Application of fat acellular matrix as drug sustained-release carrier material

Abstract

The invention discloses application of an adipose acellular matrix prepared on the basis of adipose tissue in the aspect of drug sustained release. The fat acellular matrix is prepared by performing cleaning, peroxyacetic acid virus inactivation, supercritical carbon dioxide decellularization and degreasing treatment on adipose tissues, and splitting the adipose tissues into particles with proper particle sizes. The material can be used as a drug sustained-release carrier material, has good biocompatibility, excellent drug loading and sustained-release performance and degradability, and can be widely applied to sustained release of various drugs including antibiotics, antitumor drugs and the like. The invention provides a novel and potential material source and a preparation method for the field of drug sustained release.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to a fat-free cell-free matrix prepared by processing adipose tissue, a preparation method thereof, and applications thereof in sustained drug release. Background Art

[0002] In modern medical treatment, sustained-release drug technology plays a crucial role in improving therapeutic efficacy, reducing dosing frequency, and mitigating drug toxicity and side effects. While traditional sustained-release materials, such as synthetic polymers (e.g., polylactic acid and polyglycolic acid), can achieve sustained-release effects to a certain extent, they suffer from issues such as insufficient biocompatibility, potential inflammatory reactions caused by degradation products in the body, and reliance on chemical synthesis.

[0003] Human adipose tissue is a plentiful biomaterial with unique advantages. It possesses excellent biocompatibility because it is derived from the human body and has a high affinity for human tissue. However, raw human adipose tissue cannot be used directly for sustained drug delivery due to the potential presence of pathogens such as viruses, and the presence of adipocytes and lipids that can interfere with drug loading and release.

[0004] At present, although there are some studies on the use of biomaterials for sustained drug release, the technology for sustained drug release using human adipose tissue, a special source that has undergone specific processing, is still not perfect. This invention aims to fill the gap in this field. Summary of the Invention

[0005] One purpose of the present invention is to provide a new drug sustained-release carrier material based on human adipose tissue. By performing a series of treatments on human adipose tissue, the defects of its original state are overcome, making it a material with good biocompatibility, suitable particle size and excellent drug sustained-release performance.

[0006] Another object of the present invention is to expand the application of the above-mentioned acellular matrix based on human adipose tissue in the field of sustained drug release.

[0007] In a first aspect of the present invention, a method for preparing a decellularized adipose tissue matrix as a drug sustained-release carrier material is provided, wherein the particle size of the decellularized adipose tissue matrix is ​​0.1 to 100 μm, and the decellularized adipose tissue matrix is ​​prepared by the following method, comprising:

[0008] (s1) providing washed adipose tissue;

[0009] (s2) performing viral inactivation on the adipose tissue obtained in step (s1);

[0010] (s3) treating the virus-inactivated tissue obtained in step (s2) with supercritical carbon dioxide;

[0011] (s4) performing particle size control treatment on the tissue treated with supercritical carbon dioxide obtained in step (s3) to obtain the defatted and decellularized matrix; wherein the particle size control treatment is selected from the group consisting of microfluidics, liquid nitrogen cryo-grinding, ultrasonic fragmentation, or a combination thereof.

[0012] In another preferred embodiment, the particle size of the adipose decellularized matrix is ​​1 to 80 μm, preferably 20 to 50 μm, for example, 1 μm, 5 μm, 10 μm, 20 μm, 30 μm, 50 μm, and 60 μm.

[0013] In another preferred embodiment, the adipose tissue is adipose tissue of a mammal, such as human, pig, sheep, or cattle.

[0014] In another preferred embodiment, the adipose tissue is human adipose tissue.

[0015] In another preferred embodiment, the adipose tissue is homologous adipose tissue or allogeneic adipose tissue.

[0016] In another preferred embodiment, the adipose decellularized matrix has a porous structure.

[0017] In another preferred embodiment, the porosity of the adipose decellularized matrix is ​​10% to 80%, preferably 30% to 60%.

[0018] In another preferred embodiment, the adipose decellularized matrix does not contain cells and does not contain lipid droplets.

[0019] In another preferred embodiment, the lipid droplets are oil droplets released after the adipocytes are broken.

[0020] In another preferred embodiment, the “free from fat droplets” means that in the fat extract, the volume of oil droplets accounts for less than 1% of the total liquid, preferably less than 0.5%, and more preferably less than 0.1%.

[0021] In another preferred embodiment, the cells are selected from the group consisting of endothelial cells, adipose stem cells, macrophages, and stromal cells.

