acetic acid) to obtain a dehydroabietic acid derivative. The characterization results of the dehydroabietic acid derivative were the same as those in Preparation Example 1.

[0064] Preparation Example 3

[0065] The preparation steps of dehydroabietic acid derivatives are as follows:

[0066] (1) In a nitrogen atmosphere, lithium aluminum tetrahydride (450 mmol) was added to 550 mL of tetrahydrofuran, the temperature of the reaction system was controlled to 3° C., dehydroabietic acid (100 mmol) was added in 5 equal portions in batches, stirred for 10 min after the addition was completed, and the temperature was gradually raised to room temperature. The mixture was reacted at room temperature for 6 h. The reaction system was cooled to 0° C. in an ice-water bath, and water was slowly added to quench the reaction (the temperature during the quenching process was controlled at 10° C.) until no obvious gas was released. The ice-water bath was removed, and the mixture was heated to room temperature and stirred for 10 min. After that, 1 M hydrochloric acid solution was added to adjust the pH of the mixture to 7. The mixture was concentrated to remove most of the tetrahydrofuran. The aqueous phase after removal of most of the tetrahydrofuran was extracted 3 times with ethyl acetate. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by distillation to obtain dehydroabietic acid reduced alcohol. The characterization results of the dehydroabietic acid reduced alcohol were the same as those in Preparation Example 1.

[0067] (2) Dehydroabietic acid reducing alcohol (100 mmol), 2-bromo-1-ethanesulfonic acid (110 mmol) and potassium carbonate (300 mmol) were added to 300 mL of N,N-dimethylformamide and reacted at 80° C. for 12 h. The reaction solution was diluted with a mixed organic solution (prepared by mixing dichloromethane and isopropanol in a volume ratio of 4:1), and then a saturated aqueous ammonium chloride solution was added and stirred evenly. The mixture was separated using a separatory funnel. The saturated ammonium chloride solution was then extracted with the mixed organic solution. The mixed organic phase was washed with saturated brine, concentrated under reduced pressure, and purified by column chromatography (90% n-heptane: 9% ethyl acetate: 1% acetic acid) to obtain a dehydroabietic acid derivative. The characterization results of the dehydroabietic acid derivative were the same as those in Preparation Example 1.

[0068] Example 1

[0069] A surgical anti-adhesion fluid containing icodextrin is composed of 1200 mL of component A and 300 mL of component B, which are stored separately and mixed before use. Component A comprises 50 g / L icodextrin, 0.32 g / L calcium chloride, 0.064 g / L magnesium chloride, and 2.1 g / L sodium lactate, with water for injection as the solvent. Component B comprises 27 g / L sodium chloride aqueous solution and 11.5 g / L sodium bicarbonate aqueous solution.

[0070] The method for preparing the above-mentioned anti-adhesion liquid for surgery containing icodextrin comprises the following steps:

[0071] a. Inject 1100L of 50°C water for injection into preparation tank A, start stirring (150r / min), then add 60kg of icodextrin, 0.3855kg of calcium chloride, 0.0765kg of magnesium chloride, and 2.52kg of sodium lactate, stirring while adding the materials to dissolve them. After stirring for 25 minutes, add 0.1mol / L dilute hydrochloric acid solution and stir for 15 minutes, adjust the pH value to 4.09, add water for injection to 1200L, and filter the solution through 0.45μm and 0.22μm filter cartridges for 25 minutes to obtain component A.

[0072] b. Pour 180L of 50°C water for injection into preparation tank B, start stirring (150r / min), then add 8.1kg of sodium chloride and 3.46kg of sodium bicarbonate, stirring while adding the materials to dissolve them. After stirring for 25 minutes, measure the pH value to 8.19, add water for injection to 300L, and filter the liquid through 0.45μm and 0.22μm polyethersulfone filter elements for 20 minutes to obtain component B.

[0073] c. Use non-PVC film to make a double-cavity liquid medicine bag, which is divided into chamber A and chamber B by cold welding. The liquid in preparation tank A is divided into chamber A, and the liquid in preparation tank B is divided into chamber B. The filling temperature is 30-70°C. After filling, the product is inspected by light and high-voltage discharge for leaks, and then vacuum-packed with high-barrier outer film. The vacuum packaging parameters are 3 to 1.8S; sterilize at 121°C for 15min, with a pressure coefficient of 4.5, a temperature rise gradient of 10°C / min, and an out-of-cabinet temperature of 60°C, ensuring that the sterilization index F0 value (standard sterilization time) is ≥12. Mix chambers A and B when using.

[0074] d. The finished product is tested according to quality standards. The pH of the mixed drug solution is 7.06, the insoluble particles below 5 μm are 12 / mL, the endotoxin is less than 0.1 EU / mL, the peptidoglycan is less than 0.5 ng / mL, and the osmotic pressure is 269 mOsm / L.

