Drug-loaded patch

Abstract

The invention relates to the field of new biological materials, in particular to a drug-loaded patch. The drug-loaded patch is obtained by packaging fat-soluble drugs in an acellular matrix; the drug-loaded patch at least comprises a region A and a region B, and the overlapping rate of the edge of the region A and the edge of the patch is less than or equal to 90%; the area A at least loads fat-soluble drugs, and the concentration of any fat-soluble drug on the area A is 300-1300 [mu] g / cm < 2 >; the drug-loaded patch is of a multi-layer structure, and the multi-layer structure comprises a fat-soluble drug layer and at least two acellular matrix layers. According to the obtained drug-loaded patch, based on a non-coating drug-loaded slow-release system of a lamination process, slow release of fat-soluble drugs which are released too fast in vivo in the initial stage on a non-crosslinked acellular matrix is achieved, the fat-soluble drugs are not released too fast in the initial stage of implantation, cooperative and continuous release is achieved along with the wound healing and recovery process, and the effect of healing the wound is achieved. Meanwhile, tissue regeneration is guided, degradation is synchronously achieved, the occurrence probability of adhesion is reduced, growth of granulation tissue of the wound surface is promoted, and the technical effect that 1 + 1 > 2 is achieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological new materials, in particular to a drug-loaded patch. BACKGROUND

[0002] In the field of medical devices, biological materials exhibit unique advantages over synthetic polymers: their natural degradable characteristics significantly reduce the risk of immune rejection, and decellularized matrix (ECM) as a representative biological material not only has excellent biocompatibility, but also realizes high adaptation to the dynamic microenvironment of cells by retaining the natural three-dimensional structure of tissues. However, this degradable characteristic also brings challenges - the degradation rate of materials such as ECM is usually several times faster than that of synthetic polymers, resulting in premature release of encapsulated drugs (especially lipid-soluble drugs) before reaching an effective therapeutic concentration in the lesion tissue. This contradiction highlights the importance of regulating the degradation kinetics of biological materials and the release rate of drugs.

[0003] Patent No. CN107320786B provides a slow-release anti-infection composite soft tissue repair material and its preparation method, which records that arranging anti-infection components at intervals of the bacteriostatic circle radius on the biological material layer can achieve slow release of the anti-infection components on the biological material. However, this document does not further study the stability of drug release rate and drug loading of this scheme, and it cannot be determined whether this arrangement of anti-infection components at intervals of the bacteriostatic circle radius on the biological material is suitable for large-scale batch production. SUMMARY

[0004] Lipid-soluble drugs have a very fast initial release rate in the body, which allows the drug to be quickly distributed in the body. In the prior art, the slow release of rifamycin is usually achieved by adjusting the drug loading or embedding the drug in polymer microspheres, but this drug loading adjustment or drug embedding has very strict requirements on the product process, and the final product has large deviations in drug loading and drug release, making it difficult to mass-produce related products.

[0005] The present application provides a drug-loaded patch that does not need to control the drug loading or specially set the shape and pattern of the lipid-soluble drug loading A area. Under the premise that the lipid-soluble drug has a specific initial concentration (i.e., the concentration of the lipid-soluble drug in the A area is 300-1300 μg / cm 2 ), a specific limitation on the position of the A area edge (i.e., the overlap rate of the A area edge and the patch edge is less than 100%) can achieve slow release of the lipid-soluble drug, reduce the initial release rate of the lipid-soluble drug in the body, keep the release rate within 40% during the initial release period (4h), and control the slow release to be completed within 3-5 days. In addition, the drug loading and drug release of the product can remain stable, which is especially suitable for large-scale batch production.

[0006] The first aspect of the present application provides a drug-loaded patch, which is obtained by encapsulating a fat-soluble drug with a decellularized matrix; the drug-loaded patch comprises at least two regions, i.e., region A and region B, wherein the overlap rate of the edge of region A with the edge of the patch is ≤90%; region A at least loads a fat-soluble drug, and the concentration of any one fat-soluble drug on region A is 300-1300 μg / cm 2 ; the drug-loaded patch is a multi-layer structure comprising a fat-soluble drug layer and at least two decellularized matrix layers.

[0007] Further, the overlap rate of the edge of region A with the edge of the patch is ≤86%.

[0008] Further, the overlap rate of the edge of region A with the edge of the patch is ≥0.

[0009] Illustratively, the overlap rate of the edge of region A with the edge of the patch is >0.

