An icodextrin-containing anti-adhesion solution for abdominal surgery or laparoscopic surgery and a method for preparing the same

Abstract

The application belongs to the technical field of surgical anti-adhesion liquids, and particularly relates to an anti-adhesion liquid containing icodextrin for abdominal surgery or laparoscopic surgery and a preparation method thereof. The anti-adhesion liquid is composed of component A and component B, and the component A and the component B are stored separately. The component A is a solution with water as a solvent, and includes 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 a solvent, and includes 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. The anti-adhesion liquid can clean a wound surface and prevent tissue and organ adhesion.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of anti-adhesion solution for surgery, in particular to an anti-adhesion solution for abdominal surgery or laparoscopic surgery containing icodextrin and a preparation method thereof. BACKGROUND

[0002] Postoperative adhesion is a common and disturbing complication after surgery, especially after gynecological abdominal surgery. These adhesions can not only cause female infertility, sterility, but also cause intestinal adhesion, intestinal obstruction, and various complications such as chronic abdominal pain, pelvic pain, etc., increasing the difficulty of reoperation in the pelvic and abdominal cavity, and a large number of complications related to postoperative adhesion bring more and more burden and considerable cost to surgeons and the healthcare system. Therefore, it is necessary to fully assess the risk of adhesion before surgery and take effective anti-adhesion measures during surgery to reduce the incidence of adhesion complications.

[0003] In traditional abdominal adhesion prevention measures, various materials of anti-adhesion isolation film are placed in the parts prone to adhesion, such as a hyaluronic acid-carboxymethyl cellulose film; there are also anti-adhesion gel barriers used in parts prone to adhesion, such as hyaluronic acid, chitosan and other macromolecular materials and polyethylene glycol-based , etc. These means can effectively prevent and reduce the occurrence of adhesion. However, in recent years, with the popularization of laparoscopic technology and its significant advantages, traditional open surgery is more and more replaced by laparoscopic surgery. In these surgeries, the anti-adhesion operation with film isolation and gel isolation as means has certain limitations when cooperating with laparoscopic surgery. Compared with the above, liquid floating type anti-adhesion products are more suitable for combination with laparoscopic surgery to carry out postoperative anti-adhesion operation.

[0004] Because of its unique colloid osmotic properties and macromolecular volume, icodextrin cannot diffuse directly through the peritoneal wall into the intraperitoneal microvessels and be absorbed by the body, so it can maintain its clearance efficiency in the abdominal cavity for a long time. The glycosidic bond of icodextrin is usually metabolized by amylase, which does not exist 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 of time, the icodextrin solution present in the peritoneal cavity can separate the damaged surfaces and minimize the contact between organs during the critical period of adhesion formation. FDA approved Adept 4% containing icodextrin as an anti-adhesion product after surgery. But this product still has some obvious defects. First, compared with icodextrin peritoneal dialysis solution, there are two significant differences in the clinical application mode 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, relying on colloid osmotic pressure to play a clinical role, except for a small part that is absorbed, most of it will not come into contact with the tissues in the abdominal cavity, while icodextrin as an anti-adhesion product needs to be infused into the abdominal cavity, directly contacting various organs and tissues in the abdominal cavity; (2) Icodextrin peritoneal dialysis solution has a residence time of 8-12 hours in use, and the longest is not more than 16 hours, and then it is excluded from the outside, while as an anti-adhesion product, icodextrin anti-adhesion fluid needs to stay in the abdominal cavity for at least 4 days or more until it is completely absorbed into the systemic circulation through the lymphatic system. The above clinical use mode puts forward higher biocompatibility requirements for icodextrin anti-adhesion products: (1) The composition and concentration level should be closer to the physiological body fluid of the human body; (2) Higher biocompatibility, especially pH compatibility, in order to reduce its irritability to abdominal cavity tissues and organs.

[0005] Secondly, icodextrin is more stable in a slightly acidic condition in liquid preparation. After high-temperature sterilization, icodextrin will produce a large amount of degradation products in an environment with a pH above 5.5, including 5-hydroxymethyl furfural, formaldehyde, acetaldehyde, methylglyoxal, glyoxal, 3-deoxyglucosulose ketone, etc. These glucose degradation products (GDPs) have varying degrees of cytotoxicity and can cause apoptosis of various epithelial cells in the peritoneum after long-term contact. Therefore, in order to ensure the stability of icodextrin, the pH of the product needs to be reduced, and the stable pH of icodextrin in the liquid state is between 3.5 and 5.0, but a too low pH will directly irritate the organs and tissues in the abdominal cavity, so in order to balance the stability and compatibility of icodextrin, the pH of the product is usually adjusted to between 5.0 and 5.5. But research shows that this pH range still has a negative impact on the survival of human peritoneal mesothelial cells, and long-term contact can accelerate their apoptosis, with a much higher rate than that of neutral pH solutions.

[0006] The marketed product Adept 4% cannot solve the stability problem after adding sodium bicarbonate in the formula, cannot reach the higher compatibility level of human body fluid components, and cannot solve the stability problem of icodextrin in the formula under neutral pH condition, and the glucose degradation product is also significantly higher than the physiological tolerance level.

[0007] Therefore, the present application provides a liquid anti-adhesion liquid after mixing, which has a component and pH closer to the physiological state of body fluid, prolongs the time of the liquid anti-adhesion liquid in the abdominal cavity, improves the effectiveness and safety, and optimizes the production process to improve the stability of icodextrin under high-temperature sterilization conditions. SUMMARY

[0008] In order to overcome the shortcomings of the prior art, the surgical anti-adhesion liquid containing icodextrin is suitable for various abdominal cavity surgeries in clinical application, can be used for cleaning surgical wounds during abdominal cavity surgeries, and can be used for abdominal cavity perfusion after various abdominal cavity surgeries to prevent postoperative adhesion of organs and tissues in the abdominal cavity.

[0009] One of the purposes of the present application is to provide a surgical anti-adhesion liquid containing icodextrin.

[0010] One of the purposes of the present application is achieved by adopting the following technical solutions:

[0011] A surgical anti-adhesion liquid containing icodextrin is composed of component A and component B, and component A and component B are stored separately.

[0012] The component A is a solution with water as a solvent, and includes 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 a solvent, and includes 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] Further, the component A is a solution with water as a solvent, and includes 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 a solvent, and includes 25-40 g / L sodium chloride and 10-20 g / L sodium bicarbonate.

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

[0016] Further, the anti-adhesion liquid is composed of two chambers of A component and B component, which are separated by virtual welding, and are penetrated by extrusion before use, and are mixed uniformly by shaking.

[0017] Further, the volume ratio of the A component and the B component is 1:0.1-0.5; the pH of the A component is 3.5-4.5, and the pH of the B component is 7.0-9.0; after mixing of the A component and the B component, the pH of the mixed liquid is 6.5-7.5.