[0022] In another preferred embodiment, the “cell-free” means that the average number of cells in 1 ml of fat extract is ≤1, preferably ≤0.5, more preferably ≤0.1, or 0.

[0023] In another preferred embodiment, the adipose decellularized matrix has one or more characteristics selected from the following group:

[0024] (1) The adipose decellularized matrix has a sustained release property after loading the drug, and the sustained release time can reach more than 24 hours, preferably more than 48 hours, and more preferably more than 72 hours;

[0025] (2) The adipose decellularized matrix can be gradually degraded and absorbed.

[0026] In another preferred embodiment, the drug is one or more of a small molecule compound drug, a protein drug, and a nucleic acid drug.

[0027] In another preferred embodiment, the drug is selected from the following group: antibiotics, anti-tumor drugs, analgesics, angiotensin-converting enzyme inhibitors, beta-blockers, anti-epileptic drugs, and anti-Parkinson's disease drugs.

[0028] In another preferred embodiment, the drug is selected from the following group: one or more of antibiotics, anti-tumor drugs, analgesics, hormone drugs, cardiovascular drugs, and nervous system drugs.

[0029] In another preferred embodiment, the drug is selected from the following group: cephalosporins, quinolone antibiotics, doxorubicin, paclitaxel, cisplatin, morphine, and nonsteroidal anti-inflammatory drugs.

[0030] In another preferred embodiment, the drug is selected from the following group: ibuprofen, doxorubicin, and cefuroxime.

[0031] In another preferred embodiment, in step (s1), the cleaning includes cleaning with a cleaning agent.

[0032] In another preferred embodiment, the cleaning agent is selected from the following group: deionized water, physiological saline, and PBS buffer.

[0033] In another preferred embodiment, in step (s1), the cleaning includes soaking and / or rinsing.

[0034] In another preferred embodiment, in step (s1), the volume mass ratio of the cleaning agent to the adipose tissue is 1 to 10 mL / g, preferably 2 to 8 mL / g.

[0035] In another preferred embodiment, the washed fat tissue is free of blood, broken tissue and other impurities attached to the surface of the fat tissue.

[0036] In another preferred embodiment, in step (s1), the cleaning comprises flushing the adipose tissue with 2 to 10 volumes of physiological saline for 2 to 5 times.

[0037] In another preferred embodiment, in step (s2), the virus inactivation includes mixing the adipose tissue obtained in step (s1) with a disinfectant to inactivate the virus.

[0038] In another preferred embodiment, the disinfectant is a peroxide solution.

[0039] In another preferred embodiment, the peroxide is selected from the group consisting of hydrogen peroxide, peracetic acid, chlorine dioxide, ozone, or a combination thereof.

[0040] In another preferred embodiment, the solvent of the peroxide solution is water and / or alcohol.

[0041] In another preferred embodiment, the concentration of the peroxide solution is 0.1-1 wt%, for example, 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, or 0.8 wt%.

[0042] In another preferred embodiment, in step (s2), the mass ratio of the adipose tissue obtained in step (s1) to the disinfectant is 1:1 to 1:20, preferably 1:2 to 1:20, more preferably 1:5 to 1:20, for example 1:8, 1:10.

[0043] In another preferred embodiment, the mixing comprises soaking the adipose tissue obtained in step (s1) in a disinfectant.

[0044] In another preferred embodiment, in step (s2), the virus inactivation is carried out at 10-30°C, preferably 18-25°C.

[0045] In another preferred embodiment, in step (s2), the virus inactivation time is 30 to 60 minutes, such as 45 minutes or 50 minutes.

[0046] In another preferred embodiment, in step (s2), the virus inactivation further includes a post-processing step of washing to remove the disinfectant.

[0047] In another preferred embodiment, the washing includes soaking and / or rinsing with a detergent 2 to 15 times.

[0048] In another preferred embodiment, the cleaning agent is selected from the following group: deionized water, physiological saline, and PBS buffer.

[0049] In another preferred embodiment, the volume mass ratio of the cleaning agent to the adipose tissue is 1 to 10 mL / g, preferably 2 to 8 mL / g.

[0050] In another preferred embodiment, the flushing frequency is 2 to 15 times.

[0051] In another preferred embodiment, in step (s2), the virus inactivation comprises soaking the adipose tissue obtained in step (s1) in a disinfectant in a water bath at 18 to 25° C. for 30 to 60 minutes, and then rinsing with a detergent for 2 to 15 times.