[0010] Illustratively, the overlap rate of the edge of region A with the edge of the patch is >5%.

[0011] Further, the overlap rate of the edge of region A with the edge of the patch is ≥10%.

[0012] Further, the overlap rate of the edge of region A with the edge of the patch is ≥15%.

[0013] Illustratively, 5% < the overlap rate of the edge of region A with the edge of the patch < 100%.

[0014] Illustratively, 5% < the overlap rate of the edge of region A with the edge of the patch ≤ 86%.

[0015] Illustratively, 15% ≤ the overlap rate of the edge of region A with the edge of the patch ≤ 86%.

[0016] Illustratively, the overlap rate of the edge of region A with the edge of the patch is 15%, 16%, 18%, 18.5%, 19%, 25%, 31%, 31.65%, 32%, 50%, 75%, 76%, 76.47%, 77%, 80%, 85.71%, 86%, etc.

[0017] Further, on any side length of the drug-loaded patch, the geometric overlap rate of region A with the side length of the patch is <100%.

[0018] The geometric overlap rate refers to the length geometric overlap rate.

[0019] The geometric overlap rate = the length of the edge of region A covering on a side length of the patch / the length of the side length of the patch × 100%.

[0020] Further, the drug-loaded patch at least comprises one side length with a geometric overlap rate with region A >0.

[0021] For example, the drug-loaded patch can be a quadrilateral, a pentagon, a hexagon, or an N-sided polygon.

[0022] For example, when the drug-loaded patch is a circle, the drug-loaded patch has one side length (i.e., the circumference), and the geometric overlap ratio of the circumference of the drug-loaded patch with the A region is less than 100% and greater than 0.

[0023] For example, when the drug-loaded patch is a semicircle, the drug-loaded patch has two side lengths (i.e., the arc and the diameter).

[0024] For example, when the drug-loaded patch is a sector, the drug-loaded patch has three side lengths (i.e., the arc and two radii).

[0025] For example, the shape of the drug-loaded patch can be adjusted according to the shape of the implantation site.

[0026] For example, the drug concentration of the liposoluble drug in the A region is 400-1200 μg / cm 2 .

[0027] For example, the drug concentration of the liposoluble drug in the A region is 410-1200 μg / cm 2 .

[0028] For example, the drug concentration of the liposoluble drug in the A region is 415.8-1194.3 μg / cm 2 .

[0029] For example, the drug concentration of the liposoluble drug in the A region is 400-1100 μg / cm 2 .

[0030] For example, the drug concentration of the liposoluble drug in the A region is 415.8-1055.2 μg / cm 2 .

[0031] Further, the decellularized matrix is derived from at least one of the small intestinal submucosa, the peritoneum, the dermis, the bladder basement membrane, and the pericardium.

[0032] For example, the decellularized matrix is derived from two of the small intestinal submucosa, the peritoneum, the dermis, the bladder basement membrane, and the pericardium.

[0033] For example, the decellularized matrix is derived from at least the bladder basement membrane.

[0034] For example, the decellularized matrix is derived from the small intestinal submucosa and the bladder basement membrane.

[0035] For example, the outermost layer of the multi-layer structure of the drug-loaded patch is a decellularized matrix layer derived from the decellularized matrix of the bladder basement membrane.

[0036] Preferably, the decellularized matrix is an uncrosslinked decellularized matrix.

[0037] In the present application, the decellularized matrix is purchased from Zhuorun Medical Technology (Suzhou) Co., Ltd., which is prepared from small intestinal submucosa, peritoneum, dermis, bladder basement membrane or pericardium according to the perfusion-differential pressure method disclosed in CN106075583B.