[0018] Further, 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 oxalate solution.

[0019] The prescription amount of the exciplex dextrin in the surgical anti-adhesion liquid containing exciplex dextrin is large, although the solubility is high, in order to reduce the viscosity in the preparation process and facilitate filling, it is placed in a large chamber. Magnesium chloride and calcium chloride may form calcium carbonate and magnesium carbonate precipitates in sodium bicarbonate aqueous solution in long-term storage, so they need to be placed in different chambers. Exciplex dextrin needs to be stored in weak acid conditions, and sodium bicarbonate cannot exist stably under acidic conditions, so exciplex dextrin, magnesium chloride and calcium chloride are placed in a large chamber, and sodium bicarbonate is placed separately.

[0020] Sodium lactate aqueous solution and sodium bicarbonate aqueous solution are both alkaline, and theoretically they can be placed in the same chamber, but research shows that their stability is poor after sterilization, especially after mixing, sodium bicarbonate decomposes into carbon dioxide and overflow, which significantly increases the pH level of the mixed liquid. Sodium lactate can tolerate a lower level of pH, so further, the present application preferably places sodium lactate and exciplex dextrin in the same chamber and coexists under acidic conditions.

[0021] In the present application, referring to the composition of electrolytes in human body fluid, sodium bicarbonate is added as a component instead of a pH regulator in the prescription of the present application to keep the liquid perfused into the abdominal cavity from interfering with the acid-base balance of the human body fluid, sodium bicarbonate is added to ensure that the concentration is basically consistent with the bicarbonate level in the blood and the bicarbonate level in the body fluid, and the ratio of the two in the mixed solution can keep the anti-adhesion solution have a more stable pH level and buffering capacity. At the same time, a certain concentration of sodium lactate is retained in the formula, and the sodium lactate at this concentration level can avoid the situation of excessive use of sodium bicarbonate for the purpose of pH control, which may cause the carbon dioxide partial pressure in the body fluid to be too high.

[0022] Regarding the volume ratio of component A to component B, component A is first determined as a large chamber to facilitate the dissolution of a large amount of excipient, component A is an acidic solution, component B is an alkaline solution, and the pH of the mixture is close to neutral. Under the premise of following the principle of using less pH regulator, the stability of product quality can be met, so the optimal volume ratio of components A and B is limited by the above conditions.

[0023] Further, the A component further comprises 2-5 g / L modified yeast glucan, and the preparation process of the modified yeast glucan is as follows: yeast glucan is added into a solvent, after uniform mixing, dehydroabietic acid derivative, triethylamine and 4-dimethylaminopyridine are added for reaction, and then the reaction solution is treated to obtain the modified yeast glucan.

[0024] Further, the treatment process is to pour ice water into the reaction solution, then add anhydrous ethanol, after standing and centrifugation, the precipitate is dialyzed, and the dialysate is freeze-dried.

[0025] Further, the mass ratio of the yeast glucan, the dehydroabietic acid derivative, the triethylamine and the 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 h.

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

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

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

[0029] (1) Lithium aluminum hydride is added into tetrahydrofuran, and then dehydroabietic acid is added for reaction to prepare dehydroabietic acid reduction alcohol;

[0030] (2) adding dehydroabietyl acid reductive alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate into N,N-dimethylformamide to react, and after the reaction is completed, the reaction solution is purified to obtain a dehydroabietyl acid derivative.

[0031] Further, the molar ratio of the dehydroabietyl acid to lithium aluminum hydride in step (1) is 1:4-4.5, and the amount ratio of the dehydroabietyl acid to tetrahydrofuran is 1 mmol:5-5.5 mL; in step (1), when the dehydroabietyl acid is added, the temperature of the reaction system is -2-3℃, and the reaction time is 4-6 h.

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

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

[0034] The second object of the present application is to provide a preparation method of a surgical anti-adhesion liquid containing icodextrin, which is simple and feasible, and provides a new preparation idea for the preparation of a surgical anti-adhesion liquid.

[0035] The preparation method of the surgical anti-adhesion liquid containing icodextrin comprises the following steps:

[0036] a. According to the ratio, each raw material component is added to water, stirred uniformly, and then the pH value is adjusted to the range, and then filtered, filled and prepared into A component;

[0037] b. According to the ratio, each raw material component is added to water, stirred uniformly, and then the pH value is adjusted to the range, and then filtered, filled and prepared into B component;

[0038] c. A component and B component are filled and sterilized respectively, and A component and B component are mixed for use.

[0039] Further, when packaging, A component and B component are packaged in the same soft bag made of PVC or non-PVC material, and are divided into a large chamber and a small chamber by a virtual welding process; A component is packaged in the large chamber, and B component is packaged in the small chamber; the weak welding strength of the virtual welding part between A and B parts is 3-20 N / 15 mm, and the weak welding opening force is 300-3000 N. Before use, the virtual welding is opened by extrusion, so that the solutions in the large chamber and the small chamber are connected, and then the mixture is uniformly mixed by shaking and used.

[0040] Further, the soft bag is a non-PVC material soft bag, the loading volume of the soft bag is 500-5000ml, and the preferred specifications are 1000ml, 1500ml, 2000ml and 3000ml. Further, the osmotic pressure of the liquid in the large chamber is 50-70mOsm / L, the osmotic pressure of the liquid in the small chamber is 900-1200mOsm / L, and the osmotic pressure of the liquid after mixing of the large chamber and the small chamber is 260-310mOsm / L.

[0041] Further, the sterilization temperature is 115-121 DEG C, and the time is 10-30min; and during the packaging and filling, the C-level+A-level clean condition is used.

[0042] Further, the surgical anti-adhesion liquid containing icodextrin is suitable for various clinical abdominal surgeries, can be used for cleaning surgical wounds during abdominal surgery, and can be used for abdominal perfusion after various abdominal surgeries, so as to prevent postoperative adhesion of organs and tissues in the abdominal cavity. According to the characteristics of the surgical anti-adhesion liquid, the surgical anti-adhesion liquid is especially suitable for abdominal perfusion after various laparoscopic surgeries, and the anti-adhesion effect is achieved.

