CN115607747B

CN115607747B - Double-crosslinked network drug-loaded hydrogel and preparation method and application thereof

Info

Publication number: CN115607747B
Application number: CN202211629112.4A
Authority: CN
Inventors: 袁佩琦; 石道昆; 吴艳雪; 宗果
Original assignee: Shanghai Mingyue Medical Technology Co ltd
Current assignee: Shanghai Mingyue Medical Technology Co ltd
Priority date: 2022-12-19
Filing date: 2022-12-19
Publication date: 2023-03-21
Anticipated expiration: 2042-12-19
Also published as: CN115607747A

Abstract

The invention relates to a double-crosslinking network drug-loaded hydrogel and a preparation method and application thereof, wherein the hydrogel comprises a first crosslinking network and a drug-loaded ROS response nanogel loaded in the first crosslinking network; wherein the framework material of the first cross-linked network comprises degradable synthetic polymer units and degradable natural polymer units; the drug-loaded ROS response nanogel comprises a second cross-linking network and drug molecules encapsulated in the second cross-linking network, wherein a framework material of the second cross-linking network is a degradable natural high-molecular unit, and a skeleton of the second cross-linking network contains ROS response radicals. Related application relates to a medical hydrogel film comprising the double-crosslinked network drug-loaded hydrogel and an anti-adhesion medical device, taking a uterine cavity anti-adhesion device as an example, the medical hydrogel film can generate the breakage of ROS response radicals according to an inflammation environment, and accurately release drug molecules at inflammation positions, so that the repair of endometrium is promoted while adhesion is prevented, and the recurrence of adhesion is effectively inhibited.

Description

Double-crosslinked network drug-loaded hydrogel and preparation method and application thereof

Technical Field

The invention relates to the technical field of medical instruments, in particular to a double-crosslinked network drug-loaded hydrogel and a preparation method and application thereof, and further relates to the double-crosslinked network drug-loaded hydrogel and the preparation method thereof, a medical hydrogel film and anti-adhesion medical equipment, wherein the anti-adhesion medical equipment at least comprises uterine cavity anti-adhesion equipment.

Background

For the operations of uterine cavity, abdominal cavity and the like, postoperative re-adhesion prevention is one of the important means for improving the success rate of the operations. Taking the example of uterine cavity surgery, intrauterine adhesion is also called as "Asherman syndrome", which refers to the damage of the endometrial basement layer caused by various factors. The causes of endometrial injury include induced abortion, induced labor and cesarean section, and the like, and the clinical manifestations include abnormal menstruation, periodic abdominal pain, infertility, spontaneous abortion, placental abnormality and cervical pregnancy, etc.

Hysteroscopes are the main methods for treating intrauterine adhesion at present, and surgical instruments are usually operated under direct vision, enter the uterine cavity through a natural orifice and separate the adhesion. However, for severe intrauterine adhesion patients, the re-adhesion rate after the separation operation is as high as 62.5%, and the success rate of pregnancy is only 22.5% -33.3%. Prevention of re-adhesion after hysteroscopy is particularly important because of the high adhesion rate after surgery. At present, the method for preventing re-adhesion after the intrauterine adhesion separation in clinic mainly comprises the steps of placing an intrauterine device and a saccule bracket in the uterine cavity, or using sodium hyaluronate and the like. However, for patients with severe intrauterine adhesions, the endometrium is difficult to repair and regenerate after surgery due to the endometrial regeneration repair failure, the uterine cavity structure is difficult to repair, fertility is affected, and secondary removal is required, increasing surgical complexity and infection risk.

Therefore, development of an anti-adhesion medical device capable of effectively inhibiting adhesion recurrence of patients with severe intrauterine adhesion and effectively promoting functional regeneration of endometrium is urgently needed.

Disclosure of Invention

Based on the above, the invention aims to provide the double-crosslinked network drug-loaded hydrogel and the preparation method thereof, the medical hydrogel film and the anti-adhesion medical equipment, wherein the anti-adhesion medical equipment at least comprises a uterine cavity anti-adhesion equipment. The double-crosslinked network drug-loaded hydrogel has good biocompatibility and mechanical property, can release ROS (reactive oxygen species) response nanogel containing drug molecules in vivo, controllably release the drug molecules and promote the repair of surgical wound parts, and can remarkably reduce the recurrence rate of adhesion and promote the postoperative repair when being applied to severe postoperative adhesion patients.

In a first aspect of the present invention, there is provided a double-crosslinked network drug-loaded hydrogel comprising: a first cross-linked network and a drug-loaded ROS-responsive nanogel loaded in the first cross-linked network;

wherein the content of the first and second substances,

the framework material of the first cross-linked network comprises degradable synthetic polymer units Polyd0 and degradable natural polymer units Polyd1; in the first crosslinked network, the degradable synthetic polymer unit Polyd0 and the degradable natural polymer unit Polyd1 are each independently covalently linked to an adjacent structural unit;

the drug-loaded ROS-responsive nanogel comprises a second cross-linked network and a drug molecule non-covalently entrapped in the second cross-linked network; the skeleton material of the second cross-linked network is degradable natural polymer unit Polyd2, and the skeleton of the second cross-linked network contains ROS response groups.

In a second aspect of the present invention, a method for preparing a medical hydrogel-based membrane is provided, which uses raw materials including an ROS-responsive precursor molecule, a second crosslinkable natural polymer, a drug molecule, a crosslinkable synthetic polymer, and a first crosslinkable natural polymer, and the method for preparing a double-crosslinked network drug-loaded hydrogel includes the following steps:

mixing the ROS response precursor molecule with the second cross-linkable natural macromolecule, and performing cross-linking reaction to form a second cross-linking network to prepare ROS response nanogel containing ROS response groups;

swelling the ROS response nanogel in an aqueous solution containing the drug molecules, and loading the drug molecules in the second cross-linked network to prepare a drug-loaded ROS response nanogel;

then preparing the double-crosslinked network medical hydrogel by adopting a method shown in the following mode one or mode two:

the first method is as follows: mixing the crosslinkable synthetic polymer and the first crosslinkable natural polymer, and performing a crosslinking reaction to form a first crosslinking network to prepare composite hydrogel; then swelling the composite hydrogel in a mixed solution containing the drug-loaded ROS response nanogel, and loading the drug-loaded ROS response nanogel in a first cross-linking network of the composite hydrogel to prepare the double-cross-linking network drug-loaded hydrogel;

the second method comprises the following steps: mixing the crosslinkable synthetic polymer, the first crosslinkable natural polymer and the drug-loaded ROS response nanogel, and carrying out crosslinking reaction to form a first crosslinking network so as to prepare the double crosslinking network drug-loaded hydrogel;

wherein, the first and the second end of the pipe are connected with each other,

the second cross-linkable natural polymer comprises a degradable natural polymer unit Polyd2 and at least two carbon-carbon double bonds;

the ROS-responsive precursor molecule comprises an ROS-responsive group and at least two carbon-carbon double bonds;

the cross-linkable synthetic polymer comprises a degradable synthetic polymer unit Polyd0 and at least two carbon-carbon double bonds, and the first cross-linkable natural polymer comprises a degradable natural polymer unit Polyd1 and at least two carbon-carbon double bonds;

the degradable synthetic polymer unit Polyd0, the degradable natural polymer unit Polyd1, the degradable natural polymer unit Polyd2, the ROS-responsive group and the drug molecule are as defined in the first aspect of the invention.

In a third aspect of the invention, a medical hydrogel film is provided, comprising a drug-loaded hydrogel base film, and optionally a positioning structure;

wherein the drug-loaded hydrogel base membrane comprises the double-crosslinked network drug-loaded hydrogel of the first aspect of the invention or the double-crosslinked network drug-loaded hydrogel prepared by the preparation method of the second aspect of the invention;

when medical hydrogel membrane includes location structure, location structure integral type set up in two cross-linked network medicine carrying hydrogel membrane side.

In a fourth aspect of the invention, an anti-adhesion medical device is provided, which comprises at least one of the double cross-linked network drug-loaded hydrogel of the first aspect of the invention, the double cross-linked network drug-loaded hydrogel prepared by the preparation method of the second aspect of the invention, and the medical hydrogel film of the third aspect of the invention; the anti-adhesion medical equipment at least comprises a uterine cavity anti-adhesion equipment.

In some embodiments, the uterine cavity adhesion prevention device is a medical device that prevents uterine cavity adhesion and promotes endometrial repair.

In a fifth aspect of the invention, the application of the double cross-linked network drug-loaded hydrogel according to the first aspect of the invention, the double cross-linked network drug-loaded hydrogel prepared by the preparation method according to the second aspect of the invention, or the medical hydrogel film according to the third aspect of the invention as an anti-adhesion medical device or in the preparation of an anti-adhesion medical device is provided, wherein the anti-adhesion medical device at least comprises a uterine cavity anti-adhesion device.

Taking the uterine cavity operation as an example, the inventor of the application discovers that a patient with severe uterine cavity adhesion has serious damage to the endometrial basal layer, although the hysteroscope recovers the anatomical structure of the uterine cavity and has various treatment modes for preventing the re-adhesion, the defect of regeneration and repair of the endometrium after the operation exists, and the inventor speculates that the defect is one of the factors for causing the recurrence of the adhesion of the patient with severe uterine cavity adhesion. Based on this, the treatment thinking that promotes functional regeneration of endometrium when the application proposes antiseized, and then provides the two cross-linked network medicine carrying hydrogel, medical hydrogel membrane and antiseized even medical equipment (including but not limited to the antiseized equipment of palace chamber) that can be used to at this treatment mode, when using the antiseized equipment of palace chamber that the application provided, can promote the restoration of endometrium when antiseized, effectively restrain the adhesion recurrence.

The double-crosslinked-network drug-loaded hydrogel provided by the invention comprises a first crosslinked network which takes degradable synthetic polymer Polyd0 and degradable natural polymer (marked as degradable natural polymer Polyd 1) as main framework materials, good mechanical properties are provided by the suitable degradable synthetic polymer Polyd0, excellent biocompatibility is provided by the suitable degradable natural polymer Polyd1, stress reaction is reduced, and meanwhile, a good environment is provided for cell growth. And the grid of the first cross-linked network is also loaded with ROS (reactive oxygen species) response nanogel, the ROS response nanogel adopts degradable natural polymers (marked as degradable natural polymers Poly 2) to provide main framework materials to form a second cross-linked network, so that the drug-loaded nanogel is endowed with excellent biocompatibility and low stress reaction, the framework of the second cross-linked network in the ROS response nanogel contains ROS response groups, and the second cross-linked network also contains drug molecules, so that the drug molecules can be controllably released under the action of multiple mechanisms of ROS response, diffusion and double cross-linked network degradation. The double-crosslinked network drug-loaded hydrogel is applied to postoperative severe adhesion patients, can obviously reduce the recurrence rate of adhesion, and promotes postoperative repair. Taking a drug molecule as a chemotactic factor SDF-1 alpha as an example, in an inflammatory environment, on one hand, the drug-loaded ROS response nanogel can be released from the first cross-linking network under the action of self diffusion movement and degradation of the first cross-linking network, and on the other hand, along with the breakage of ROS response radicals in the drug-loaded ROS response nanogel, the drug molecule can be released under the action of multiple mechanisms of ROS response, diffusion and degradation of the second cross-linking network, so that the chemotactic factor can be controllably and accurately released at an inflammation part, and the utilization rate of released drugs is improved; the released chemotactic factor can recruit the homing of stem cells, directionally migrate to the damaged wound surface and promote the repair of inflammatory parts. The application provides a two crosslinked network medicine carrying hydrogel when being used for preventing adhesion, can also play the effect of promoting the restoration, effectively restrain the adhesion recurrence. The current commercial anti-adhesion materials focus on the improvement of the anti-adhesion effect per se, and no solution is found from the intrinsic mechanism of adhesion recurrence.

The double-crosslinked network drug-loaded hydrogel has suitable mechanical properties, and can not bring uncomfortable foreign body sensation to patients. The intrauterine device and the saccule support can cause foreign body sensation to patients, and the use experience is not good.

The framework materials of the first cross-linking network and the second cross-linking network of the double cross-linking network drug-loaded hydrogel are degradable materials, so that the double cross-linking network drug-loaded hydrogel can be degraded in vivo without being taken out, and the increase of the complexity of the operation and the risk of infection can be avoided. The current commercial anti-adhesion material can not be degraded, needs to be taken out for the second time, and increases the operation complexity.

The double-crosslinked network drug-loaded hydrogel and the medical hydrogel membrane containing the same can be used as anti-adhesion medical equipment or used as raw materials to further prepare anti-adhesion medical equipment, the anti-adhesion medical equipment comprises but is not limited to uterine cavity anti-adhesion equipment, the anti-adhesion effect of a hydrogel material can be exerted, the drug-loaded ROS response nanogel loaded in the first crosslinked network can be released from the first crosslinked network under the action of a single mechanism or a double mechanism of diffusion and/or degradation of the first crosslinked network, furthermore, the breakage of ROS response groups can be realized by utilizing the ROS environment of a post-operation inflammation part, so that drug molecules (such as chemotactic factors) can be released controllably and accurately under the action of multiple mechanisms of ROS response, diffusion and degradation of the second crosslinked network, the higher the ROS concentration is, the stronger the responsiveness is, the larger the release amount of the drug molecules is, the repair of the inflammation part can be effectively promoted, and the adhesion recurrence can be effectively inhibited. In the anti-adhesion uterine cavity device, the medical hydrogel membrane can be in a folded or curled structure in the initial state, is pressed and held in a conveying system, is pushed into the body through the uterine cavity, is unfolded in the body, physically isolates the damaged endometrium, plays an anti-adhesion role, does not bring uncomfortable foreign body sensation to a patient, and has experience feeling remarkably superior to implanted materials such as an intrauterine device, a balloon stent and the like.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more fully understand the present application and the advantages thereof, the drawings used in the description of the embodiments will be briefly introduced below. It is obvious that the drawings in the following description are only some embodiments of the application, and that other drawings can be derived from these drawings by a person skilled in the art without inventive effort. It is also to be noted that the drawings are drawn in simplified form and are provided solely for the purpose of facilitating and distinctly facilitating the description of the invention. The various dimensions of each component shown in the figures are arbitrarily illustrated, may be precision or may not be drawn to scale. For example, the dimensions of the elements in the figures may be exaggerated where appropriate to improve clarity. The various features of the drawings are not necessarily to scale unless specifically indicated. The present invention is not limited to each size of each component.

