Application of copper-based organic framework materials in the preparation of contraceptive preparations, contraceptive preparations and methods

Abstract

The present application provides an application of a copper-based organic skeleton material in the preparation of a contraceptive preparation, a contraceptive preparation and a method, and relates to the field of biomedicine technology. The copper-based organic skeleton material is a copper-based bioactive material that has a killing effect on the trophoblast cells of the embryo. The copper-based organic skeleton material acts on the trophoblast cells of the embryo to prevent the normal implantation of the embryo; the trophoblast cells are damaged by the released divalent copper ions, and the divalent copper ions are oxidized into monovalent copper ions on the surface of the trophoblast cells; the divalent copper ions and the monovalent copper ions are transported together into the trophoblast cells; in the trophoblast cells, the divalent copper ions and the monovalent copper ions induce a Fenton-like reaction, increase the level of intracellular reactive oxygen species, cause trophoblast cell damage, damage the mitochondria of the trophoblast cells, and then mediate the death of the trophoblast cells, thereby avoiding the clinical side effects and side effects of copper-containing contraceptive devices on the body, and at the same time avoiding interference with hormone homeostasis and endocrine balance in the body.

Description

Technical Field

[0001] The present application relates to the field of biomedical technology, and in particular to the application of copper-based organic framework materials in the preparation of contraceptive preparations, contraceptive preparations and methods. Background Art

[0002] Unexpected pregnancy is an important social public health issue, which has adverse effects on women's bodies, families and society. Abortion can cause great physical and psychological impacts on women. In addition, the medical expenses required for abortion are also a huge cost for families and society.

[0003] Among the effective contraceptive methods currently used for women, the copper intrauterine device (Cu-IUD) is still a widely used, low-cost, long-term contraceptive medical device in China and even around the world. The Cu-IUD is placed in the uterus to inactivate sperm by releasing copper ions and cause uterine inflammation to interfere with embryo implantation, thereby achieving a better contraceptive effect. However, the long-term placement of the Cu-IUD in the human uterus can cause various adverse reactions in women, including irregular bleeding, non-bacterial inflammatory reactions, irregular menstruation, irregular pain, etc.

[0004] Therefore, there is an urgent need for a contraceptive method that can avoid various adverse reactions caused by the above-mentioned copper intrauterine contraceptive device to women. Summary of the invention

[0005] The present application provides an application of a copper-based organic framework material in the preparation of a contraceptive preparation, a contraceptive preparation and a method to solve the above-mentioned problems existing in existing contraceptive methods.

[0006] In a first aspect of the present application, there is provided an application of a copper-based organic skeleton material in the preparation of a contraceptive preparation, wherein the copper-based organic skeleton material is a copper-based bioactive material that has a killing effect on the trophoblast cells of the embryo, and the copper-based organic skeleton material uses an organic matter as an organic ligand of the copper-based organic skeleton, and the organic matter is any one of vitamin B13, vitamin B3 or tannic acid; the copper-based organic skeleton material acts on the trophoblast cells of the embryo to prevent the normal implantation of the embryo; the copper-based organic skeleton material damages the trophoblast cells by releasing divalent copper ions, and the divalent copper ions are oxidized into monovalent copper ions on the surface of the trophoblast cells; the divalent copper ions and the monovalent copper ions are transported together into the trophoblast cells; in the trophoblast cells, the divalent copper ions and the monovalent copper ions induce a Fenton-like reaction, increase the level of intracellular reactive oxygen species, cause trophoblast cell damage, damage the mitochondria of the trophoblast cells, and then mediate the death of the trophoblast cells.

[0007] The copper-based organic skeleton material releases copper ions to kill the trophoblast cells of the embryo, preventing the trophoblast cells of the embryo from completing the invasion of the uterus, thereby preventing the normal implantation and growth and development of the embryo. The copper-based organic skeleton material releases copper ions to damage the mitochondrial organelles of the trophoblast cells and mediate the death of the trophoblast cells through oxidative stress.

[0008] Specifically, a copper-based organic framework material is used as a carrier material for copper ions, trophoblast HTR8 / SVneo cells are used as model cells, and a rat pregnancy model is used as an animal model.

[0009] In an optional embodiment of the present application, the copper-based organic skeleton material is a vitamin B13-copper organic skeleton material, which decomposes and releases divalent copper ions in the microenvironment, and the divalent copper ions are reduced to monovalent copper ions on the cell membrane surface. After the divalent copper ions and the monovalent copper ions are taken up by embryonic trophoblast cells, the divalent copper ions and the monovalent copper ions induce a Fenton-like reaction in the cells, resulting in an increase in the production of reactive oxygen species and an oxidative stress reaction in the cells; the oxidative stress reaction causes the depolarization of the mitochondrial membrane in the embryonic trophoblast cells, increased mitochondrial membrane permeability, mitochondrial damage, and mediates the death of the trophoblast cells. The above-mentioned microenvironment refers to the uterine microenvironment. The above-mentioned reactive oxygen species are H2O2, ·O 2- , OH, etc.

[0010] In an optional embodiment of the present application, the copper-based organic framework material is vitamin B13 Cu-MOF, and the copper-based organic framework material decomposes in a microenvironment to release vitamin B13 monomers. The vitamin B13 monomers have good biocompatibility and are used to reduce the side effects and toxicity of the copper-based organic framework material to the body.

[0011] In an optional embodiment of the present application, the concentration range of the contraceptive preparation when used is greater than 10 μg mL -1 , less than 100 μg mL -1 , can be selected as 10 μg mL -1 , 35 μg mL -1 , 65 μg mL -1 , 95 μg mL -1 , 100 μg mL -1 Those skilled in the art can make settings according to their needs, and this application does not limit them, and only actual use shall prevail.

[0012] In an optional embodiment of the present application, during the application process, the release rate of the copper-based organic framework material is controlled by pH response or temperature change, or the release rate of the copper-based organic framework material in the contraceptive preparation is controlled by controlling the degradation rate of the contraceptive preparation itself.

[0013] Among them, the mechanism of pH-responsive controlled release is: the microenvironment of the uterus is acidic, and the copper-based organic framework material can decompose and release copper ions under acidic conditions; according to the pH value, the decomposition rate of the copper-based organic framework material contraceptive preparation can be adjusted, thereby achieving controlled release of copper ions from the copper-based organic framework material.

[0014] The mechanism of controlled release due to temperature changes is: the microenvironment temperature of the uterus is a dynamically changing process and is regulated by the physiological cycle; according to the change in temperature, the decomposition rate of the copper-based organic skeleton contraceptive preparation can be adjusted, and the increase in temperature can increase the decomposition rate of the copper-based organic skeleton material, thereby achieving controlled release of copper ions from the copper-based organic skeleton material.