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

[0044] 1. The present application provides a surgical anti-adhesion liquid containing icodextrin, which is mainly used for preventing adhesion after various laparoscopic surgeries. The anti-adhesion liquid contains icodextrin and modified yeast glucan and other components, which jointly act on the components in the anti-adhesion liquid, realize the cleaning effect on the wound surface, and prevent postoperative adhesion. Specifically, the modified yeast glucan introduced in the present application is mainly modified by using dehydroabietic acid derivatives, and the sulfonic acid group of the dehydroabietic acid derivatives reacts with the hydroxyl group in the yeast glucan to form a sulfonate. On the one hand, the water solubility of the yeast glucan is improved, the hydrophilic groups of the yeast glucan, such as sulfonate groups and hydroxyl groups, are crosslinked with icodextrin to form a stable crosslinked system, thereby improving the stability of icodextrin, and on the other hand, the unique hydrophobic tricyclic skeleton and rigid structure of dehydroabietic acid can effectively improve the retention time of the yeast glucan in the body, thereby prolonging the duration of the synergistic effect of the yeast glucan and icodextrin, continuously isolating the surface of the damaged tissue, reducing the direct contact between the uterus and the tissue after the operation, inhibiting the deposition of fibrin and the formation of adhesion, and realizing the effective and safe postoperative anti-adhesion effect.

[0045] 2. The present application adopts a double-chamber packaging form, and stores the acidic A component and the basic B component containing icodextrin separately by virtual welding, mixes them before clinical use, so as to realize the neutral pH value (6.5-7.5), thereby inhibiting the degradation of traditional icodextrin (pH value is controlled in the range of 5.0-5.5 and is directly used) to generate harmful GDP substances, reducing the irritability of low pH value to blood vessels and tissues, and improving the biocompatibility of the product.

[0046] 3. The application provides a preparation method of the surgical anti-adhesion liquid containing icodextrin, which adopts general large-volume infusion product production process and GMP management requirements. In the production process, key process control parameters are further refined according to product characteristics, such as feeding temperature, feeding sequence, liquid precision filtration cycle time, pH control range, control of opening force of false welding part of finished product, vacuum degree of high barrier film packaging (affecting the level and uniformity of heat penetration in the sterilization process), sterilization condition control and the like (to ensure the sterility guarantee level of large-volume liquid), and the comprehensive control of the above process parameters ensures the quality reliability of the product.

[0047] 4. The anti-adhesion liquid provided by the application provides a new preparation idea for preparing the surgical anti-adhesion liquid, and the anti-adhesion liquid can be used for intraoperative lavage and irrigation in various abdominal surgeries (open / laparoscopic) and postoperative abdominal perfusion, reduces and prevents postoperative adhesion. BRIEF DESCRIPTION OF DRAWINGS

[0048] Figure 1 It is a physical map of the surgical anti-adhesion liquid of the application;

[0049] Figure 2 It is an infrared spectrum of the modified yeast glucan of the application;

[0050] Figure 3 It is a biocompatibility result graph of the anti-adhesion liquid of the application;

[0051] Figure 4 It is a detection condition graph of methanal and acetaldehyde of the sample of example 2 of the application after sterilization;

[0052] Figure 5 It is a detection condition graph of 2-k-DG, 3-DG, GO, MGO and other impurities in the sample of example 2 of the application. DETAILED DESCRIPTION

[0053] In the following, the application is further described in combination with the drawings and specific embodiments, and it should be noted that, under the premise of not conflicting, each embodiment described below or each technical feature can be combined to form a new embodiment. The specific conditions not mentioned in the embodiments are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used, such as the conventional products obtained through the market channel, are not specifically mentioned.

[0054] The icodextrin of the application has a molecular weight of 23-40 kDa, which 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 the dehydroabietic acid derivative is as follows:

[0057]

[0058] (1) Under the nitrogen atmosphere, lithium aluminum hydride (450 mmol) was added into 500 mL of tetrahydrofuran, and the temperature of the reaction system was controlled to 0°C. Dehydroabietic acid (100 mmol) was added in 5 batches, and after the addition was completed, it was stirred for 10 min, and gradually warmed to room temperature, and reacted at room temperature for 5 h. The reaction system was cooled to 0°C with an ice water bath, and water was slowly added to quench the reaction (the temperature was controlled to 10°C during the quenching process). After the ice water bath was removed, the mixture was warmed to room temperature, stirred for 10 min, and then 1M hydrochloric acid solution was added to adjust the pH of the mixture to 7. Most of the tetrahydrofuran was removed by concentration, and the water phase after removing most of the tetrahydrofuran was extracted with ethyl acetate for 3 times. The organic phase was combined and washed with saturated brine, then dried with anhydrous sodium sulfate, and concentrated under negative pressure. The dehydroabietic acid reduction alcohol was obtained by distillation purification. The characterization results of the dehydroabietic acid reduction alcohol are as follows: 1 HNMR (C 20 H 30 O, 300 MHz, DMSO-d6): δ 7.03 (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 reduction 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 isopropyl alcohol at a volume ratio of 4:1), and then stirred uniformly after adding saturated ammonium chloride aqueous solution. The mixture was separated by a separatory funnel, and the saturated ammonium chloride solution was extracted with the above-mentioned mixed organic solution. The mixed organic phase was washed with saturated brine, concentrated under negative pressure, and purified by column chromatography (90% n-heptane: 9% ethyl acetate: 1% acetic acid) to obtain the dehydroabietic acid derivative. The characterization results of the dehydroabietic acid derivative are as follows: 1 HNMR (C 22 H 34O4S, 300 MHz, DMSO-d6): δ 8.6 (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 [M-H] - The above results demonstrate that the target product is obtained.

[0060] Preparation Example 2

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

[0062] (1) Under a nitrogen atmosphere, lithium aluminum hydride (400 mmol) was added to 500 mL of tetrahydrofuran, and the temperature of the reaction system was controlled to -2°C. Dehydroabietic acid (100 mmol) was added in five portions, and after the addition was completed, the system was stirred for 10 min, and gradually warmed to room temperature. The reaction was carried out at room temperature for 4 h. The reaction system was cooled to 5°C with an ice water bath, and water was slowly added to quench the reaction (the temperature was controlled to 10°C during the quenching process). After the ice water bath was removed, the system was warmed to room temperature, stirred for 10 min, and then 1M hydrochloric acid solution was added to adjust the pH of the system to 7. Most of the tetrahydrofuran was removed by concentration, and the aqueous phase was extracted with ethyl acetate three times. The organic phases were combined and washed with saturated brine, dried with anhydrous sodium sulfate, and concentrated under negative pressure. The dehydroabietic acid reduction alcohol was obtained by distillation purification. The characterization results of the dehydroabietic acid reduction alcohol were the same as those of Preparation Example 1.

[0063] (2) Dehydroabietic acid reduction 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 the reaction was carried out at 60°C for 18 h. The reaction solution was diluted with a mixed organic solution (prepared by mixing dichloromethane and isopropyl alcohol at a volume ratio of 4:1), and then saturated ammonium chloride aqueous solution was added and stirred uniformly. The mixture was separated by a separatory funnel, and the saturated ammonium chloride solution was extracted with a mixed organic solution. The combined organic phase was washed with saturated brine, concentrated under negative pressure, and purified by column chromatography (90% n-heptane:9% ethyl acetate:1% acetic acid) to obtain the dehydroabietic acid derivative. The characterization results of the dehydroabietic acid derivative were the same as those of Preparation Example 1.

