In-situ curable reagent composition, Anti-adhesion hydrogel, preparation method therefor and use thereof

Abstract

The present application relates to the technical field of biomaterials, and relates to an in-situ curable reagent composition, an anti-adhesion hydrogel, a preparation method therefor and a use thereof. The in-situ curable reagent composition provided in the present application comprises a polyethylene glycol aldehyde-based derivative solution and a polylysine solution. The mass percentage content of a polyethylene glycol aldehyde-based derivative in the polyethylene glycol aldehyde-based derivative solution is 5-20%; the mass percentage content of polylysine in the polylysine solution is 4-10%; and the molar ratio of aldehyde groups in the polyethylene glycol aldehyde-based group derivative to amino groups in the polylysine is 0.05-0.2:1. The composition provided in the present application is composed of the polyethylene glycol aldehyde-based derivative and the polylysine which have good biocompatibility. The two components can be formed into a hydrogel by means of a Schiff base reaction. The gelation time and degradation time of the obtained hydrogel are adapted to clinical requirements. The characteristics of injectability and in-situ gelation are suitable for different uterine cavity structures, and the postoperative uterine cavity adhesion prevention effect is excellent.

Description

In-situ curable reagent composition, anti-adhesion hydrogel and preparation method and application thereof

[0001] The present application claims priority to the Chinese patent application No.CN202411263713.7, filed on September 10, 2024, entitled "In-situ curable composition, anti-adhesion hydrogel and preparation method and application thereof", the whole content of which is incorporated herein by reference. TECHNICAL FIELD

[0002] The present application belongs to the technical field of biomaterials, and specifically relates to an in-situ curable reagent composition, anti-adhesion hydrogel and preparation method and application thereof. BACKGROUND

[0003] Intrauterine adhesions (IUA) is a common gynecological disease that seriously harms the reproductive function and has poor treatment effect, and seriously affects the reproductive physiology and physical and mental health of women. Literature reports that the incidence of IUA caused by multiple artificial abortions and curettage is as high as 25% to 30%, and has become the main cause of reduced menstrual flow and secondary infertility. At present, there is no effective treatment method for severe IUA to restore reproductive function and menstrual physiology. (Chinese Journal of Obstetrics and Gynecology, 2015, 50(12): 881-887.)

[0004] Transcervical resection of adhesion (TCRA) is the most commonly used surgical method for treating intrauterine adhesions, but the postoperative intrauterine re-adhesion rate is as high as 62.5%, and the pregnancy success rate is only 22.5% to 33.3% (Chinese Journal of Obstetrics and Gynecology, 2015, 50(12): 881-887.). In addition to TCRA surgery, drug therapy is also used to treat intrauterine adhesions, but all have certain side effects or risks, causing great burden to the patient's body, bringing great inconvenience and economic burden to the patient.

[0005] In addition to surgery and drugs, the current clinical will also use the products to prevent intrauterine adhesion, such as balloon, intrauterine stent, etc., especially after TCRA operation, to prevent the occurrence of intrauterine adhesion again. Its working principle is mainly to form a physical barrier, so as to prevent the contact of endometrium in the process of self-repair, and prevent the occurrence of adhesion. But the balloon, stent and other products do not have conformability, and have a compression effect on the endometrium during use, affecting the normal repair of the endometrium. Moreover, the stent and balloon are not degradable, and need to be taken out of the uterus by an additional traction device after a certain period of time, and the placement time usually needs one week, so it is easy to cause infection. In some cases, sodium hyaluronate gel can be used as an isolation product. Although such gel has good injectability, degradability and biocompatibility, under the natural pressure contraction of the uterine cavity, the retention time of sodium hyaluronate gel in the uterus is short, and it cannot effectively provide enough time for physical isolation.