Wherein like reference numerals refer to like parts in the following description.

Fig. 1 is a schematic flow chart of a preparation process of a double-crosslinked network drug-loaded hydrogel according to an embodiment of the present application; the method comprises the following steps of (A) preparing ROS response nanogel by using carbon-carbon double-bonded hyaluronic acid (HA 2) and ketone thiol with double-bonded ends as raw materials, and further swelling drug-loaded molecules to obtain drug-loaded ROS response hydrogel; (B) The preparation method comprises the steps of crosslinking carbon-carbon double-bonded hyaluronic acid (HA 1) and double-end carbon-carbon double-bonded PLGA to prepare PLGA/HA1 composite hydrogel, and then swelling in a liquid phase containing the drug-loaded ROS response hydrogel to prepare the double-crosslinked network drug-loaded hydrogel;

FIG. 2 is a schematic flow chart illustrating the preparation of a double-crosslinked network drug-loaded hydrogel according to an embodiment of the present disclosure; the method comprises the following steps of (A) preparing ROS response nanogel by using carbon-carbon double-bonded hyaluronic acid (HA 2) and ketone thiol with double-bonded ends as raw materials, and further swelling drug-loaded molecules to obtain drug-loaded ROS response hydrogel; (B) The double-crosslinked network drug-loaded hydrogel is prepared by crosslinking carbon-carbon double-bonded hyaluronic acid HA1, double-carbon double-bonded PLGA and drug-loaded ROS responsive hydrogel;

fig. 3 is a schematic structural view of a hydrogel film for chinese medical use according to an embodiment of the present application.

Description of reference numerals: 100, medical hydrogel film; 130, drug-loaded hydrogel basement membrane; 140, a positioning structure.

Detailed Description

The present invention will be described in further detail with reference to the drawings, embodiments and examples. It should be understood that these embodiments and examples are given solely for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention, which is provided for the purpose of providing a more thorough understanding of the present disclosure. It is also understood that the present invention may be embodied in many different forms and is not limited to the embodiments and examples described herein, and that various changes and modifications may be effected therein by one of ordinary skill in the art without departing from the spirit and scope of the invention and equivalents thereof. Furthermore, in the following description, numerous specific details are set forth in order to provide a more thorough understanding of the present invention, and it is to be understood that the present invention may be practiced without one or more of these details.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments and examples only and is not intended to be limiting of the invention.

Unless otherwise indicated or contradicted, terms or phrases used herein have the following meanings:

the term "and/or", as used herein, is intended to include any one of two or more of the associated listed items, as well as any and all combinations of the associated listed items, including any two of the associated listed items, any more of the associated listed items, or all combinations of the associated listed items. It should be noted that when at least three items are connected by at least two conjunctive combinations selected from "and/or", "or" and/or ", it should be understood that in this application, the technical solutions unquestionably include the technical solutions all connected by" logical and ", and also unquestionably include the technical solutions all connected by" logical or ". For example, "A and/or B" includes three side-by-side schemes of A, B and combinations of A and B. For example, a reference to "a, and/or, B, and/or, C, and/or, D" includes any one of a, B, C, and D (i.e., all connected by "logical or"), any and all combinations of a, B, C, and D (i.e., any two or any three of a, B, C, and D), and any four combinations of a, B, C, and D (i.e., all connected by "logical and").

In this application, when at least three features are connected by at least two conjunctions selected from "and/or", "or" and "," and/or ", this is equivalent to the expression" having one or more features ", such as" TA, and/or, TB, and/or, TC, and/or, TD "is equivalent to" having one or more of the following features: TA, TB, TC and TD ".

The present invention relates to "plural", etc., and indicates that it is 2 or more in number, unless otherwise specified. For example, "one or more" means one or two or more.

In the present specification, the term "suitable" in "any suitable manner", and the like shall be construed as being capable of implementing the technical solution of the present invention, solving the technical problem of the present invention, and achieving the intended technical effect of the present invention.

The terms "preferably", "better" and "better" are used herein only to describe better embodiments or examples, and it should be understood that the scope of the present invention is not limited by these terms. If multiple 'preferences' appear in one technical scheme, if no special description exists, and no contradiction or mutual restriction exists, each 'preference' is independent.

In the present invention, "further", "still further", "specifically" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of the present invention.

In the present invention, "optionally", "optional" and "optional" refer to the presence or absence, i.e., to any one of two juxtapositions selected from "present" and "absent". If multiple optional parts appear in one technical scheme, if no special description exists, and no contradiction or mutual constraint relation exists, each optional part is independent.

In the present invention, the terms "first", "second", "third", "fourth", etc. in the terms of "first aspect", "second aspect", "third aspect", "fourth aspect", etc. are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or quantity, nor as implying that importance or quantity indicating the technical feature being indicated. Also, "first," "second," "third," "fourth," etc. are used for non-exhaustive enumeration of description purposes only and should not be construed as a closed limitation to the number.

In the present invention, the technical features described in the open type include a closed technical solution composed of the listed features, and also include an open technical solution including the listed features.

In the present invention, a range of values (i.e., a range of values) is included, and unless otherwise stated, the distribution of values that are selectable within the range of values is considered to be continuous and includes both the endpoints (i.e., the minimum and maximum) of the range of values and each value between the endpoints. Unless otherwise specified, when a numerical range refers to integers only within the numerical range, the inclusion of both endpoints of the range, and each integer between the endpoints, is equivalent to the direct recitation of each integer. Where multiple numerical ranges are provided to describe a feature or characteristic, the numerical ranges may be combined. In other words, unless otherwise indicated, all numerical ranges disclosed herein are to be understood to include any and all subranges subsumed therein. The "numerical value" in the numerical range may be any quantitative value such as a number, a percentage, a ratio, or the like. "numerical range" is intended to broadly encompass any type of numerical range, including percentage ranges, proportional ranges, ratio ranges, and the like.

The temperature parameter in the present invention is not particularly limited, and is allowed to be constant temperature treatment or to vary within a certain temperature range. It will be appreciated that the described thermostatic process allows the temperature to fluctuate within the accuracy of the instrument control. Allowing fluctuations in the range of, for example,. + -. 5 deg.C,. + -. 4 deg.C,. + -. 3 deg.C,. + -. 2 deg.C, + -. 1 deg.C.

In the present invention, the term "room temperature" generally means 4 ℃ to 35 ℃, preferably 20 ℃. + -. 5 ℃. In some embodiments of the invention, "room temperature" refers to 10 ℃ to 30 ℃. In some embodiments of the invention, "room temperature" refers to 20 ℃ to 30 ℃.

In the present invention, the units relating to the data range, if only with units following the right end point, indicate that the units of the left end point and the right end point are the same. For example, 3 to 5h indicate that the units of the left end point "3" and the right end point "5" are all h (hours).

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. The citation referred to herein is incorporated by reference in its entirety for all purposes unless otherwise in conflict with the present disclosure's objectives and/or technical solutions. Where a citation is referred to herein, the definition of a reference in the document, including features, terms, nouns, phrases, etc., that is relevant, is also incorporated by reference. In the present invention, when the citation is referred to, the cited examples and preferred embodiments of the related art features are also incorporated by reference into the present application, but the present invention is not limited to the embodiments. It should be understood that where the citation conflicts with the description herein, the application will control or be adapted in accordance with the description herein.

Taking the uterine cavity operation as an example, the inventor of the application discovers that a patient with severe uterine cavity adhesion has serious damage to the endometrial basal layer, although the hysteroscope recovers the anatomical structure of the uterine cavity and has various treatment modes for preventing the re-adhesion, the defect of regeneration and repair of the endometrium after the operation exists, and the inventor speculates that the defect is one of the factors for causing the recurrence of the adhesion of the patient with severe uterine cavity adhesion. Based on this, the treatment thinking that promotes functional regeneration of endometrium when the application proposes antiseized, and then provides two networking medicine carrying hydrogel, medical hydrogel membrane and antiseized even medical equipment (including but not limited to the antiseized even equipment of palace chamber) that can be used to this treatment mode, uses the antiseized even equipment of palace chamber that the application provided, can promote the restoration of endometrium when antiseized, effectively restrain the adhesion recurrence.

wherein the content of the first and second substances,

the framework material of the first cross-linked network comprises degradable synthetic polymer units Polyd0 and degradable natural polymer units Polyd1; preferably, in the first crosslinked network, the degradable synthetic polymer unit poly 0 and the degradable natural polymer unit poly 1 are each independently covalently linked to an adjacent structural unit;

the drug-loaded ROS-responsive nanogel comprises a second cross-linked network and a drug molecule non-covalently entrapped in the second cross-linked network; the skeleton material of the second cross-linked network is degradable natural polymer unit poly 2, and the skeleton of the second cross-linked network contains Reactive Oxygen Species (ROS) response groups.

The double-crosslinked network drug-loaded hydrogel provided by the invention comprises a first crosslinked network which takes degradable synthetic polymer Polyd0 and degradable natural polymer (marked as degradable natural polymer Polyd 1) as main framework materials, good mechanical properties are provided through the suitable degradable synthetic polymer Polyd0, excellent biocompatibility is provided through the suitable degradable natural polymer Polyd1, stress reaction is reduced, and a good environment is provided for cell growth, ROS (reactive oxygen species) response nanogel is loaded in a grid of the first crosslinked network, the ROS response nanogel provides main framework materials through the degradable natural polymer (marked as degradable natural polymer Polyd 2) to form a second crosslinked network, the drug-loaded nanogel is endowed with excellent biocompatibility and low stress reaction, a framework of the second crosslinked network in the ROS response nanogel contains ROS response groups, and the second crosslinked network also contains drug molecules, so that the drug molecules can be controllably released under the multiple mechanisms of ROS response, diffusion and double-crosslinked network degradation. The double-crosslinked network drug-loaded hydrogel is applied to postoperative severe adhesion patients, can obviously reduce the recurrence rate of adhesion, and promotes postoperative repair. Taking the drug molecule as the chemotactic factor SDF-1 alpha as an example, in an inflammatory environment, on one hand, the drug-loaded ROS response nanogel can be released from the first cross-linking network under the degradation action of diffusion and the first cross-linking network, and on the other hand, along with the breakage of ROS response radicals in the drug-loaded ROS response nanogel, the drug molecule can be released under the multiple mechanism actions of ROS response, diffusion and second cross-linking network degradation, so that the chemotactic factor can be controllably and accurately released at an inflammation position, the utilization rate of the released drug is improved, and the released chemotactic factor can recruit the homing of stem cells, directionally migrate to an injured wound surface and promote the repair of the inflammation position. The hydrogel material has excellent anti-adhesion performance, and when the double-crosslinked network drug-loaded hydrogel provided by the application is used for preventing adhesion, the double-crosslinked network drug-loaded hydrogel can also play a role in promoting repair and effectively inhibiting adhesion recurrence. The current commercial anti-adhesion materials focus on the improvement of the anti-adhesion effect per se, and no solution is found from the intrinsic mechanism of adhesion recurrence.

In the present invention, the "hydrogel" refers to a highly hydrophilic gel having a three-dimensional network structure, which rapidly swells in water and can retain a large volume of water without dissolving in the swollen state.

In the present invention, "degradable" means, without other limitations, that breaking of chemical bonds can occur. Further, unless otherwise specified, it means that cleavage of chemical bonds can occur in an organism. The conditions under which degradation occurs are related to the type of responsive group of the stimulating factor. For example, the ROS-responsive group may undergo cleavage of a chemical bond in the ROS environment resulting in degradation of the compound of interest.

In the present invention, "ROS" refers to reactive oxygen species. ROS are chemically active oxygen-containing molecules including: hydrogen peroxide (H) ₂ O ₂ ) Singlet oxygen (1O) ₂ ) Superoxide (O) ₂ ⁻ ) Superoxide radicalAnion (O) ₂ · ⁻ ) And a hydroxyl radical (. OH). ROS are natural by-products of cellular oxygen metabolism. Under normal physiological conditions, ROS are involved in vital activities such as cell signaling, cell cycle, cell proliferation, and the like. In abnormal cases, ROS levels are elevated in cells, such as mitochondria in inflamed tissues, which are much higher than in normal cells.

In the present invention, the "ROS-responsive group" refers to a group that undergoes chemical bond cleavage upon encountering ROS. The ROS-responsive group in the present invention is not particularly limited, and any currently known group that can undergo chemical bond cleavage in the ROS environment can be included in the scope of the "ROS-responsive group" in the present invention. Types of ROS-responsive groups may include, but are not limited to, sulfur-containing species, selenium-containing species, tellurium-containing species, boron-containing species.

In the present invention, "polymer" refers to a substance produced by polymerization of monomer molecules, and its structure includes at least 3 repeating units formed of monomer molecules. In the present invention, the average molecular weight of the polymer is more than 1000 Da, unless otherwise specified.

In the present invention, "polymer" and "macromolecule" each independently refer to a substance having a molecular weight greater than 1000 Da, including but not limited to natural macromolecules.