[0064] Preparation Example 3

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

[0066] (1) In a nitrogen atmosphere, lithium aluminum hydride (450 mmol) was added into 550 mL of tetrahydrofuran, and the temperature of the reaction system was controlled to 3°C. Dehydroabietic acid (100 mmol) was added in 5 batches, and after the addition was completed, the system was stirred for 10 min, and gradually warmed to room temperature, and reacted at room temperature for 6 h. The reaction system was cooled to 0°C with an ice water bath, and water was slowly added to quench the reaction (the temperature was controlled to 10°C during the quenching process). After the ice water bath was removed, the system was warmed to room temperature, stirred for 10 min, and then 1M hydrochloric acid solution was added to adjust the pH of the system to 7. Most of the tetrahydrofuran was removed by concentration, and the water phase was extracted with ethyl acetate three times. The organic phase was combined and washed with saturated brine, dried with anhydrous sodium sulfate, and concentrated under negative pressure. The dehydroabietic acid reduction alcohol was obtained by distillation purification. The characterization results of the dehydroabietic acid reduction alcohol were the same as those of Preparation Example 1.

[0067] (2) Dehydroabietic acid reduction 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 isopropyl alcohol at a volume ratio of 4:1), and then saturated ammonium chloride aqueous solution was added and stirred uniformly. The mixture was separated by a separatory funnel, and the saturated ammonium chloride solution was extracted with a mixed organic solution. The combined organic phase was washed with saturated brine, concentrated under negative 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 of Preparation Example 1.

[0068] Example 1

[0069] A surgical anti-adhesion liquid containing icodextrin, which is composed of 1200 mL of component A and 300 mL of component B, the component A and the component B are stored separately, and the component A and the component B are mixed before use; the 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, and the solvent is water for injection; the component B comprises 27 g / L sodium chloride aqueous solution and 11.5 g / L sodium bicarbonate aqueous solution.

[0070] The preparation method of the above-mentioned surgical anti-adhesion liquid containing icodextrin, comprising the following steps:

[0071] a. Into the preparation tank A, 1100 L of 50℃ water for injection was injected, and the stirring (150 r / min) was started, then 60 kg of Eudragit L30D-55, 0.3855 kg of calcium chloride, 0.0765 kg of magnesium chloride, 2.52 kg of sodium lactate were put in, and the stirring was kept on to dissolve the materials. After 25 min, 0.1 mol / L dilute hydrochloric acid solution was added, and the stirring was kept on for 15 min. The pH value was adjusted to 4.09. The injection water was added to 1200 L. The solution was filtered through 0.45 μm and 0.22 μm filter cartridges for 25 min, and then the A component was obtained.

[0072] b. Into the preparation tank B, 180 L of 50℃ water for injection was injected, and the stirring (150 r / min) was started, then 8.1 kg of sodium chloride and 3.46 kg of sodium bicarbonate were put in, and the stirring was kept on to dissolve the materials. After 25 min, the pH value was measured to be 8.19. The injection water was added to 300 L. The solution was filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges for 20 min, and then the B component was obtained.

[0073] c. The non-PVC film was used to prepare a double-chamber solution bag, which was divided into A chamber and B chamber by virtual welding. The solution in the preparation tank A was divided and filled into the A chamber, and the solution in the preparation tank B was divided and filled into the B chamber. The filling temperature was 30-70℃. The product after filling and sealing was subjected to lamp inspection and high-voltage discharge leak detection, and then was vacuum packaged with high-barrier outer film. The vacuum packaging parameters were 3 to 1.8 S. The product was subjected to moist heat sterilization at 121℃ for 15 min, with a pressure coefficient of 4.5, a temperature rising gradient of 10℃ / min, and an outlet cabinet temperature of 60℃. The sterilization index F0 value (standard sterilization time) was ≥12, and then the product was obtained. When used, the A chamber and the B chamber were mixed.

[0074] d. The finished product was detected according to the quality standard. The pH value of the mixed solution was 7.06, the number of insoluble particles below 5 μm was 12 per mL, the endotoxin was less than 0.1 EU / mL, the peptidoglycan was less than 0.5 ng / mL, and the osmotic pressure was 269 mOsm / L.

[0075] The appearance and size of the folded sample of the sample of Example 1 are shown in Figure 1 .

[0076] Example 2

[0077] A surgical anti-adhesion solution containing Eudragit L30D-55, which is composed of 1260 mL of A component and 240 mL of B component. The A component and the B component are stored separately, and the A component and the B component are mixed before use. The A component includes 47.62 g / L of Eudragit L30D-55, 4 g / L of modified yeast glucan, 0.31 g / L of calcium chloride, 0.061 g / L of magnesium chloride, and 2 g / L of sodium lactate, and the solvent is water for injection. The B component includes 33.75 g / L of sodium chloride aqueous solution and 14.38 g / L of sodium bicarbonate aqueous solution.

[0078] wherein the modified yeast glucan is prepared by the following steps:

[0079] 3 g of yeast glucan was added to 40 mL of N, N-dimethylformamide at 13°C, and after being mixed evenly, 8 g of the dehydroabietyl acid derivative of Preparation Example 1, 9 g of triethylamine, and 9 g of 4-dimethylaminopyridine were added, and the reaction was performed at 50°C for 3 h. After the reaction was completed, the reaction solution was poured into 95 mL of ice water, and then 180 mL of anhydrous ethanol was added, followed by standing and centrifugation. The precipitate was dialyzed with deionized water for 4 days, and the dialysis solution was freeze-dried to obtain the modified yeast glucan.

[0080] The preparation method of the surgical anti-adhesion solution containing icodextrin described above comprises the following steps:

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

[0082] b. 180 L of 50°C water for injection was injected into a preparation tank B, and stirring was started (150 r / min), and then 8.1 kg of sodium chloride and 3.45 kg of sodium bicarbonate were added while stirring to dissolve them. After stirring for 25 min, the pH value was measured to be 8.19. Injection water was added to 240 L, and the solution was filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges for 20 min to obtain the B component.

[0083] c. A non-PVC film double-chamber solution bag was used, which was divided into A and B chambers by virtual welding. The solution in the preparation tank A was divided and filled into the A chamber, and the solution in the preparation tank B was divided and filled into the B chamber. The filling temperature was 30-70°C. The product after filling and sealing was subjected to lamp inspection and high-voltage discharge leak detection, and then vacuum packaging was performed using a high-barrier outer packaging film with a vacuum packaging parameter of 3 to 1.8 S. The product was subjected to moist heat sterilization at 121°C for 15 min with a pressure coefficient of 4.5 and a temperature rise gradient of 10°C / min, and the cabinet outlet temperature was 60°C to ensure that the sterilization index F0 value (standard sterilization time) was ≥12. Thus, the product was obtained, and the A and B chambers were mixed for use.