[0038] Generally speaking, the higher the drug loading of the biomaterial, the higher the initial concentration of the drug, the faster the release rate at the initial stage of drug release, and the longer the drug release period. However, the inventors have found through a large amount of creative labor that for the non-crosslinked decellularized matrix system of the present application which contains a fat-soluble drug, the fat-soluble drug in the A zone has an initial concentration range of 300-1300 μg / cm 2 , the overlap rate of the edge of the A zone and the edge of the patch is less than 100%, the release rate at the initial stage (4h) of release of the fat-soluble drug is always maintained within 40%, and the release period is maintained at 3-5 days for complete release. For the present scheme, if the initial concentration of the fat-soluble drug in the A zone is reduced to 300 μg / cm 2 or increased to 1300 μg / cm 2 , the release rate at the initial stage (4h) of release of the fat-soluble drug will be accelerated, the release rate will be much greater than 40%, or the release period of the fat-soluble drug will be significantly shortened. Moreover, the release rate at the initial stage (4h) of release of the fat-soluble drug and the release period are not substantially related to the drug loading, for example, the concentration of the fat-soluble drug (total drug loading / patch area) is increased from about 100 μg / cm 2 to about 600 μg / cm 2 , the technical effects of the release rate at the initial stage (4h) of release being always maintained within 40% and the release period being maintained at 3-5 days for complete release are still achieved. The above-mentioned sustained release effect and the release period of 3-5 days are particularly evident when the decellularized matrix is derived from small intestinal submucosa and bladder basement membrane.

[0039] For example, the fat-soluble drug is selected from one or more of fat-soluble vitamin drugs, hormone drugs, antibiotic drugs, and antitumor drugs.

[0040] Preferably, the fat-soluble drug is selected from one or two of fat-soluble vitamin drugs, hormone drugs, antibiotic drugs, and antitumor drugs.

[0041] For example, the fat-soluble drug is selected from one of fat-soluble vitamin drugs, hormone drugs, antibiotic drugs, and antitumor drugs.

[0042] Preferably, the fat-soluble drug is an antibiotic drug and / or an antitumor drug.

[0043] Further, the fat-soluble drug is an antibiotic drug or an antitumor drug.

[0044] Illustratively, the vitamin drug is selected from one or more of vitamin A, vitamin D, vitamin E, and vitamin K.

[0045] Illustratively, the hormone drug is a glucocorticoid and / or a sex hormone, wherein the glucocorticoid is selected from one or more of dexamethasone, methylprednisolone, betamethasone, and prednisone; and the sex hormone is selected from one or more of testosterone, dihydrotestosterone, estradiol, and progesterone.

[0046] Illustratively, the antibiotic drug is selected from one or more of rifamycin, tetracycline, a macrolide antibiotic, a quinolone antibiotic, and a nitroimidazole antibiotic.

[0047] Illustratively, the antibiotic drug is selected from rifamycin and / or tetracycline.

[0048] Illustratively, the antitumor drug is selected from one or more of paclitaxel, vincristine, lomustine, and cyclophosphamide.

[0049] The fat-soluble drug, such as rifamycin in the fat-soluble antibiotic, generally has a color (such as red, orange, yellow, etc.); or, when the fat-soluble drug layer is prepared, other pharmaceutical excipients with specific colors are added, so that the obtained drug-loaded patch has an A zone that can be more intuitively distinguished by color.

[0050] Illustratively, the drug-loaded patch is obtained by encapsulating at least two fat-soluble drugs in the decellularized matrix.

[0051] Illustratively, the drug-loaded patch is obtained by encapsulating at least two fat-soluble antibiotic drugs in the decellularized matrix.

[0052] Illustratively, the drug-loaded patch is obtained by encapsulating tetracycline and rifamycin in the decellularized matrix.

[0053] Further, if the drugs encapsulated in the patch are at least two fat-soluble drugs encapsulated in the decellularized matrix, especially two fat-soluble antibiotics of tetracycline and minocycline, the drugs in the system can be released more stably, and unexpectedly have a certain impact on the mechanical properties of the drug-loaded patch, which is manifested as the uniformity of the tensile strength of the patch being more stable.

[0054] Illustratively, the tetracycline is selected from one or more of minocycline hydrochloride, minocycline, doxycycline hydrochloride, doxycycline, and tigecycline.

[0055] Illustratively, the rifamycin is selected from one or more of rifampicin, rifapentine, and rifabutin.

[0056] Further, the B region of the drug-loaded patch is substantially free of the same fat-soluble drug loaded in the A region.

[0057] For example, when the drug-loaded patch encapsulates rifamycin only by the acellular matrix, the A region of the drug-loaded patch loads rifamycin, and the fat-soluble drug layer is a rifamycin drug layer, then the B region of the drug-loaded patch is substantially free of rifamycin, or a small amount of rifamycin is loaded in the B region at the boundary between the B region and the A region.

[0058] For example, when the drug-loaded patch encapsulates rifamycin and tetracycline by the acellular matrix, and the fat-soluble drug layer simultaneously includes a rifamycin layer and a tetracycline layer, the A region of the drug-loaded patch loads a large amount of rifamycin and part of tetracycline, then the B region of the drug-loaded patch is substantially free of rifamycin and tetracycline, or a small amount of rifamycin and tetracycline is loaded in the B region at the boundary between the B region and the A region.