In the present invention, the "natural polymer" refers to a polymer substance present in an animal, a plant or other organism. The molecular chain of the natural polymer may include a segment composed of a repeating unit, such as hyaluronic acid, chitosan, and the like. The molecular chain of the natural polymer may not include a segment composed of a repeating unit, such as some proteins, nucleic acids, etc.

In the present invention, unless otherwise specified, "polymer unit" and "macromolecular unit" are relative to the entire molecule. An intact molecule comprising "polymer units" comprises, in addition to the polymer units, other structural units, which are connected intramolecularly (e.g., covalently) to adjacent structural units. The complete molecule containing the "macromolecule unit" includes other structural units besides the macromolecule unit, and intramolecular connection (such as covalent bond connection) exists between the macromolecule unit and the adjacent structural units. An example of a polymer unit is degradable synthetic polymer unit poly d0. Examples of the polymer unit include degradable natural polymer unit Polyd1 and degradable natural polymer unit Polyd2.

In the present invention, "load" means stably entrapping. The acting force for realizing stable entrapment can be a physical constraint action of a cross-linked network, a chemical action or a combination of the two; the chemical action can be covalent connection or non-covalent connection (such as hydrogen bond, hydrophilic-hydrophobic interaction and other non-covalent connection, such as physical embedding action and the like).

In the present invention, "crosslinked network" refers to a three-dimensional spatial network. The molecule having a crosslinked structure has a plurality of segments linked together by crosslinking points. Due to the constraint of the cross-linking points, the segments cannot move freely. Molecules with a crosslinked network are insoluble in solvents and only swell.

In the present invention, the "first cross-linked network" provides a cross-linked backbone of a double cross-linked network drug-loaded hydrogel, comprising a large mesh with a relatively large mesh size, so as to be able to accommodate the drug-loaded ROS-responsive nanogel.

In the invention, the 'second cross-linked network' provides a cross-linked skeleton of the drug-loaded ROS response nanogel, the overall size of the cross-linked network is relatively small, and the cross-linked network can be stably loaded in a large grid of the first cross-linked network in a nanoscale (such as 300 to 700nm).

In the present invention, unless otherwise specified, "non-covalent" refers to a chemical linkage system in which "non-covalent" may or may not exist with respect to "covalent linkage". The linking means of "non-covalent" is not particularly limited, and some non-limiting examples include hydrogen bonding, hydrophilic-hydrophobic interaction, and the like. Examples of "no chemical linkage present" are physical entrapment etc.

In the present invention, "drug molecules non-covalently entrapped in the second crosslinked network" means that drug molecules are entrapped in the second crosslinked network in a manner other than covalent bonding, either in a "non-covalent" chemical bonding manner or in the absence of chemical bonding.

In the present invention, "natural degradable polymer" refers to a degradable natural polymer. All contain natural degradable polymer in the framework material of first crosslinked network and second crosslinked network of this application, natural degradable polymer in the first crosslinked network can be write as Polyd1, and natural degradable polymer in the second crosslinked network can be write as Polyd2. The degradable natural polymers Polyd1 and Polyd2 in the two crosslinked networks can be the same or different, the types can be the same or different, and the molecular weights can be the same or different. In some embodiments, both the Polyd1 and the Polyd2 are different in type and molecular weight. In other embodiments, the Polyd1 and Polyd2 are of the same type, such as being units of any of hyaluronic acid, chitosan, or collagen, but may be of the same or different molecular weights. In other embodiments, the molecular weights of Polyd1 and Polyd2 are substantially the same (with a deviation of not more than. + -. 10% based on the amount of Polyd 1), and the species may be the same or different.

In the invention, the drug molecules in the drug-loaded ROS response nanogel are encapsulated in the double cross-linked networks, and the release behavior of the drug molecules is influenced by multiple actions of ROS response groups, the degradation action of the second cross-linked network, the size of the drug-loaded ROS response nanogel, the degradation action of the first cross-linked network and the like. By adjusting various parameters such as the type and the content of ROS response groups, the type of the second cross-linked network, the type of the first cross-linked network, the content of the degradable units, the mass ratio of the second cross-linked network to the first cross-linked network and the like, the drug molecules can be released in a better mode in a slow release manner, can be released accurately and fully at the focus part, can also avoid burst release from starting to release, and can be better matched with the repair cycle of the focus part.

In the present invention, the degradable synthetic polymer in the first crosslinked network may be denoted as poly d0. In some embodiments, the degradable synthetic polymer Polyd0 has a number average molecular weight between two adjacent cross-links of the first cross-linked network selected from 5 to 20 kDa. Can be controlled by controlling the molecular weight of the starting materials (crosslinkable synthetic polymers).

The size of the first cross-linked network can be adjusted by changing the molecular weight of the reaction raw materials (including the first cross-linkable natural macromolecule and the cross-linkable synthetic polymer) and the ratio of the first cross-linked network to the cross-linkable synthetic polymer). The mesh size of the second crosslinked network can be adjusted by changing the molecular weight of the reaction raw materials (including the second crosslinkable natural macromolecule and the ROS-responsive precursor molecule) and the ratio of the two.

Covalent bonds may or may not be present between the second crosslinked network and the first crosslinked network.

In some embodiments, the drug-loaded ROS-responsive nanogel is non-covalently loaded in the first cross-linked network, at which point there is no chemical bonding between the drug-loaded ROS-responsive nanogel and the first cross-linked network. At this point, the drug-loaded ROS-responsive nanogel can be released from the first cross-linked network through a dual mechanism of diffusion and degradation of the first cross-linked network.

In some embodiments, the drug-loaded ROS-responsive nanogel is loaded into the first crosslinked network by covalent linkage. At this point, the drug-loaded ROS-responsive nanogel can be released from the first cross-linked network under a first cross-linked network degradation mechanism.

In some embodiments, the mass ratio of the second crosslinked network to the first crosslinked network is (0.1 to 2): 2 to 10, and further can be 1: (2 to 7). The mass ratio of the second crosslinked network to the first crosslinked network may also be selected from the following interval consisting of any one ratio or any two ratios: 1.

In some embodiments, the mass percentage of the first cross-linked network in the double cross-linked network drug-loaded hydrogel is selected from 0.5% to 10%, and may also be selected from an interval consisting of any one or two of the following percentages: 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, etc., by dry weight.

In the present invention, "on a dry weight basis" means the mass of the free solvent is not included. For example, water is not included in the hydrogel.

In some embodiments, the drug-loaded ROS-responsive nanogel is present in the double-crosslinked network drug-loaded hydrogel in an amount of 0.5% to 30% by weight based on dry weight. The weight percentage of the drug-loaded ROS response nanogel in the double-crosslinked network drug-loaded hydrogel can be selected from the interval consisting of any one or two of the following percentages: 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 15%, 20%, 25%, 30%, etc.

The drug-loading rate in the double-crosslinking-network drug-loaded hydrogel is related to the parameters such as the mass ratio of the second crosslinking network to the first crosslinking network, the mass percentage of the first crosslinking network in the double-crosslinking-network drug-loaded hydrogel, the mass percentage of the drug-loaded ROS response nanogel in the double-crosslinking-network drug-loaded hydrogel, and the like, and the drug-loading rate can be controlled by controlling the feeding ratio of corresponding raw materials, so that the cost can be well controlled under the condition of meeting the drug quantity requirement of a wound repair part.

In some embodiments, the average particle size of the drug-loaded ROS-responsive nanogel is selected from 300 to 700 nm. The size of the drug-loaded ROS response nanogel is matched with the size of the grid of the first cross-linking network, so that the grid structure of the first cross-linking network can accommodate the drug-loaded ROS response nanogel and can limit the motion space of the drug-loaded ROS response nanogel to a certain extent, and the drug-loaded ROS response nanogel can be stably kept in the drug-loaded hydrogel of the double cross-linking network and cannot be easily leaked from the first cross-linking network. When meeting ROS environment, the drug-loaded ROS responds to the breakage of ROS radicals in the nanogel, so that drug molecules are released from the second cross-linked network, and can also be freely diffused in the first cross-linked network to be released from the double cross-linked network drug-loaded hydrogel.

In some embodiments, the ROS-responsive group may be selected from one or more of the following groups: ketone thiol (-S-C (CH)) ₃ ) ₂ -S-), thioether linkage (-S-), monoselenic linkage (-Se-), diselenic linkage (-Se-Se-), tellurium (Te-), oxalic acidEster group (-OC (= O) -C (= O) -O-), thiazolinone group (C: (C))

) Boronic ester groups (e.g. of

) Boronic acid groups (e.g. -B (OH) ₂ ) And proline oligomeric chains.

In some embodiments, the proline oligomeric chain has a structure such as

The formula is shown in the specification, wherein n is an integer selected from 3 to 8. In some of these embodiments, the number of proline units in the proline oligomeric chain is selected from 3 to 7 (e.g., 3, 4, 5, or 7). In some of these embodiments, n =7.

Examples of ROS-responsive groups are-Ar-OC (= O) -C (= O) -O-, where Ar is an arylene group, such as phenylene for example, and further such as 1, 4-phenylene for example.

As used herein, the term "arylene" refers to a divalent residue derived from an aromatic group by the loss of one more hydrogen atom from the aromatic ring, having two monovalent radical centers. The arylene group can be a monocyclic arylene group, or a fused ring arylene group, or a polycyclic arylene group, at least one of which for polycyclic ring species is an aromatic ring system. Suitable examples include, but are not limited to, arylene groups derived from the following aromatic rings: benzene, biphenyl, naphthalene, anthracene, phenanthrene, perylene, triphenylene, and derivatives thereof.

As used herein, the term "aryl" refers to a monovalent aromatic hydrocarbon radical derived from an aromatic ring by the loss of a hydrogen atom from the aromatic ring, i.e., by the direct formation of a monovalent attachment site on the ring. The aryl group can be a monocyclic aryl group, or a fused ring aryl group, or a polycyclic aryl group, at least one of which is an aromatic ring system for polycyclic ring species. Suitable examples include, but are not limited to, aryl groups derived from the following aromatic hydrocarbons: benzene, biphenyl, naphthalene, anthracene, phenanthrene, perylene, triphenylene, and hydrocarbon derivatives thereof.

Examples of ROS-responsive groups are also arylboronic acid ester groups, furtherPhenylboronate groups, non-limiting examples being

。

Examples of ROS-responsive groups are also arylboronic acid groups, further phenylboronic acid groups, non-limiting examples being-Ar-B (OH) ₂ Wherein Ar is an arylene group, such as phenylene, further exemplified by, for example, 1, 4-phenylene.

In some embodiments, the ROS-responsive group is selected from one or more of the following groups: keto thiol (-S-C (CH) ₃ ) ₂ -S-, thioether-linkage (-S-), monoselenium-linkage (-Se-), diselenium-linkage (-Se-Se-), divalent tellurium (-Te-), oxalate group (-OC (= O) -C (= O) -O-), aryloxalate group (-Ar-OC (= O) -C (= O) -O-), thiazolinone-group (-Se-) (Te-)

) Boronic ester groups (e.g. of

) Arylboronic acid ester groups (e.g. aryl boronic acid ester groups)

) Boronic acid group (-B (OH) ₂ ) Aryl boronic acid group (-Ar-B (OH) ₂ ) And proline oligomeric chains (e.g. of

Wherein n is an integer selected from 3 to 8, such as 3, 4, 5, 6, 7 or 8); ar in the above groups is independently the same or different, and is further independently a phenyl group, and is further independently a1, 4-phenylene group.

In the present invention, the drug molecules may be selected from macromolecular drugs. At the moment, the drug molecules can be stably encapsulated in the second cross-linked network under the condition that covalent bonds are not generated with the cross-linked network, and the diffusion effect is well inhibited when the cross-linked network is not degraded and/or ROS response groups are not broken, so that the bioavailability of the drug molecules is improved. In addition, the double-crosslinking network drug-loaded hydrogel can be stored under the water-containing condition, and at the moment, the leakage of drug molecules can be avoided in the storage process by utilizing the constraint effect of the double-crosslinking network on the drug molecules, so that the storage stability of the product is ensured.

In some embodiments, the drug molecule comprises one or more of a chemokine and an estrogen. In some embodiments, the drug molecule is selected from one or more of a chemokine and an estrogen.

When the medicine molecules comprise chemotactic factors, the regeneration and repair of endometrium can be promoted along with the release of the chemotactic factors, and when the double-crosslinked network drug-loaded hydrogel is applied to patients with severe intrauterine adhesion, the recurrence rate of adhesion can be obviously reduced, and the repair of endometrium can be promoted. The chemokine can be, but is not limited to, SDF-1 α.

When the drug molecules comprise estrogen, the composition can promote rapid regeneration of endometrium, reduce anticoagulation effect, and reduce or even prevent occurrence of uterine cavity secondary adhesion caused by basal layer exposure, bleeding and other reasons after operation.

In some embodiments, the phase mass content of the drug molecule in the drug-loaded ROS-responsive nanogel is 0.001 to 0.05. Mu.g/g, and may be 0.001. Mu.g/g, 0.002. Mu.g/g, 0.004. Mu.g/g, 0.005. Mu.g/g, 0.006. Mu.g/g, 0.008. Mu.g/g, 0.01. Mu.g/g, 0.02. Mu.g/g, 0.03. Mu.g/g, 0.04. Mu.g/g, 0.05. Mu.g/g, and the like, on a dry basis, and may be selected from a content interval consisting of any two of the above values.