[0006] SUMMARY

[0007] The application provides a reagent composition capable of in-situ curing, comprising a polyethylene glycol aldehyde derivative solution and a polylysine solution; the mass percentage content of the polyethylene glycol aldehyde derivative in the polyethylene glycol aldehyde derivative solution is 5-20%; the mass percentage content of the polylysine in the polylysine solution is 4-10%; and the molar ratio of aldehyde groups in the polyethylene glycol aldehyde derivative to amino groups in the polylysine is 0.05-0.2:1. BRIEF DESCRIPTION OF DRAWINGS

[0008] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the drawings needed in the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0009] Fig. 1 is a real object diagram of the initial state of the adhesion test of the in-situ curable composition described in Example 1 and pigskin;

[0010] Fig. 2 is a real object diagram of the adhesion test of the in-situ curable composition described in Example 1 and pigskin immersed in PBS buffer for 30 min;

[0011] Fig. 3 is a real object diagram of the adhesion test of the in-situ curable composition described in Example 1 and pigskin immersed in PBS buffer for 5 h. DETAILED DESCRIPTION

[0012] The application provides a reagent composition capable of in-situ curing, comprising a polyethylene glycol aldehyde derivative solution and a polylysine solution;

[0013] The mass percentage of the polyethylene glycol aldehyde derivative in the polyethylene glycol aldehyde derivative solution is 5-20%; the mass percentage of the polylysine in the polylysine solution is 4-10%; and the molar ratio of the aldehyde group in the polyethylene glycol aldehyde derivative to the amino group in the polylysine is 0.05-0.2:1.

[0014] In the present application, all raw material components are commercially available products well known to those skilled in the art, unless otherwise specified.

[0015] In the present application, the mass percentage of the polyethylene glycol aldehyde derivative in the polyethylene glycol aldehyde derivative solution is 5-20%, and in some embodiments, the mass percentage of the polyethylene glycol aldehyde derivative in the polyethylene glycol aldehyde derivative solution can be 5%, 10%, 15% or 20%; the polyethylene glycol aldehyde derivative is preferably a multi-arm polyethylene glycol capped with an aldehyde group; the aldehyde group includes one or more of unsubstituted aldehyde group, alkane aldehyde group and aromatic aldehyde group, and in some embodiments, the aldehyde group can be unsubstituted aldehyde group or aromatic aldehyde group; the multi-arm polyethylene glycol capped with an aldehyde group preferably has 4-8 arms, and in some embodiments, the multi-arm polyethylene glycol capped with an aldehyde group has 4 arms, 6 arms or 8 arms; and the number average molecular weight of the polyethylene glycol aldehyde derivative is preferably 5000-40000 Da, and in some embodiments, the number average molecular weight of the polyethylene glycol aldehyde derivative can be 5000 Da, 10000 Da, 20000 Da, 30000 Da or 40000 Da.

[0016] In the present application, the solvent of the polyethylene glycol aldehyde derivative solution is preferably a phosphate buffer; the phosphate buffer is preferably a sodium dihydrogen phosphate buffer; the concentration of sodium dihydrogen phosphate in the sodium dihydrogen phosphate buffer is preferably 0.01-0.15 M (mol / L); and the pH value of the polyethylene glycol aldehyde derivative solution is preferably 3-6, and in some embodiments, the pH value of the polyethylene glycol aldehyde derivative solution can be 3, 4, 5 or 6; when the pH value of the polyethylene glycol aldehyde derivative solution is not within the above range, it is also preferred to include reducing the pH value of the polyethylene glycol aldehyde derivative solution to the above range; and the reagent for reducing the pH value is preferably aqueous phosphoric acid, aqueous acetic acid or aqueous hydrochloric acid.

[0017] In the present application, the mass percentage of polylysine in the polylysine solution is 4-10%, and in some embodiments, the mass percentage of polylysine in the polylysine solution can be 4%, 6%, 8% or 10%; the polylysine is preferably ε-polylysine; the ε-polylysine preferably includes linear ε-polylysine and / or hyperbranched ε-polylysine; the volume ratio of the polyethylene glycol aldehyde derivative solution and the polylysine solution is preferably 1-10:1-10, and in some embodiments, the volume ratio of the polyethylene glycol aldehyde derivative solution and the polylysine solution can be 1:1, 3:5, 4:6, 1:8, 7:1 or 8:3.