[0052] In another preferred embodiment, in step (s3), the supercritical carbon dioxide treatment comprises placing the virus-inactivated tissue obtained in step (s2) into a supercritical carbon dioxide treatment device, wherein the parameters of the supercritical carbon dioxide treatment include one or more of the following:

[0053] (1) The pressure is 15 to 30 MPa, preferably 20 to 25 MPa;

[0054] (2) The temperature is 30-40°C, preferably 33-37°C;

[0055] (3) The carbon dioxide flow rate is 0.05 to 0.3 L / min, preferably 0.09 to 0.1 L / min;

[0056] (4) The time is 12-48 hours, preferably 16-24 hours.

[0057] In another preferred embodiment, in step (s4), the microfluidic technology comprises: passing the tissue treated with supercritical carbon dioxide obtained in step (s3) through a microchannel to obtain the decellularized extract.

[0058] In another preferred embodiment, the size of the microchannel is 0.1-1 mm, preferably 0.2-0.8 mm, for example 0.5 mm.

[0059] In another preferred embodiment, the flow rate of the tissue passing through the microchannel is 0.5-2 ml / min, for example, 1 ml / min.

[0060] In another preferred embodiment, in step (s4), the liquid nitrogen cryo-grinding comprises: immersing the tissue treated with supercritical carbon dioxide obtained in step (s3) in liquid nitrogen, and grinding the tissue after it is completely frozen to obtain the decellularized extract.

[0061] In another preferred embodiment, the liquid level of liquid nitrogen is 2 to 5 times, preferably 2 to 3 times, the volume of the tissue treated with supercritical carbon dioxide obtained in step (s3).

[0062] In another preferred embodiment, the grinding speed is 5000-15000 rpm, preferably 5000-8000 rpm.

[0063] In another preferred embodiment, the grinding is performed 2 to 8 times, preferably 2 to 5 times.

[0064] In another preferred embodiment, the grinding time for each time is independently 2 to 10 minutes, preferably 2 to 5 minutes.

[0065] In another preferred embodiment, the method comprises the following steps:

[0066] (s1) obtaining human adipose tissue from a human fat collection route, and rinsing the adipose tissue 3-5 times with 0.9% saline solution under a sterile environment, with each rinse volume of 5-10 times the weight of the adipose tissue, to remove visible impurities, blood, and broken tissue;

[0067] (s2) immersing the cleaned adipose tissue in a 0.2%-0.5% peracetic acid solution at 18-25°C for 30-60 minutes to inactivate viruses, followed by rinsing the adipose tissue 8-10 times with 0.9% sterile saline, with each rinse volume being 8-12 times the weight of the adipose tissue;

[0068] (s3) placing the virus-inactivated adipose tissue in a supercritical carbon dioxide treatment device at a pressure of 20-25 MPa, a temperature of 33-37° C., a carbon dioxide flow rate of 0.09-0.1 L / min, and a time of 16-24 hours to achieve decellularization and fat removal;

[0069] (s4) Using microfluidics or liquid nitrogen cryogenic grinding technology, the decellularized and degreased tissue is lysed into particles with a size range of 1-100 μm.

[0070] In another preferred embodiment, the flow rate of the microfluidic technology is controlled at 0.5-2 ml / min.

[0071] In another preferred embodiment, liquid nitrogen grinding is performed by immersing the sample in liquid nitrogen by adding liquid nitrogen, maintaining the liquid nitrogen level at 2-3 times the volume of the sample, grinding at a speed of 5000-8000 rpm, each grinding time of 2-5 minutes, and grinding 2-5 times.

[0072] In a second aspect of the present invention, a fat-decellularized matrix is ​​provided. The particle size of the fat-decellularized matrix is ​​0.1 to 100 μm, and the fat-decellularized matrix is ​​prepared by the following method, comprising:

[0073] (s1) providing washed adipose tissue;

[0074] (s2) performing viral inactivation on the adipose tissue obtained in step (s1);

[0075] (s3) treating the virus-inactivated tissue obtained in step (s2) with supercritical carbon dioxide;

[0076] (s4) performing particle size control treatment on the tissue treated with supercritical carbon dioxide obtained in step (s3) to obtain the defatted and decellularized matrix; wherein the particle size control treatment is selected from the group consisting of microfluidics, liquid nitrogen cryo-grinding, ultrasonic fragmentation, or a combination thereof.

[0077] In another preferred embodiment, the adipose decellularized matrix and steps (s1) to (s4) are independently as described in the first aspect of the present invention.

[0078] In a third aspect of the present invention, a sustained-release drug is provided, comprising:

[0079] (a) the adipose decellularized matrix according to the second aspect of the present invention; and

[0080] (b)Medications.