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

[0085] Example 3

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

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

[0088] 3 g of yeast glucan is added to 30 mL of N,N-dimethylformamide at 10°C, 7.5 g of the dehydroabietyl acid derivative prepared in Preparation Example 2, 8 g of triethylamine, and 8 g of 4-dimethylaminopyridine are added after uniform mixing, and the reaction is carried out at 40°C for 4 h, after the reaction is completed, the reaction solution is poured into 90 mL of ice water, 150 mL of anhydrous ethanol is added, and then standing and centrifugation are carried out, the precipitate is dialyzed with deionized water for 3 days, and the dialysate is freeze-dried to obtain the modified yeast glucan.

[0089] The preparation method of the above-mentioned surgical anti-adhesion liquid containing icodextrin comprises the following steps:

[0090] a. 1000 L of 50°C water for injection is injected into a preparation tank A, stirring is started (150 r / min), and then 60 kg of icodextrin, 3.6 kg of modified yeast glucan, 0.3855 kg of calcium chloride, 0.0765 kg of magnesium chloride, and 2.52 kg of sodium lactate are added while stirring to dissolve them, 0.1 mol / L dilute hydrochloric acid solution is added after stirring for 25 min, stirring is carried out for 15 min to adjust the pH value to 4.12, water for injection is added to 1200 L, and the liquid is filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges in sequence for 25 min to obtain the A component.

[0091] b. 180 L of 50°C water for injection is injected into a preparation tank B, stirring is started (150 r / min), and then 8.1 kg of sodium chloride and 3.45 kg of sodium bicarbonate are added while stirring to dissolve them, the temperature is lowered to 25°C after stirring for 25 min, carbon dioxide gas is introduced until the pH value is 7.39, water for injection is added to 300 L, and the liquid is filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges for 20 min to obtain the B component.

[0092] c. The double-chambered liquid medicine bag is made of non-PVC film, and is divided into chamber A and chamber B by virtual welding. The liquid medicine in preparation tank A is divided and filled into chamber A, and the liquid medicine in preparation tank B is divided and filled into chamber B. The filling temperature is 30-70°C. The product after filling and sealing is subjected to lamp inspection and high-voltage discharge leak detection, and then is vacuum packaged by using a high-barrier outer packaging film. The vacuum packaging parameters are 3-1.8S. The product is subjected to moist heat sterilization at 115°C for 30 min, with a pressure coefficient of 4.5, a temperature rising gradient of 10°C / min, and an outlet cabinet temperature of 60°C, so as to ensure that the sterilization index F0 value (standard sterilization time) is ≥12. The product is packaged to obtain a finished product, and A chamber and B chamber are mixed during use.

[0093] d. The finished product is detected according to the quality standard. The pH of the mixed liquid medicine is 6.84, the number of insoluble microparticles below 5um is 12 / ml, the endotoxin is lower than 0.1 EU / ml, the peptidoglycan is lower than 0.5 ng / ml, and the osmotic pressure is 272 mOSM / L.

[0094] Example 4

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

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

[0097] 3 g of yeast glucan is added to 40 mL of N, N-dimethylformamide at 15°C, 9 g of the dehydroabietyl acid derivative prepared in Preparation Example 3, 10 g of triethylamine, and 10 g of 4-dimethylaminopyridine are added after uniform mixing, and the reaction is carried out at 60°C for 4 h. After the reaction is completed, the reaction solution is poured into 100 mL of ice water, 200 mL of anhydrous ethanol is added, and then standing and centrifugation are carried out. The precipitate is dialyzed with deionized water for 5 days, and the dialysate is freeze-dried to obtain the modified yeast glucan.

[0098] The preparation method of the above-mentioned surgical anti-adhesion liquid containing icodextrin comprises the following steps:

[0099] a. Into the preparation tank A, 800 L of 50℃ water for injection was injected, and stirring was started (150 r / min), then 40 kg of Eudragit®, 4.25 kg of modified yeast glucan, 0.257 kg of calcium chloride, 0.051 kg of magnesium chloride, 1.68 kg of sodium lactate were added, and stirring was started while the materials were being dissolved, 0.1 mol / L dilute hydrochloric acid solution was added after 25 min of stirring, and stirring was continued for 15 min, the pH value was adjusted to 3.80, and water for injection was added to 850 L, and the liquid was filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges in sequence for 25 min, and A component was obtained.

[0100] b. Into the preparation tank B, 100 L of 50℃ water for injection was injected, and stirring was started (150 r / min), then 5.4 kg of sodium chloride and 2.3 kg of sodium bicarbonate were added, and stirring was started while the materials were being dissolved, and after 25 min of stirring, the temperature was lowered to 25℃, sodium hydroxide was added, the pH value was adjusted to 8.97, and water for injection was added to 150 L, and the liquid was filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges for 20 min, and B component was obtained.

[0101] c. A non-PVC film double-chamber liquid medicine bag was used, which was divided into A chamber and B chamber by virtual welding, the liquid medicine in the preparation tank A was divided and filled into the A chamber, and the liquid medicine in the preparation tank B was divided and filled into the B chamber, the filling temperature was 30-70℃, the product after filling and sealing was subjected to lamp inspection and high-voltage discharge leak detection, then vacuum packaging was performed using a high-barrier outer packaging film, the vacuum packaging parameters were 3 to 1.8 S; the product was subjected to moist heat sterilization at 121℃ for 15 min, the pressure coefficient was 4.5, the temperature rise gradient was 10℃ / min, the cabinet temperature was 60℃, the sterilization index F0 value (standard sterilization time) was ≥12, and the product was obtained after boxing and packaging, and when used, the A chamber and the B chamber were mixed.

[0102] d. The product was detected according to the quality standard, the pH value of the mixed liquid medicine was 6.98, the insoluble particles below 5 μm were 12 / ml, the endotoxin was lower than 0.1 EU / ml, the peptidoglycan was lower than 0.5 ng / ml, and the osmotic pressure was 288 mOsm / L.

[0103] Comparative Example 1

[0104] Comparative Example 1 and Example 2 were basically the same, except that the modified yeast glucan was replaced with an equal amount of water-soluble yeast glucan.

[0105] The preparation method of the water-soluble yeast glucan is as follows: 1.4 parts of concentrated sulfuric acid is added dropwise into 1 part of n-propanol, stirring while dropping, and then 0.5 parts of yeast glucan is added into the above reaction solution, and the reaction is carried out at-6 ℃ for 2 h. After the reaction is completed, centrifugation is carried out at-10 ℃, and then the liquid in the reaction system is removed. The precipitate is washed with pre-cooled n-propanol for 3 times, and then the precipitate is dissolved in deionized water, centrifuged, and the precipitate is removed. Finally, the supernatant is freeze-dried to obtain the water-soluble yeast glucan.