[0075] The appearance of the sample in Example 1 and the actual picture of the folded large and small chambers are shown in Figure 1 shown.

[0076] Example 2

[0077] A surgical anti-adhesion fluid containing icodextrin is composed of 1260 mL of component A and 240 mL of component B, which are stored separately and mixed before use. Component A comprises 47.62 g / L icodextrin, 4 g / L modified yeast glucan, 0.31 g / L calcium chloride, 0.061 g / L magnesium chloride, and 2 g / L sodium lactate, with water for injection as the solvent. Component B comprises 33.75 g / L sodium chloride aqueous solution and 14.38 g / L sodium bicarbonate aqueous solution.

[0078] The preparation steps of modified yeast glucan are as follows:

[0079] 3 g of yeast glucan was added to 40 mL of N,N-dimethylformamide at 13° C. and mixed thoroughly. 8 g of the dehydroabietic acid derivative prepared in Preparation Example 1, 9 g of triethylamine, and 9 g of 4-dimethylaminopyridine were then added. The mixture was reacted at 50° C. for 3 h. After completion of the reaction, the reaction solution was poured into 95 mL of ice water. 180 mL of anhydrous ethanol was added, and the mixture was allowed to stand and centrifuged. The precipitate was dialyzed against deionized water for 4 days, and the dialyzate was freeze-dried to obtain modified yeast glucan.

[0080] The method for preparing the above-mentioned anti-adhesion liquid for surgery containing icodextrin comprises the following steps:

[0081] a. Inject 1100L of 50°C water for injection into preparation tank A, start stirring (150r / min), then add 60kg of icodextrin, 5.04kg of modified yeast beta-glucan, 0.3855kg of calcium chloride, 0.0765kg of magnesium chloride, and 2.52kg of sodium lactate, stir while adding the materials to dissolve them, stir for 25min, add 0.1mol / L dilute hydrochloric acid solution and stir for 15min, adjust the pH value to 4.09, add water for injection to 1260L, and filter the solution through 0.45μm and 0.22μm filter cartridges for 25min to obtain component A.

[0082] b. Pour 180L of 50°C water for injection into preparation tank B, start stirring (150r / min), then add 8.1kg of sodium chloride and 3.45kg of sodium bicarbonate, stirring while adding the materials to dissolve them. After stirring for 25 minutes, measure the pH value to 8.19, add water for injection to 240L, and filter the liquid through 0.45μm and 0.22μm polyethersulfone filter cartridges for 20 minutes to obtain component B.

[0083] c. Use non-PVC film to make a double-cavity liquid medicine bag, which is divided into chamber A and chamber B by cold welding. The liquid in preparation tank A is divided into chamber A, and the liquid in preparation tank B is divided into chamber B. The filling temperature is 30-70°C. After filling, the product is inspected by light and high-voltage discharge for leaks, and then vacuum-packed with high-barrier outer film. The vacuum packaging parameters are 3 to 1.8S; sterilize at 121°C for 15min, with a pressure coefficient of 4.5, a temperature rise gradient of 10°C / min, and an out-of-cabinet temperature of 60°C, ensuring that the sterilization index F0 value (standard sterilization time) is ≥12. Mix chambers A and B when using.

[0084] d. The finished product is tested according to quality standards. The pH of the mixed drug solution is 7.06, the insoluble particles below 5 μm are 12 / mL, the endotoxin is less than 0.1 EU / mL, the peptidoglycan is less than 0.5 ng / mL, and the osmotic pressure is 275 mOsm / L.

[0085] Example 3

[0086] A surgical anti-adhesion fluid containing icodextrin is composed of 1200 mL of component A and 300 mL of component B, which are stored separately and mixed before use. Component A comprises 50 g / L icodextrin, 3 g / L modified yeast glucan, 0.32 g / L calcium chloride, 0.064 g / L magnesium chloride, and 2.1 g / L sodium lactate, with water for injection as the solvent. Component B comprises 27 g / L sodium chloride aqueous solution and 11.5 g / L sodium bicarbonate aqueous solution.

[0087] The preparation steps of modified yeast glucan are as follows:

[0088] 3 g of yeast glucan was added to 30 mL of N,N-dimethylformamide at 10° C. and mixed thoroughly. 7.5 g of the dehydroabietic acid derivative prepared in Preparation Example 2, 8 g of triethylamine, and 8 g of 4-dimethylaminopyridine were then added. The mixture was reacted at 40° C. for 4 h. After completion of the reaction, the reaction solution was poured into 90 mL of ice water. 150 mL of anhydrous ethanol was added, and the mixture was allowed to stand and centrifuged. The precipitate was dialyzed against deionized water for 3 days, and the dialyzate was freeze-dried to obtain modified yeast glucan.