[0015] The mechanism of the degradation of contraceptive preparations themselves is as follows: uterine mucus is a hydrogel-like liquid containing various ions, which can promote the decomposition and release of copper ions of copper ion carriers in copper-based organic framework contraceptive preparations. The decomposition rate of contraceptive preparations can be controlled by changing the physical and chemical properties of contraceptive preparations, thereby controlling the release rate of copper ions.

[0016] In the second aspect of the present application, a contraceptive preparation is provided, the contraceptive preparation is composed of a copper-based organic framework material and a carrier, the copper-based organic framework material is synthesized from a copper salt material and an organic ligand, and the carrier is an organic high molecular polymer; the above-mentioned organic high molecular polymer can be any one of thermosensitive hydrogel, sodium alginate, chitosan, polylactic acid, agarose, liposome, polyethylene glycol, gelatin, acrylic acid or other organic high molecular materials that can form gel. In the present application, thermosensitive hydrogel is preferably used as the carrier of the copper-based organic framework material in the contraceptive preparation.

[0017] The copper salt material is any one of copper nitrate, copper chloride, copper sulfate, copper acetate and hydrates thereof; the organic ligand is any one of vitamin B13, vitamin B3 and tannic acid. The organic ligands as the raw materials for the synthesis of the copper-based organic framework material may include organic compounds such as nicotinic acid, orotic acid and tannic acid.

[0018] In an optional embodiment of the present application, the copper-based organic framework material damages the organelles of trophoblast cells and mediates the death of the trophoblast cells by releasing copper ions; and the carrier is used to maintain the slow release of copper ions in the copper-based organic framework material.

[0019] Since the embodiment of the present application can be administered after the formation of embryonic trophoblast cells, or before embryonic invasion, it acts on the embryonic trophoblast cells, preventing the embryonic trophoblast cells from completing the invasion of the uterus, restricting the growth of embryonic trophoblast cells, and achieving the purpose of contraception. Therefore, the copper-based organic skeleton material in the embodiment of the present application selects a copper-based organic skeleton material that has a killing effect on embryonic trophoblast cells; based on the above-mentioned pH response and temperature response controlled release or controlling the degradation rate of the contraceptive preparation itself, the mechanism of controlling the sustained release of copper ions in the copper-based organic skeleton material in the contraceptive preparation is controlled.

[0020] Optionally, the copper base in the copper-based organic framework material is derived from an inorganic metal material, and the inorganic metal material includes a single-component or multi-component metal-doped copper-based organic framework material, a single-component or multi-component metal compound-doped copper-based organic framework material;

[0021] The central ligand constituting the copper-based organic framework material is an organic matter that is friendly to the body and has good biocompatibility; the copper element constituting the copper-based organic framework material is a life element.

[0022] In an optional embodiment of the present application, the synthesis method of the copper-based organic framework material includes solvent thermal, liquid phase synthesis method, solid phase synthesis method, ultrasound-assisted method, high temperature-assisted method, surfactant method, and microwave oven-assisted method.

[0023] In an optional embodiment of the present application, the contraceptive preparation is administered before the embryo invades the uterus; the administration method of the contraceptive preparation includes a minimally invasive uterine cavity administration method on the back and a minimally invasive uterine cavity administration method on the abdomen; the administration method of the contraceptive preparation also includes an intrauterine cavity perfusion administration method. The contraceptive preparation is administered after the formation of the embryonic trophoblast and before invasion; the administration method of the contraceptive preparation is preferably an intrauterine cavity perfusion administration method.

[0024] After the fertilized egg is formed, the embryo's trophoblast cells are formed, and the embryo invades the uterus during the embryo's trophoblastic invasion. At this time, the embryo has not yet begun to invade the uterus. Therefore, administration after the embryo's trophoblastic cells are formed and before the embryo invades, can act on the embryo's trophoblastic cells, causing damage or even death to the embryo's trophoblastic cells, resulting in restricted embryo growth and achieving the purpose of contraception.

[0025] The above-mentioned contraceptive preparation can be administered to animals by minimally invasive dosing on the back, minimally invasive dosing on the abdomen, and intrauterine perfusion. Due to the special environment of the uterine cavity, in the present embodiment, intrauterine perfusion is preferred to directly administer the above-mentioned contraceptive preparation into the uterine cavity. In the present embodiment, in future clinical applications, intrauterine perfusion is also preferred to directly administer the above-mentioned contraceptive preparation into the uterine cavity.

[0026] In the third aspect of the present application, a non-therapeutic contraceptive method is provided, the method comprising: placing the contraceptive preparation described in any one of the second aspects in the uterine cavity based on intrauterine administration, so that the copper-based organic skeleton material kills the embryonic trophoblast cells based on the sustained release of copper ions, thereby preventing the normal implantation of embryonic cells; the copper-based organic skeleton material is a vitamin B13-copper-based organic skeleton material, which releases copper ions and vitamin B13 monomers at the same time.

[0027] This application includes the following advantages:

[0028] 1) The present application uses copper-based organic skeleton materials to prepare contraceptive preparations, which can avoid excessive reactive oxygen species from damaging the uterine cavity to a certain extent, avoid the side effects and adverse reactions of clinical copper-containing contraceptive devices on the body, and avoid interfering with the homeostasis of hormones and the balance of endocrine in the body;

[0029] 2) The present application provides an application of a copper-based organic framework material in the preparation of a contraceptive preparation, a contraceptive preparation and a contraceptive method, which transfers the contraceptive object from the human body (i.e., the mother) to the blastocyst in the cell state, thereby minimizing systemic or local damage to the human body;

[0030] 3) The copper element of the copper-based organic framework material used in the contraceptive preparation of the present application is an essential element for human life, and is also an effective contraceptive component of clinical copper-containing IUDs. In addition to playing a contraceptive role, it can play a vital role in the normal function of life, thereby reducing the toxicity of the copper-based organic framework material;

[0031] 4) Vitamin B13, the organic ligand of the copper-based organic framework material used in the contraceptive preparation of the present application, is an organic substance that is friendly to the human body and has good tissue compatibility. It plays a vital role in the normal function of life and can reduce the toxicity of the copper-based organic framework material;

[0032] 5) The contraceptive preparation proposed in the present application can be administered via the intrauterine route, thereby avoiding blood and organ metabolic circulation. The natural self-clearance function of the physiological cycle can also avoid the risk of copper-based organic framework materials accumulating in the uterine cavity or even in the body. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the description of the embodiments of the present application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0034] Figure 1 A schematic diagram of a contraceptive method provided in an embodiment of the present application in which a copper-based organic framework material acts on embryonic trophoblast cells;

[0035] Figure 2 This is a characterization result diagram of a copper-based organic framework material provided in an example of the present application;

[0036] Figure 3 This is a diagram showing the cell compatibility results of a copper-based organic framework material provided in an example of the present application;

[0037] Figure 4 This is a graph showing the copper ion release results of a copper-based organic framework material provided in an example of the present application;