[0106] Comparative Example 2

[0107] The comparative example 2 and the example 2 are basically the same, and the difference is that the sodium lactate in the A component is replaced by the B component.

[0108] The comparative example 2 provides a preparation method of a surgical anti-adhesion liquid containing icodextrin, which comprises the following steps:

[0109] a. 1100 L of 50 ℃ water for injection is injected into a preparation tank A, and stirring (150 r / min) is started, and then 60 kg of icodextrin, 5.04 kg of modified yeast glucan, 0.3855 kg of calcium chloride, and 0.0765 kg of magnesium chloride are added, and stirring is carried out while adding the materials to make them dissolved. After stirring for 25 min, 0.1 mol / L dilute hydrochloric acid solution is added, and stirring is carried out for 15 min to adjust the pH value to 4.59. Water for injection is added to 1260 L, and the liquid is filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges in sequence for 25 min to obtain the A component.

[0110] b. 180 L of 50 ℃ water for injection is injected into a preparation tank B, and stirring (150 r / min) is started, and then 8.1 kg of sodium chloride, 2.52 kg of sodium lactate, and 3.45 kg of sodium bicarbonate are added, and stirring is carried out while adding the materials to make them dissolved. After stirring for 25 min, hydrochloric acid is added to adjust the pH value to 7.19. Water for injection is added to 240 L, and the liquid is filtered through 0.45 μm and 0.22 μm polyether sulfone filter cartridges in sequence for 20 min to obtain the B component.

[0111] c. A double-chamber liquid bag made of non-PVC film is used, which is divided into A chamber and B chamber by virtual welding. The liquid in the preparation tank A is divided and filled into the A chamber, and the liquid in the preparation tank B is divided and filled into the B chamber. The filling temperature is 30-70 ℃. The product after filling and sealing is subjected to lamp inspection and high-voltage discharge leak detection, and then vacuum packaging is carried out by using a high-barrier outer packaging film. The vacuum packaging parameters are 3 to 1.8 S. The product is subjected to moist heat sterilization at 121 ℃ for 15 min, with a pressure coefficient of 4.5, a temperature rising gradient of 10 ℃ / min, and an outlet cabinet temperature of 60 ℃. The sterilization index F0 value (standard sterilization time) is ≥12. The finished product is obtained by packaging, and the A chamber and the B chamber are mixed during use.

[0112] d.The finished product was detected according to the quality standard, the pH of the mixed drug solution was 8.16, the insoluble microparticles below 5um were 12 per ml, the endotoxin was lower than 0.1 EU / ml, the peptidoglycan was lower than 0.5 ng / ml, and the osmotic pressure was 269 mOSM / L.

[0113] Test Example 1

[0114] The modified yeast glucan prepared in Preparation Example 1 of the present application was subjected to infrared testing, and the results are shown in Figure 2 .

[0115] From the infrared spectrum of Figure 2 , it can be seen that the modified yeast glucan has absorption peaks at 3228 cm -1 , 1250 cm -1 , and 796 cm -1 , and an isopropyl absorption peak at 1375 cm -1 , and characteristic peaks of three substitutions of benzene rings at 850 cm -1 and 680 cm -1 . The above results show that the dehydroabietic acid derivative has been successfully grafted onto the yeast glucan.

[0116] Test Example 2

[0117] Biocompatibility test of the dehydroabietic acid derivative:

[0118] Logarithmic growth phase mouse fibroblasts L929 were resuspended with DMEM complete medium, inoculated into a 96-well plate at a density of 1×10 4 cells / mL, 100 μL was added to each well, and cultured in a 37℃, 5% CO2 cell incubator for 24 h. After the cells were completely adherent, the following groups were set up: dehydroabietic acid derivative group: 100 μL of a solution of the dehydroabietic acid derivative at a concentration of 3 μg / mL 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 24 h of continuous culture, the 96-well plate was removed, and the morphological characteristics of the cells in each group were observed under an inverted microscope, and the results are shown in Figure 3 , 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] From the infrared spectrum of Figure 3The results show that the cell samples treated by the dehydroabietic acid derivative and the blank control group have high similar growth state, the cells maintain normal fibroblast morphological characteristics of spindle or polygon, the cytoplasm is transparent and the boundary is clear, and no obvious cell shrinkage, membrane integrity damage and other abnormal phenomena are observed, which indicates that the dehydroabietic acid derivative has no obvious cytotoxicity to the mouse fibroblast L929, exhibits excellent in-vitro biological compatibility, and preliminarily verifies the biological safety of the dehydroabietic acid derivative as the material of the anti-adhesion fluid.

[0120] Test Example 3

[0121] Human peritoneal mesothelial cells (CP-H180 cells) are used as the detection material of cell compatibility, and 2x10 4 cells / mL cell suspension is prepared, 180 μL of the cell suspension is added to each well of a 96-well plate for seeding, a total of 4 groups of 6 repeated wells, and the plate is cultured at 37 DEG C and 5% CO2 for 24 hours. After the cells adhere, the culture medium is aspirated and replaced with the same volume of the sample to be tested, which is normal saline, the products of Examples 1-4 and the products of Comparative Examples 1-2. After 24 hours of culture, the cell activity is detected by using the CCK8 method (Cell Counting Kit-8). The cell culture medium containing 10% cck8 liquid is prepared, and then the supernatant in each well is carefully aspirated, 100 μL of 10% CCK8 culture medium is added, and the plate is further cultured for 4 hours. The absorbance value of the sample is measured at 450 nm by using an enzyme-labeled instrument, and the cell activity is compared and analyzed according to the absorbance value, and the results are shown in Table 1.

[0122] Table 1: Results of cell proliferation experiment of CP-H180 cells treated with different samples for 24 hours

[0123] Drug 450 nm absorbance average Physiological 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 product of the present application has no effect on the proliferation and growth of human peritoneal mesothelial cells, and has good biocompatibility, which indicates that the pH value of the product obtained by the present application is appropriate, and the irritability of low pH value to blood vessels, tissues, abdominal organs and the like is solved. Compared with normal saline, the biocompatibility of the product is improved to different degrees.

[0125] Test Example 4

[0126] The HPLC method is used to detect the degradation product GDPs of icodextrin in the products of Examples 1 and 2 and Adept 4%, and the detection results are shown in Table 2, Figure 4 and Figure 5 is the test result graph of the product of Example 2.