In some embodiments, the phase mass content of the drug molecule in the double cross-linked network drug-loaded hydrogel is 0.0005 to 0.01 μ g/g, and may be 0.0005 μ g/g, 0.001 μ g/g, 0.0015 μ g/g, 0.002 μ g/g, 0.0025 μ g/g, 0.003 μ g/g, 0.0035 μ g/g, 0.004 μ g/g, 0.0045 μ g/g, 0.005 μ g/g, 0.0055 μ g/g, 0.006 μ g/g, 0.007 μ g/g, 0.008 μ g/g, 0.009 μ g/g, 0.01 μ g/g, and the like by dry weight, and may be selected from a content interval consisting of any two of the above values.

In some embodiments, the double cross-linked network drug-loaded hydrogel has one or more of the following characteristics:

covalent bonding or non-covalent bonding exists between the second crosslinked network and the first crosslinked network;

the mass ratio of the second crosslinking network to the first crosslinking network is (0.1 to 2) to (2 to 10);

the weight percentage of the drug-loaded ROS response nanogel in the double-crosslinked network drug-loaded hydrogel is 0.5-30 percent, and the drug-loaded ROS response nanogel is calculated by dry weight;

the average grain diameter of the drug-loaded ROS response nanogel is selected from 300 to 700 nm;

the drug molecules are selected from one or more of chemokines and estrogens; and

the relative mass content of the drug molecules in the double-crosslinked network drug-loaded hydrogel is 0.0005 to 0.01 mu g/g by dry weight;

the above features can be further combined in the technical scheme of the double-crosslinked network drug-loaded hydrogel according to any suitable manner.

In some embodiments, the mass ratio between the degradable synthetic polymer unit Polyd0, the degradable natural polymer unit Polyd1 and the degradable natural polymer unit Polyd2 is (2 to 7): 1: (0.1 to 1). Can be reasonably adjusted by controlling the corresponding raw material dosage ratio.

In some embodiments, in the second crosslinked network, the degradable natural polymer units poly d2 are selected from one or more of hyaluronic acid units (HA units), collagen units, and chitosan units.

In some embodiments, the molecular weight of the degradable natural polymer units poly d2 in the second crosslinked network may be selected from 4 to 8 kDa, and may also be selected from a numerical range (in terms of number average molecular weight) consisting of any one or two of the following molecular weights: 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa and the like.

In some embodiments, the ROS-responsive groups in the second crosslinked network independently comprise one or more of the following groups: ketothioketal groups, thioether linkages, monoseleno linkages, diseleno linkages, tellurium bivalences, oxalate groups, thiazolinone groups, borate groups, boronic acid groups and proline oligomeric chains, and further wherein the number of proline units (n) in the proline oligomeric chain is as defined above.

In some embodiments, the amount of the ROS-responsive group-containing substance per gram of the second crosslinked network is from 0.001 to 0.05mol, and further may be from 0.005 to 0.05 mol. The amount of ROS-responsive species contained per gram of the second crosslinked network may also be selected from any one or any two of the following ranges of values: 0.002 mol, 0.005 mol, 0.006 mol, 0.007 mol, 0.008 mol, 0.01 mol, 0.02 mol, 0.03 mol, 0.04 mol, 0.05mol, and the like. The amount of the ROS response group-containing substance in each gram of the second crosslinking network directly influences the content of the ROS response group in the double crosslinking network drug-loaded hydrogel, and further influences the responsiveness to the abnormal level of ROS in vivo. By reasonably controlling this content, the drug release properties, including but not limited to release rate, release period, etc., can be better adjusted.

in the second crosslinking network, the degradable natural polymer units Polyd2 are selected from one or more of hyaluronic acid units, collagen units and chitosan units;

in the second cross-linked network, the number average molecular weight of the degradable natural polymer units Polyd2 is independently selected from 4 to 8 kDa;

in the second crosslinked network, the ROS-responsive groups independently comprise one or more of the following groups: ketothioketal group, thioether bond, mono-selenium bond, di-selenium bond, divalent tellurium, oxalate group, thiazolinone group, borate group, boric acid group and proline oligomer chain, wherein the number of proline units in the proline oligomer chain is selected from 3 to 7; and

the amount of the ROS-responsive group-containing substance per gram of the second crosslinked network is 0.001 to 0.05mol (alternatively, 0.005 to 0.05 mol);

In some embodiments, the degradable synthetic polymer units poly 0 in the first crosslinked network comprise one or more of polyester units, furtherThe polyester unit is an aliphatic polyester unit, and further the repeating unit of the polyester unit is [ -C (= O) -U [ - ] _R -O-]Wherein, any one of U _R Independently selected from C _1-10 Alkylene groups. In some embodiments, any one of U _R Independently selected from C _1-10 Alkylene, which may be methylene (-CH) ₂ -), ethylene (e.g. -CH) ₂ CH ₂ -、-CH(CH ₃ ) -), propylene (e.g. -CH) ₂ CH ₂ CH ₂ -、-CH ₂ CH(CH ₃ )-、-CH(CH ₃ )CH ₂ -、-C(CH ₃ ) ₂ -), butylene (e.g., n-butyl, etc.), pentylene (e.g., n-pentyl, etc.), hexylene, heptylene, octylene, nonylene, decylene. In some embodiments, any one of U _R Independently selected from C _1-6 An alkylene group. In some embodiments, any one of U _R Independently selected from methylene-CH ₂ -、-CH(CH ₃ ) And n-pentyl, in which case the degradable synthetic polymer unit Polyd0 is selected from one or more of polylactic acid units (PLA units), poly (lactic-glycolic acid) units (PLGA units, i.e. lactic-glycolic acid copolymer units) and polycaprolactone units (PCL units). U shape _R When methylene, polyd0 corresponds to polyglycolide units; u shape _R is-CH (CH) ₃ ) When, poly d0 corresponds to polylactic acid (i.e. polylactide) units; u shape _R Is methylene and-CH (CH) ₃ ) In combination of (a), poly d0 corresponds to poly (lactic-glycolic acid) units; u shape _R In the case of n-pentyl group, polyd0 corresponds to polycaprolactone units.

In some embodiments, the degradable synthetic polymer units poly 0 in the first crosslinked network comprise one or more of polylactic acid units (PLA units), poly (lactic-glycolic acid) units (PLGA units), and polycaprolactone units (PCL units).

In some embodiments, in the first crosslinked network, the degradable natural polymer units Polyd1 are selected from one or more of hyaluronic acid units, collagen units, and chitosan units.

In some embodiments, the degradable natural polymers Polyd1, polyd2 are each independently selected from one or more of hyaluronic acid units, collagen units, and chitosan units.

In some embodiments, the degradable synthetic polymer units poly 0 in the first crosslinked network have a molecular weight independently selected from 10 to 30 kDa, in number average molecular weight. The number average molecular weight of the degradable synthetic polymer unit poly 0 may also be selected from the following numerical intervals consisting of any one or two molecular weights: 10 kDa, 12 kDa, 15 kDa, 16 kDa, 18 kDa, 20 kDa, 25 kDa, 26 kDa, 28 kDa, 30 kDa and the like. By adjusting the molecular weight of the degradable synthetic polymer units poly 0, the degradation properties of the first cross-linked network can be better controlled, thereby better adjusting the drug release properties.

In some embodiments, the molecular weight of the degradable natural polymer units poly 1 in the first crosslinked network may be selected from 4 to 8 kDa, and may also be selected from a numerical range (in terms of number average molecular weight) consisting of any one or two of the following molecular weights: 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa and the like.

In some embodiments, the mass ratio of the degradable synthetic polymer unit Polyd0 to the degradable natural polymer unit Polyd1 in the first cross-linked network is (2 to 7): 1. the mass ratio of the degradable synthetic polymer units poly 0 to the degradable natural polymer units poly 1 may be selected from any one or two of the following ranges: 1, 2.5. By controlling the mass ratio of the degradable synthetic polymer units Polyd0 to the degradable natural polymer units Polyd1 in a proper range, different requirements for degradable properties and mechanical properties can be better balanced.

In some embodiments, the covalent bonding between degradable synthetic polymer unit poly 0 and degradable natural polymer unit poly 1 in the first crosslinked network is selected from one or more of the following: carbon-carbon single bonds, ester bonds, and amide bonds.

in the first crosslinked network, the degradable synthetic polymer units poly 0 are selected from one or more of polyester units; further can be selected from one or more of polylactic acid unit, poly (lactic acid-glycolic acid) unit and polycaprolactone unit;

in the first cross-linked network, the degradable natural polymer units Polyd1 are selected from one or more of hyaluronic acid units, collagen units and chitosan units;

in the first crosslinking network, the molecular weight of a degradable synthetic polymer unit Polyd0 is independently selected from 10 to 30 kDa, and the number average molecular weight of a degradable natural polymer unit Polyd1 is independently selected from 4 to 8 kDa; further optionally, in the second cross-linked network, the number average molecular weight of the degradable natural polymer units Polyd2 is independently selected from 4 to 8 kDa;

in the first cross-linked network, the mass ratio of the degradable synthetic polymer units Polyd0 to the degradable natural polymer units Polyd1 is (2 to 7): 1; and

in the first crosslinked network, the covalent bond connection between the degradable synthetic polymer unit poly 0 and the degradable natural polymer unit poly 1 is selected from one or more of the following: carbon-carbon single bonds, ester bonds, and amide bonds;

In some embodiments, the double-crosslinked network drug-loaded hydrogel contains 30-95% by mass of water. The mass percentage of the water in the double-crosslinked network drug-loaded hydrogel can be selected from the interval consisting of any one or two of the following percentages: 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, etc.

In a second aspect of the present invention, a method for preparing a double-crosslinked network drug-loaded hydrogel is provided, which can be used for preparing the double-crosslinked network drug-loaded hydrogel of the first aspect of the present invention.

In some embodiments, a method for preparing a double-crosslinked network drug-loaded hydrogel is provided, which uses a raw material comprising an ROS-responsive precursor molecule, a second crosslinkable natural macromolecule, a drug molecule, a crosslinkable synthetic polymer, and a first crosslinkable natural macromolecule, and the method for preparing the double-crosslinked network drug-loaded hydrogel comprises the following steps:

s100: mixing the ROS response precursor molecule with a second cross-linkable natural macromolecule, and carrying out cross-linking reaction to form a second cross-linking network so as to prepare ROS response nanogel containing ROS response groups;

s200: swelling the ROS response nanogel in an aqueous solution containing drug molecules, and loading the drug molecules in a second cross-linked network to prepare a drug-loaded ROS response nanogel; in some preferred embodiments, the ROS response nanogel is swelled in an aqueous reagent (such as water), then swelled in an aqueous solution containing drug molecules, and the drug molecules are loaded in the second cross-linked network to prepare the drug-loaded ROS response nanogel, and at this time, the loading of the drug molecules can be rapidly completed by utilizing the internal and external concentration difference of the drug molecules;

s300: then, a first cross-linked network is constructed by adopting a method shown in the following mode I or mode II to prepare the medical hydrogel with the double cross-linked network:

mode one (S300 a): mixing the crosslinkable synthetic polymer with the first crosslinkable natural polymer, and carrying out crosslinking reaction to form a first crosslinking network so as to prepare the composite hydrogel; swelling the composite hydrogel in a mixed solution containing the drug-loaded ROS response nanogel, and loading the drug-loaded ROS response nanogel in a first crosslinking network of the composite hydrogel to prepare the double-crosslinking network drug-loaded hydrogel;

mode two (S300 b): mixing a crosslinkable synthetic polymer, a first crosslinkable natural polymer and the drug-loaded ROS response nanogel, and carrying out crosslinking reaction to form a first crosslinking network so as to prepare a double-crosslinking-network drug-loaded hydrogel;

wherein the content of the first and second substances,

the ROS response precursor molecule comprises an ROS response group and at least two carbon-carbon double bonds;

In step S100, the amount of the ROS-responsive precursor molecule substance added per gram of the second crosslinkable natural polymer may be 0.0005 to 0.05mol, and may be selected from any one of the following or a numerical range formed by any two of the following: 0.002 mol, 0.005 mol, 0.006 mol, 0.007 mol, 0.008 mol, 0.01 mol, 0.02 mol, 0.03 mol, 0.04 mol, 0.05mol, and the like. Non-limiting examples also include 0.001 to 0.01 mol, 0.005 to 0.05mol, and the like.

In step S200, the drug-loaded ROS response nanogel has a drug molecule content of 0.001 to 0.05 μ g/g per unit mass, such as 0.001 μ g/g, 0.002 μ g/g, 0.004 μ g/g, 0.005 μ g/g, 0.006 μ g/g, 0.008 μ g/g, 0.01 μ g/g, 0.02 μ g/g, 0.03 μ g/g, 0.04 μ g/g, 0.05 μ g/g, and the like, and may be selected from any two of the above-mentioned values, based on dry weight. In some embodiments, the chemokine content per unit mass of the drug-loaded ROS-responsive nanogel is 0.0053 μ g/g, on a dry weight basis.

In the first mode, the weight ratio of the crosslinkable synthetic polymer to the first crosslinkable natural polymer is (2 to 7): 1, can also be selected from the interval consisting of any one or two of the following ratios: 1, 2.5. Therefore, the mass ratio of the degradable synthetic polymer units Polyd0 to the degradable natural polymer units Polyd1 can be reasonably controlled in a proper range, and different requirements on the degradable property and the mechanical property are well balanced.

In some embodiments, the concentration of the drug-loaded ROS-responsive nanogel in the mixed solution containing the ROS-responsive nanogel is 2 to 10 mg/mL, and further can be 4 to 6 mg/mL. Non-limiting examples are 2 mg/mL, 3 mg/mL, 4 mg/mL, 5mg/mL, 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, 10 mg/mL, and the like.

In some embodiments, when the composite hydrogel is swelled by using the mixed solution containing the drug-loaded ROS response nanogel, the volume-to-mass ratio of the mixed solution to the composite hydrogel is (2-5): (0.1 to 0.5).