[0018] In the present application, the solvent of the polylysine solution is borate buffer, phosphate buffer or water; the water is preferably physiological saline or sterile water for injection; the pH value of the polylysine solution is 5-10, and in some embodiments, the pH value of the polylysine solution can be 5, 6, 7, 8, 9 or 10; when the pH value of the polylysine solution is not within the above range, it is also preferable to adjust the pH value of the polylysine solution to the above range; the reagent for adjusting the pH value is preferably an aqueous sodium hydroxide solution, an aqueous potassium hydroxide solution or an aqueous sodium carbonate solution.

[0019] The in-situ curable composition provided by the present application is composed of a synthetic polymer material polyethylene glycol aldehyde derivative with good biocompatibility and a natural polymer polylysine, and the two can form a hydrogel through a Schiff base reaction. Since both components are low-viscosity two-component, they can be cured in-situ in the uterine cavity, with a suitable curing speed, effectively conforming to the uterine cavity structure of different sizes and shapes, and achieving more reliable physical isolation; the hydrogel has biomechanical matching and low swelling performance, does not cause compression to the endometrium, and does not affect the repair process of the endometrium; the hydrogel has good physical stability and will not be discharged from the uterine cavity in advance, providing sufficient uterine cavity retention for a long enough time to ensure that adhesion does not occur; the gel time window and gel degradation time window of the obtained hydrogel adapt to the actual clinical use requirements, and the in-vivo degradation time does not affect the menstrual cycle of women.

[0020] Moreover, the hydrogel has good biocompatibility, can reduce the stimulation to the human body, and avoids the side effects and risks in the existing treatment methods. Secondly, by adjusting the parameters of the two components, such as concentration, molar ratio and pH, the gel time, swelling degree, biocompatibility and other properties of the hydrogel in-vivo and in-vitro can be adjusted. This adjustment method can be adjusted according to the clinical requirements, so that the gel time window and in-vivo and in-vitro degradation period of the hydrogel adapt to the physiology and menstrual cycle of women, improve the treatment effect, shorten the treatment period, and reduce the economic and psychological burden of patients.

[0021] The application further provides a preparation method of the composition.

[0022] The polyethylene glycol aldehyde derivative solution and the polylysine solution are prepared respectively to obtain the composition.

[0023] In the application, after the polyethylene glycol aldehyde derivative solution and the polylysine solution are prepared, the pH value is adjusted and sterilization treatment is performed in sequence; the pH value adjusting condition is consistent with the above technical solution, and will not be repeated here; the sterilization treatment is preferably high-pressure steam sterilization treatment; the temperature of the high-pressure steam sterilization treatment is preferably 121-126°C, and in some embodiments, the temperature of the high-pressure steam sterilization treatment can be 121°C, 123°C or 125°C; the time of the high-pressure steam sterilization treatment is preferably 15-30 min, and in some embodiments, the time of the high-pressure steam sterilization treatment can be 15 min, 20 min or 30 min.

[0024] The application further provides an anti-adhesion hydrogel, and the preparation raw material of the anti-adhesion hydrogel is the composition according to the above technical solution or the composition obtained by the preparation method according to the above technical solution.

[0025] In the application, the pH value of the anti-adhesion hydrogel is preferably 6-8, and in some embodiments, the pH value of the anti-adhesion hydrogel can be 6, 7 or 8; the initial Young's modulus is preferably 20-150 kPa, and in some embodiments, the initial Young's modulus of the anti-adhesion hydrogel can be 20 kPa, 50 kPa, 80 kPa, 100 kPa, 120 kPa or 150 kPa; the swelling rate is preferably -50%-50%, and in some embodiments, the swelling rate of the anti-adhesion hydrogel can be -50%, -20%, 5%, 25% or 50%; the residence time in the uterine environment or the simulated uterine environment is preferably 3-7 d, and in some embodiments, the residence time of the anti-adhesion hydrogel in the uterine environment can be 3 d, 4 d, 5 d, 6 d or 7 d.