[0089] The method for preparing the above-mentioned anti-adhesion liquid for surgery containing icodextrin comprises the following steps:

[0090] a. Pour 1000L of 50°C water for injection into preparation tank A, start stirring (150r / min), then add 60kg of icodextrin, 3.6kg of modified yeast beta-glucan, 0.3855kg of calcium chloride, 0.0765kg of magnesium chloride, and 2.52kg of sodium lactate, stirring while adding the materials to dissolve them. After stirring for 25 minutes, add 0.1mol / L dilute hydrochloric acid solution and stir for 15 minutes, adjust the pH value to 4.12, add water for injection to 1200L, and filter the liquid through 0.45μm and 0.22μm polyethersulfone filter cartridges for 25 minutes to obtain component A.

[0091] b. Pour 180L of 50°C water for injection into preparation tank B, start stirring (150r / min), then add 8.1kg of sodium chloride and 3.45kg of sodium bicarbonate, stir while adding the materials to dissolve them, stir for 25 minutes, cool to 25°C, pass carbon dioxide gas to a pH value of 7.39, add water for injection to 300L, filter the liquid through 0.45μm and 0.22μm polyethersulfone filter cartridges for 20 minutes, and then obtain component B.

[0092] c. Use non-PVC film to make a double-cavity liquid medicine bag, which is divided into chamber A and chamber B by cold welding. The liquid in preparation tank A is divided into chamber A, and the liquid in preparation tank B is divided into chamber B. The filling temperature is 30-70°C. After filling, the product is inspected by light and high-voltage discharge for leaks, and then vacuum-packed with high-barrier outer film. The vacuum packaging parameters are 3-1.8S; sterilize at 115°C with wet heat for 30min, a pressure coefficient of 4.5, a temperature rise gradient of 10°C / min, and an output temperature of 60°C, ensuring that the sterilization index F0 value (standard sterilization time) is ≥12. The finished product is packaged in boxes and mixed with chambers A and B before use.

[0093] d. The finished product is tested according to quality standards. The pH of the mixed drug solution is 6.84, the insoluble particles below 5 μm are 12 / ml, the endotoxin is less than 0.1 EU / ml, the peptidoglycan is less than 0.5 ng / ml, and the osmotic pressure is 272 mOSM / L.

[0094] Example 4

[0095] A surgical anti-adhesion fluid containing icodextrin is composed of 850 mL of component A and 150 mL of component B, which are stored separately and mixed before use. Component A comprises 47.06 g / L icodextrin, 5 g / L modified yeast glucan, 0.302 g / L calcium chloride, 0.06 g / L magnesium chloride, and 1.976 g / L sodium lactate, with water for injection as the solvent. Component B comprises 36 g / L sodium chloride aqueous solution and 15.33 g / L sodium bicarbonate aqueous solution.

[0096] The preparation steps of modified yeast glucan are as follows:

[0097] 3 g of yeast glucan was added to 40 mL of N,N-dimethylformamide at 15° C. and mixed thoroughly. Then, 9 g of the dehydroabietic acid derivative prepared in Preparation Example 3, 10 g of triethylamine, and 10 g of 4-dimethylaminopyridine were added. The mixture was reacted at 60° C. for 4 h. After completion of the reaction, the reaction solution was poured into 100 mL of ice water. 200 mL of anhydrous ethanol was added, and the mixture was allowed to stand and centrifuged. The precipitate was dialyzed against deionized water for 5 days, and the dialyzate was freeze-dried to obtain modified yeast glucan.

[0098] The method for preparing the above-mentioned anti-adhesion liquid for surgery containing icodextrin comprises the following steps:

[0099] a. Inject 800L of 50°C water for injection into preparation tank A, start stirring (150r / min), then add 40kg of icodextrin, 4.25kg of modified yeast beta-glucan, 0.257kg of calcium chloride, 0.051kg of magnesium chloride, and 1.68kg of sodium lactate, stirring while adding the materials to dissolve them. After stirring for 25 minutes, add 0.1mol / L dilute hydrochloric acid solution and stir for 15 minutes, adjust the pH value to 3.80, add water for injection to 850L, and filter the liquid through 0.45μm and 0.22μm polyethersulfone filter cartridges for 25 minutes to obtain component A.

[0100] b. Pour 100L of 50°C water for injection into preparation tank B, start stirring (150r / min), then add 5.4kg of sodium chloride and 2.3kg of sodium bicarbonate, stir while adding the materials to dissolve them, stir for 25 minutes, cool to 25°C, add sodium hydroxide, adjust the pH value to 8.97, add water for injection to 150L, ​​and filter the liquid through 0.45μm and 0.22μm polyethersulfone filter cartridges for 20 minutes to obtain component B.