In some embodiments, the mixed solution containing the ROS-responsive nanogel includes an aqueous solvent, and further, the aqueous reagent is water.

In the second mode, the weight ratio of the crosslinkable synthetic polymer, the first crosslinkable natural polymer and the drug-loaded ROS response nanogel can be (2 to 7): 1: (0.05 to 1). The weight ratio of the crosslinkable synthetic polymer to the first crosslinkable natural polymer may be selected from any of the above examples. The weight ratio of the first cross-linkable natural polymer to the drug-loaded ROS-responsive nanogel can be selected from any one or two of the following intervals: 1. By controlling the weight ratio of the cross-linkable synthetic polymer, the first cross-linkable natural polymer and the drug-loaded ROS to the nanogel, the ratio of the degradable synthetic polymer unit Polyd0 to the degradable natural polymer unit Polyd1 to the degradable natural polymer unit Polyd2 can be better adjusted, and the mass ratio of the second cross-linked network to the first cross-linked network can be better adjusted, so that the drug release behavior can be better improved, and better mechanical properties can be provided.

In some embodiments, the molecular weight of the crosslinkable synthetic polymer may be selected from 10 to 30 kDa in number average molecular weight. Non-limiting examples are 10 kDa, 15 kDa, 20 kDa, 25 kDa, 30 kDa, etc., and can also be selected from the numerical range consisting of any two molecular weights mentioned above.

In some embodiments, the molecular weight of the first crosslinkable natural polymer may be selected from 4 to 8 kDa, and may be selected from a numerical range (in terms of number average molecular weight) consisting of any one or two of the following molecular weights: 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa and the like.

In some embodiments, the molecular weight of the second crosslinkable natural polymer may be selected from 4 to 8 kDa, and may be selected from a numerical range (in terms of number average molecular weight) composed of any one or two of the following molecular weights: 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa and the like.

In some embodiments, ROS-responsive nanogels comprising ROS-responsive groups may be prepared using a method comprising step S100: mixing ketone thiol with double terminal double bonds and hyaluronic acid HA2 with double carbon-carbon bonds in a water/isopropanol mixed solvent, stirring and mixing in the presence of sodium bicarbonate, adding potassium persulfate, and carrying out a crosslinking reaction in a nitrogen atmosphere at a reaction temperature of 65-75 ℃ for 3-5 h to obtain HA nanogel containing ketone thiol groups; optionally, the amount of the double-ended double-bond ketone thiol compound added in each gram of the carbon-carbon double-bonded hyaluronic acid HA2 is 0.001 to 0.01 mol; optionally, the volume ratio of water to isopropanol in the water/isopropanol mixed solvent is 1 (0.1 to 0.2).

In step S100, non-limiting examples of reaction temperatures include 65 ℃, 66 ℃, 67 ℃, 68 ℃, 69 ℃, 0 ℃, 71 ℃, 72 ℃, 73 ℃, 74 ℃, 75 ℃ and the like; non-limiting examples of reaction times are 3 h, 3.5 h, 4h, 4.5 h, 5h, etc.

In step S100, the amount of the double-ended ketal thiol added per gram of the carbon-carbon double-bonded hyaluronic acid HA2 may be selected from 0.0005 to 0.05mol, and may be selected from any one of the following numerical intervals or a numerical interval consisting of any two of the following: 0.002 mol, 0.005 mol, 0.006 mol, 0.007 mol, 0.008 mol, 0.01 mol, 0.02 mol, 0.03 mol, 0.04 mol, 0.05mol, and the like. Non-limiting examples also include 0.001 to 0.01 mol, 0.005 to 0.05mol, and the like.

In some embodiments, the drug-loaded ROS-responsive nanogel can be prepared by a method comprising step S200 as follows: swelling the HA nano gel containing the ketothiol group in an aqueous solution with the final concentration of a pharmaceutical factor of 0.005 to 0.1 mu g/mL, and stirring at room temperature (such as 20 to 30 ℃) for a certain time (such as 8 to 15 hours, further such as 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours and the like) to prepare the drug-loaded ROS response nano gel. In some embodiments, the drug-loaded ROS-responsive nanogel can be prepared by a method comprising step S200 as follows: swelling HA nano gel containing ketothiol groups in an aqueous reagent (such as water), adding a pharmaceutical factor with the final concentration of 0.005-0.1 mu g/mL, and stirring at room temperature (such as 20-30 ℃) for a certain time (such as 8-15 h, further such as 8 h, 9 h, 10 h, 11 h, 12h, 13 h, 14 h, 15 h and the like) to prepare the drug-loaded ROS response nano gel. In step S200, the final concentration of the pharmaceutical agent may be 0.005 to 0.1. Mu.g/mL, such as, but not limited to, 0.005. Mu.g/mL, 0.01. Mu.g/mL, 0.02. Mu.g/mL, 0.03. Mu.g/mL, 0.04. Mu.g/mL, 0.05. Mu.g/mL, 0.06. Mu.g/mL, 0.07. Mu.g/mL, 0.08. Mu.g/mL, 0.09. Mu.g/mL, 0.1. Mu.g/mL, and the like.

In some embodiments, the method of the first embodiment may include the following step S300a: mixing double-ended carbon-carbon double-bonded PLGA and carbon-carbon double-bonded hyaluronic acid HA1 with a PBS buffer solution with the pH value of 7.2-7.6, adding potassium persulfate and tetramethylethylenediamine, and carrying out crosslinking reaction under the protection of oxygen and nitrogen at the reaction temperature of 65-75 ℃ for 20-40 min to prepare PLGA/HA1 composite hydrogel; soaking and rinsing PLGA/HA1 composite hydrogel in a PBS buffer solution; adding the rinsed PLGA/HA1 composite hydrogel into a mixed solution containing 2-10 mg/mL of drug-loaded ROS response nanogel for swelling (the PLGA/HA1 composite hydrogel can be subjected to swelling after being lyophilized in advance), wherein the swelling time is 4-8 h, and preparing the double-crosslinked network drug-loaded hydrogel; optionally, the weight ratio of the double-ended carbon-carbon double-bonded PLGA to the carbon-carbon double-bonded hyaluronic acid HA1 is (2 to 7): 1, and may also be selected from any suitable examples of the weight ratio of the crosslinkable synthetic polymer to the first crosslinkable natural polymer described above.

In step S300a, non-limiting examples of the reaction temperature include 65 ℃, 66 ℃, 67 ℃, 68 ℃, 69 ℃, 0 ℃, 71 ℃, 72 ℃, 73 ℃, 74 ℃, 75 ℃ and the like; non-limiting examples of reaction times are 20 min, 25 min, 30min, 35 min, 40 min, and the like.

In some embodiments, in step S300a, in the mixed solution of the drug-loaded ROS-responsive nanogel, the concentration of the drug-loaded ROS-responsive nanogel may be 2 to 10 mg/mL, such as, by way of non-limiting example, 2 mg/mL, 3 mg/mL, 4 mg/mL, 5mg/mL, 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, 10 mg/mL, and the like, and may also be selected from a concentration interval consisting of any two of the above concentrations.

In some embodiments, the method of mode two may include the following step S300b: mixing double-ended carbon-carbon double-bonded PLGA, carbon-carbon double-bonded hyaluronic acid HA1 and ROS (reactive oxygen species) carrying drug with PBS (phosphate buffer solution) with the pH of 7.2-7.6, adding potassium persulfate and tetramethylethylenediamine, and carrying out crosslinking reaction under the protection of oxygen and nitrogen at the reaction temperature of 65-75 ℃ for 20-40 min to prepare double-crosslinked network drug-carrying hydrogel; optionally, the weight ratio of the double-ended carbon-carbon double-bonded PLGA, the carbon-carbon double-bonded hyaluronic acid HA1 and the drug-loaded ROS response nanogel is (2 to 7): 1: (0.05 to 1), and can be selected from any suitable examples of the weight ratio of the crosslinkable synthetic polymer, the first crosslinkable natural polymer and the drug-loaded ROS-responsive nanogel.

In step S300b, non-limiting examples of the reaction temperature include 65 ℃, 66 ℃, 67 ℃, 68 ℃, 69 ℃, 0 ℃, 71 ℃, 72 ℃, 73 ℃, 74 ℃, 75 ℃ and the like; non-limiting examples of reaction times are 20 min, 25 min, 30min, 35 min, 40 min, and the like.

In some embodiments, the carbon-carbon double-bonded hyaluronic acid HA2 is prepared by a method comprising the following steps: reacting hyaluronic acid and methacrylic anhydride under the conditions of pH 7.8-8.2 and ice bath for 18-30 h (such as 18 h, 20 h, 24h, 25 h, 26 h, 30 h and the like), precipitating, collecting a solid phase, dialyzing in water, and freeze-drying; wherein, 0.01 to 0.05mol (such as 0.01 mol, 0.02 mol, 0.03 mol, 0.04 mol, 0.05mol and the like) of methacrylic anhydride is added into each gram of hyaluronic acid.

In some embodiments, the carbon-carbon double-bonded hyaluronic acid HA1 is prepared by a method comprising the following steps: reacting hyaluronic acid and methacrylic anhydride under the conditions of pH 7.8-8.2 and ice bath for 18-30 h (such as 18 h, 20 h, 24h, 25 h, 26 h, 30 h and the like), precipitating, collecting a solid phase, dialyzing in water, and freeze-drying; wherein, 0.01 to 0.05mol (such as 0.01 mol, 0.02 mol, 0.03 mol, 0.04 mol, 0.05mol and the like) of methacrylic anhydride is added into each gram of hyaluronic acid.

In some embodiments, the double-ended carbon-carbon double-bonded PLGA is prepared by a method comprising: double-end hydroxylated PLGA and single-end carbon-carbon double-bonded polyethylene glycol HO-PEG-CH = CH ₂ Reacting in the presence of a solvent, dimethylaminopyridine and EDC & HCl at a reaction temperature of 45-55 ℃ (such as 45 ℃, 46 ℃, 48 ℃,50 ℃, 52 ℃, 55 ℃ and the like) for 16-30 h (such as 18 h, 20 h, 24h, 25 h, 26 h, 30 h and the like), dialyzing in water, and drying; optionally, the number average molecular weight of the polyethylene glycol with single-end carbon-carbon double bond is selected from 1500 to 2500 Da.

In some embodiments, the molecular weight of the double-ended hydroxylated PLGA may be selected from 10 to 28 kDa, in number average molecular weight. Non-limiting examples are 10 kDa, 15 kDa, 20 kDa, 25 kDa, 28 kDa, etc., and can also be selected from the numerical range consisting of any two molecular weights mentioned above.

In some embodiments, the molecular weight of the polyethylene glycol with single terminal carbon-carbon double bond can be selected from 1500 to 2500 Da (i.e. 1.5 to 2.5 kDa) in terms of number average molecular weight. Non-limiting examples are 1.5 kDa, 2 kDa, 2.5 kDa, etc., and can also be selected from the numerical range consisting of any two molecular weights mentioned above.

Furthermore, the dosage of the single-ended carbon-carbon double-bonded polyethylene glycol is excessive, so that the ratio of the dosage of the single-ended carbon-carbon double-bonded polyethylene glycol to the dosage of the double-ended hydroxylated PLGA is larger than 2, and the double ends of the PLGA are ensured to be modified with carbon-carbon double bonds.

In some embodiments, the prepared double cross-linked network drug-loaded hydrogel is as defined in any one of the embodiments of the first aspect of the invention. In step S200, the drug molecule content in the double-crosslinked network drug-loaded hydrogel per unit mass is 0.0005 to 0.01 μ g/g, some non-limiting examples include 0.0005 μ g/g, 0.001 μ g/g, 0.0015 μ g/g, 0.002 μ g/g, 0.0025 μ g/g, 0.003 μ g/g, 0.0035 μ g/g, 0.004 μ g/g, 0.0045 μ g/g, 0.005 μ g/g, 0.0055 μ g/g, 0.006 μ g/g, 0.007 μ g/g, 0.008 μ g/g, 0.009 μ g/g, 0.01 μ g/g, and the like, and may be selected from a content interval formed by any two of the above values, and is dry weight basis. In some embodiments, the chemokine content per unit mass of the double cross-linked network drug-loaded hydrogel is 0.00078 μ g/g, dry weight basis.

In some embodiments, the carboxyl-terminated ketal thiol has the formula

Wherein two Z are the same and are both C _1-3 An alkylene group. In some embodiments, Z is methylene or 1, 2-ethylene.

In some embodiments, the carboxyl-terminated ketal thiol is prepared by a method comprising: reacting acetone and mercaptocarboxylic acid molecules SH-Z-COOH at the temperature of 55-65 ℃ for 48-96h, wherein the molar ratio of the acetone to the mercaptocarboxylic acid molecules is 2:1.

in some embodiments, both the carbon-carbon double-bonded hyaluronic acid HA1 and the carbon-carbon double-bonded hyaluronic acid HA2 are prepared by a method comprising the following steps: reacting hyaluronic acid and methacrylic anhydride under the conditions of pH 8-9 and ice bath for 18-30 hours, precipitating, collecting a solid phase, dialyzing in water, and freeze-drying.

In some embodiments, when the first cross-linked network is constructed in the manner shown in the first manner (S300 a), the ROS-responsive group is ketansercaptan, the degradable synthetic polymer unit Polyd0 is PLGA, and both the degradable natural polymer unit Polyd1 and the degradable natural polymer unit Polyd2 are hyaluronic acid, for example, the method for preparing the double cross-linked network drug-loaded hydrogel can refer to fig. 1. In the figure 1, (A) comprises the steps of preparing ROS response nanogel by using carbon-carbon double-bonded hyaluronic acid (HA 2) and double-end double-bonded ketalized thiol as raw materials, and further swelling a drug-loaded molecule to obtain a drug-loaded ROS response hydrogel; (B) The method comprises the steps of crosslinking carbon-carbon double-bonded hyaluronic acid (HA 1) and double-carbon double-bonded PLGA to prepare PLGA/HA1 composite hydrogel, and then swelling the composite hydrogel in a liquid phase containing the drug-loaded ROS response hydrogel to prepare the double-crosslinked network drug-loaded hydrogel.