[0026] The hydrogel provided by the application can be directly applied to the uterine cavity by injection and the like, which is convenient, easy to operate, reduces the complexity of surgical treatment, and is suitable for the uterine cavity structures of different individual women, so that the individual differences in postoperative uterine cavity re-adhesion can be prevented. Meanwhile, the hydrogel can be gradually degraded in the body, and does not need to be removed by secondary surgery, thereby reducing the complexity of treatment and the risk of infection and improving the life quality of patients.

[0027] The application further provides a preparation method of the anti-adhesion hydrogel according to the above technical solution, comprising the following steps:

[0028] The polyethylene glycol aldehyde derivative solution and the polylysine solution are mixed, and solidified to obtain the anti-adhesion hydrogel.

[0029] In the present application, the mixing method is preferably double mixing syringe mixing; the solidification time is preferably 20-150s, and in some embodiments, the solidification time can be 20s, 30s, 50s, 80s, 100s, 120s or 150s; the solidification is preferably performed in the uterine cavity; the device for injecting the uterine cavity is preferably a single-lumen catheter; the single-lumen catheter is preferably a single-lumen catheter with a cervical obturator.

[0030] The present application also provides the use of the composition of the above technical solution, the composition obtained by the preparation method of the above technical solution, the anti-adhesion hydrogel of the above technical solution or the anti-adhesion hydrogel obtained by the preparation method of the above technical solution in the preparation of a uterine cavity anti-adhesion preparation.

[0031] In the present application, the use of the composition or anti-adhesion hydrogel in the preparation of a uterine cavity anti-adhesion preparation is preferably direct use or as a drug carrier.

[0032] The hydrogel provided by the present application is used in the preparation of a uterine cavity anti-adhesion preparation, and can carry drugs as a carrier and be injected into the uterine cavity. After the hydrogel is solidified in situ, the drug is released slowly in the uterine cavity, directly acts on the endometrium, more effectively promotes the repair of the endometrium, and reduces the systemic side effects caused by oral administration of the drug.

[0033] In order to further illustrate the present application, the composition which can be solidified in situ, the anti-adhesion hydrogel, the preparation method and the application thereof provided by the present application are described in detail below in combination with the drawings and examples, but they should not be understood as limiting the scope of protection of the present application.

[0034] Example 1

[0035] A polyethylene glycol aldehyde derivative (PEG-CHO) solution is prepared with a phosphate buffer (0.02M sodium dihydrogen phosphate), and a polylysine (PLL) solution is prepared with purified water, i.e. a crosslinking agent solution.

[0036] A 10% phosphoric acid aqueous solution is used to adjust the pH value of the polyethylene glycol aldehyde derivative solution to 3.5-3.8, and a 10M sodium hydroxide aqueous solution is used to adjust the pH value of the polylysine solution to 7.5-8.5. The pH values of the solutions in different groups are shown in Table 1.

[0037] The polyethylene glycol aldehyde derivative solution and the polylysine solution are subjected to high-pressure steam sterilization treatment, and the sterilization conditions are 121℃ and 15min;

[0038] Preparation of the gel using double injection device: equal volume of polyethylene glycol aldehyde derivative solution and polylysine solution were loaded into two syringes of double injection device consisting of double mixing assembly, the two solutions were mixed uniformly in the injection needle by pushing the double injection device at the same rate, the mixture was injected into the uterine cavity using single lumen catheter with cervical obturator, after waiting for a certain time, the catheter was pulled out after the solidification was completed, and the anti-adhesion gel was formed in situ in the uterine cavity. The preparation conditions and test results of the gel performance are shown in Table 1.

[0039] The test method of the gel performance mainly includes the following steps:

[0040] Gel time test: 200 μL of polyethylene glycol aldehyde derivative solution and 200 μL of polylysine solution were taken in a clean test tube, vortex mixed, and the time was recorded at the same time. The time when the reaction solution had no fluidity was the gel time.

[0041] Gel 24h extraction liquid pH test: 200 μL of polyethylene glycol aldehyde derivative solution and 200 μL of polylysine solution were taken in a clean test tube, vortex mixed, and the time was recorded at the same time. The time when the reaction solution had no fluidity was the gel time.