[0127] Table 2

[0128] Degradation impurities GDPs species Example 1 product Example 2 product Market product Adept 4% Formaldehyde Methanal (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-Deoxyglucosone 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 show that the above-mentioned compatibility and packaging form of the present application solve the shortcomings of the marketed product Adept 4% that cannot add bicarbonate in the formula due to stability problems and incompatibility, and at the same time, because the pH level of the environment in which icodextrin is placed is changed, the level of various degradation impurities generated after sterilization of icodextrin is significantly reduced, thereby improving the biocompatibility of the product.

[0130] Test Example 5

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

[0132] The test was carried out according to the method recommended in the national standard GB / T 16886.11-2021 "Biological evaluation of medical devices Part 11: Systemic toxicity test". ICR mice weighing 20-22 g were selected, provided by Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd., and the feeding temperature was controlled at 20-22°C, and the relative humidity was controlled at 40-50%. The light control was 12h light 12h dark, and the adaptive feeding was 6 days. The above-mentioned mice were divided into control group and example group, 5 in each group.

[0133] Each animal in the example group was injected with the anti-adhesion liquid obtained in Example 2 intraperitoneally, and the injection dose was 50 mL / kg; the control group mice were injected with the same amount of 0.9% sodium chloride injection. The body weight of the animals was measured immediately before injection, and after injection, the immediate biological response of the animals (dyspnea, convulsions or prone phenomenon) was observed, and the general state and toxicity response of the animals in the test group and the control group were observed and recorded at 4h, 24h, 48h and 72h, and the animal body weight was weighed and recorded at the initial, 24h, 48h and 72h, and the animal toxicity response observation results after injection.

[0134] Under the conditions of this test, the body weight of the animals in the control group and the example group did not change significantly, and no convulsions or prone animals were observed, indicating that the anti-adhesion liquid sample of Example 2 had no acute systemic toxicity reaction.

[0135] Test Example 6

[0136] Test animals: ordinary level adult New Zealand albino rabbits, 5-6 months old, sexually mature body weight greater than 2.5 kg, female (not pregnant and not pregnant), a total of 30, provided by Tongxiang Yinhai Animal Husbandry Professional Cooperative, production license: SCXK(Zhe)2023-0002. Raising in the laboratory animal experiment building of Zhejiang Province Medical Device Inspection Research Institute Xuelin Street, providing Jiangsu Province Synergy Pharmaceutical Biological Engineering Co., Ltd. rabbit feed every day, controlling the feeding temperature at 20-22℃, the relative humidity is controlled at 40%-50%. Light control 12h light 12h dark, adaptive feeding for 6 days.

[0137] Construction of New Zealand self-hybrid rabbit pelvic adhesion model: New Zealand albino rabbits were fasted for 12h before surgery and allowed to drink water. Then, 0.3mL / kg of a mixture of Xylazine, Domitor and Dormicum (1:1:1) was injected intramuscularly to anesthetize the rabbits. The abdomen was shaved and disinfected with iodophor for 5 times.

[0138] The rabbits were placed in a supine position before surgery, and a longitudinal incision was made in the middle of the abdomen, about 2cm from the pubic bone, about 4cm long. The skin layer was roughly the same as that of humans and was very thin. The uterus was found to be double, pink, and the uterine horn was long. The oviduct was at the end of the uterine horn, wrapping the ovary, which was white. The uterus and its surrounding organs were observed and recorded, and attention was paid to the presence or absence of adhesions.

[0139] The uterine horn was exposed, and mechanical trauma was performed on both sides of the uterine horn about 4cm long. The serosal surface of the double horns was wiped with sterile gauze for about 5 minutes until subserosal hemorrhage and punctate hemorrhage occurred. Before the abdominal wall was closed, the animals were treated as follows:

[0140] Example 1 group: first wash the wound with 20mL of the anti-adhesion liquid prepared in Example 1, then intraperitoneally inject 20mL of the anti-adhesion liquid prepared in Example 1, 10 in each group;

[0141] Example 2 group: first wash the wound with 20mL of the anti-adhesion liquid prepared in Example 2, then intraperitoneally inject 20mL of the anti-adhesion liquid prepared in Example 2, 10 in each group;

[0142] Example 3 group: first wash the wound with 20mL of the anti-adhesion liquid prepared in Example 3, then intraperitoneally inject 20mL of the anti-adhesion liquid prepared in Example 3, 10 in each group;

[0143] Example 4 group: first wash the wound with 20mL of the anti-adhesion liquid prepared in Example 4, then intraperitoneally inject 20mL of the anti-adhesion liquid prepared in Example 4, 10 in each group;

[0144] Comparative Example 1 group: first wash the wound with 20mL of the anti-adhesion liquid prepared in Comparative Example 1, then intraperitoneally inject 20mL of the anti-adhesion liquid prepared in Comparative Example 1, 10 in each group;

[0145] The adhesion prevention liquid prepared in Comparative Example 2 was used to flush the wound and then 20 mL of the adhesion prevention liquid prepared in Comparative Example 2 was perfused into the abdominal cavity, 10 rabbits in each group.

[0146] The Ringer's lactate injection was used to flush the wound and then 20 mL of the Ringer's lactate injection was perfused into the abdominal cavity, 10 rabbits in each group.

[0147] The blank control group was not treated, 10 rabbits in each group.

[0148] The abdominal wall of each group of animals was continuously sutured with 2 / 0 non-absorbable suture, and the skin was intermittently sutured with 2 / 0 non-absorbable suture, and then sterilized with iodophor and covered with sterile dressing. After the operation, the rabbits were given intramuscular injection of 200,000 units of penicillin sodium in one side of the gluteus muscle, and were placed in a warm place, and attention was paid to keep warm and keep the respiratory tract unobstructed. Water can be taken 12 hours after the operation, and food can be taken the next day; within 3 days after the operation, 200,000 IU of penicillin was injected intramuscularly twice a day to prevent infection; the activity and food intake of the animals were observed and recorded every day after the operation.

[0149] On the 7th day after the operation, the New Zealand white rabbits were subjected to a second laparotomy operation, and the abdominal wall was cut in a "U" shape at a distance of 3 cm from the surgical incision to fully expose the pelvic abdominal organs and peritoneum. The adhesion between the uterus and the surrounding organs, other abnormal conditions in the pelvic cavity, such as the nature of ascites, etc. were observed and recorded.

[0150] The adhesion grade and score were determined according to the test scheme provided by the client, and the grading standard was determined and scored, and the grading standard is shown in Table 3. Three experimenters qualitatively and quantitatively evaluated each experimental animal by parameters including adhesion range, adhesion position and strength, and the score results were recorded in Table 4.