In some embodiments, when the first cross-linked network is constructed in the manner shown in the second manner (S300 b), the ROS-responsive group is ketansercaptan, the degradable synthetic polymer unit poly 0 is PLGA, and both the degradable natural polymer unit poly 1 and the degradable natural polymer unit poly 2 are hyaluronic acid, for example, and the method for preparing the double cross-linked network drug-loaded hydrogel can refer to fig. 2. In fig. 2, (a) comprises preparing ROS-responsive nanogel from carbon-carbon double-bonded hyaluronic acid (HA 2) and double-bonded ketothiol at both ends, and further swelling drug-loaded molecules to obtain drug-loaded ROS-responsive hydrogel; (B) The double-crosslinked network drug-loaded hydrogel is prepared by crosslinking carbon-carbon double-bonded hyaluronic acid HA1, double-carbon double-bonded PLGA and drug-loaded ROS response hydrogel.

further, when medical hydrogel membrane includes location structure, location structure integral type set up in two cross-linked network medicine carrying hydrogel membrane side.

In some embodiments, the drug-loaded hydrogel-based membrane has two opposing surfaces.

In some embodiments, the drug-loaded hydrogel base film has two opposite trapezoidal surfaces, and further, the upper dimension of the trapezoid is selected from 0.5cm to 1cm, the lower dimension of the trapezoid is selected from 1.5cm to 2cm, and the height of the trapezoid is selected from 2.0cm to 2.5cm.

In some embodiments, the drug-loaded hydrogel-based membrane has a thickness selected from the group consisting of 300 μm to 600 μm. The thickness of the drug-loaded hydrogel base film can also be selected from any one or two of the following intervals: 300 μm, 350 μm, 400 μm, 450 μm, 500 μm, 550 μm, 600 μm, and the like.

In some embodiments, the drug-loaded hydrogel base film has two opposing trapezoidal surfaces with the positioning structure disposed on the top or bottom side of the two opposing trapezoidal surfaces.

In some embodiments, the medical hydrogel film has the structure shown in fig. 3. At this time, the medical hydrogel film 100 includes a medical hydrogel base film 130 and a positioning structure 140. At this time, the positioning structure 140 is disposed on the upper bottom side (narrower side) of the trapezoidal surface of the medical hydrogel base film 130.

In a fourth aspect of the invention, an anti-adhesion medical device is provided, which comprises at least one of the double-crosslinked network drug-loaded hydrogel of the first aspect of the invention, the double-crosslinked network drug-loaded hydrogel prepared by the preparation method of the second aspect of the invention, and the medical hydrogel film of the third aspect of the invention; further, the anti-adhesion medical device at least comprises a uterine cavity anti-adhesion device.

The double-crosslinked-network drug-loaded hydrogel and the medical hydrogel membrane containing the double-crosslinked-network drug-loaded hydrogel provided by the invention can be used as an anti-adhesion medical device or used as a raw material for further preparing the anti-adhesion medical device, the anti-adhesion medical device comprises but is not limited to a uterine cavity anti-adhesion device, the anti-adhesion effect of a hydrogel material can be exerted, the drug-loaded ROS response nanogel loaded in the first crosslinked network can be released from the first crosslinked network under the action of a single mechanism or a double mechanism of diffusion and/or degradation of the first crosslinked network, furthermore, the breakage of ROS response groups can be realized by utilizing the ROS environment of a postoperative inflammation part, so that drug molecules (such as chemokines) can be released controllably and accurately under the action of multiple mechanisms of ROS response, diffusion and degradation of the second crosslinked network, the higher the ROS concentration is, the stronger the responsiveness is, the larger the release amount of the drug molecules is, the repair of the inflammation part can be effectively promoted, and the adhesion recurrence can be effectively inhibited.

In some embodiments, the uterine cavity adhesion prevention device is formed by winding or folding any one of the double-crosslinked network drug-loaded hydrogel of the first aspect of the invention, the double-crosslinked network drug-loaded hydrogel prepared by the preparation method of the second aspect of the invention, and the medical hydrogel film of the third aspect of the invention.

In some embodiments, the uterine cavity adhesion prevention device is in a cylindrical structure or a multilayer structure.

In a fifth aspect of the present invention, the application of the double-crosslinked network drug-loaded hydrogel of the first aspect of the present invention, the double-crosslinked network drug-loaded hydrogel prepared by the preparation method of the second aspect of the present invention, or the medical hydrogel film of the third aspect of the present invention as an anti-adhesion medical device or in the preparation of an anti-adhesion medical device is provided, further, the anti-adhesion medical device at least comprises a uterine cavity anti-adhesion device.

Prevent the palace chamber adhesion as the example, among the antiseized even equipment of palace chamber that this application provided, the initial condition of hydrogel membrane can be for folding/coiled structure, press and hold in conveying system, through palace chamber propelling movement to internal, then absorb water in internal expansion, damaged endometrium is kept apart to physics, plays antiseized effect, can not bring uncomfortable foreign body sensation for patient, experience feels and is showing superior to implant materials such as intrauterine device and sacculus support. After the intrauterine adhesion device is placed into the uterine cavity, the intrauterine adhesion device can be rapidly released in an inflammatory environment through the responsive breakage of ROS responsive radicals so as to recruit stem cells and promote endometrial repair.

In some embodiments, the uterine cavity adhesion prevention device can be unfolded into a planar sheet-like form in a use state in vivo.

Some specific examples are provided below.

Embodiments of the present invention will be described in detail with reference to examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures for the conditions not specified in the following examples, preferably with reference to the guidelines given in the present invention, may also be performed according to the experimental manual or the conventional conditions in the art, may also be performed according to the conditions suggested by the manufacturer, or may be performed according to the experimental procedures known in the art.

In the following specific examples, the measurement parameters relating to the components of the raw materials, if not specified otherwise, may be subject to slight deviations within the accuracy of the weighing. Temperature and time parameters are involved to allow for acceptable deviation of the instrument test accuracy or operational accuracy.

In the following examples, the molecular weight-index average molecular weight is referred to unless otherwise specified.

In the following examples, the room temperature is 20 to 30 ℃ unless otherwise specified.

In the following examples, the following test instruments and parameters were used unless otherwise specified.

GPC instrumentation and parameters: agilent 1200 high performance liquid chromatograph (Agilent, usa), GPC gel chromatography workstation (tokyo longzhida). Oil phase chromatographic column: TSK gel G3000HHR column (5 μm,7.8 mm. Times.300 mm); mobile phase: tetrahydrofuran (HPLC grade); column temperature: 40 ℃; water phase chromatographic column: TSKgel G3000SWXL type (300 mm. Times.7.8 mm) high performance gel exclusion chromatography column with 0.05 mol/L phosphate buffer solution (pH 6.8) as mobile phase, flow rate: 1.0 mL/min; sample introduction amount: 100. mu L; a differential refractive detector.

¹ H NMR nuclear magnetic instrument and parameters: the NMR spectrometer was of the JEOL-FX60Q type, ¹ the H NMR operating frequency was 59.75MHz, with TMS (tetramethylsilane) as internal standard. Solvents for testing the double-ended carbon-carbon double-bonded ketone thiol and the methacrylated hyaluronic acid are respectively deuterated chloroform and deuterated water.

Example 1 stepwise crosslinking-swelling entrapment where the ROS-responsive group is a ketoconstricting thiol group

1.1. Synthesis of double-ended carbon double-bonded ketals:

(1) Distilled water, acetone and thioglycolic acid were mixed in a volume ratio of 9.

(2) Removing the solvent by rotary evaporation to obtain a solid powdery sample, washing with distilled water, centrifuging, removing the supernatant, repeating for 3-5 times, and vacuum drying to remove residual water to obtain double-end carboxylated ketone mercaptan;

(3) 2g of the double-terminal carboxylated ketal thiol was dissolved in methylene chloride, 0.03g of Dicyclohexylcarbodiimide (DCC) and 0.05g of Dimethylaminopyridine (DMAP) were added in this order, and after stirring at room temperature for 30 minutes, 5g of 3-buten-1-ol was added. And reacting for 12 hours at room temperature. And (4) performing rotary evaporation on the solvent, and purifying by using a silica gel column to obtain the double-ended carbon-carbon double-bonded ketone thiol.

According to ¹ H NMR test results confirm that the double-ended carbon-carbon double-bonded ketal thiol is successfully synthesized according to a characteristic peak at a chemical shift of 5.0 ppm.

1.2. Synthesis of methacrylated hyaluronic acid (HAMA, also denoted as HA 2):

sodium hyaluronate (HA-Na) 1g (molecular weight 6 kDa) was dissolved in 100mL of deionized water, and 8 mL of methacrylic anhydride was added. After fully and uniformly mixing, adjusting the pH value to 8.0-9.0 by using a NaOH solution, and reacting for 24 hours in an ice-water bath environment. Precipitating the reacted solution with 0 deg.C anhydrous ethanol for 3 times, collecting the lower layer precipitate, dissolving in distilled water, dialyzing for 72 hr, and lyophilizing.

According to ¹ According to the H NMR test result, according to the characteristic peak at the chemical shift of 5.5 ppm, the methacrylic acid hyaluronic acid (HAMA) is successfully synthesized, and the carbon-carbon double bond grafting rate of the HAMA is 100% -150%. The double bond grafting rate is the grafting rate of double bonds on the HA sugar unit, and the ratio of the methyl hydrogen peak area positioned on the double bond grafting unit to the hydrogen area of the main chain of the HA sugar unit is calculated according to an NMR nuclear magnetic spectrum, so that the double bond grafting rate is obtained. NMR spectrum shows that a plurality of carbon-carbon double bonds are grafted in one molecule of HAMA.

1.3. Synthesis of drug-loaded ROS-responsive nanogels (ROS-responsive nanogels containing SDF-1 α):

(1) 50mg of HAMA,28mg (0.0833 mmol) of a double-ended carbon-carbon double-bonded ketalthiol, 60mg of sodium bicarbonate were added to a three-necked flask, and 20mL of deionized water and 3mL of isopropanol were added. After stirring and reacting for 30min at 70 ℃, 20mg of potassium persulfate is added, and the reaction is carried out for 4h at 70 ℃ under the protection of nitrogen. The precipitate was washed three times with distilled water. Drying at 50 ℃ to obtain the nanogel containing the ketal and thiol groups.

The nanogel size was approximately 535 nm as measured by dynamic light scattering.

(2) And swelling the nanogel containing the ketotropic thiol group in distilled water, adding SDF-1 alpha with the final concentration of 0.01 mu g/mL, and stirring at room temperature for 12h to obtain the ROS response nanogel containing the SDF-1 alpha.

The drug loading can be measured by an ELISA enzyme-linked reaction kit: the drug-loaded ROS-responsive nanogel had a drug molecule (chemokine) content of 0.0053 μ g/g, dry weight basis, per unit mass.

1.4. Double-carbon double-bonded poly (lactic-co-glycolic acid) (double-carbon double-bonded PLGA, CH = CH) ₂ -PLGA-CH=CH ₂ ) The preparation of (1):

(1) PLGA was synthesized using conventional methods. Respectively weighing 1.07g of L-lactide and 0.875g of glycolide, and adding a catalyst SnCl ₂ The melt polycondensation reaction was initiated with propylene glycol (0.01 g) at 165 ℃ for 10 hours. After the reaction, the reactant was dissolved in toluene, centrifuged at 5000rpm, the lower precipitate was taken out, and the solution and centrifugation were repeated three times, and the lower precipitate was vacuum-dried to obtain white powder, and finally 1.8g of double-ended hydroxylated PLGA was obtained.

The number average molecular weight was 15 kDa by GPC.

(2) 1g of PLGA (15 kDa in number average molecular weight, hydroxylated at both ends) and 10.4g of PEG with a carboxyl group at one end and a carbon-carbon double bond at the other end (HOOC-PEG-CH = CH) ₂ ) (molecular weight 2000 Da) was dissolved in 100mL of dimethyl sulfoxide (DMSO). 1.1g of 4-Dimethylaminopyridine (DMAP) and 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl) are subsequently added. Reacting at 50 ℃ and stirring for 24 hours, adding the reaction solution into a dialysis bag with the molecular weight cutoff of 10000Da, dialyzing with deionized water for 4 to 5 days, and then performing rotary evaporation and drying to obtain double-ended carbon-carbon double-bonded PLGA, which is recorded as CH = CH ₂ -PLGA-CH=CH ₂ 。

GPC showed that the molecular weight was about 19 kDa, and it was found that double-ended carbon-carbon double-bonded PLGA was successfully synthesized.

1.5. Preparation of ROS-loaded PLGA/HA composite hydrogel responding to nanogel

(1) 300mg of CH = CH ₂ -PLGA-CH=CH ₂ Dissolving the mixed solution and 60mg of MAHA in 2mL of PBS (pH = 7.4) solution respectively, adding 50mg of potassium persulfate (APS) and 5 muL of Tetramethylethylenediamine (TEMED), introducing nitrogen into the mixed solution to remove oxygen, pouring the mixed solution into a mold, placing the mold into a 70 ℃ oven, standing for 30min to form gel, and obtaining carbon-carbon double bond cross-linkingA linked PLGA/HA composite hydrogel. And taking out the gel, and then putting the gel into a PBS buffer solution for soaking and rinsing for 3 times to remove unreacted monomers and impurities.