[0038] Figure 5 A schematic diagram of a copper-based organic framework material provided in an embodiment of the present application inducing a Fenton-like reaction in embryonic trophoblast cells;

[0039] Figure 6 A schematic diagram of a copper-based organic framework material provided in an example of the present application mediating trophoblast cell death through oxidative stress;

[0040] Figure 7 This is a schematic diagram of the contraceptive effect of a copper-based organic framework material in an animal body provided in an embodiment of the present application;

[0041] Figure 8 A schematic diagram of the characterization and contraceptive effect of a thermosensitive hydrogel-loaded copper-based organic framework material provided in the examples of the present application;

[0042] Fig. 9 This is a schematic diagram of the safety of a thermosensitive hydrogel-loaded copper-based organic framework material in an animal body provided in an example of the present application. DETAILED DESCRIPTION

[0043] The following will be combined with the accompanying drawings provided in the embodiments of the present application to clearly and completely describe the technical solution provided in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0044] The current idea of ​​contraceptive measures is to hinder the production of egg cells or hinder the combination of egg cells and sperm. This idea is based on the fact that most of the contraceptive measures currently used in clinical practice (for example: oral hormonal contraceptives, skin patches, subcutaneous patches, vaginal rings, IUDs, implants, and tubal ligation) will cause harm to the human body. Therefore, the embodiments of the present application change the idea and perform contraceptive treatment at the embryo level to avoid the side effects of current clinical contraceptive methods on the human body.

[0045] The normal process of embryo implantation is as follows: after the fertilized egg is formed, the embryo's trophoblast cells are formed, the embryo's trophoblast cells complete the invasion of the uterus, and finally grow and develop smoothly.

[0046] In recent years, copper-based organic skeleton materials have shown great application potential in biology, medicine and other fields due to their good physical and chemical properties. Therefore, the embodiment of the present application proposes a concept: after the formation of the embryo's trophoblast and before the invasion, the copper-based organic skeleton material is used to release copper ions to mediate the death of the embryo's trophoblast cells, preventing the trophoblast cells from completing the invasion of the uterus, so that the embryonic cell growth is restricted and naturally falls off, thereby achieving the purpose of contraception. Since the unimplanted embryo belongs to the cell state, the killing effect of copper ions on cells can also avoid moral and ethical concerns. The copper ions released by the copper-based organic skeleton material are also the effective contraceptive ingredients of the clinical copper-containing contraceptive device, and the embodiment of the present application has a good clinical transformation prospect. In addition, the organic ligands released by the copper-based organic skeleton material have no side effects on the body, reduce the toxicity of the copper-based organic skeleton material, and improve the biosafety of the copper-based organic skeleton material.

[0047] In order to solve the various adverse reactions caused by existing contraceptive materials, this application proposes to use vitamin copper organic skeleton to release copper ions to damage trophoblast cells to achieve the purpose of contraception. Figure 1 , Figure 1 This is a schematic diagram of a contraceptive method provided in an embodiment of the present application in which a copper-based organic framework material acts on embryonic trophoblast cells. The copper-based organic framework material shown in the figure is a vitamin B13-copper organic framework material (hereinafter abbreviated as VB13 Cu-MOF, wherein V is vitamin vitamin and MOF is a metal-organic framework Metal-organic frameworks). Figure 1 From Part A, we can see that the normal process of embryo implantation is as follows: after the fertilized egg is formed, the embryonic trophoblast cells are formed, the embryonic trophoblast cells complete the invasion of the uterus, and the embryo grows and develops smoothly. When the embryonic trophoblast cells die, the embryonic cells fall off naturally, achieving the purpose of contraception. Among them, after the contraceptive preparation is placed in the uterine cavity, because the contraceptive preparation can achieve the slow release of copper ions of VB13 Cu-MOF by pH / temperature regulation or controlling the degradation rate of the contraceptive preparation itself, when the embryonic trophoblast cells are formed, the released copper ions will act on the embryonic trophoblast cells, and the embryonic trophoblast cells will absorb the released copper ions. After the copper ions enter the embryonic trophoblast cells, they induce a Fenton-like reaction in the cells, resulting in an increase in the production of reactive oxygen species, causing the cells to be in a state of oxidative stress, causing embryonic trophoblast cell apoptosis and cell iron death, Figure 1 Part B in the middle is the fallopian tube without the copper-based organic framework material. It can be seen that the embryo develops normally without the copper-based organic framework material. Figure 1Part C in the middle is the fallopian tube under the action of the copper-based organic framework material. It can be seen that the embryonic cells are detached; and the process of embryo detachment and normal embryonic development in the fallopian tube is reversible.

[0048] Next, the application of a copper-based organic framework material in the preparation of a contraceptive preparation, the contraceptive preparation and the method proposed in the present application are described in detail in conjunction with specific embodiments.

[0049] Example 1: Characterization of copper-based organic framework materials

[0050] In the examples of the present application, raw materials of copper-based organic framework materials are purchased or synthesized, and the physical and chemical properties of the synthesized copper-based organic framework materials, such as morphology, size, elements, optics, and thermal stability, are characterized.

[0051] The raw materials for the synthesis of copper-based organic framework materials, copper salts and organic central ligands, can be purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., Shanghai McLean Biochemical Technology Co., Ltd., Merck & Co., Ltd., or other companies that are qualified to synthesize copper-based organic framework materials.

[0052] The synthesis chemical methods of copper-based organic framework materials include: solvent thermal, liquid phase synthesis, solid phase synthesis, ultrasound assisted method, high temperature assisted method, surfactant method, microwave oven assisted method, etc. In this application, the solvent thermal method is preferred to synthesize copper-based organic framework materials, and the parameter range used is as follows:

[0053] The temperature range is from room temperature to 120°C; the solvent system includes water reaction system, DMF (N, N-dimethylformamide) reaction system, organic alcohol reaction system, DMSO (dimethyl sulfoxide) reaction system, organic ether reaction system, etc., and their mixed systems; the reaction time is 0.5h to 48h; the purification system includes water reaction system, DMF (N, N-dimethylformamide) reaction system, organic alcohol reaction system, DMSO (dimethyl sulfoxide) reaction system, organic ether reaction system, etc., and their mixed systems; the freeze-drying method includes freeze-drying method, natural evaporation method, drying method, etc. The copper salt raw materials of the copper-based organic framework material include copper chloride, copper sulfate, copper acetate, etc.; the copper-based organic framework material includes mono- or poly-metal copper-based organic framework materials, single-component or multi-component metal compound copper-based organic framework materials; the central ligand of the copper-based organic framework material includes natural organic materials, artificially synthesized organic materials, such as vitamin B3, vitamin B13, tannic acid, etc.; the constituent elements of the copper-based organic framework material are all essential life elements for the human body.