[0059] For example, when the drug-loaded patch encapsulates rifamycin, tetracycline and paclitaxel by the acellular matrix, the fat-soluble drug layer simultaneously includes a rifamycin layer, a tetracycline layer and a paclitaxel layer, and the A region of the drug-loaded patch includes A1 region, A2 region and A3 region which do not overlap each other, wherein the A1 region loads rifamycin, the A2 region loads tetracycline, and the A3 region loads paclitaxel, then the B region of the drug-loaded patch is substantially free of rifamycin, tetracycline and paclitaxel, or a small amount of rifamycin, tetracycline or paclitaxel is loaded in the B region at the boundary between the B region and the A region.

[0060] For example, when the drug-loaded patch encapsulates rifamycin, tetracycline and paclitaxel by the acellular matrix, the fat-soluble drug layer simultaneously includes a rifamycin layer, a tetracycline layer and a paclitaxel layer, and the A region of the drug-loaded patch includes A1 region, A2 region and A3 region which overlap each other, wherein the A1 region loads rifamycin, the A2 region loads tetracycline, and the A3 region loads paclitaxel, then the B region of the drug-loaded patch is substantially free of rifamycin, tetracycline and paclitaxel, or a small amount of rifamycin, tetracycline or / and paclitaxel is loaded in the B region at the boundary between the B region and the A region.

[0061] For example, the fat-soluble drug is directly encapsulated into the drug-loaded patch.

[0062] For example, the fat-soluble drug is encapsulated into the drug-loaded patch in the form of a suspension of fat-soluble drug complexes.

[0063] When the fat-soluble drug is encapsulated into the drug-loaded patch in the form of a suspension of fat-soluble drug complexes, the fat-soluble drug complexes are obtained by reacting the fat-soluble drug with metal cations or liposomes.

[0064] For example, an organic solution of the fat-soluble drug is laid and / or sprayed onto the acellular matrix layer to form the fat-soluble drug layer.

[0065] The suspension of the liposoluble drug is prepared by chelating the liposoluble drug with metal cations.

[0066] The organic solution of the liposoluble drug is prepared by adding the liposoluble drug into 80wt%-95wt% aqueous ethanol solution.

[0067] Further, the concentration of the liposoluble drug in the organic solution of the liposoluble drug is 0.01-0.1g / mL.

[0068] The suspension of the liposoluble drug is prepared by chelating the liposoluble drug with metal cations.

[0069] For example, the suspension of tetracycline is prepared by chelating tetracycline with diamagnetic metal cations, wherein the diamagnetic metal cations are Ca 2+ , Mg 2+ , Ba 2+ , Sr 2+ and / or Zn 2+ . Specifically, the suspension of tetracycline is prepared by adding metal cation salt solution into tetracycline aqueous solution and adjusting pH to neutral.

[0070] For example, the suspension of paclitaxel is prepared by chelating paclitaxel with metal cations, wherein the metal cations are Cu 2+ , Pt 2+ and / or Pt 3+ . Specifically, the suspension of paclitaxel is prepared by adding metal cation salt solution into paclitaxel solution after esterification, amidation and cross-linking of paclitaxel and adjusting pH to neutral.

[0071] The drug-loaded patch of the present application is a multi-layer structure, which comprises at least a liposoluble drug layer and a decellularized matrix layer. The liposoluble drug layer is formed by spraying or / and laying the organic solution of the liposoluble drug onto the decellularized matrix layer.

[0072] That is, the preparation of the fat-soluble drug layer, in a first aspect, the fat-soluble drug organic solution or the fat-soluble drug suspension of one-time spraying can be regarded as a single-layer fat-soluble drug layer; in a second aspect, the fat-soluble drug organic solution (or the fat-soluble drug suspension) and the decellularized matrix layer on which the fat-soluble drug organic solution (or the fat-soluble drug suspension) is laid can be regarded as a fat-soluble drug layer. For example, the fat-soluble drug organic solution is laid on the decellularized matrix layer derived from the small intestinal submucosa and / or the bladder basement membrane to form a fat-soluble drug layer.