[0042] Gel 24h swelling degree test: 200 μL of polyethylene glycol aldehyde derivative solution and 200 μL of polylysine solution were taken in a clean test tube, vortex mixed, and the time was recorded at the same time. The time when the reaction solution had no fluidity was the gel time.

[0043] Table 1 Test results of preparation conditions and gel performance of the preparation method described in Example 1

[0044] As can be seen from Table 1, by different concentration ratios of polyethylene glycol-polylysine (PLL), it can be found that when the molar ratio of polyethylene glycol aldehyde end group-crosslinking agent amino increases, the 24hr swelling rate decreases. Therefore, the swelling rate of the gel can be controlled by the concentration ratio, so as to match the clinical needs. In addition, under the same crosslinking agent concentration (groups 1, 2, groups 3, 4, 5, 6), increasing the polyethylene glycol concentration can significantly improve the solidification speed.

[0045] Example 2

[0046] Hydrogels were prepared according to the preparation method provided in Example 1, the difference of preparation conditions and the test results of the gel performance are shown in Table 2.

[0047] Table 2 Preparation conditions of the preparation method described in Example 2 and test results of the performance of the obtained gel

[0048] As can be seen from Table 2, the pH of the linear PLL crosslinking agent has a great influence on the curing time, and reducing the pH can improve the curing speed.

[0049] Example 3

[0050] Hydrogels were prepared according to the preparation method provided in Example 1, the difference of preparation conditions and the test results of the gel performance are shown in Table 3.

[0051] Table 3 Preparation conditions of the preparation method described in Example 3 and test results of the performance of the obtained gel

[0052] As can be seen from Table 3, the concentration of the hyperbranched PLL has a different influence on the swelling rate than the linear PLL. When the concentration of the hyperbranched PLL is increased (the molar ratio of polyethylene glycol end group to crosslinking agent amino group is reduced), the swelling rate first increases (groups 11, 12, 13) and then decreases (groups 15, 16), but the overall change is not large, while the molar ratio of polyethylene glycol end group to crosslinking agent amino group of the linear PLL has a great influence on the swelling rate of the gel. In addition, the pH of the hyperbranched PLL crosslinking agent has a great influence on the gel time, but the rule is different from that of the linear PLL: the gel time of the hyperbranched PLL becomes faster as the pH increases (groups 12, 13, 14); while the linear PLL is the opposite.

[0053] Comparative Example 1

[0054] Hydrogels were prepared according to the preparation method provided in Example 1, the difference being that linear PLL and polyetherimide (PEI) were used as crosslinking agents, and the preparation conditions and the test results of the gel performance compared with Example 1 are shown in Table 4.

[0055] Table 4 Preparation conditions of the preparation method described in Example 1 and Comparative Example 1 and test results of the performance of the obtained gel

[0056] As can be seen from Table 4, under the condition of keeping the same molar ratio of polyethylene glycol end group to crosslinking agent amino (primary amine), the introduction of PEI can significantly accelerate the gel curing speed and reduce the 24h swelling rate of the gel. Too fast gel curing time may lead to a smaller clinical operation window. In addition, the reproductive toxicity test of the PEI material in this patent is unqualified, so the PEI material has a potential reproductive toxicity risk and is not suitable for application in the uterine cavity field.

[0057] Comparative Example 2

[0058] The hydrogel was prepared according to the preparation method provided in Example 1, except that PEI was used as the crosslinking agent, and the preparation conditions and test results of the gel performance are shown in Table 5.

[0059] Table 5 Preparation conditions of the preparation method described in Comparative Example 2 and test results of the performance of the obtained gel

[0060] As can be seen from Table 5, when only PEI is selected as the crosslinking agent, the gelation time of the gel is too fast, and the in vitro degradation rate of the gel is too fast. In addition, the PEI material in the present patent has been tested for reproductive toxicity, and the result is unqualified, so the PEI material has potential reproductive toxicity risk and is not suitable for application in the uterine cavity field.

[0061] Comparative Example 3

[0062] The hydrogel was prepared according to the preparation method provided in Example 1, except that polyethylene glycol-succinimidyl glutarate (PEG-SG) was used to prepare the polyethylene glycol solution, and methoxypolyethylene glycol-amino (PEG-NH2) was used to prepare the crosslinking agent solution, and the preparation conditions and test results of the gel performance are shown in Table 6.