[0101] c. Use non-PVC film to make a double-chamber liquid medicine bag, which is divided into chamber A and chamber B by cold welding. The liquid in preparation tank A is divided into chamber A, and the liquid in preparation tank B is divided into chamber B. The filling temperature is 30-70°C. After filling, the product is inspected by light and high-voltage discharge for leaks, and then vacuum-packed with high-barrier outer film. The vacuum packaging parameters are 3 to 1.8S; sterilize at 121°C for 15min, with a pressure coefficient of 4.5, a temperature rise gradient of 10°C / min, and a cabinet temperature of 60°C. Ensure that the sterilization index F0 value (standard sterilization time) is ≥12. The finished product is packed in boxes and mixed with chambers A and B before use.

[0102] d. The finished product is tested according to quality standards. The pH of the mixed drug solution is 6.98, the insoluble particles below 5 μm are 12 / ml, the endotoxin is less than 0.1 EU / ml, the peptidoglycan is less than 0.5 ng / ml, and the osmotic pressure is 288 mOsm / L.

[0103] Comparative Example 1

[0104] This comparative example 1 is basically the same as Example 2, except that the modified yeast glucan is replaced with an equal amount of water-soluble yeast glucan.

[0105] The preparation method of the water-soluble yeast glucan of the present invention comprises the following steps: adding 1.4 parts of concentrated sulfuric acid dropwise to 1 part of n-propanol according to parts by weight, stirring while dropping, and after uniform mixing, adding 0.5 parts of yeast glucan to the reaction solution, reacting at -6°C for 2 hours, centrifuging at -10°C after the reaction, removing the liquid in the reaction system, washing the precipitate three times with pre-cooled n-propanol, dissolving the precipitate with deionized water, centrifuging, removing the precipitate, and finally freeze-drying the supernatant to obtain the water-soluble yeast glucan.

[0106] Comparative Example 2

[0107] This comparative example 2 is basically the same as Example 2, except that the sodium lactate in component A is replaced by component B.

[0108] This comparative example 2 provides a method for preparing an anti-adhesion liquid for surgery containing icodextrin, comprising the following steps:

[0109] a. Pour 1100L of 50°C water for injection into preparation tank A, start stirring (150r / min), then add 60kg of icodextrin, 5.04kg of modified yeast beta-glucan, 0.3855kg of calcium chloride, and 0.0765kg of magnesium chloride, stirring while adding the materials to dissolve them. After stirring for 25 minutes, add 0.1mol / L dilute hydrochloric acid solution and stir for 15 minutes, adjust the pH value to 4.59, add water for injection to 1260L, and filter the solution through 0.45μm and 0.22μm polyethersulfone filter cartridges for 25 minutes to obtain component A.

[0110] b. Pour 180L of 50°C water for injection into preparation tank B, start stirring (150r / min), then add 8.1kg of sodium chloride, 2.52kg of sodium lactate and 3.45kg of sodium bicarbonate, stir while adding the materials to dissolve them, stir for 25 minutes, add hydrochloric acid to adjust the pH value to 7.19, add water for injection to 240L, and filter the liquid through 0.45μm and 0.22μm polyethersulfone filter cartridges for 20 minutes to obtain component B.

[0111] c. Use non-PVC film to make a double-chamber liquid medicine bag, which is divided into chamber A and chamber B by cold welding. The liquid in preparation tank A is divided into chamber A, and the liquid in preparation tank B is divided into chamber B. The filling temperature is 30-70°C. After filling, the product is inspected by light and high-voltage discharge for leaks, and then vacuum-packed with high-barrier outer film. The vacuum packaging parameters are 3 to 1.8S; sterilize at 121°C for 15min, with a pressure coefficient of 4.5, a temperature rise gradient of 10°C / min, and a cabinet temperature of 60°C. Ensure that the sterilization index F0 value (standard sterilization time) is ≥12. The finished product is packed in boxes and mixed with chambers A and B before use.

[0112] d. The finished product is tested according to quality standards. The pH of the mixed drug solution is 8.16, the insoluble particles below 5 μm are 12 / ml, the endotoxin is less than 0.1 EU / ml, the peptidoglycan is less than 0.5 ng / ml, and the osmotic pressure is 269 mOSM / L.

[0113] Test Example 1

[0114] The modified yeast glucan prepared in Example 1 of the present invention was subjected to infrared testing, and the results were as follows: Figure 2 shown.

[0115] Depend on Figure 2 The infrared spectrum of modified yeast glucan shows that the peak of the spectrum is 3228 cm -1 , 1250cm -1 and 796cm -1 There are absorption peaks at 1375 cm -1 There is an isopropyl absorption peak at 850cm -1 , 680cm -1 The above results indicate that the dehydroabietic acid derivatives have been successfully grafted onto yeast glucan.