[0081] In another preferred embodiment, the mass ratio of the adipose decellularized matrix to the drug is 5 to 100:1, preferably 10 to 50:1, such as 20:1 or 30:1.

[0082] In another preferred embodiment, the adipose decellularized matrix, the drug, and steps (s1) to (s4) are independently as described in the first aspect of the present invention.

[0083] In another preferred embodiment, the sustained-release drug is prepared using the method described in the fourth aspect of the present invention.

[0084] In a fourth aspect of the present invention, a method for preparing the sustained-release drug according to the third aspect of the present invention is provided, the method comprising: mixing adipose decellularized matrix with a drug solution, allowing the drug to be fully adsorbed in the matrix to obtain the sustained-release drug.

[0085] In another preferred embodiment, the ratio of the added amount of the adipose decellularized matrix to the drug is 5 to 100:1, preferably 10 to 50:1, such as 20:1 or 30:1.

[0086] In another preferred embodiment, the mass-to-volume ratio of the adipose decellularized matrix to the drug solution is 0.01-1 g / mL, preferably 0.05-0.5 g / mL.

[0087] In another preferred embodiment, the concentration of the drug in the drug solution is 1-20 mg / ml, such as 5 mg / ml, 8 mg / ml, or 10 mg / ml.

[0088] In another preferred embodiment, the mixing is performed at room temperature, preferably 15-40°C.

[0089] In another preferred embodiment, the mixing time is 12 to 96 hours, preferably 16 to 48 hours.

[0090] In a fifth aspect of the present invention, a sustained-release preparation is provided, comprising:

[0091] (a) the sustained-release drug according to the third aspect of the present invention; and

[0092] (b) Pharmaceutically acceptable excipients and carriers.

[0093] In another preferred embodiment, the sustained-release preparation is in the form of a solid preparation, a semisolid preparation, or a liquid preparation.

[0094] In another preferred embodiment, the sustained-release preparation is in the form of an injection, an implant, a gel, a patch, a powder, a tablet, a capsule, a granule, a tincture, an oral solution or a lozenge.

[0095] In another preferred embodiment, the sustained-release preparation is administered orally, by injection, local implantation or external application.

[0096] In another preferred embodiment, the sustained-release preparation is in the form of a topical administration preparation.

[0097] In another preferred embodiment, the sustained-release preparation is administered by local injection, local implantation or external application.

[0098] In the sixth aspect of the present invention, there is provided a use of the adipose decellularized matrix according to the second aspect of the present invention, the sustained-release drug according to the third aspect of the present invention, or the sustained-release preparation according to the fifth aspect of the present invention in sustained-release of drugs.

[0099] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0100] After extensive and in-depth research, the inventors have discovered for the first time a biodegradable drug-release carrier material with excellent sustained-release properties. This material is a small-particle adipose-derived acellular matrix derived from adipose tissue. This material is prepared by washing the adipose tissue, inactivating viruses with peracetic acid, decellularizing and removing oil with supercritical carbon dioxide, and then pyrolyzing it into particles of a suitable size. This material exhibits excellent biocompatibility and a suitable particle size, making it widely applicable for the sustained-release of a variety of drugs, including antibiotics and anti-tumor drugs. This is the basis for the present invention.

[0101] the term

[0102] 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.

[0103] As used herein, the terms "comprise," "include," and "contain" are used interchangeably to include not only closed definitions but also semi-closed and open definitions. In other words, the terms include "consisting of," "consisting essentially of."

[0104] As used herein, when used in reference to a specific recited value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0105] (1) Adipose decellularized matrix

[0106] As used herein, "adipose decellularized matrix" and "drug sustained-release carrier material" are used interchangeably and refer to a decellularized matrix based on adipose tissue in the present invention that has been subjected to virus inactivation, decellularization, defatting, and lysis into particles of a suitable size, which can be used as a drug sustained-release carrier and allows for sustained-release of drugs after loading the drug.

[0107] (2) Preparation method of adipose decellularized matrix material

[0108] 1. Adipose tissue collection and initial processing

[0109] Obtain human adipose tissue from a strictly ethically reviewed and legally authorized source, such as fat waste generated during liposuction surgery (ensure the patient has signed an informed consent form). On a sterile, Class 100 laminar flow-controlled operating table, quickly transfer the collected adipose tissue to a sterile container and rinse it with 0.9% saline. Gentle and thorough rinsing is essential to remove any blood, broken tissue, and other impurities adhering to the adipose tissue surface.