[0054] In the embodiments of the present application, characterization methods such as optical microscope, scanning electron microscope, transmission electron microscope, particle size instrument, X-ray diffraction spectrum, differential scanning calorimeter, thermogravimetric analysis or infrared spectrum are used to characterize the physical and chemical properties such as morphology, size, elements, optics, thermal stability, etc. of the copper-based organic framework material. The characterization methods are all general methods and will not be repeated here.

[0055] For the characterization results of copper-based organic framework materials, please refer to Figure 2 , Figure 2 This is a characterization result diagram of a copper-based organic framework material provided in an example of the present application.

[0056] In the present application, VB13 Cu-MOF was prepared by a solvothermal method. Specifically, 0.32 g of orotic acid (vitamin B13) and 0.56 g of copper nitrate hydrate were placed in a wide-mouth glass bottle, and a solvent system (a solvent reaction system prepared by a volume ratio of DMF and water of 1:1) was added, heated to 90 degrees Celsius, reacted for 30 minutes, and then purified (washed with water, DMF, and a mixed solution of water and DMF) and freeze-dried (freeze-drying method) to obtain a sample (VB13Cu-MOF).

[0057] like Figure 2 As shown, Figure 2 Part A is the suspension of VB13Cu-MOF synthesized by solvent thermal method, and the suspension is a light dark green suspension. Figure 2 Part B in the middle shows the morphology of VB13 Cu-MOF characterized by transmission electron microscopy, which shows that VB13 Cu-MOF is a regular block. Figure 2 Part C in the middle is the result of statistical analysis of the size of VB13 Cu-MOF using a particle size analyzer. The average particle size of VB13Cu-MOF is 3.88μm and the PdI value is 0.21. Figure 2 Part D in the middle is the result of analyzing the thermal stability of VB13Cu-MOF using a differential scanning calorimeter. The highest heat-resistant temperature (peak value) of VB13 Cu-MOF is 277.2°C. Figure 2 Part E in the middle is the result of analyzing the thermal stability of VB13 Cu-MOF using a thermogravimetric analyzer. The peak temperature of VB13 Cu-MOF is 320.9°C. Figure 2 The F part is the result of analyzing the chemical shifts of different types of protons in VB13 Cu-MOF using a hydrogen spectrum nuclear magnetic resonance spectrometer. The chemical shifts of each type of proton in VB13 Cu-MOF have strong peaks at 0.2759HZ, 542.04HZ, 752.4HZ, 933.2HZ and 4359.1HZ. Figure 2Part G in the middle is the result of elemental analysis of VB13 Cu-MOF using an X-ray diffractometer. The diffraction peaks are approximately located at 9.58 2θ, 14.36 2θ, 19.22 2θ, 20.65 2θ, 22.83 2θ, 26.67 2θ, 27.71 2θ, 29.11 2θ, 32.02 2θ, 32.81 2θ, 37.38 2θ, 39.89 2θ, 43.86 2θ, 45.99 2θ, 53.58 2θ, 56.77 2θ and 75.06 2θ, and there are no impurity peaks. Figure 2 The H part is the result of detecting the infrared absorption spectrum of VB13 Cu-MOF using an infrared spectrometer. There are obvious infrared absorption peaks at the horizontal axis wavelengths of 3232.5nm, 3117.3nm, 2979.9nm, 2819.8nm, 1647.3nm, 1478.1nm, 1425.1nm, 1374.0nm, 1305.0nm, 1238.5nm, 1013.8nm, 880.3nm, 799.3nm, 553.5nm and 451.3nm. The position and intensity of the peaks are consistent with the characteristics of VB13 Cu-MOF. Figure 2 Part I shows the results of ultraviolet absorption spectrum detection of VB13 Cu-MOF using an ultraviolet absorption spectrometer. VB13Cu-MOF has a typical ultraviolet absorption peak near 288nm. The spectral characterization results show that the VB13 Cu-MOF suspension has good dispersibility and no foreign matter.

[0058] Example 2: Cytocompatibility of copper-based organic framework materials

[0059] CCK8 reagent was used to test the different concentrations of VB13 Cu-MOF (0.31 μg mL -1 , 0.63 μg mL -1 , 1.25 μg mL -1 ) were evaluated for cytocompatibility with embryonic trophoblast HTR8 / SVneo cells and human endometrial stromal (HESC) cells within 24 hours. The results of this evaluation can be found in Figure 3 , Figure 3 This is a diagram showing the cell compatibility results of a copper-based organic framework material provided in an example of the present application. Figure 3 Part A in the middle shows the cell compatibility results of different concentrations of VB13 Cu-MOF with embryonic trophoblast HTR8 / SVneo cells and human endometrial stromal (HESC) cells at 24 hours. Figure 3 Part B shows the staining results of live cells and dead cells. Figure 3 Part C in the middle shows the results of VB13 promoting the proliferation of HTR8 / Svneo cells. Figure 3Part D shows the results of VB13 promoting HESC cell proliferation.

[0060] from Figure 3 As can be seen in part A, the cell compatibility of VB13 Cu-MOF for the two cell lines is positively correlated with time and concentration. -1 After treatment with VB13 Cu-MOF, the survival rate of HTR8 / SVneo cells was 36.58±3.07%, while that of HESC cells was 42.59±7.85%; -1 The survival rate of HTR8 / SVneo cells was 26.73±2.93% and that of HE SC cells was 34.57±7.37% when exposed to 1.25 μg mL -1 In the presence of VB13 Cu-MOF, the survival rate of HTR8 / SVneo cells was 6.84±1.11%, while that of HESC cells was 17.92±3.53%. Figure 3 As shown in part B, the number of cells emitting red fluorescence in HTR8 / SVneo cells is greater than that in HESC cells, while the number of cells emitting green fluorescence in HESC cells is greater than that in HTR8 / SVneo cells. Within 24 hours, the same concentration of VB13Cu-MOF is more toxic to HTR8 / SVneo cells than to HESC cells, indicating that HTR8 / SVneo cells are more sensitive to VB13 Cu-MOF than normal human endometrial somatic cells and are more susceptible to the effects of MOF materials.

[0061] from Figure 3 It can be seen from parts C and D that VB13, as an organic ligand of VB13 MOF material, has the effect of promoting cell proliferation, and can bring VB13 MOF material higher biocompatibility and in vivo decomposition products that are friendly to the body. Copper ions are also an effective contraceptive component of Cu-IUD that affects sperm motility in clinical practice. Unlike Cu-IUD, the copper organic skeleton not only releases copper ions to play a contraceptive role, but also releases vitamin monomers that are friendly to the body, and has good tissue biocompatibility and low toxicity.