[0116] Test Example 2

[0117] Biocompatibility test of dehydroabietic acid derivatives:

[0118] Mouse fibroblast L929 cells in logarithmic growth phase were resuspended in DMEM complete medium and cultured at a rate of 1×10 4 Cells were seeded into 96-well plates at a density of 100 μL / mL, and 100 μL was added to each well. The cells were cultured in a cell culture incubator at 37°C and 5% CO2 for 24 h. After the cells were completely attached, the following groups were set up: dehydroabietic acid derivative group: 100 μL of a 3 μg / mL dehydroabietic acid derivative solution prepared with DMEM complete medium was added; positive control group: 100 μL of DMEM complete medium containing 5% phenol was added; blank control group: 100 μL of DMEM complete medium was added. After further culture for 24 h, the 96-well plate was removed and the morphological characteristics of the cells in each group were observed under an inverted microscope. The results are shown in Figure 2. Figure 3 As shown, Figure 3 a is the cell compatibility diagram of the dehydroabietic acid derivative group, Figure 3 b is the cell compatibility diagram of the blank control group, Figure 3 c is the cell compatibility diagram of the positive control group.

[0119] Depend on Figure 3The results showed that the cell samples treated with the dehydroabietic acid derivatives showed a highly similar growth state to the blank control group. The cells maintained the spindle-shaped or polygonal morphology of normal fibroblasts, with transparent cytoplasm and clear boundaries. No obvious abnormal phenomena such as cell shrinkage and membrane integrity damage were observed. This indicates that the dehydroabietic acid derivatives of the present invention have no obvious cytotoxicity to mouse fibroblast L929 cells and exhibit excellent in vitro biocompatibility, preliminarily verifying their biosafety as materials for anti-adhesion fluids.

[0120] Test Example 3

[0121] Human peritoneal mesothelial cells (CP-H180 cells) were used as the test material for cell compatibility. 2×10 4 cells / mL cell suspension, 180 μL cell suspension was added to each well of a 96-well plate, with 4 groups of 6 replicate wells in each group, and cultured for 24 hours at 37°C and 5% CO2. After the cells adhered, the culture medium was sucked out and replaced with the same volume of sample to be tested, which were normal saline, the products of Examples 1 to 4, and the products of Comparative Examples 1-2. After 24 hours of culture, the cell activity was detected using the CCK8 method (Cell Counting Kit-8 cell counting method). A cell culture medium containing 10% cck8 solution was prepared, and the supernatant of each well was carefully removed. 100 μL 10% CCK8 culture medium was added and the culture was continued for 4 hours. The absorbance of the sample was measured at 450 nm on a microplate reader, and the viability of the cells was compared and analyzed according to the absorbance values. The results are shown in Table 1.

[0122] Table 1 Results of cell proliferation experiments on CP-H180 cells treated with different samples for 24 h

[0123] drug Average absorbance at 450nm Normal saline 0.252 Example 1 0.285 Example 2 0.301 Example 3 0.296 Example 4 0.299 Comparative Example 1 0.282 Comparative Example 2 0.271

[0124] The present invention adopts a dual-chamber packaging form, wherein the acidic part A and the alkaline part B containing icodextrin are stored separately by cold welding and mixed before clinical use. The product of the present invention has no effect on the proliferation and growth of human peritoneal mesothelial cells and has good biocompatibility, indicating that the pH value of the product obtained by the present invention is appropriate, and the irritation of low pH value to blood vessels, tissues, abdominal organs, etc. is solved. Compared with normal saline, the biocompatibility of the product is improved to varying degrees.

[0125] Test Example 4

[0126] The levels of GDPs, a degradation product of icodextrin, in the products of Examples 1 and 2 and Adept 4% were determined by HPLC. The test results are shown in Table 2. Figure 4 and Figure 5 This is a test result diagram of the product in Example 2.

[0127] Table 2

[0128] Types of degraded impurities GDPs Example 1 Product Example 2 product Marketed product: Adept 4% Formaldehyde (μg / ml) 0.009735 0.09967 0.15091 Acetaldehyde (μg / ml) 0.33512 0.35485 3.72230 3-Deoxyglucosone 3-DG (μg / ml) Not detected Not detected 3.0717 2-Glucuronone 2-k-DG (μg / ml) 0.0766 0.0758 0.0803 Glyoxal GO (μg / ml) 0.0851 0.0842 0.1409 Methylglyoxal MGO (μg / ml) 0.0423 0.0455 0.0800 Maximum unknown single impurity (μg / ml) 0.6751 0.6865 0.7795 Total unknown impurities (μg / ml) 1.0321 1.0932 1.8387 5-Hydroxymethylfurfural (absorbance) 0.015 0.012 0.017

[0129] The above results indicate that the above-mentioned combination and packaging form of the present invention overcome the drawbacks of the marketed product Adept 4%, which cannot include bicarbonate in the formulation due to stability issues and incompatibility. Furthermore, by changing the pH level of the environment in which icodextrin is located, the levels of various degradation impurities generated after icodextrin sterilization are significantly reduced, thereby improving the biocompatibility of the product.