[0073] In the present application, the fat-soluble drug layer can be continuously stacked or / and discontinuously stacked, as long as the fat-soluble drug on the A area in the whole drug-loaded patch has a defined local unit concentration, that is, the drug concentration of the fat-soluble drug on the A area is 300-1300 μg / cm 2 .

[0074] Further, for the whole drug-loaded patch, the encapsulated drug amount of each fat-soluble drug of the drug-loaded patch is 130-600 μg / cm 2 .

[0075] Further, for the whole drug-loaded patch, the encapsulated drug amount of each fat-soluble drug of the drug-loaded patch is 130-600 μg / cm 2 , such as 133.4-574.2 μg / cm 2 .

[0076] Further, the ratio of the transverse tensile strength and the longitudinal tensile strength of the drug-loaded patch is 1: (0.8-1.25).

[0077] For example, when the fat-soluble drug organic solution or the fat-soluble drug suspension of one-time spraying is regarded as a single-layer fat-soluble drug layer, the preparation of the drug-loaded patch at least includes the following steps: (1) Preparation of a decellularized matrix layer: laying a decellularized matrix with water; (2) Preparation of a fat-soluble drug layer: continuing to spray n (n≥1, and n is an integer) layers of fat-soluble drug organic solution or fat-soluble drug suspension on the decellularized matrix layer obtained in step (1), so that the fat-soluble drug organic solution or the fat-soluble drug suspension is distributed in a patterned manner on the decellularized matrix layer obtained in step (1); wherein the patterned distribution refers to a distribution mode in which the sprayed bodies are arranged in a predetermined pattern by a specific method.

[0078] For example, the patterned distribution can be a regular pattern distribution and / or an irregular pattern distribution, such as a repeated pattern distribution and / or a non-repeated pattern distribution, such as a zebra pattern distribution, a circular area distribution and / or a polygonal area distribution, such as a field-shaped distribution and / or a back-shaped distribution, etc.

[0079] In an embodiment, the predetermined pattern is a gap between the at least two independent shape units, that is, the A region of the drug-loaded patch is composed of at least two independent shape units, each of which is separated by a physical gap, and the A region as a whole does not form a continuous single entity. In this embodiment, the physical gap is a gap with a width greater than 0, and the outer contour lines of each of the independent shape units do not intersect in space.

[0080] (3) molding: the decellularized matrix layer and the fat-soluble drug layer are stacked in any order and any number of layers, and it is ensured that the upper and lower two outer surfaces of the drug-loaded patch are both decellularized matrix layers, the overlapping rate of the edge of the A region and the edge of the drug-loaded patch is less than 100%, and the fat-soluble drug concentration of the A region at this time is 300-1300 μg / cm 2 ; and the drug-loaded patch is obtained after the molding process.

[0081] Generally, after molding, the total area shape presented by all the fat-soluble drug layers of the drug-loaded patch is recorded as the A region of the drug-loaded patch.

[0082] In an embodiment, when the fat-soluble drug organic solution (or suspension) and the decellularized matrix on which it is laid are regarded as a fat-soluble drug layer, the preparation method of the drug-loaded patch at least includes: (1) preparation of a decellularized matrix material layer: laying a decellularized matrix with water; (2) preparation of a fat-soluble drug layer: laying a decellularized matrix with a fat-soluble drug organic solution (or suspension), repeating and stacking n (n≥1, and n is an integer) times, and then performing a molding process to cut into an arbitrary shape of a gap unit, and one or more gap units form a fat-soluble drug layer; (3) molding: the decellularized matrix layer and the fat-soluble drug layer are stacked in any order and any number of layers, and it is ensured that the upper and lower two outer surfaces of the drug-loaded patch are both decellularized matrix layers, and the drug-loaded patch is obtained after the molding process.

[0083] In an embodiment, the gap between the gap units in step (2) is a gap between the at least two independent shape units, that is, the A region of the drug-loaded patch is composed of at least two independent shape units, each of which is separated by a physical gap, and the A region as a whole does not form a continuous single entity. In this embodiment, the physical gap is a gap with a width greater than 0, and the outer contour lines of each of the independent shape units do not intersect in space.