[0110] 2. Virus inactivation step

[0111] After initial treatment, the adipose tissue is immersed in a precisely formulated peracetic acid solution with a mass fraction of 0.2%-0.5%. This process is performed in a constant temperature water bath maintained at a constant temperature of 18-25°C, and the treatment time is strictly controlled within 30-60 minutes. Peracetic acid effectively inactivates viruses that may be present in adipose tissue, including but not limited to enveloped and non-enveloped viruses such as hepatitis B virus, hepatitis C virus, and HIV. Following treatment, the tissue is immediately rinsed with a large amount of sterile saline at a mass fraction of 0.9% to completely remove any residual peracetic acid and prevent it from adversely affecting subsequent processing and material properties.

[0112] 3. Supercritical carbon dioxide treatment

[0113] The virus-inactivated adipose tissue is transferred to a supercritical carbon dioxide treatment system. In this system, the pressure is 20-25 MPa, the temperature is 33-37°C, the carbon dioxide flow rate is 0.09-0.1 L / min, and the treatment time is 16-24 hours. In a supercritical state, carbon dioxide has a density similar to that of a liquid and a diffusion coefficient similar to that of a gas, allowing it to effectively penetrate into the adipose tissue. This unique physical property allows carbon dioxide to destroy the cell membrane structure of fat cells, prompting the release of cellular contents and their discharge with the carbon dioxide flow, while simultaneously separating the oil components from the tissue, achieving the dual effects of decellularization and oil removal.

[0114] 4. Particle size control and material molding

[0115] After the adipose tissue is treated with supercritical carbon dioxide, microfluidics or ultrasonic crushing technology is used to control the particle size. When using microfluidics, precise control of particle size is achieved by adjusting the size and flow rate of the microchannel (the flow rate is controlled at 0.5-2 ml / min). If liquid nitrogen freezing grinding technology is used, the sample is immersed in liquid nitrogen by adding liquid nitrogen, and the liquid nitrogen level is maintained at 2-3 times the sample volume. The grinding speed is 5000-8000 rpm, each grinding time is 2 to 5 minutes, and grinding is performed 2 to 5 times. After treatment, the adipose tissue is cracked into particles with a particle size range of 1-100 μm, more preferably 20-50 μm. These particles constitute the final drug sustained-release carrier material, and their particle size has a key influence on the drug load and sustained-release performance.

[0116] The particle size of acellular adipose-derived matrix particles directly determines their specific surface area. A reduction in particle size leads to a significant increase in the specific surface area of ​​the particles. A larger specific surface area means that the particle surface can provide more drug binding sites (such as exposed functional groups on collagen, fibronectin, and laminin, as well as micropores and nanopores within the matrix), thereby significantly increasing the maximum loading capacity and loading efficiency of the target drug (especially drugs loaded via adsorption or surface binding mechanisms) per unit mass of carrier material.

[0117] Particle size influences loading capacity (providing more binding sites) and initial release (increasing exposed area) by determining specific surface area. Particle size regulates overall release rate and sustained-release duration by determining the average diffusion path length of drug molecules (a longer path results in slower release). Particle size influences the accessibility and utilization efficiency of pores within the matrix, thereby affecting loading depth and the complexity of drug diffusion. However, too small a particle size can lead to faster in vivo degradation, accelerating drug release and negating the sustained-release effect.

[0118] (3) Characteristics of drug sustained-release carrier materials

[0119] 1. Biocompatibility

[0120] The drug-release carrier material prepared by the present invention is derived from human adipose tissue. After processing, it retains good compatibility with human tissues while effectively reducing immunogenicity. During implantation or use in the body, it does not trigger significant immune rejection reactions and can coexist harmoniously with surrounding tissues, creating a stable environment for drug release.

[0121] 2. Drug loading and sustained release performance

[0122] The material has a porous structure with a porosity ranging from 30% to 60%. This porous structure provides numerous adsorption sites and storage space for drug molecules, enabling effective drug loading through various methods such as physical adsorption and encapsulation. In in vitro experiments simulating in vivo environments, the material demonstrated excellent sustained-release properties for a variety of drugs, with sustained-release periods lasting up to 24-72 hours. The drug molecules are slowly released from the material into the surrounding environment, achieving a long-lasting effect, reducing dosing frequency, and enhancing therapeutic efficacy.

[0123] 3. Degradability

[0124] Under the physiological environment of the body, the drug-release carrier material is gradually degraded and absorbed. As the material degrades, the drug is continuously released, and the degradation products are processed by the body's normal metabolic pathways, avoiding potential adverse reactions caused by residual substances in the body and eliminating the need for secondary surgery to remove them.