[0063] Table 6 Preparation conditions of the preparation method described in Comparative Example 3 and test results of the performance of the obtained gel

[0064] As can be seen from Table 6, the gelation time of the PEG-SG+PEG-NH2 gel is too fast, and the in vitro swelling of the gel is too large, which may cause compression to the inside of the uterine cavity.

[0065] As can be seen from the above examples and comparative examples, compared with the comparative examples, the gelation time of the hydrogel provided in the present patent is adequate, giving the doctor more adequate operation time and not blocking the injection catheter; the swelling rate of the hydrogel is lower, and the compression to the uterine cavity is smaller; the mechanical properties are good; and the biocompatibility is good.

[0066] Test Example 1

[0067] Gel in vitro degradation test: 200 μL of polyethylene glycol aldehyde derivative solution and 200 μL of polylysine solution were taken in a clean test tube, vortexed to form a gel, and the initial mass m0of the gel was recorded. 4 mL of PBS buffer with pH = 7.4 was added to the test tube, which was placed in a water bath shaker at 37°C and 40 rpm for in vitro degradation. The PBS was replaced regularly, and the mass mof the gel after degradation was recorded regularly. n The degradation degree of the gel on the nth day was calculated according to the formula: 100%*(m n -m0) / m0. The in vitro degradation test results are shown in Tables 7 and 8.

[0068] Table 7 Test results of in vitro degradation experiment of gels obtained in Examples 1-2

[0069] Table 8 Test results of in vitro degradation experiment of gels obtained in Example 3

[0070] As can be seen from Table 7, the faster the degradation rate, the lower the molar ratio of polyethylene glycol aldehyde end group-crosslinking agent amino group; the higher the pH of linear PLL crosslinking agent, the faster the degradation rate. As can be seen from Table 8, compared with linear PLL crosslinking agent, the degradation rate of hyperbranched PLL crosslinked gel is faster.

[0071] Test Example 2

[0072] In vitro cytotoxicity study: tested according to the standard GB / T 16886.5-2017. Four concentrations (100%, 75%, 50%, 25%) of test product leaching solution were prepared with MEM medium (containing 10% fetal bovine serum), as well as blank, 100% negative control and positive control, leaching time 72 hours, leaching ratio 0.2g:1mL. Under the condition of 37C, 5% CO2, respectively, with test product leaching solution, blank and the rest two controls, the semi-confluent monolayer cells of L-929 type (NIH3T3) mouse fibroblasts were cultured in 96-well plates. After 72 hours, MTT colorimetric determination, reading on the enzyme marker at 570nm and 650nm, calculating the survival rate of cells. The survival rate of cells was calculated according to the following formula: survival rate % = R / R0x100. Wherein, R: the average optical density of detection group, positive control group and negative control group; R0: the average optical density of blank group. Test results are all >70% indicating qualified. The test results of in vitro degradation experiment are shown in Table 9.

[0073] Table 9 Test results of in vitro cytotoxicity of gels obtained in Examples 1-2

[0074] As can be seen from Table 9, the polyethylene glycol derivative material, polylysine material and the gel formed therefrom have low cytotoxicity and are good biocompatible materials.

[0075] Test Example 3

[0076] Mechanical property test: the gel obtained in Example 1 was placed in a polytetrafluoroethylene mold to form a cylindrical gel sample with a diameter of 9mm and a height of 6mm. The sample was placed on a universal testing machine for testing, with a maximum sensor pressure of 50N, a compression speed of 0.6mm / min, and a maximum pressure set until the set maximum pressure was reached. Parallel test was performed for 3 times, and the average value was taken. The mechanical property test results are shown in Table 10.

[0077] Table 10 Results of in vitro cytotoxicity test of gels obtained in Example 1

[0078] As can be seen from Table 10, according to the compression modulus data, the gels obtained in Groups 1 and 3 have a compression modulus of about 30-260 kPa within a strain of 0-30%, and have good compression resistance. The good mechanical properties of the gels can enable them to resist the compression force of the uterine cavity and not be discharged from the uterus.