[0151] Table 3: Adhesion degree grading evaluation standard

[0152]

[0153] Table 4: Pelvic adhesion grade and score of each group

[0154]

[0155]

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

[0157] All of the New Zealand albino rabbits in the postoperative blank control group had adhesion, and the adhesion of the uterus itself and its contralateral side accounted for more than 70%, and the adhesion of the uterus and the bladder accounted for more than 20%, mainly in grades III and IV. The adhesion of the sodium lactate Ringer's injection group was similar to that of the blank control group, and all of the rabbits had adhesion, and the adhesion of the uterus itself and its contralateral side accounted for more than 90%, and the adhesion was dense and difficult to separate, mainly in grades III and IV. The adhesion-preventing liquid containing icodextrin in the postoperative example 1-3 group had been absorbed, and the adhesion-free condition accounted for more than 60%, and the adhesion was mainly loose membrane adhesion, and the adhesion range was less, and it was easier to separate compared with the blank control group, mainly in grade 0, which indicated that the adhesion range and the strength of the toughness separation of the test sample group were significantly better than those of the blank control group and the sodium lactate Ringer's injection group. The test showed that the surgical adhesion-preventing liquid containing icodextrin could effectively reduce the formation of adhesion, and the postoperative adhesion-preventing effect was significant.

[0158] The total score of the adhesion grade of the postoperative comparative example 1 group (the modified yeast glucan was replaced with an equal amount of water-soluble yeast glucan) was obviously higher than that of the example 1-4 group, and the total score of example 1 was also slightly higher than that of the example 2-4 group. This is because the addition of the modified yeast glucan can not only improve the water solubility of the yeast glucan, but also can cross-link with icodextrin through the hydrophilic groups of sulfonate and hydroxyl groups to improve the stability of icodextrin. Moreover, dehydroabietic acid belongs to tricyclic diterpenes, and its unique hydrophobic tricyclic skeleton and rigid structure can effectively improve the retention time of the yeast glucan in the body, and then improve the duration of the synergistic effect of the yeast glucan and icodextrin, continuously isolate the damaged tissue surface, reduce the direct contact between the uterus and the tissue, inhibit the deposition of fibrin and the formation of adhesion, and achieve an effective and safe adhesion-preventing effect.

[0159] In addition, the total score of the adhesion grade of the comparative example 2 group was obviously higher than that of the example 1-3 group, which indicated that the compatibility stability of the bicarbonate and the lactate in the application was poor. The compatibility and double-chamber packaging form of the application solved the shortcomings that the marketed product Adept 4% cannot add bicarbonate in the formula due to stability problems and compatibility contraindications, inhibited the degradation of traditional process icodextrin (pH is controlled in the range of 5.0-5.5 and is directly used) to generate harmful GDPs substances, solved the irritability of low pH to blood vessels, tissues, abdominal organs and the like, and then improved the adhesion-preventing effect.

[0160] The above-described embodiments are only preferred embodiments of the application, and cannot be used to limit the protection scope of the application. Any non-essential changes and replacements made by those skilled in the art on the basis of the application all belong to the protection scope of the application.

Claims

1. An adhesion-preventing solution for celiac surgery or laparoscopic surgery containing isomalt, characterized in that, Consists of A component and B component, A component and B component are stored separately; the A component is a solution with water as solvent, including 20-75g / L Eudragit L30D-55, 1~3g / L sodium lactate, 0.1~0.5g / L calcium chloride, 0.05~0.08g / L magnesium chloride; the B component is a solution with water as solvent, including 25~40g / L sodium chloride, 10~20g / L sodium bicarbonate; The volume ratio of the A component and B component is 1:0.1~0.8; the pH of the A component is 3.0~5.0, and the pH of the B component is 6.0~9.0; The A group further comprises 2-5 g / L modified yeast glucan, wherein the modified yeast glucan is prepared by adding yeast glucan into a solvent, mixing uniformly, adding dehydroabietic acid derivative, triethylamine and 4-dimethylaminopyridine for reaction, and then performing post-treatment on the reaction solution to obtain the modified yeast glucan; and the structural formula of the dehydroabietic acid derivative is . The preparation process of the dehydroabietic acid derivative includes the following steps: (1) adding lithium aluminum hydride into tetrahydrofuran, then adding dehydroabietic acid for reaction to prepare dehydroabietic acid reduction alcohol; (2) adding dehydroabietic acid reduction alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate into N,N-dimethylformamide for reaction, and then preparing dehydroabietic acid derivative through purification after the reaction is completed.

2. The adhesion-preventing solution for celiac surgery or laparoscopic surgery containing isoxaglate according to claim 1, characterized by, The mass ratio of the yeast glucan, dehydroabietic acid derivative, triethylamine and 4-dimethylamino pyridine is 1:(2.5~3):(2.6~3.5):(2.6~3.5), and the concentration of the yeast glucan in the solvent is 0.075~0.1g / mL; the solvent is N,N dimethylformamide; the reaction temperature is 40~60℃, and the reaction time is 2~4h.

3. The anti-adhesion solution for celiac surgery or laparoscopic surgery containing isoxaglate according to claim 1, characterized by, In step (1), the molar ratio of the dehydroabietic acid and lithium aluminum hydride is 1:4~4.5, and the dosage ratio of the dehydroabietic acid and tetrahydrofuran is 1mmol:5~5.5mL; in step (1), when the dehydroabietic acid is added, the temperature of the reaction system is -2~3℃, and the reaction time is 4~6h.

4. The anti-adhesion solution for celiac surgery or laparoscopic surgery according to claim 1, wherein the excipient is a mixture of 50% by weight of the excipient of the excipient of claim 1 and 50% by weight of the excipient of the excipient of claim 2. In step (2), the molar ratio of the dehydroabietic acid reduction alcohol, 2-bromo-1-ethanesulfonic acid and inorganic carbonate is 1:(1~1.1):(2.5~3), and the concentration of the dehydroabietic acid reduction alcohol in N,N-dimethylformamide is 0.3~0.5mol / L.

5. The anti-adhesion solution for celiac surgery or laparoscopic surgery according to claim 1, wherein the excipient is a mixture of 50% by weight of the excipient of the excipient of claim 1 and 50% by weight of the excipient of the excipient of claim 2. In step (2), the inorganic carbonate is potassium carbonate or sodium carbonate, the reaction temperature is 60~80℃, and the reaction time is 12~18h.

6. The method of preparing the adhesive barrier solution for celiac surgery or laparoscopic surgery containing isomalt according to any one of claims 1 to 5, characterized in that, Including the following steps: a. adding each raw material component into water according to the ratio, stirring uniformly, adjusting the pH value to the range, then filtering and filling to prepare the A component; b. adding each raw material component into water according to the ratio, stirring uniformly, adjusting the pH value to the range, then filtering and filling to prepare the B component; c. filling and sterilizing the A component and B component respectively, and mixing the A component and B component for use.

7. The method for preparing the anti-adhesion fluid for celiac surgery or laparoscopic surgery containing isoxaglate according to claim 6, characterized by, The sterilization temperature is 115~121℃, and the sterilization time is 10~30min.

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