[0062] Example 3: Copper ion release results of copper-based organic framework materials

[0063] Gene sequencing-RNA transcriptome analysis, gas chromatography-mass spectrometry (GCMS), liquid chromatography-mass spectrometry (LCMS), gas chromatography-liquid chromatography coupled (GCLC), electrospray ionization mass spectrometry (ESI-MS), matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), secondary ion mass spectrometry (SIMS)-cell metabolome analysis, qPCR analysis of the relative expression of related genes, Western Blot (protein immunoblotting) analysis of the release of copper ions and the expression of related genes and proteins of copper ion transporters, the results of this example can be found in Figure 4 , Figure 4 This is a graph showing the copper ion release results of a copper-based organic framework material provided in an embodiment of the present application. Figure 4 Part A in the middle shows the ion release of VB13 Cu-MOF. Figure 4 Part B shows the gene expression of cellular copper ion transporter. Figure 4 Part C in the middle shows the protein expression results of CTR1 / SLC31A1 and CTR1 / SLC31A1. Figure 4 Part D in the middle shows the changes of copper ions in HTR8 / Svneo cells. Figure 4 Part E in the middle shows the performance of cell endocytosis. Figure 4 Part F in the middle shows the performance of cell phagocytic behavior.

[0064] Depend on Figure 4 As shown in part A, within 24 hours, VB13 Cu-MOF released Cu in RPMI-1640 complete medium. 2+ The level showed a trend of increasing. At 24 hours, Cu 2+ The release concentration was 1.14±0.11μg mL -1 ; VB13 Cu-MOF releases Cu within 8 hours 2+ The speed is faster, after 8h Cu 2+ The release rate of Cu in the extracellular environment tends to be slow. 2+ Increased copper transporters and corresponding gene expressions in cells are upregulated. CTR1 / SLC31A1 is a monovalent copper ion transporter on the cell membrane. Divalent copper ions are reduced to monovalent copper ions on the cell membrane and transported to the intracellular matrix by the CTR1 / SLC31A1 transporter. DMT1 / SLC11A2 is a divalent metal transporter 1 (DMT1), which is a proton-coupled transmembrane metal ion transporter in mammals. It can transport divalent copper ions from the extracellular environment to the intracellular matrix. Figure 4 As shown in part B, the extracellular environment Cu 2+The increase in copper in the cells caused an up-regulation in the gene expression of CTR1 / SLC31A1 and CTR1 / SLC31A1 within 8 hours. The reasons for the down-regulation of gene expression after 8 hours may be: on the one hand, the gene is used to translate into the corresponding protein, which is hydrolyzed after the translation is completed; on the other hand, the intracellular copper overload affects the normal metabolism of the cell, thereby affecting the synthesis of related genes; finally, the intracellular copper overload also inhibits the expression and synthesis of the copper transporter gene. Figure 4 As can be seen in the middle C part, the expression of CTR1 / SLC31A1 transporter and DMT1 / SLC11A2 transporter showed an up-regulated trend, indicating that the extracellular environment of Cu 2+ Continuously transported into the cell, leading to intracellular copper overload. Figure 4 The results in part D also showed that compared with the control group at 0h, the intracellular Cu 2+ Levels increased, reaching a maximum of 0.23 ± 0.05 μg mL -1 In addition, at 24 h, intracellular Cu 2+ The possible reason for the decrease in the level of Cu at 12 h is that more cells died at 24 h, and the cell membrane permeability increased, resulting in the partial intracellular Cu 2+ Loss.

[0065] Depend on Figure 4 It can be seen from parts E and F that under the action of copper-based organic framework materials, the biological regulation processes involving cell endocytosis and cell phagocytosis did not undergo significant changes, further indicating that VB13 Cu-MOF acts on HTR8 / SVneo cells by releasing copper ions.

[0066] Example 4: Copper-based organic framework materials induce Fenton-like reaction in embryonic trophoblast cells, leading to increased ROS components

[0067] The intracellular Fenton reaction refers to the conversion of H2O2 into . OH process. However, other metal cations, such as Cu 2+ , Cu + 、Ag + , Mn 2+ etc., involved in catalyzing the conversion of intracellular H2O2 into . The process of OH is called Fenton-like reaction. Copper is considered to be an efficient Fenton-like catalyst. + The maximum reaction rate with H2O2 (104M -1 s -1 ) is significantly higher than Fe 2+ (63M -1 s -1 ).

[0068] According to the results of the cellular copper homeostasis experiment, monovalent copper ions and divalent copper ions are continuously transported into the cell through copper transporters, resulting in intracellular copper overload, which in turn triggers the generation of a Fenton-like reaction. The cellular copper homeostasis experiment was conducted using HTR8 / SVneo cells. The experimental results are available at Figure 5 , Figure 5 This is a schematic diagram of a copper-based organic framework material provided in an embodiment of the present application inducing a Fenton-like reaction in embryonic trophoblast cells, Figure 5 The A part is the superoxide anion (·O 2- ) changes, Figure 5 Part B shows the changes in intracellular H2O2 levels. Figure 5 Part C shows the changes in the level of hydroxyl radicals (·OH).

[0069] like Figure 5 As shown in part A, due to the active transport of copper ions and the redox reaction in the cell, a large amount of oxygen (O2) is reduced to superoxide anions (·O 2- ), so O 2- As a reaction product, it increases first and then decreases. 2- The reason for the later reduction is that 2- It is further reduced to hydrogen peroxide (H2O2) under the catalysis of superoxide dismutase (SOD). Figure 5 As shown in part B, compared with the control group at 0h, the intracellular H2O2 level increased at 4h, 8h, 12h and 24h. After 4h, the intracellular H2O2 level decreased for several reasons: First, Cu transported by CTR1 / SLC31A1 + It undergoes a redox reaction with H2O2 to generate hydroxyl radicals (·OH) and hydroxyl ions (OH - ); second, the reduced glutathione in the intracellular redox regulation system also undergoes a redox reaction with H2O2 to generate oxidized glutathione, ·OH and OH - ·OH as the reduction product of H2O2, such as Figure 5 As shown in part C, the content level in the cell showed a clear increasing trend.

[0070] Example 5: Copper-based organic frameworks mediate trophoblast cell death via oxidative stress

[0071] VB13 Cu-MOF material decomposes copper ions in complete culture medium and enters the cytoplasm through active transport of copper transporters on the cell membrane of trophoblast HTR8 / SVneo. Overload of copper ions of different valence states in the cell triggers Fenton-like reaction and excessive ROS production. Excessive ROS leads to damage of trophoblast HTR8 / SVneo cells, including cell oxidative stress, mitochondrial damage, DNA damage and cell cycle arrest. Oxidative stress, organelle damage and DNA breakage will further lead to the death of trophoblast cells. The experimental results of copper-based organic framework materials mediating trophoblast cell death through oxidative stress can be found in Figure 6 , Figure 6 This is a schematic diagram of a copper-based organic framework material provided in an example of the present application mediating trophoblast cell death through oxidative stress, Figure 6 Part A in the middle shows the staining images of trophoblast HTR8 / SVneo cells treated with VB13 Cu-MOF at different time points. Figure 6 Part B shows the proteomic results of trophoblast HTR8 / SVneo cells treated with VB13 Cu-MOF. Figure 6 Part C in the middle is the differentially expressed genes involved in regulating cell apoptosis. Figure 6 Part D in the middle is the key regulatory genes of cellular oxidative stress with significant differences. Figure 6 Part E in the middle is the protein-protein interaction network of key regulatory genes. Figure 6 Part F in the middle shows the expression of key regulatory genes of cell apoptosis and ferroptosis.