[0125] (IV) Application of drug sustained-release carrier materials

[0126] The drug sustained-release carrier material of the present invention can be widely used in the preparation of sustained-release drugs of various drug types, and can ultimately be made into various sustained-release preparations.

[0127] 1. Antibiotic sustained release

[0128] For the treatment of localized infections, antibiotics (such as cephalosporins and quinolones) can be loaded onto the material and made into implants or injections. Implanted into the infected area or injected around the lesion, the material can continuously release antibiotics over a long period of time, maintaining effective local drug concentrations, thereby effectively killing pathogens and reducing the side effects of systemic medication and the development of drug-resistant bacteria.

[0129] 2. Sustained release of anti-tumor drugs

[0130] In tumor treatment, chemotherapy drugs (such as paclitaxel and cisplatin) are combined with materials to produce sustained-release drugs, and then combined with necessary excipients to form formulations. Through local implantation or intratumoral injection, the drugs are slowly released within the tumor tissue, increasing the drug concentration at the tumor site and enhancing the killing effect on tumor cells. At the same time, the concentration of chemotherapy drugs in normal tissues is reduced, reducing the toxic side effects on normal tissues (such as the bone marrow, gastrointestinal tract, liver and kidney function, etc.).

[0131] 3. Sustained-release analgesics

[0132] For patients with chronic pain, analgesics (such as morphine and nonsteroidal anti-inflammatory drugs) can be loaded into the material and made into patches or implants. This can achieve slow release of the analgesics, maintain stable blood drug concentrations, effectively relieve pain, and reduce the pain of frequent drug administration and the risk of drug addiction.

[0133] 4. Application in other pharmaceutical fields

[0134] In the treatment of cardiovascular diseases, it can be used for the sustained release of drugs such as angiotensin-converting enzyme inhibitors and beta-blockers; in the treatment of neurological diseases, it can be used for the sustained release of anti-epileptic drugs, anti-Parkinson's disease drugs, etc., providing more optimized dosing plans for the treatment of these diseases.

[0135] (5) Sustained-release preparations

[0136] The adipose decellularized matrix of the present invention can be prepared into a preparation after loading the drug to perform sustained drug release.

[0137] The sustained-release preparation comprises the adipose decellularized matrix of the present invention, or the sustained-release drug of the present invention, and a pharmaceutically acceptable carrier.

[0138] In the present invention, the dosage form of the sustained-release preparation includes (but is not limited to) injections, implants, gels or patches.

[0139] The term "pharmaceutically acceptable carrier" refers to one or more compatible solid, semisolid, or liquid fillers suitable for human or animal use and possessing sufficient purity and low toxicity. "Compatibility" refers to the ability of the components of a pharmaceutical composition to be compatible with the active ingredient of the drug, as well as with each other, without significantly reducing the drug's efficacy.

[0140] It should be understood that in the present invention, the carrier is not particularly limited and can be selected from materials commonly used in the art, or prepared by conventional methods, or purchased from the market. Some examples of pharmaceutically acceptable carriers include vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), buffers, chelating agents, thickeners, pH regulators, transdermal enhancers, colorants, flavorings, stabilizers, antioxidants, preservatives, antibacterial agents, pyrogen-free water, etc.

[0141] Examples of antioxidants include tocopherol and its ester derivatives, ascorbic acid, ascorbyl stearate, nordihydroguaiaretic acid, butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), citric acid, 2-mercaptobenzimidazole, and ethylenediaminetetraacetic acid. These antioxidants may be used alone or in combination of two or more.

[0142] The skin penetration enhancer is not particularly limited as long as it is a compound generally recognized to have an absorption promoting effect on the skin. Examples thereof include fatty alcohols such as isostearyl alcohol, fatty acids such as capric acid, fatty acid derivatives such as propylene glycol monolaurate, isopropyl myristate, isopropyl palmitate, and lauric acid diethanolamide, and glycols such as propylene glycol and polyethylene glycol. The skin penetration enhancer may be used alone or in combination of two or more.

[0143] The preservative is not particularly limited, but preferred examples include ethyl paraoxybenzoate, propyl paraoxybenzoate, and butyl paraoxybenzoate. The filler is not particularly limited, but preferred examples include calcium carbonate, magnesium carbonate, silicates (such as aluminum silicate, calcium silicate, and magnesium silicate), and cellulose derivatives (such as hydroxypropyl cellulose, hydroxypropyl methylcellulose, and carboxymethyl cellulose).