[0084] In step (3) of the preparation method of the drug-loaded patch, the number of layers and the stacking order of the acellular matrix layer and the fat-soluble drug layer are not limited, for example, the order from bottom to top is 1 layer of acellular matrix layer, 1 layer of fat-soluble drug layer and 1 layer of acellular matrix layer; for example, the order from bottom to top is 8 layers of acellular matrix layer, 2 layers of fat-soluble drug layer and 6 layers of acellular matrix layer; for example, the order from bottom to top is 1 layer of acellular matrix layer, 1 layer of fat-soluble drug layer, 4 layers of acellular matrix layer, 1 layer of fat-soluble drug layer and 1 layer of acellular matrix layer; for example, the order from bottom to top is 2 layers of acellular matrix layer, 2 layers of fat-soluble drug layer, 4 layers of acellular matrix layer, 1 layer of fat-soluble drug layer and 1 layer of acellular matrix layer, and the like. Although the stacking order of the acellular matrix layer and the fat-soluble drug layer is not limited, the outermost layer of the drug-loaded patch must be the acellular matrix layer.

[0085] Even if the number of layers is increased, the mechanical uniformity (the transverse tensile strength and the longitudinal tensile strength always tend to be 1:1) and the mechanical stability of the drug-loaded patch will not be reduced, and there is no risk of application due to instability of the patch caused by pressure, swelling and other environments.

[0086] The second aspect of the present application provides a drug-loaded patch for preparing an anti-pollution wound material or an anti-tumor material.

[0087] When the drug-loaded patch is applied to the preparation of an anti-pollution wound material, the drug-loaded patch is applied to a drug and instrument used in a high-infection period of surgical operation, wherein the high-infection period of surgical operation usually refers to within three days after surgical operation. Further, the drug-loaded patch is applied to the preparation of a temporary abdominal closure material for abdominal cavity opening. The drug-loaded patch of the present application is used as a temporary abdominal closure material, and does not need to be removed when a certain abdominal closure is determined. Since the fat-soluble drug of the patch has been basically released within 3-5 days, it is not necessary to consider the influence of long-term implantation of the drug on the human body.

[0088] Advantages: (1) The drug-loaded patch obtained by the present application realizes the slow release of the fat-soluble drug with initial in vivo release too fast on the non-crosslinked acellular matrix, so that the fat-soluble drug is not released too fast in the initial implantation period, but is released continuously along with the wound healing recovery process, simultaneously guides tissue regeneration and degradation, reduces the probability of adhesion, promotes the growth of granulation tissue on the wound surface, and has a technical effect of "1+1>2"; (2) Based on the specific structure that the overlapping rate of the edge of region A and the edge of the patch is less than 100%, and in cooperation with the specific local concentration of the fat-soluble drug in region A, the release rate of the fat-soluble drug in the initial release period (4h) is always maintained within 40%, the release period is maintained for 3-5 days, and the product has high stability, i.e., the drug loading amount stability, the release stability and the mechanical uniformity stability, which is beneficial to large-scale production. Attached Figure Description

[0089] Figure 1 (a) is a schematic diagram of the drug-loaded patch obtained in Example 1. Figure 1 (b) is a photograph of the drug-loaded patch obtained in Example 1; Figure 2 This is a schematic diagram of the drug-loaded patches obtained in Example 4 and Comparative Examples 1-2, wherein... Figure 2 (a) is a schematic diagram of the drug-loaded patch obtained in Comparative Example 1. Figure 2 (b) is a schematic diagram of the drug-loaded patch obtained in Comparative Example 2. Figure 2 (c) is a schematic diagram of the drug-loaded patch obtained in Example 4; Figure 3 This is a schematic diagram of the drug-loaded patch obtained in Examples 5-6. Figure 3 (a) is a schematic diagram of the drug-loaded patch obtained in Example 5. Figure 3 (b) is a schematic diagram of the drug-loaded patch obtained in Example 6; Figure 4 This is a schematic diagram of the drug-loaded patch obtained in Comparative Example 4. Detailed Implementation

[0090] In this invention, all raw materials used are commercially available, and the decellularized matrix is ​​from Zhuoran Medical Technology (Suzhou) Co., Ltd. It is prepared from the submucosa of the small intestine, peritoneum, dermis, bladder basement membrane or pericardium according to the perfusion-pressure difference method described in patent CN106075583B.

[0091] Example 1 This example provides a drug-loaded patch, which is obtained by encapsulating a lipid-soluble rifamycin antibiotic (specifically rifampin) in a decellularized matrix. Figure 1 Drug-loaded patches include region A ( Figure 1 (a) dark area) and area B ( Figure 1 (a) The light-colored part). The drug-loaded patch is a square with a side length of 10 cm. The drug-loaded patch has a multi-layer structure, which includes 4 layers of lipid-soluble drug and 5 layers of decellularized matrix.