[0130] Test Example 5

[0131] Acute in vivo toxicity test of anti-adhesion fluid:

[0132] The test was conducted according to the method recommended in the national standard GB / T16886.11-2021 "Biological Evaluation of Medical Devices Part 11: Systemic Toxicity Test". ICR mice weighing 20-22g were selected and provided by Zhejiang Weitonglihua Experimental Animal Technology Co., Ltd. The breeding temperature was controlled at 20℃-22℃ and the relative humidity was controlled at 40%-50%. The light control was 12 hours light and 12 hours dark, and the adaptive breeding was carried out for 6 days. The above mice were divided into a control group and an example group, with 5 mice in each group.

[0133] Each animal in the Example group was intraperitoneally injected with the anti-adhesion solution obtained in Example 2 at a dose of 50 mL / kg; mice in the control group were injected with an equivalent volume of 0.9% sodium chloride injection. Animal body weights were measured immediately before injection. Following injection, the animals' immediate biological reactions (such as dyspnea, convulsions, or prone position) were observed. The general condition and toxic reactions of the animals in the experimental and control groups were observed and recorded 4, 24, 48, and 72 hours after injection. Animal body weights were measured and recorded at the initial, 24, 48, and 72-hour intervals, and toxic reactions were observed.

[0134] Under the experimental conditions, there was no significant change in the body weight of the animals in the control group and the example group, and no convulsions or prone positions were observed in the animals, indicating that the anti-adhesion liquid sample of Example 2 had no acute systemic toxicity reaction.

[0135] Test Example 6

[0136] Experimental animals: 30 adult, female, nulliparous, non-pregnant New Zealand albino rabbits, 5-6 months of age, sexually mature and weighing more than 2.5 kg, provided by Tongxiang Yinhai Animal Husbandry Cooperative, production license: SCXK(Zhe)2023-0002. They were housed in the Xuelin Street Laboratory Animal Laboratory Building of the Zhejiang Institute of Medical Device Inspection and Research, and provided daily with rabbit feed from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd. The temperature was maintained at 20°C-22°C, and the relative humidity was maintained at 40%-50%. A 12-hour light-on / 12-hour dark cycle was maintained, and the animals were acclimated for 6 days.

[0137] To establish a pelvic adhesion model in New Zealand albino rabbits, the rabbits were fasted for 12 hours before surgery, with no food or water withdrawal. Weight was recorded and recorded. A mixture of Zotai 50, Domitor, and Suminxin (1:1:1) was then injected intramuscularly at 0.3 mL / kg to anesthetize the rabbits. The abdomen was shaved and disinfected five times with iodine.

[0138] Before surgery, the patient lies supine. A midline longitudinal incision approximately 4 cm long is made, 2 cm from the pubic bone. The abdomen is advanced gradually (the skin is thin, similar to that of humans). The uterus is located. It is double, pink, and has long horns. The fallopian tubes are located at the distal ends of the horns, surrounding the ovaries, which appear white. Observe and record the condition of the uterus and surrounding organs, noting the presence of adhesions.

[0139] The uterine horns were exposed and mechanical trauma was performed on each side of the uterine horns, approximately 4 cm in length. The serosal surface of the horns was wiped with sterile gauze for approximately 5 minutes until subserous bleeding and punctate hemorrhage occurred. Before the abdominal wall was closed, the animals were treated as follows:

[0140] Example 1 group: first rinse the wound with 20 mL of the anti-adhesion liquid prepared in Example 1 and then perfuse the peritoneal cavity with 20 mL of the anti-adhesion liquid prepared in Example 1, 10 rats in each group;

[0141] Example 2 group: the wound was first flushed with 20 mL of the anti-adhesion liquid prepared in Example 2, and then 20 mL of the anti-adhesion liquid prepared in Example 2 was perfused into the abdominal cavity. There were 10 rats in each group.

[0142] Group 3: The wound was first flushed with 20 mL of the anti-adhesion solution prepared in Example 3, and then the peritoneal cavity was perfused with 20 mL of the anti-adhesion solution prepared in Example 3. There were 10 rats in each group.

[0143] Group 4: The wound was first flushed with 20 mL of the anti-adhesion solution prepared in Example 4, and then the peritoneal cavity was perfused with 20 mL of the anti-adhesion solution prepared in Example 4. There were 10 rats in each group.