[0144] Typically, in addition to the active ingredient, the liquid dosage form may contain an inert diluent commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers, and suspending agents.

[0145] The material dosage form should be compatible with the mode of administration. The material of the present invention can also be used with other synergistic therapeutic agents (including before, during or after use). When using the material of the present invention, the medicine of a safe and effective amount is applied to the desired object (such as a human or non-human mammal), and the safe and effective amount is usually at least about 10 micrograms / kg body weight, and in most cases is no more than about 8 milligrams / kg body weight, preferably the dosage is about 10 micrograms / kg body weight-about 1 milligram / kg body weight. Of course, the specific dosage should also consider factors such as route of administration, patient health status, etc., and these are all within the skill range of skilled physicians.

[0146] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which no specific conditions are specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0147] Example 1

[0148] Adipose tissue collection and washing: 150 g of adipose tissue was collected from patients who had voluntarily donated the tissue (with signed informed consent) in the hospital liposuction operating room. The tissue was quickly transferred to a sterile container in the aseptic operating room and rinsed three times with 750 ml of 0.9% saline solution to remove visible impurities and blood.

[0149] Virus inactivation: Soak the cleaned adipose tissue in a 0.3% peracetic acid solution in a 20°C water bath for 45 minutes. Rinse with 0.9% sterile saline nine times, 1200 ml each time, to ensure complete removal of the peracetic acid.

[0150] Supercritical carbon dioxide treatment: The virus-inactivated adipose tissue is placed in a supercritical carbon dioxide treatment device with a pressure of 20-25 MPa, a temperature of 33-37°C, a carbon dioxide flow rate of 0.09-0.1 L / min, and a time of 16-24 hours to achieve decellularization and oil removal.

[0151] Particle size control: Microfluidic technology is used with a microfluidic channel size of 0.5 mm. The flow rate is set to 1 ml / min to lyse the treated tissue to obtain particles with a size of 20-30 μm, which are drug sustained-release carrier materials.

[0152] Example 2

[0153] 200 g of adipose tissue was collected from liposuction surgery and rinsed with normal saline 4 times, 1000 ml each time.

[0154] Soak in 0.4% peracetic acid solution at 22°C for 50 minutes, then rinse with normal saline 10 times, 1500 ml each time.

[0155] Place in a supercritical carbon dioxide device with a pressure of 20-25 MPa, a temperature of 33-37°C, a carbon dioxide flow rate of 0.09-0.1 L / min and a time of 16-24 hours.

[0156] Use liquid nitrogen cryo-grinding, immerse the sample in liquid nitrogen by adding liquid nitrogen, maintain the liquid nitrogen level at 2-3 times the sample volume, grind at a speed of 5000-8000 rpm, grind for 2-5 minutes each time, and grind 2-5 times.

[0157] Example 3: Antibiotic sustained-release test (taking cefuroxime as an example)

[0158] 12 g of the drug sustained-release carrier material prepared in Example 1 was placed into 120 ml of 8 mg / ml cefuroxime solution, and stirred at room temperature for 24 hours to allow the drug to be fully adsorbed on the material.

[0159] The cefuroxime-loaded material was made into an implant and implanted into the infection site of a pre-established rabbit local infection model (Staphylococcus aureus infection).

[0160] On days 1, 2, 3, 5, 7, and 10 after implantation, tissue fluid from the infected area was collected and the cefuroxime concentration in the tissue fluid was measured using high-performance liquid chromatography (HPLC). The results showed that the implant prepared with the material of the present invention continuously released cefuroxime over 10 days, effectively controlling the infection. In contrast, the control group (conventional cefuroxime injection) showed that the drug concentration dropped to a low level within 3-4 days, failing to effectively suppress the infection.

[0161] Example 4: Antitumor Drug Sustained Release Test (Taking Doxorubicin as an Example)

[0162] 8 g of the drug sustained-release carrier material prepared in Example 2 was mixed with 80 ml of a 5 mg / ml doxorubicin solution and stirred for 30 hours to load doxorubicin onto the material.

[0163] The doxorubicin-loaded material was made into an implantable sustained-release drug and implanted into the tumor site of tumor-bearing mice (breast cancer model).

[0164] The efficacy of the sustained-release drug was evaluated by regularly observing changes in tumor volume in mice (measured every three days), measuring doxorubicin concentrations in the blood (measured weekly), and recording mouse survival. The results showed that compared with traditional intravenous doxorubicin injection, the sustained-release drug of the present invention significantly inhibited tumor growth, reduced peak doxorubicin concentrations in the blood by approximately 60%, reduced toxicity to normal tissues such as the heart and bone marrow, and prolonged mouse survival by approximately 30%.