(2) Freeze-drying the PLGA/HA composite hydrogel prepared in the first step, and then soaking the PLGA/HA composite hydrogel in a mixed solution of 2mL of ROS-responsive nanogel carrying medicine (ROS-responsive nanogel containing SDF-1 alpha) and water (the concentration of the ROS-responsive nanogel carrying SDF-1 alpha is about 5 mg/mL) at room temperature for swelling for 5h to obtain the PLGA/HA hydrogel carrying the ROS-responsive nanogel carrying medicine, namely the double-crosslinked network medicine-carrying hydrogel in the application.

The drug loading can be measured by an ELISA enzyme-linked reaction kit: the content of drug molecules (chemotactic factors) in the double-crosslinked network drug-loaded hydrogel per unit mass is 0.00078 mu g/g, which is calculated by dry weight.

1.6. Preparation of roll film

The hydrogel is cut by scissors or a blade to obtain the drug-loaded hydrogel film, namely the medical hydrogel film.

Some samples are shaped as shown in fig. 3, the upper and lower surfaces are both trapezoids with an upper bottom of 1cm, a lower bottom of 2cm and a height of 2.5cm, and a positioning structure is reserved on the upper and lower sides (i.e. the narrower side of the trapezoids).

The material is coiled into a cylinder shape by a tool and is placed in a conveying system.

Example 2 stepwise crosslinking-swelling entrapment method with ROS-responsive groups as diselenyl groups

The double-ended carboxylated ketothiole was changed to a double-ended carboxylated diselenide compound in example 1, where the ROS-responsive group was changed from a ketothiole group to a diselenide bond, and the remaining steps were identical to example 1.

The synthesis of the double-terminal carboxylated diselenide compound is shown below. 1g of selenocysteine hydrochloride and 3.2g of triethylamine are weighed out and dissolved in 60mL of dichloromethane, a 100mL three-necked flask is added, nitrogen is introduced, and magnetic stirring is carried out for about 15min under an ice water bath. 1.5g of methacryloyl chloride was slowly added dropwise over a half hour period to a three-necked flask. After reacting for 2 hours in an ice-water bath, heating to room temperature and reacting for 24 hours in a nitrogen atmosphere. The product is washed by deionized water and saturated brine alternately for three times, is dried by adding anhydrous sodium sulfate for more than 2h, is rotated at 40 ℃ and is distilled under reduced pressure to remove the organic solvent, and then is recrystallized in a mixed solvent of ethyl acetate and n-hexane with the volume ratio of 1. The double-terminal carboxylated diselenide compound was further converted into a double-terminal double-bonded diselenide compound by the same operation as in example 1.

Example 3 one-pot Cross-linking drug coating, ROS-responsive group is a ketoconstricting thiol group

The ROS-responsive nanogel that was finally swollen into the first crosslinked network hydrogel in example 1 was changed to be crosslinked after being blended with the reaction components of the first crosslinked network, and the remaining steps were identical to example 1.

3.1. Synthesis of a double-ended ketomercaptol the same as in example 1;

3.2. synthesis of methacrylated hyaluronic acid (HAMA) as in example 1;

3.3. synthesis of ROS-responsive nanogels containing SDF-1 α as in example 1;

3.4. double-carbon double-bonded poly (lactic acid-glycolic acid) (CH = CH) ₂ -PLGA-CH=CH ₂ ) Was prepared as in example 1;

3.5. preparing the PLGA/HA composite hydrogel loaded with the ROS response nanogel:

300mg of CH = CH ₂ -PLGA-CH=CH ₂ And uniformly dispersing 60mg of MAHA and 10mg of ROS in 2mL of PBS (pH = 7.4) solution, then adding 50mg of potassium persulfate (APS) and 5 muL of Tetramethylethylenediamine (TEMED), introducing nitrogen into the mixed solution to remove oxygen, pouring the mixed solution into a mold, placing the mold into an oven at 70 ℃ and standing for 30min to form gel, and thus obtaining the carbon-carbon double bond crosslinked PLGA/HA composite hydrogel. And taking out the composite gel, putting the composite gel into PBS buffer solution for soaking and rinsing for 3 times, and removing unreacted monomers and impurities. The PLGA/HA hydrogel loaded with the drug-loaded ROS response nanogel, namely the double-crosslinked network drug-loaded hydrogel in the application, is obtained.

3.6. The procedure for the roll film preparation was the same as in example 1.

Example 4 one-pot Cross-linking drug coating method, ROS-responsive group is diseleno

A double-crosslinked network drug-loaded hydrogel was prepared in substantially the same manner as in example 3, except that the double-terminal carboxylated ketothiol was changed to a double-terminal carboxylated diselenide compound in example 3, and the ROS-responsive group was changed from a ketothiol group to a diselenide bond. Wherein the synthesis and modification of the diselenide compound are in accordance with example 2.

Comparative example 1.

Substantially the same procedure as in example 1 was employed except that the chain length of the double-ended carbon-double-bonded PLGA was changed.

Synthesis of double-carbon-double-bonded ketothiolates, methacrylated hyaluronic acid (HAMA), ROS-responsive nanogels containing SDF-1. Alpha. The same method as 1.1 to 1.3 in example 1 was used.

Poly (lactic-co-glycolic acid) (double-ended carbon-carbon double-bonded PLGA, CH = CH) was prepared in substantially the same manner as in 1.4 of example 1 ₂ -PLGA-CH=CH ₂ ) (ii) a Except that the dosage of the propylene glycol is as follows: initiation was carried out by adding twice the equivalent of propylene glycol as in example 1.

The number average molecular weight of the double-end carbon double-bonded PLGA is 7500 Da through GPC test.

The double cross-linked network drug-loaded hydrogel and the rolled film were prepared in the same manner as 1.5 and 1.6 in example 1.

Comparative example 2.

Synthesis of double-ended carbon-carbon double-bonded ketal thiol, methacrylated hyaluronic acid (HAMA), and SDF-1. Alpha. Containing ROS-responsive nanogels was carried out in the same manner as in 1.1 to 1.3 of example 1.

Poly (lactic-co-glycolic acid) (double-ended carbon-carbon double-bonded PLGA, CH = CH) was prepared in substantially the same manner as in 1.4 of example 1 ₂ -PLGA-CH=CH ₂ ) (ii) a Except that the dosage of the propylene glycol is as follows: initiation was carried out by adding 0.5 times the equivalent of propylene glycol relative to example 1.

The number average molecular weight of the double-ended carbon-carbon double-bonded PLGA is 27000 Da through GPC test.

Comparative example 3.

Essentially the same as in example 2, except that the chain length of the double-ended carbon-carbon double-bonded PLGA was changed.

Synthesis of a double-terminal-bonded diselenide compound, methacrylated hyaluronic acid (HAMA), and an SDF-1. Alpha. Containing ROS-responsive nanogel were carried out in the same manner as in example 2, respectively.

Poly (lactic-co-glycolic acid) (double-ended carbon-carbon double-bonded PLGA, CH = CH) was prepared in substantially the same manner as in example 2 ₂ -PLGA-CH=CH ₂ ) (ii) a Except that the dosage of the propylene glycol is as follows: initiation was carried out by adding twice the equivalent of propylene glycol as in example 2.

Double cross-linked network drug loaded hydrogels and wound films were prepared in the same manner as 1.5 and 1.6 in example 1.

Comparative example 4.

Poly (lactic-co-glycolic acid) (double-ended carbon-carbon double-bonded PLGA, CH = CH) was prepared in substantially the same manner as in example 2 ₂ -PLGA-CH=CH ₂ ) (ii) a Except that the dosage of the propylene glycol is as follows: initiation was carried out by adding 0.5 times the equivalent of propylene glycol relative to example 2.

The number average molecular weight of the double-terminal carbon double-bonded PLGA was 28000 Da by GPC.

Comparative example 5 a blank hydrogel of a nested cross-linked network was prepared and then drug loading was performed by swelling (i.e. a blank hydrogel of a nested cross-linked network was formed by first loading a second cross-linked network, which did not contain a drug, on the first cross-linked network, followed by swelling of the drug loading).

Synthesis of double-ended carbon-double-bonded ketalized thiol and methacrylated hyaluronic acid (HAMA) was carried out in the same manner as in example 1.

The ROS-responsive nanogels were prepared without drug using the method of example 1.3: 50mg of HAMA,28mg (0.0833 mmol) of a two-terminal-carbon double-bonded ketalized thiol, 60mg of sodium bicarbonate were placed in a three-necked flask, and 20mL of deionized water and 3mL of isopropanol were added. After stirring and reacting for 30min at 70 ℃, 20mg of potassium persulfate is added, and the reaction is carried out for 4h at 70 ℃ under the protection of nitrogen. The precipitate was washed three times with distilled water. Drying at 50 ℃ to obtain blank nanogel containing the ketal thiol groups (blank refers to non-encapsulated drug molecules) which is recorded as ROS response blank nanogel.

Preparation of double-ended carbon-carbon double-bonded poly (lactic acid-glycolic acid), preparation of PLGA/HA composite hydrogel loaded with ROS-responsive blank nanogel (denoted as "blank double-crosslinked-network hydrogel"), and preparation of the same method as in example 1 were used.

Swelling blank double-crosslinked network hydrogel (PLGA/HA composite hydrogel loaded with ROS response blank nanogel) in distilled water, adding SDF-1 alpha with the final concentration of 0.01 mu g/mL, stirring at room temperature for reaction for 12h, and obtaining the composite gel loaded with the SDF-1 alpha.

After cleaning and balancing, the drug loading is tested by an ELISA enzyme-linked reaction kit, the drug loading data can not be tested, and the surface drug content is extremely low and is below the detection limit.

A roll film was prepared in the same manner as 1.6 in example 1.

Comparative example 6 microgel coated System with Single crosslinked network (with first crosslinked network only, using PLGA/HA composite hydrogel coating)

Synthesis of methacrylated hyaluronic acid (HAMA) and preparation of poly (lactic acid-glycolic acid) having double carbon-carbon double bonds Using the same method as in example 1.

Gel synthesis of single cross-linked network: 300mgCH = CH ₂ -PLGA-CH=CH ₂ And uniformly dispersing 60mg of MAHA into 2mL of PBS (pH = 7.4) solution, then adding 50mg of potassium persulfate (APS) and 5 muL of Tetramethylethylenediamine (TEMED), introducing nitrogen into the mixed solution to remove oxygen, pouring the mixed solution into a mold, placing the mold into a 70 ℃ oven, and standing for 30min to form gel, thereby obtaining the carbon-carbon double bond crosslinked PLGA/HA composite hydrogel. And taking out the composite gel, putting the composite gel into PBS buffer solution for soaking and rinsing for 3 times, and removing unreacted monomers and impurities.

And (3) replacing and swelling the PLGA/HA composite gel in PBS containing SDF-1 alpha with the final concentration of 0.01 mu g/mL, and stirring at room temperature for reaction for 24 hours to obtain the composite gel loaded with the SDF-1 alpha.

Comparative example 7 nanogel-coated System with Single crosslinked network (with second crosslinked network only, coated with HA hydrogel)

Synthesis of methacrylated hyaluronic acid (HAMA) the same procedure as in example 1 was followed.

(1) ROS-responsive nanogels containing SDF-1 α were synthesized as in example 1, with the same particle size.

(2) The method for responding to nanogel loaded SDF-1 alpha by ROS is the same as that in example 1, and the finally detected drug loading rate is 0.0053 mu g/g.

Test analysis method

1. Test method of nanogel size: the nanogel size was measured using a Zetasizer Nano S laser particle sizer (Malvern, uk) and dispersed in PBS at 0.01% concentration, based on the clear solution.

Test samples: including ROS-responsive nanogels containing ROS-responsive radicals, drug-loaded ROS-responsive nanogels, and the like.

2. Property of drug loading

The drug loading in the gel (mass percent, dry weight) was measured as follows.

10mg of the drug-loaded rice gel was placed in 2mL of 10% H ₂ O ₂ The cells were soaked for 5 hours, and after filtration, the supernatant was collected and the concentration was measured according to the SDF-1. Alpha. ELISA (Enzyme Linked Immunosorbent Assay) protease-Linked reaction kit instructions.

A sample to be tested: nanogel (drug-loaded ROS response nanogel), and a drug-loaded hydrogel membrane (double-crosslinked network drug-loaded hydrogel, SDF-1 alpha-loaded composite gel, and the like).

3. Anti-adhesion effect

The water contact angle test instrument and the method comprise the following steps: the contact angle measuring instrument is a model JCW-360 analyzer from chender experimental facilities ltd. A drop of water (5. Mu.L) was dropped on the gel surface, and then the contact angle was measured by the dynamic water contact angle method. The test procedure was as follows: after dropping the water drop on the surface of the gel, the water drop was photographed for 1 min, and the dynamic change of the contact angle was observed. And taking 150 photos in the first 10 s at the photographing speed of 10 photos/s and in the second 50s at the photographing speed of 1 photo/s, then calculating the contact angle of a water drop on each photo in a simulation mode, and finally averaging the contact angle values of the 150 photos to obtain the water contact angle value of the sample.

4. Release behavior

Four parallel samples of 1g composite gel blocks were prepared and placed in 2mL of 1% H ₂ O ₂ The supernatants were filtered at 1h, 5h, 12h and 24h, respectively, and SDF-1 α concentration was measured according to the SDF-1 α ELISA (Enzyme Linked Immuno-Sorbent Assay) protease-Linked reaction kit instructions.

Test results and analysis

The test results can be found in table 1.

In comparative examples 1 to 7, the drug-loaded hydrogel films were significantly lower in drug-loaded amount than in the examples of the present invention (examples 1 to 5). The following reasons are presumed to be included.