[0079] Test Example 4

[0080] Tissue adhesion test: First, the pigskin was soaked in PBS buffer with pH = 7.4 for 24 h, the surface moisture of the pigskin was wiped dry, the composition in Group 3 of Example 1 was injected directly on the pigskin to form a gel, and then the gel was soaked in PBS buffer with pH = 7.4, and the gel shedding was observed. The results of the tissue adhesion test are shown in Figures 1-3, wherein Figures 1-3 correspond to the initial shape of the gel, the shape after soaking for 30 min, and the shape after soaking for 5 h, respectively.

[0081] As can be seen from Figures 1-3, according to the state of the gel after different soaking times and the effect of the gel adhering to the pigskin, the present gel has a certain tissue adhesion effect.

[0082] Test Example 5

[0083] Intrauterine biocompatibility and degradation period test: rats and rabbits were selected as experimental objects, and the gels obtained in Example 1 and Example 2 were used to observe the degradation of the gels in the uterine cavity at regular intervals. The results of the degradation test are shown in Table 11.

[0084] Table 11 Results of intrauterine biocompatibility and degradation period test of gels obtained in Examples 1-2

[0085] As can be seen from Table 11, the gels have a suitable degradation time in the uterine cavity of rats and rabbits, and do not cause recurrence of uterine adhesion due to too fast degradation of the gels, nor do they cause physical and mental discomfort to women and affect the normal physiological cycle of women due to too slow degradation of the gels.

[0086] Test Example 6

[0087] Reproductive toxicity test: the potential toxicity of the gel obtained in Example 2 on embryo development was evaluated by in vitro routine culture of mouse embryos. Experimental method: the test was carried out according to GB / T 16886.3-2019 "Biological evaluation of medical devices Part 3: Tests for genetic toxicity, carcinogenicity and reproductive toxicity". The extraction solution was prepared by using M16 culture medium at an extraction ratio of 0.1 g: 1 mL, 37°C, 72h, 40rpm extraction conditions, and the embryos were cultured by the microdrop method. The collected mouse embryos were randomly divided into each test group of pre-equilibrated microdrops, and cultured in a 37°C, 5% CO2, saturated humidity incubator for 96h. The number of mouse embryos in each test group was 20-25. The embryo division was recorded in the incubator at 24h, 48h, 72h. After 96h, the blastocyst morphology was observed, the number of blastocysts was recorded, and the blastocyst formation rate was calculated. Blastocyst formation rate calculation: blastocyst formation rate (%) = blastocyst number / (1-cell embryo number) * 100%, blastocyst formation rate ≥ 80% is qualified. The test results are shown in Table 12.

[0088] Table 12 Reproductive toxicity test results of the gel obtained in Example 2

[0089] As can be seen from Table 12, the gel obtained in group 9 has a blastocyst formation rate of 88% in the reproductive toxicity test, indicating that polyethylene glycol derivative materials, polylysine materials and gels formed therefrom will not affect the reproductive function and development process of mammals, and are predicted to have no adverse effects on parental reproductive function such as reproduction, pregnancy, delivery, lactation, and on the development of offspring embryos-fetus, postnatal development.

[0090] As can be seen from the above examples and test examples, the present patent proposes a uterine cavity anti-adhesion hydrogel composed of synthetic polymer material polyethylene glycol derivative and natural polymer polylysine with good biocompatibility, which forms a hydrogel through Schiff base reaction. By adjusting the parameters of the two components, such as concentration, molar ratio, pH, etc., the in vitro and in vivo gelation time, gel pH, swelling degree, biocompatibility and other properties of the hydrogel are studied, and the reproductive toxicity is studied according to the "Guiding Principles for the Registration and Review of Absorbable Anti-adhesion Products for Abdominal and Pelvic Surgery", which can ensure the safety of the hydrogel and avoid causing new harm to the patient. The gelation time window and gel degradation time window proposed in the present patent adapt to the actual clinical use requirements, and the injectability and in-situ gelation characteristics are suitable for the uterine cavity structure of different individual women. The hydrogel not only prevents the formation of postoperative uterine cavity re-adhesion, but also does not affect the menstrual cycle of women during in vivo degradation.