[0072] 1.25 μg mL -1 After 0h, 4h, 8h, 12h and 24h of VB13 Cu-MOF treatment of trophoblast HTR8 / SVneo cells, double staining of dead cells and live cells was performed. Green fluorescence represents live cells, and red fluorescence represents dead cells. Figure 6 In the middle A part, at 0h, the cells were not dead and all showed green fluorescence. As time extended to 24h, the green fluorescence decreased and the red fluorescence increased, indicating that the number of cell deaths continued to increase under the action of high levels of copper ions released by VB13 Cu-MOF.

[0073] To determine the potential cell death mechanism, untreated trophoblast HTR8 / SVneo cells were used as blank control and VB13 Cu-MOF-treated trophoblast HTR8 / SVneo cells were analyzed by RNA-Seq. Figure 6As shown in Part B, Wiki pathway enrichment analysis is a commonly used data analysis tool for RNA-Seq. Its enrichment analysis results show that apoptosis and ferroptosis are the potential mechanisms of VB13 Cu-MOF-mediated trophoblast HTR8 / SVneo cell death. In addition, Nanomaterial induced apoptosis is one of the Wiki pathway enrichment analysis pathways. Compared with the control group, there is no significant difference in the treatment group (p value>0.05), indicating that cell damage and death are caused by copper ions released by VB13 Cu-MOF. From the chord diagram ( Figure 6 (Part C) The differentially expressed genes involved in the regulation of cell apoptosis include ACTG1, BBC3, BCL2A1, BID, BIRC 2, BIRC3, CASP9, CTSB, CTSK, CTSL, CTSV, CTSW, CYCS, DAB2IP, DDIT3, DFFB, EIF2AK3, ERN1, FADD, FOS, GADD 45A, NFKB1, NGF, PARP2, PARP4, PIK3CB, TUBA4A and Bax; the differentially expressed genes involved in the regulation of cell apoptosis include ACSL3, ACSL5, CP, FTH1, FTL, GCLM, HMOX1, MAP1LC3B, MAP1LC3B2, PRNP, SAT1, SLC11A2, SLC39A14, SLC3A2, SLC40A1 and SLC7A11. The applicant conducted cluster analysis on differentially regulated genes related to ROS metabolism, cellular oxidative stress, mitochondrial damage, DNA damage, cell cycle arrest, apoptosis and ferroptosis, such as Figure 6 As shown in D, the key regulatory genes SOD2, HSPA1A and part of HSPA1B for cellular oxidative stress, the key regulatory genes BCL2L1 and BCL2A1 for mitochondrial damage, the key regulatory genes Bax and CYCS for cell apoptosis, and the key regulatory genes GPX4, SAT1 and HMOX1 for ferroptosis all have obvious significant differences. A protein-protein interaction network was constructed for these key genes, as shown in Figure 1. Figure 6As shown in part E, in the interaction network, the thickness of the connecting line is proportional to the degree of association between genes. From the perspective of the protein-protein interaction network, SOD2, HSPA1A, HSPA1B, BCL2L1, BCL2A1, Bax, CYCS, GPX4, SAT1 and HMOX1 are closely connected to each other, and are directly or indirectly related to each other, indicating that VB13 Cu-MOF materials mediate trophoblast HTR8 / SVneo cell death through copper ions. There are multiple synergistic mechanisms. In order to further verify the accuracy of gene expression in the cluster analysis heat map, key regulatory genes with significant differences in cell oxidative stress and mitochondrial damage, such as BCL2L1, B CL2A1, SOD2, HSPA1A and HSPA1B, were selected for RT-PCR analysis. Figure 6 As shown in parts F and G, these genes showed similar expression trends at 0, 4, 8, 12, and 24 hours, for example, BCL2L1 expression was downregulated while other genes were upregulated, indicating that the RNA-Seq analysis results have a more accurate reference.

[0074] Example 6: Contraceptive effect of copper-based organic framework materials in animals

[0075] In order to further verify the contraceptive effect of copper-based organic framework materials in animals, this application uses experimental animals Wistar female rats to establish an animal pregnancy model to verify the contraceptive effect of copper-based organic framework materials; the animal pregnancy model is a Wistar female rat in a fertilized state after successful mating; the drug administration method is minimally invasive surgery on the back of rats. Figure 7 , Figure 7 This is a schematic diagram of the contraceptive effect of a copper-based organic framework material in an animal body provided in an example of the present application.

[0076] Figure 7 Part A is a schematic diagram of the process of administering VB13 Cu-MOF to an animal pregnancy model for contraception. On the 10th day after fertilization of rats, the contraceptive effect of the copper-based organic framework material was verified by palpating the abdomen to see if there was a grainy feeling of the embryo and performing minimally invasive abdominal surgery. The results of this part can be found in Figure 7 Part A shows the development process of the embryo in the uterus of Wistar rats after pregnancy, including the formation of the fertilized egg on the first day, the formation of the embryonic trophoblast around the fourth day, and the embryo implantation on the fifth to fifth day. Finally, on the tenth day, the normally growing embryo can be clearly observed in the rat's uterus, confirming that the female rat is successfully pregnant.

[0077] Figure 7Part B shows the contraceptive effect of different concentrations of VB13 Cu-MOF in the animal pregnancy model. The contraceptive effect of different concentrations of VB13 Cu-MOF in vivo is used to determine the in vivo administration concentration of VB13 Cu-MOF. Figure 7 As shown in Part B, on the 4th day after the successful pregnancy of Wistar female rats, the rats were anesthetized and injected with different concentrations (35 μg mL -1 , 65 μg mL -1 , 95 μg mL -1 )20μL VB13 Cu-MOF, the purpose is to allow VB13 Cu-MOF to cause acute cytotoxicity to the embryonic trophoblast cells formed on day 5.5, prevent the trophoblast cells from invading the rat endometrium, and thus exert a contraceptive effect; at the same time, water solvent was injected into the right uterus of the rat as a blank control group.