[0092] The preparation method of the drug-loaded patch is as follows: (1) Preparation of decellularized matrix layer 1: Decellularized matrix derived from bladder basement membrane was laid with water; (2) Preparation of the liposoluble drug layer 1 (regarding the organic solution of rifampicin sprayed at one time as a single layer of liposoluble drug layer): 2 layers of organic solution of rifampicin were continuously sprayed on the decellularized matrix layer 1 obtained in step (1), so that the organic solution of rifampicin was distributed in a zebra pattern (a total of 10 strips) on the decellularized matrix layer 1 obtained in step (1), each strip had a length L of 0.25 cm in contact with the edge of the patch, and each strip had the same direction, an angle of 45° with the horizontal direction, and was parallel to each other, and there was a gap between each strip.

[0093] (3) Preparation of the liposoluble drug layer 2 (regarding the organic solution of rifampicin sprayed at one time as a single layer of liposoluble drug layer): 2 layers of organic solution of rifampicin were continuously sprayed on the liposoluble drug layer 1 obtained in step (2), so that the organic solution of rifampicin was distributed in a zebra pattern (a total of 10 strips) on the decellularized matrix layer 1 obtained in step (1), each strip had a length L of 0.25 cm in contact with the edge of the patch, and each strip had the same direction, an angle of 45° with the horizontal direction, and was parallel to each other, and there was a gap between each strip.

[0094] The angle between the strip direction of the liposoluble drug layer 2 and the strip direction of the liposoluble drug layer 1 was 45°.

[0095] The organic solution of rifampicin was prepared by adding rifampicin to 91wt% ethanol aqueous solution, and the concentration of rifampicin in the organic solution of rifampicin was 0.01g / mL. (4) Forming: 3 layers of decellularized matrix layer 1, 1 layer of liposoluble drug layer 1, 1 layer of liposoluble drug layer 2 and 2 layers of decellularized matrix layer 1 were sequentially stacked from bottom to top, and then laminated to obtain the drug-loaded patch.

[0096] Example 2

[0097] This example provides a drug-loaded patch, which is different from example 1 in that the drug-loaded patch is obtained by encapsulating a liposoluble antitumor drug (specifically paclitaxel) with a decellularized matrix. In the preparation process, the preparation of the liposoluble drug layer 1 and the liposoluble drug layer 2 is obtained by spraying the organic solution of paclitaxel.

[0098] The organic solution of paclitaxel was prepared by adding paclitaxel to 91wt% ethanol aqueous solution. The concentration of paclitaxel in the organic solution of paclitaxel was 0.01g / mL.

[0099] Example 3

[0100] This example provides a drug-loaded patch, which is different from example 1 in that the drug-loaded patch is obtained by encapsulating a liposoluble antibiotic (specifically minocycline hydrochloride) with a decellularized matrix. In the preparation process, the preparation of the liposoluble drug layer 1 and the liposoluble drug layer 2 is obtained by spraying the suspension of minocycline hydrochloride.

[0101] Minocycline hydrochloride suspension was prepared by adding 85 μL of 0.5 g / mL calcium chloride aqueous solution into 10 mL of 0.01 g / mL minocycline hydrochloride aqueous solution (Ca 2+ is the antimagnetic metal cation), and adjusting the pH to 7.4 with 0.1 mol / L sodium hydroxide aqueous solution (i.e. the suspension of tetracycline antibiotics is prepared by adding minocycline hydrochloride and adding Ca 2+ in a molar mass ratio of 1:1.75).

[0102] Example 4

[0103] This example provides a drug-loaded patch, which is obtained by encapsulating a fat-soluble rifamycin antibiotic (specifically, rifampicin) with a decellularized matrix. As shown in Figure 2 (c), the drug-loaded patch includes an A region (dark part of Figure 2 (c)) and a B region (light part of Figure 2 (c)), and the overlap between the edge of the A region and the edge of the drug-loaded patch is 15%. The drug-loaded patch is a square with a side length of 5 cm.