In comparative examples 1 and 3, it is presumed that the relatively small molecular weight and short segment length of PLGA in the first crosslinked network result in relatively small mesh size of the first crosslinked network, and thus may result in relatively small amount of nanogel that can be accommodated in the first crosslinked network.

In comparative examples 2 and 4, it is presumed that the relatively large molecular weight and long chain length of PLGA in the first crosslinked network results in a relatively large mesh size of the first crosslinked network, and thus, it may result in that the nanogel is not easily stably bound in the mesh of the first crosslinked network.

The hydrogel film of comparative example 5 was almost not loaded with drugs, presumably because nanogels were wrapped in the microgrids in the first cross-linked network, making it difficult to diffuse the drugs into nanogels with smaller lattices by means of swelling, and also resulting in failure to improve drug release behavior by the response property of ROS groups in nanogels. In the examples of the present application, the drug is first entrapped in the lattice of the nanogel, and the drug molecules can be relatively stably entrapped in the lattice of the nanogel either by being nested in the first cross-linked network by swelling (examples 1,2 and 5) or by being entrapped in the first cross-linked network in situ with the cross-linking reaction of the first cross-linked network (examples 3 and 4).

Comparative example 6 a microgel system with a single cross-linked network was used with very low drug loading.

Comparative example 7 a nanogel system using a single cross-linked network, although having a certain drug loading, has a severe burst release phenomenon and is not suitable for clinical use.

In addition, in comparative examples 1 to 4 and 6, the release period of the drug is short, the release amount exceeds 50% when the drug is released for 1 hour, the release amount reaches more than 70% when the release time is 5 hours, the release period is short, and no sustained release effect is achieved.

The water contact angle is a test means for representing the hydrophilicity and the hydrophobicity of the material, and for the anti-adhesion material, the more hydrophilic the material, the more adhesion-preventing effect is achieved. A smaller water contact angle indicates a more hydrophilic material. Referring to table 1, it can be seen that the contact angles of the examples 1 to 4 and the comparative examples 1 to 7 are all less than 90 degrees, which indicates that the material is relatively hydrophilic, wherein the water contact angles of the comparative examples 2 and 4 are 67 degrees and 72 degrees respectively, and the material is relatively more hydrophobic compared with other groups, possibly due to the fact that polyester chain segments used in the two groups of materials are relatively long and the hydrophobicity is relatively high.

TABLE 1 test results

In Table 1, "drug loading in the nanogel" indicates the relative mass content of the drug in the nanogel, which is measured in units of μ g/g in terms of dry weight, that is, micrograms of drug contained in each gram of nanogel. The drug loading capacity in the drug-loaded hydrogel film refers to the relative mass content of the drug in the drug-loaded hydrogel film, and the unit is mu g/g by dry weight, namely microgram of the drug contained in each gram of the drug-loaded hydrogel film. The "cumulative release" at various time points represents the micrograms of drug released cumulatively at a certain time point per gram of drug-loaded hydrogel membrane.

The technical features of the above embodiments and examples can be combined in any suitable manner, and for the sake of brevity, all possible combinations of the technical features of the above embodiments and examples are not described, however, as long as there is no contradiction between the combinations of the technical features, the combinations of the technical features should be considered to be within the scope of the description in the present specification.

The above examples only show some embodiments of the present invention, so as to facilitate the detailed and detailed understanding of the technical solutions of the present invention, but not to be construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Furthermore, it should be understood that various changes or modifications can be made by those skilled in the art after reading the above teachings of the present invention, and equivalents obtained thereby also fall within the scope of the present invention. It should also be understood that the technical solutions provided by the present invention, which are obtained by logical analysis, reasoning or limited experiments, are within the scope of the present invention as set forth in the appended claims. Therefore, the protection scope of the present invention should be subject to the content of the appended claims, and the description and the drawings can be used for explaining the content of the claims.

Claims

1. A double-crosslinked network drug-loaded hydrogel, which is characterized by comprising: a first cross-linked network and a drug-loaded ROS-responsive nanogel loaded in the first cross-linked network;

wherein the content of the first and second substances,

the framework material of the first cross-linked network comprises degradable synthetic polymer units Polyd0 and degradable natural polymer units Polyd1; in the first crosslinked network, the degradable synthetic polymer unit Polyd0 and the degradable natural polymer unit Polyd1 are each independently covalently linked to an adjacent structural unit; in the first cross-linked network, the degradable synthetic polymer unit Polyd0 is selected from one or more of polylactic acid unit, lactic acid-glycolic acid copolymer unit and polycaprolactone unit; in the first cross-linked network, the degradable natural polymer units Polyd1 are selected from one or more of hyaluronic acid units, collagen units and chitosan units;

the drug-loaded ROS-responsive nanogel comprises a second cross-linked network and a drug molecule non-covalently entrapped in the second cross-linked network; the skeleton material of the second cross-linked network is degradable natural polymer unit Polyd2, and the skeleton of the second cross-linked network contains ROS response groups; in the second cross-linked network, the degradable natural polymer units Polyd2 are selected from one or more of hyaluronic acid units, collagen units and chitosan units;

the drug molecule includes a chemokine.

2. The double cross-linked network drug-loaded hydrogel of claim 1, wherein one or more of the following are provided:

(ii) covalent or non-covalent linkages between the second crosslinked network and the first crosslinked network;

the mass ratio of the second cross-linked network to the first cross-linked network is (0.1 to 2) to (2 to 10);

the weight percentage of the drug-loaded ROS response nanogel in the double-crosslinking-network drug-loaded hydrogel is 0.5-30 percent based on dry weight;

the drug molecule further comprises an estrogen; and

the relative mass content of the drug molecules in the double-crosslinking-network drug-loaded hydrogel is 0.0005 to 0.01 mu g/g by dry weight.

3. The double-crosslinked network drug-loaded hydrogel of claim 1 or 2, characterized by one or more of the following group:

in the second crosslinked network, the ROS-responsive groups independently comprise one or more of the following groups: ketothioketal groups, thioether bonds, mono-selenium bonds, di-selenium bonds, divalent tellurium, oxalate groups, thiazolinone groups, borate groups, boronic acid groups and proline oligomeric chains, wherein the number of proline units in the proline oligomeric chains is selected from 3 to 7; and

and the amount of the ROS-responding group-containing substance in each gram of the second crosslinking network is 0.0005 to 0.05 mol.

4. The double-crosslinked network drug-loaded hydrogel of claim 1 or 2, characterized by one or more of the following group:

in the first cross-linked network, the number average molecular weight of the degradable synthetic polymer units Polyd0 is independently selected from 10 to 30 kDa, and the number average molecular weight of the degradable natural polymer units Polyd1 is independently selected from 4 to 8 kDa;

in the first crosslinked network, the covalent bond connection between the degradable synthetic polymer unit poly 0 and the degradable natural polymer unit poly 1 is selected from one or more of the following: carbon-carbon single bonds, ester bonds, and amide bonds.

5. The preparation method of the double-crosslinked network drug-loaded hydrogel according to any one of claims 1 to 4, wherein raw materials comprising an ROS-responsive precursor molecule, a second crosslinkable natural polymer, a drug molecule, a crosslinkable synthetic polymer and a first crosslinkable natural polymer are adopted, and the preparation method of the double-crosslinked network drug-loaded hydrogel comprises the following steps:

swelling the ROS response nanogel in an aqueous solution containing the drug molecules, and loading the drug molecules in the second cross-linked network to prepare a drug-loaded ROS response nanogel; the average grain diameter of the drug-loaded ROS response nanogel is selected from 300 to 700 nm;

then, a first cross-linked network is constructed by adopting a method shown in the following mode I or mode II to prepare the medical hydrogel with the double cross-linked network:

the method I comprises the following steps: mixing the crosslinkable synthetic polymer with the first crosslinkable natural polymer, and carrying out crosslinking reaction to form the first crosslinking network to prepare the composite hydrogel; then swelling the composite hydrogel in a mixed solution containing the drug-loaded ROS response nanogel, and loading the drug-loaded ROS response nanogel in a first cross-linking network of the composite hydrogel to prepare the double cross-linking network drug-loaded hydrogel;

the second method comprises the following steps: mixing the crosslinkable synthetic polymer, the first crosslinkable natural polymer and the drug-loaded ROS response nanogel, and carrying out a crosslinking reaction to form a first crosslinking network so as to prepare the double-crosslinking-network drug-loaded hydrogel;

wherein the content of the first and second substances,

the degradable synthetic polymer unit Polyd0, the degradable natural polymer unit Polyd1, the degradable natural polymer unit Polyd2, the ROS-responsive group and the drug molecule are as defined in any one of claims 1 to 4.

6. The method of claim 5, wherein the ROS-responsive nanogel containing ROS-responsive groups is prepared by a method comprising: mixing ketone thiol with double terminal double bonds and hyaluronic acid HA2 with double carbon-carbon bonds in a water/isopropanol mixed solvent, stirring and mixing in the presence of sodium bicarbonate, adding potassium persulfate, and carrying out a crosslinking reaction in a nitrogen atmosphere at a reaction temperature of 65-75 ℃ for 3-5 h to obtain HA nanogel containing ketone thiol groups; wherein the amount of the substance of the double-ended double-bond ketal thiol added in each gram of the carbon-carbon double-bonded hyaluronic acid HA2 is 0.005 to 0.05 mol; the volume ratio of water to isopropanol in the water/isopropanol mixed solvent is 1 (0.1 to 0.2);

the drug-loaded ROS response nanogel is prepared by adopting a method comprising the following steps: swelling the HA nano gel containing the ketothiol group in an aqueous reagent, adding the drug factor with the final concentration of 0.005-0.1 mu g/mL, and stirring at 20-30 ℃ for 8-15 h to prepare the drug-loaded ROS response nano gel;

the method in the first mode comprises the following steps: mixing double-ended carbon-carbon double-bonded PLGA and carbon-carbon double-bonded hyaluronic acid HA1 with a PBS buffer solution with the pH of 7.2-7.6, adding potassium persulfate and tetramethylethylenediamine, and carrying out a crosslinking reaction under the protection of oxygen and nitrogen at the reaction temperature of 65-75 ℃ for 20-40 min to prepare PLGA/HA1 composite hydrogel; soaking and rinsing the PLGA/HA1 composite hydrogel in a PBS buffer solution; adding the rinsed PLGA/HA1 composite hydrogel into a mixed solution containing 2-10 mg/mL of the drug-loaded ROS response nanogel for swelling for 4-8 h to prepare the double-crosslinked network drug-loaded hydrogel; wherein the weight ratio of the double-ended carbon-carbon double-bonded PLGA to the carbon-carbon double-bonded hyaluronic acid HA1 is (2 to 7): 1;

the method shown in the second mode comprises the following steps: mixing double-end carbon-carbon double-bonded PLGA, carbon-carbon double-bonded hyaluronic acid HA1 and the drug-loaded ROS response nanogel with a PBS buffer solution with the pH of 7.2-7.6, adding potassium persulfate and tetramethylethylenediamine, and carrying out crosslinking reaction under the protection of oxygen removal and nitrogen at the reaction temperature of 65-75 ℃ for 20-40 min to prepare the double-crosslinked network drug-loaded hydrogel; wherein the weight ratio of the double-ended carbon-carbon double-bonded PLGA, the carbon-carbon double-bonded hyaluronic acid HA1 and the drug-loaded ROS response nanogel is (2 to 7): 1: (0.05 to 1).

7. The method according to claim 6,

the carbon-carbon double-bonded hyaluronic acid HA2 is prepared by adopting a method comprising the following steps: reacting hyaluronic acid and methacrylic anhydride under the conditions of pH 7.8 to 8.2 and ice bath for 18 to 30 hours, precipitating, collecting a solid phase, dialyzing in water, and freeze-drying; wherein 0.01 to 0.05mol of methacrylic anhydride is added into each gram of hyaluronic acid;

the carbon-carbon double-bonded hyaluronic acid HA1 is prepared by adopting a method comprising the following steps: reacting hyaluronic acid and methacrylic anhydride under the conditions of pH 7.8-8.2 and ice bath for 18-30 h, precipitating, collecting a solid phase, dialyzing in water, and freeze-drying; wherein, 0.01 to 0.05mol of methacrylic anhydride is added into each gram of hyaluronic acid;

the double-ended carbon-carbon double-bonded PLGA is prepared by a method comprising the following steps: double-end hydroxylated PLGA and single-end carbon-carbon double-bonded polyethylene glycol HO-PEG-CH = CH ₂ Reacting in the presence of a solvent, dimethylaminopyridine and EDC & HCl at the temperature of 45-55 ℃ for 16-30 h, dialyzing in water, and drying; wherein the number average molecular weight of the polyethylene glycol with the single-ended carbon-carbon double bond is selected from 1500 to 2500 Da.

8. The medical hydrogel film is characterized by comprising a drug-loaded hydrogel base film and a positioning structure or not;

wherein the drug-loaded hydrogel basement membrane comprises the double-crosslinked network drug-loaded hydrogel of any one of claims 1 to 4 or the double-crosslinked network drug-loaded hydrogel prepared by the preparation method of any one of claims 5 to 7;

9. An anti-adhesion medical device, which is characterized by comprising at least one of the double-crosslinked network drug-loaded hydrogel of any one of claims 1 to 4, the double-crosslinked network drug-loaded hydrogel prepared by the preparation method of any one of claims 5 to 7, and the medical hydrogel film of claim 8; the anti-adhesion medical equipment at least comprises a uterine cavity anti-adhesion equipment.

10. An adhesion-preventing medical device according to claim 9, wherein the uterine cavity adhesion-preventing medical device is a medical device for preventing uterine cavity adhesion and promoting endometrial repair.