[0144] Comparative Example 1 group: first rinse the wound with 20 mL of the anti-adhesion liquid prepared in Comparative Example 1 and then perfuse the wound with 20 mL of the anti-adhesion liquid prepared in Comparative Example 1, 10 rats in each group;

[0145] Comparative Example 2 group: first rinse the wound with 20 mL of the anti-adhesion liquid prepared in Comparative Example 2, then perfuse the wound with 20 mL of the anti-adhesion liquid prepared in Comparative Example 2, 10 rats in each group;

[0146] Sodium lactate Ringer's injection group: first rinse the wound with 20 mL of sodium lactate Ringer's injection and then perfuse 20 mL of sodium lactate Ringer's injection into the abdominal cavity, 10 rats in each group;

[0147] Blank control group: no treatment, 10 mice in each group.

[0148] In each group, the abdominal wall of the animals was sutured continuously with 2 / 0 non-absorbable sutures, and the skin was sutured interrupted with 2 / 0 non-absorbable sutures. After disinfection with iodine, a sterile dressing was applied. Following surgery, 200,000 units of penicillin sodium were injected intramuscularly into the gluteus maximus muscle of the rabbit. The rabbit was placed in a warm place to maintain warmth and placed in a lateral position to maintain airway patency. Water was allowed 12 hours after surgery and food was allowed the following day. Within 3 days after surgery, 200,000 IU of penicillin were injected intramuscularly twice daily to prevent infection. The animals' activity and food intake were observed and recorded daily.

[0149] On the 7th day after surgery, the New Zealand Baihua rabbits underwent a second laparotomy. The abdominal wall was cut open in a U-shape 3 cm away from the surgical incision to fully expose the pelvic and abdominal organs and peritoneum. The adhesions between the uterus and the surrounding organs, and other abnormal conditions in the pelvic and abdominal cavity, such as the characteristics of ascites, were observed and recorded.

[0150] Adhesion grading and scoring were determined according to the test protocol provided by the client. The grading criteria are shown in Table 3. Three experimenters blindly evaluated each animal using qualitative and quantitative parameters, including adhesion extent, location, and intensity. The scoring results are recorded in Table 4.

[0151] Table 3 Adhesion degree grading standard

[0152]

[0153] Table 4 Pelvic adhesion grades and scores in each group

[0154]

[0155]

[0156] Note: Total score = grade score × number of rabbits.

[0157] Adhesion all occurs in all New Zealand albino rabbits of blank control group after operation, and uterus itself and adhesion with the opposite side account for more than 70%, and adhesion of uterus to bladder accounts for more than 20%, mainly with grade III and grade IV. Adhesion situation in sodium lactate Ringer's injection group is similar to that in blank control group, and all rabbits all appear adhesion, and uterus itself and adhesion with the opposite side account for more than 90%, and adhesion is dense, and is difficult to be separated with grade III and grade IV by blunt force. The anti-adhesion liquid containing icodextrin of embodiment 1-3 group peritoneal perfusion after operation has been absorbed, and no adhesion situation accounts for more than 60%, and adhesion is mainly with looser membranous adhesion, and adhesion range is less, and is easier to separate than blank control group, and is mainly with grade 0, illustrates that the test sample group is significantly better than blank control group and sodium lactate Ringer's injection group aspect adhesion range and toughness separation strength, and test shows that the operation anti-adhesion liquid containing icodextrin can effectively reduce the formation of adhesion, and the anti-adhesion effect after operation is remarkable.

[0158] The total adhesion grade score of Comparative Example 1 (modified yeast glucan was replaced with an equal amount of water-soluble yeast glucan) after surgery was significantly higher than that of Examples 1-4, and the total score of Example 1 was also slightly higher than that of Examples 2-4. This is because the addition of modified yeast glucan not only improves the water solubility of yeast glucan, but also its hydrophilic sulfonate and hydroxyl groups can cross-link with icodextrin to improve the stability of icodextrin. In addition, dehydroabietic acid belongs to the tricyclic diterpenoid class of compounds. Its unique hydrophobic tricyclic skeleton and rigid structure can effectively increase the retention time of yeast glucan in the body, thereby increasing the duration of its synergistic effect with icodextrin, continuously isolating the damaged tissue surface, reducing direct contact between the uterus and tissue, inhibiting fibrin deposition and adhesion formation, and achieving an effective and safe anti-adhesion effect.

[0159] In addition, the total adhesion grade score of Comparative Example 2 was significantly higher than that of Examples 1-3, indicating that the compatibility of bicarbonate and lactate in the present invention is relatively poor. The combination and dual-chamber packaging of the present invention address the shortcomings of the marketed product Adept 4%, which cannot include bicarbonate in the formulation due to stability issues and incompatibility. This not only inhibits the degradation of icodextrin (controlled within the pH range of 5.0 to 5.5 for direct use) in conventional processes to produce harmful GDPs, but also addresses the irritation of low pH to blood vessels, tissues, abdominal organs, etc., thereby improving the anti-adhesion effect.