[0165] Example 5: Analgesic sustained-release test (taking ibuprofen as an example)

[0166] 10 g of the drug sustained-release carrier material prepared in Example 1 was mixed with 100 ml of a 10 mg / ml ibuprofen solution and stirred for 20 hours to load the ibuprofen onto the material.

[0167] The ibuprofen-loaded material was made into a patch and applied to the joints of a rat arthritis pain model.

[0168] The sustained-release effect was evaluated by observing behavioral changes in the rats (such as joint mobility and the frequency of joint licking and biting) and measuring blood ibuprofen concentrations. The results showed that the patch continuously released ibuprofen over 48 hours, effectively alleviating joint pain in the rats. The ibuprofen concentration in the blood fluctuated minimally, demonstrating a more sustained and stable analgesic effect compared to oral ibuprofen sustained-release capsules.

[0169] The above examples fully demonstrate the feasibility and advantages of the human adipose tissue-based sustained-release drug carrier material prepared by the present invention in various drug sustained-release applications. In practical applications, the material preparation parameters and drug loading methods can be further optimized according to the specific drug type and treatment needs to achieve better therapeutic effects.

Claims

1. A method for preparing a decellularized adipose tissue matrix as a drug sustained-release carrier material, wherein the adipose tissue matrix has a particle size of 0.1 to 100 μm and is prepared by the following method, comprising: (s1) providing washed adipose tissue; (s2) performing viral inactivation on the adipose tissue obtained in step (s1); (s3) treating the virus-inactivated tissue obtained in step (s2) with supercritical carbon dioxide; (s4) performing particle size control treatment on the tissue treated with supercritical carbon dioxide obtained in step (s3) to obtain the defatted and decellularized matrix; wherein the particle size control treatment is selected from the group consisting of microfluidics, liquid nitrogen cryo-grinding, ultrasonic fragmentation, or a combination thereof.

2. The use according to claim 1, characterized in that The particle size of the adipose decellularized matrix is ​​1 to 80 μm.

3. The use according to claim 1, characterized in that The adipose decellularized matrix has a porous structure, and the porosity of the adipose decellularized matrix is ​​10% to 80%, preferably 30% to 60%.

4. The use according to claim 1, wherein The adipose decellularized matrix has one or more characteristics selected from the following group: (1) The adipose decellularized matrix has a sustained release property after loading the drug, and the sustained release time can reach more than 24 hours, preferably more than 48 hours, and more preferably more than 72 hours; (2) The adipose decellularized matrix can be gradually degraded and absorbed.

5. The use according to claim 1, characterized in that The drug is one or more of a small molecule compound drug, a protein drug, and a nucleic acid drug.

6. The use according to claim 1, wherein In step (s3), the supercritical carbon dioxide treatment comprises placing the virus-inactivated tissue obtained in step (s2) into a supercritical carbon dioxide treatment device, wherein the parameters of the supercritical carbon dioxide treatment include one or more of the following: (1) The pressure is 15 to 30 MPa, preferably 20 to 25 MPa; (2) The temperature is 30-40°C, preferably 33-37°C; (3) The carbon dioxide flow rate is 0.05 to 0.3 L / min, preferably 0.09 to 0.1 L / min; (4) The time is 12-48 hours, preferably 16-24 hours.

7. A fat-free cell matrix, characterized in that: The particle size of the adipose decellularized matrix is ​​0.1 to 100 μm, and the adipose decellularized matrix is ​​prepared by the following method, which includes: (s1) providing washed adipose tissue; (s2) performing viral inactivation on the adipose tissue obtained in step (s1); (s3) treating the virus-inactivated tissue obtained in step (s2) with supercritical carbon dioxide; (s4) performing particle size control treatment on the tissue treated with supercritical carbon dioxide obtained in step (s3) to obtain the defatted and decellularized matrix; wherein the particle size control treatment is selected from the group consisting of microfluidics, liquid nitrogen cryo-grinding, ultrasonic fragmentation, or a combination thereof.

8. A sustained-release drug comprising: (a) the adipose decellularized matrix according to claim 7; and (b)Medications.

9. A sustained-release preparation comprising: (a) The sustained-release drug according to claim 8; and (b) pharmaceutically acceptable excipients or carriers.

10. The sustained-release preparation according to claim 9, wherein The dosage form of the sustained-release preparation is injection, implant, gel, patch, powder, tablet, capsule, granule, tincture, oral solution or lozenge.