[0078] Figure 7 Part C in the middle is a uterine section diagram of the contraceptive effect of Wistar rats. Figure 7 The D part is Figure 7 HE staining of uterine tissue sections at the red line of the uterus of the middle rat. The in vivo contraceptive results showed that compared with the right uterus of the Wistar female rats, which was full of normally developed embryos (the total number of normal embryos on the right side of the two rats was 14), the left uterus was treated with 35 μg mL - 1 After VB13Cu-MOF injection, a small number of embryos still appeared (the total number of normal embryos on the left side of the two rats was 7), which indicates that the contraceptive efficiency of VB13 Cu-MOF at this concentration is not high in vivo. Figure 7 As shown in the rightmost image in Part B, the dosage concentration of the left uterus of Wistar rats was 95 μg mL-1 VB13 Cu-MOF. Due to the high dosage concentration, a small amount of VB13Cu-MOF remained in the endometrial epithelial cells (see Figure 7 The left uterus of rats without any embryos showed abnormal decidualization and swelling, which was different from the normal decidualization of embryo implantation. In addition, during the menstrual period, endometrial shedding occurs in the superficial area of ​​the uterus, which means that the VB13 Cu-MOF remaining in the endometrial epithelial cells is theoretically cleared out of the body by the uterus. In addition, the real picture in the middle ( Figure 7 Part B) shows that compared with the blank control in which the right side of the rat uterus was full of normally developed embryos (a total of 14 normal embryos in the right side of the two rats), the right side of the uterus injected with 65 μg mL -1No developing embryos were observed after VB13 Cu-MOF (the total number of normal embryos on the left side of the two rats was 0), and the uterus had normal appearance and no uterine decidualization reaction. -1 The contraceptive effect was significant, and there was no inflammatory cell infiltration in the uterine cell structure ( Figure 7 The optimal concentration of VB13 Cu-MOF for contraception in Wistar female mice is 65 μg mL -1 It can not only play a good contraceptive effect in animals, but also does not affect the normal macroscopic morphology of the rat uterus and does not cause local infiltration of lymphocytes in the uterus.

[0079] Figure 7 Part E is a schematic diagram of the process of administering VB13 Cu-MOF to an animal pregnancy model. Administration of VB13 Cu-MOF at different time points is also one of the most important factors affecting the contraceptive effect in vivo. Figure 7 Part F in the middle shows the results of administering VB13Cu-MOF to the animal pregnancy model at different time points. Figure 7 As shown in the middle F part, after Wistar female mice were pregnant, 65 μg mL -1 VB13 Cu-MOF, and then samples were taken on the 6th day after administration to observe the in vivo contraceptive effect of different administration times. Figure 7 As can be seen in the middle F part, there are normally growing embryos on both sides of the two rats. The reason is that the uterine cavity of SD female rats is a semi-open channel. Before the implantation of the embryonic trophoblast, that is, before the 5th to 5.5th day after the formation of the fertilized egg, VB13 Cu-MOF has been excreted from the body due to the daily activities of the rats. Therefore, the administration on the second day of pregnancy and fertilization of the female rats has no contraceptive effect. On the contrary, when the drug was administered on the 4th day, a significant contraceptive effect was still achieved. When the drug was administered on the 6th day, VB13 Cu-MOF also prevented the further development of the rat embryos, resulting in abnormal embryonic development and the appearance of resorbed fetuses. See Figure 7 In the middle G part, Figure 7Part G in the middle is the HE staining result of the tissue section of the rat uterus. Administration on the 2nd and 4th days did not affect the macroscopic morphology of the uterus and the cell structure of the endometrium; but when administered on the 6th day, VB13 Cu-MOF caused abnormal development of the embryo that had been implanted in the uterus, and the embryo was affected by the uneven distribution of MOF materials, causing the uterus to undergo different degrees of decidualization, and blood clots appeared in the uterine cavity. In general, the best time for administration should be the fourth day after the formation of the fertilized egg, because this time point is the period of embryonic trophoblast formation and rapid proliferation; while administration on the second day had no contraceptive effect due to the short retention time of VB13 Cu-MOF in the rat uterus. Although administration on the 6th day prevented the further growth and development of the embryo, it also caused congestion and different degrees of decidualization in the rat uterus.

[0080] In order to explore whether the use of a carrier to load the copper-based organic framework material has an impact on the contraceptive time and effect of the copper-based organic framework material, the following is a verification and explanation in combination with Example 7 and Example 8.

[0081] Example 7: Characterization and contraceptive effect of thermosensitive hydrogel loaded copper-based organic framework material

[0082] Organic polymers are used as drug delivery carriers for copper-based organic skeleton materials. Through the effects of controlled and sustained release, the contraceptive time of copper-based organic skeleton materials can be prolonged and the toxicity of copper-based organic skeleton materials can be reduced. The loading materials of copper-based organic skeleton materials include thermosensitive hydrogels, sodium alginate, chitosan, polylactic acid, agarose, liposomes, polyethylene glycol, gelatin, acrylic acid, and other organic polymer materials that can form gels; the drug loading methods of copper-based organic skeleton materials include direct loading and in-situ synthesis.

[0083] This embodiment uses thermosensitive hydrogel to load copper-based organic skeleton materials to achieve the purpose of controlled and sustained release of the copper-based organic skeleton materials. This is because thermosensitive hydrogel is a gel with good biocompatibility and does not affect cell proliferation.

[0084] Thermossensitive hydrogels loaded with copper-based organic framework materials achieve sustained release of copper-based organic framework materials in the following ways: pH-responsive controlled release: the uterus is in an acidic environment, and the thermosensitive hydrogel material can decompose and release drugs under acidic conditions. The decomposition rate of the material can be adjusted according to the pH value, thereby controlling the release of copper ions of the copper-based organic framework material; controlled release due to changes in temperature in the uterine microenvironment, where higher temperatures lead to faster release of copper ions; material self-degradation: uterine mucus is a hydrogel-like liquid containing various ions, which can chemically corrode the thermosensitive hydrogel material and decompose and release the copper ions of the copper-based organic framework material. The rate of chemical corrosion of the thermosensitive hydrogel can be controlled by changing the physical and chemical properties of the thermosensitive hydrogel, thereby controlling the release of copper ions from the copper-based organic framework material.

[0085] The contraceptive effect of VB13 Cu-MOF@TH (Thermosensitive hydrogel) in Wistar female mice was further determined. Figure 8 , Figure 8 Schematic diagram of the characterization and contraceptive effect of a thermosensitive hydrogel-loaded copper-based organic framework material provided in the examples of the present application.

[0086] Figure 8 Part A shows the process of administering VB13 Cu-MOF@TH to the animal pregnancy model. VB13 Cu-MOF@TH was minimally invasively injected into the left uterus of Wistar rats on days 2, 4, and 6 after the formation of the fertilized egg, while the right uterus was injected with pure thermosensitive hydrogel as a blank control. In the VB13 Cu-MOF@TH thermosensitive hydrogel, the concentration of VB13 Cu-MOF was 65 μg mL -1 The reason is that the previous experimental results showed that the appropriate concentration of VB13 Cu-MOF for contraception in Wistar female mice is 65 μg mL -1 (Example 6).