[0091] Although the above examples have made a detailed description of the present application, it is only a part of the examples of the present application, not all the examples, and other examples can be obtained under the premise of no creativity according to the present examples, which all belong to the protection scope of the present application.

Claims

1. An in situ curable reagent composition characterized in that, The polyethylene glycol aldehyde derivative solution and the polylysine solution; The mass percentage of the polyethylene glycol aldehyde derivative in the polyethylene glycol aldehyde derivative solution is 5-20%; the mass percentage of the polylysine in the polylysine solution is 4-10%; and the molar ratio of the aldehyde groups in the polyethylene glycol aldehyde derivative to the amino groups in the polylysine is 0.05-0.2:

1.

2. The reagent composition according to claim 1, characterized by The polyethylene glycol aldehyde derivative is an aldehyde group-terminated multi-arm polyethylene glycol; the aldehyde groups include one or more of unsubstituted aldehyde groups, alkane aldehyde groups and aromatic aldehyde groups; the aldehyde group-terminated multi-arm polyethylene glycol has 4-8 arms; and the number average molecular weight of the polyethylene glycol aldehyde derivative is 5000-40000 Da.

3. The reagent composition of claim 1, wherein The polylysine is ε-polylysine; the ε-polylysine includes linear ε-polylysine and / or hyperbranched ε-polylysine; The volume ratio of the polyethylene glycol aldehyde derivative solution to the polylysine solution is 1-10:1-10.

4. The reagent composition according to any one of claims 1 to 3, characterized in that, The pH value of the polyethylene glycol aldehyde derivative solution is 3-6; and the solvent of the polyethylene glycol aldehyde derivative solution is a phosphate buffer solution; The pH value of the polylysine solution is 5-10; and the solvent of the polylysine solution is a borate buffer solution, a phosphate buffer solution or water.

5. A process for the preparation of the reagent composition according to any one of claims 1 to 4, characterized in that, The method comprises the following steps: The polyethylene glycol aldehyde derivative solution and the polylysine solution are prepared respectively to obtain the reagent composition.

6. The production method according to claim 5, characterized by, After the polyethylene glycol aldehyde derivative solution and the polylysine solution are prepared, the method further comprises a sterilization treatment; the sterilization treatment is high-pressure steam sterilization treatment, the temperature is 121-126°C, and the time is 15-30 min.

7. An anti-blocking hydrogel, characterized in that, The raw material for preparing the anti-adhesion hydrogel is the reagent composition of any one of claims 1-4 or the reagent composition obtained by the preparation method of claim 5 or 6.

8. The anti-blocking hydrogel according to claim 7, wherein The anti-adhesion hydrogel has a pH value of 6-8, an initial Young's modulus of 20-150 kPa, a swelling rate of -50%-50%, and a residence time in a uterine environment or a simulated uterine environment of 3-7 d.

9. A method of preparing the anti-blocking hydrogel according to claim 7 or 8, characterized in that, The method comprises the following steps: The polyethylene glycol aldehyde derivative solution and the polylysine solution are mixed to perform solidification, thereby obtaining the anti-adhesion hydrogel.

10. The method of claim 9, wherein, The mixing is performed by double mixing syringes.

11. The preparation method according to claim 9, characterized in that, The solidification time is 20-150 s.

12. Use of the reagent composition of any one of claims 1-4, the reagent composition obtained by the preparation method of claim 5 or 6, the anti-adhesion hydrogel of claim 7 or 8, or the anti-adhesion hydrogel obtained by the preparation method of any one of claims 9-11 in the preparation of an intrauterine anti-adhesion preparation.

13. Use according to claim 12, characterized in that, The reagent composition or the anti-adhesion hydrogel is used as a drug carrier.

14. Use according to claim 13, characterized in that, The preparation method of the intrauterine anti-adhesion preparation comprises the following steps: The reagent composition and a drug are injected into a uterine cavity to perform in-situ solidification, thereby obtaining the intrauterine anti-adhesion preparation.

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