[0160] The above embodiments are only preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and replacements made by technicians in this field on the basis of the present invention fall within the scope of protection required by the present invention.

Claims

1. An anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin, characterized in that: The invention comprises component A and component B, which are stored separately; component A is a solution with water as solvent, comprising 20-100 g / L icodextrin, 0-20 g / L sodium lactate, 0.1-0.6 g / L calcium chloride, 0.03-0.15 g / L magnesium chloride, and 0-60 g / L sodium chloride; component B is a solution with water as solvent, comprising 0-54 g / L sodium chloride, 10-20 g / L sodium bicarbonate, and 0-19 g / L sodium lactate; The volume ratio of the component A to the component B is 1:0.1-0.9; the pH of the component A is 3.0-6.5, and the pH of the component B is 4.5-9.

0.

2. The anti-adhesion liquid for laparoscopic or laparoscopic surgery containing icodextrin according to claim 1, characterized in that: The invention comprises component A and component B, which are stored separately; component A is a solution with water as solvent, comprising 20-75 g / L icodextrin, 1-3 g / L sodium lactate, 0.1-0.5 g / L calcium chloride, and 0.05-0.08 g / L magnesium chloride; component B is a solution with water as solvent, comprising 25-40 g / L sodium chloride and 10-20 g / L sodium bicarbonate; The volume ratio of the component A to the component B is 1:0.1-0.8; the pH of the component A is 3.0-5.0, and the pH of the component B is 6.0-9.

0.

3. The anti-adhesion liquid for laparoscopic or laparoscopic surgery containing icodextrin according to claim 2, characterized in that: The A component also includes 2-5 g / L modified yeast glucan. The preparation process of the modified yeast glucan is as follows: yeast glucan is added to a solvent, mixed evenly, and then a dehydroabietic acid derivative, triethylamine, and 4-dimethylaminopyridine are added to react, and the reaction solution is post-treated to obtain the modified yeast glucan.

4. The anti-adhesion liquid for laparoscopic or laparoscopic surgery containing icodextrin according to claim 3, characterized in that: The mass ratio of the yeast glucan, dehydroabietic acid derivative, triethylamine, and 4-dimethylaminopyridine is 1:(2.5-3):(2.6-3.5):(2.6-3.5), and the concentration of the yeast glucan in the solvent is 0.075-0.1 g / mL; the solvent is N,N-dimethylformamide; the reaction temperature is 40-60°C, and the reaction time is 2-4 hours.

5. The anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin according to claim 3, characterized in that: The structural formula of the dehydroabietic acid derivative is The preparation process of the dehydroabietic acid derivative comprises the following steps: (1) adding lithium aluminum tetrahydride to tetrahydrofuran, and then adding dehydroabietic acid to react to obtain dehydroabietic acid reduced alcohol; (2) Adding dehydroabietic acid reducing alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate into N,N-dimethylformamide for reaction, and purifying the reaction solution after the reaction is completed to obtain a dehydroabietic acid derivative.

6. The anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin according to claim 5, characterized in that: In step (1), the molar ratio of dehydroabietic acid to lithium aluminum tetrahydride is 1:4-4.5, and the amount ratio of dehydroabietic acid to tetrahydrofuran is 1 mmol:5-5.5 mL. In step (1), when dehydroabietic acid is added, the temperature of the reaction system is -2-3°C, and the reaction time is 4-6 hours.

7. The anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin according to claim 5, characterized in that: In step (2), the molar ratio of the dehydroabietic acid reducing alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate is 1:(1-1.1):(2.5-3); and the concentration of the dehydroabietic acid reducing alcohol in N,N-dimethylformamide is 0.3-0.5 mol / L.

8. The anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin according to claim 5, characterized in that: In step (2), the inorganic carbonate is potassium carbonate or sodium carbonate, the reaction temperature is 60-80° C., and the reaction time is 12-18 h.

9. The method for preparing the anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin according to any one of claims 1 to 8, characterized in that: The steps include: a. Add the raw material components to water according to the ratio, stir evenly, adjust the pH value to the range, and then filter and fill to obtain component A; b. Add the raw material components to water according to the ratio, stir evenly, adjust the pH to the range, filter and fill to obtain component B; c. Fill component A and component B separately and sterilize them. Mix component A and component B when using.

10. The method for preparing the anti-adhesion liquid for abdominal surgery or laparoscopic surgery containing icodextrin according to claim 9, characterized in that: The sterilization temperature is 115-121° C., and the sterilization time is 10-30 minutes.

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