[0087] Figure 8 Part B shows the contraceptive effect of different concentrations of VB13 Cu-MOF@TH in the animal pregnancy model. Minimally invasive injection of VB13Cu-MOF@TH on the 2nd and 4th days successfully prevented embryo implantation in the left uterus of female mice (the total number of embryos growing normally in the uterus was 0), while the thermosensitive hydrogel did not affect the growth of embryos in the right uterus (the total number of embryos growing normally in the uterus was 35). There was a significant difference in the number of embryos growing normally in the left and right uteri, indicating that the thermosensitive hydrogel carrier can prolong the contraceptive effect of VB13 Cu-MOF in Wistar female mice. Minimally invasive injection of VB13 Cu-MOF@TH on the 6th day, although the embryos were able to grow and develop in the left uterus of female mice (the total number of embryos growing normally in the uterus was 8), the number of embryos that could develop normally was less than that in the right uterus injected with pure hydrogel only (the total number of embryos growing normally in the uterus was 16), indicating that administration on the 6th day can also prevent embryo implantation, but the effect is not obvious and there is a certain contraceptive risk.

[0088] Figure 8 Part C in the middle is a uterine section diagram of the contraceptive effect of Wistar rats. Figure 8 The D part is Figure 8 HE staining of uterine tissue sections at the red line of the uterus of rats in the middle. Figure 8In the enlarged area of ​​HE staining, the cell morphology and structure of endometrial epithelial cells and endometrial stroma are intact, without lymphocyte infiltration and abnormal aggregation ( Figure 8 In general, compared with the bare VB13Cu-MOF, the thermosensitive hydrogel could prolong the retention time and action time of VB13 Cu-MOF, and improve the tissue compatibility of VB13Cu-MOF, without abnormal rat uterine decidualization.

[0089] Example 8: Safety of Thermosensitive Hydrogel Loaded Copper-based Organic Framework Materials in Animals

[0090] The safety of the drug in the pregnant rat model was investigated by drug administration, including inflammatory factors in the blood, estrogen levels in serum, blood copper ion concentrations, cell morphology and structural changes in various metabolic organs, etc. The safety results can be found in Fig. 9 , Fig. 9 This is a schematic diagram of the safety of a thermosensitive hydrogel-loaded copper-based organic framework material in an animal body provided in an example of the present application.

[0091] Fig. 9 Part A and Part B are the changes in blood copper concentration and serum estrogen levels in female rats on the 2nd, 4th, 6th, and 8th days, respectively. There was no significant change in blood copper and estrogen levels in female rats, indicating that VB13 Cu-MOF@TH had no interference or effect on the blood copper level and estrogen in normal female rats. In addition, after injection of VB13 Cu-MOF@TH, the blood copper of pregnant female rats ( Fig. 9 Part A) and estrogen ( Fig. 9 The changes in the 2nd, 4th, 6th and 8th days (part B) were also not significant, including the blank control group and the pregnant drug group. These results show that VB13Cu-MOF@TH does not cause copper homeostasis and hormone metabolism homeostasis imbalance in pregnant or non-pregnant female rats through intrauterine administration of Wistar rats.

[0092] Similarly, interleukin-1 (IL-1) inflammatory factor was used to evaluate local and systemic inflammation after minimally invasive intrauterine injection of VB13 Cu-MOF@TH in rats. Fig. 9 Part C shows the changes in serum estrogen levels. Compared with the non-pregnancy blank control group and the pregnancy blank control group, there was no significant change in the inflammatory factors in the blood of the pregnancy treatment group, indicating that VB13Cu-MOF@TH would not cause systemic or systemic inflammation on the 2nd, 4th, 6th, and 8th days after uterine injection in female mice.

[0093] In addition, the control group, the pregnancy group, and the pregnancy group + VB13 Cu-MOF@TH administration group were subjected to histopathological analysis of the main organs, including the brain, heart, liver, spleen, kidney, and ovary, on the 8th day of injection administration. Fig. 9 Part D shows the histopathological analysis of the brain, heart, liver, spleen, kidney and ovary of the animal pregnancy model on the 8th day of injection administration. These organs did not show local inflammatory cell infiltration and aggregation, and were all normal and healthy. In general, these results show that VB13 Cu-MOF@TH has good biosafety in in vivo contraception.

[0094] As for the system embodiment, since it is basically similar to the method embodiment, the description is relatively simple, and the relevant parts can be referred to the partial description of the method embodiment.

[0095] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0096] The above is a detailed introduction to the application of a copper-based organic framework material in the preparation of contraceptive preparations, contraceptive preparations and methods provided by the present application. Specific examples are used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the method of the present application and its core idea. At the same time, for those skilled in the art, according to the idea of ​​the present application, there will be changes in the specific implementation methods and application scopes. In summary, the content of this specification should not be understood as limiting the present application.

Claims

1. An application of a copper-based organic framework material in the preparation of a contraceptive preparation, characterized in that: The copper-based organic framework material uses organic matter as an organic ligand of the copper-based organic framework, and the organic matter is vitamin B13; The copper-based organic framework material acts on the trophoblast cells of the embryo to prevent normal implantation of the embryo; The copper-based organic framework material damages trophoblast cells by releasing divalent copper ions, and the divalent copper ions are oxidized into monovalent copper ions on the surface of the trophoblast cells; The divalent copper ions and the monovalent copper ions are transported together into the trophoblast cells; In the trophoblast cells, the divalent copper ions and the monovalent copper ions induce a Fenton-like reaction, which increases the level of intracellular reactive oxygen species and causes damage to the trophoblast cells, thereby damaging the mitochondria of the trophoblast cells and mediating the death of the trophoblast cells.

2. The use of the copper-based organic framework material according to claim 1 in the preparation of contraceptive preparations, characterized in that: The contraceptive preparation is used in a concentration range of greater than 10 μg mL -1 , less than 100 μg mL -1 .

3. The use of the copper-based organic framework material according to claim 1 in the preparation of contraceptive preparations, characterized in that: The release rate of the copper-based organic framework material is controlled by pH response or temperature change, or the release rate of the copper-based organic framework material in the contraceptive preparation is controlled by controlling the degradation rate of the contraceptive preparation itself.

4. A contraceptive preparation, characterized in that: The contraceptive preparation is composed of a copper-based organic framework material and a carrier, wherein the copper-based organic framework material is synthesized from a copper salt material and an organic ligand, and the carrier is a thermosensitive hydrogel; The copper salt material is any one of copper nitrate, copper chloride, copper sulfate, copper acetate and hydrates thereof; The organic ligand is vitamin B13.

5. The contraceptive preparation according to claim 4, characterized in that The synthesis method of the copper-based organic framework material includes solvent thermal method, liquid phase synthesis method, solid phase synthesis method, ultrasound assisted method, high temperature assisted method, surfactant method and microwave oven assisted method.

Images