[0104] The preparation method of the drug-loaded patch is as follows: (1) Preparation of the decellularized matrix layer 1: use water to lay the bladder basement membrane-derived decellularized matrix; (2) Preparation of the fat-soluble drug layer (the entire small intestine submucosa-derived decellularized matrix laid with rifampicin organic solution is regarded as the fat-soluble drug layer): use rifampicin organic solution to lay the small intestine submucosa-derived decellularized matrix, repeat stacking 4 times, and then cut into square-shaped interval units with a side length of 1 cm after forming (specifically, laminating), and 3 interval units form 1 layer of fat-soluble drug layer; wherein the rifampicin organic solution is prepared by adding rifampicin into 91 wt% ethanol aqueous solution, and the concentration of rifampicin in the rifampicin organic solution is 0.03 g / mL; (3) Forming: stack 3 layers of decellularized matrix layer 1, 1 layer of fat-soluble drug layer, and 2 layers of decellularized matrix layer 1 from bottom to top, and then laminate to obtain the drug-loaded patch.

[0105] This example is similar to the preparation method of Example 1 as recorded in the patent specification of CN107320786B, and the interval units of the fat-soluble drug layer are arranged between the decellularized matrix layers at an interval of the radius of the inhibition zone, and the overlap around the inhibition zone ensures that there is no omission on the surface of the patch, as shown in 2(c).

[0106] Example 5

[0107] This example provides a drug-loaded patch, which is different from Example 1 in that the overlap rate between the edge of the A region of the drug-loaded patch and the edge of the patch is 50%, as shown inFigure 3 (a), the drug-loaded patch has different A areas (b), dark regions) and B areas (c), light regions) distribution. The drug-loaded patch is a square with a side length of 10 cm, and the lipid-soluble drug layer has 4 strips, each strip has an overlapping length of d1=2.5 cm with the edge of the patch. Figure 3 Figure 3 (a), the drug-loaded patch has different A areas (b), dark regions) and B areas (c), light regions) distribution. The drug-loaded patch is a square with a side length of 10 cm, and the lipid-soluble drug layer has 4 strips, each strip has an overlapping length of d1=2.5 cm with the edge of the patch.

[0108] Specifically, the preparation method of the lipid-soluble drug layer is as follows: continue to spray 4 layers of rifampicin organic solution on the decellularized matrix layer 1 obtained in step (1), so that the rifampicin organic solution presents a zebra pattern distribution (a total of 4 strips) on the decellularized matrix layer 1 obtained in step (1), each strip has a length d1=2.5 cm in contact with the edge of the patch, and each strip has the same direction, the angle with the horizontal direction is 45°, and they are parallel to each other, and there is a gap between each strip.

[0109] Example 6

[0110] This example provides a drug-loaded patch, which is different from example 4 in that the overlapping rate of the A area edge of the drug-loaded patch with the edge of the patch is 86%, as shown in Figure 3 (a), the drug-loaded patch has different A areas (b), dark regions) and B areas (c), light regions) distribution. The drug-loaded patch is a square with a side length of 10 cm, and the lipid-soluble drug layer has 4 strips, each strip has an overlapping length of d1=2.5 cm with the edge of the patch. Figure 3 Figure 3 (a), the drug-loaded patch has different A areas (b), dark regions) and B areas (c), light regions) distribution. The drug-loaded patch is a square with a side length of 10 cm, and the lipid-soluble drug layer has 4 strips, each strip has an overlapping length of d1=2.5 cm with the edge of the patch. Figure 3 (b), d2=0.25 cm.

[0111] Example 7

[0112] This example provides a drug-loaded patch, which is different from example 1 in that the decellularized matrix is derived from bladder basement membrane and small intestinal submucosa.

[0113] The preparation method of the drug-loaded patch is as follows: (1) Preparation of decellularized matrix layer 1: lay the decellularized matrix derived from bladder basement membrane with water; (2) Preparation of decellularized matrix layer 2: lay the decellularized matrix derived from small intestinal submucosa with water; (3) Preparation of lipid-soluble drug layer 1 and lipid-soluble drug layer 2: same as example 1.

[0114] (4) Forming: stack 1 layer of decellularized matrix layer 1, 2 layers of decellularized matrix layer 2, 1 layer of lipid-soluble drug layer 1, 1 layer of decellularized matrix layer 2 and 1 layer of decellularized matrix layer 1 from bottom to top, and then laminate to obtain

[0115] Example 8​​

[0116] This example provides a drug-loaded patch that differs from Example 5 in that the total number of layers in the drug-loaded patch is increased.

[0117] The specific preparation method is as follows: after preparing decellularized matrix layer 1, lipid-soluble drug layer 1, and lipid-soluble drug layer 2 according to the method in Example 5, 7 layers of decellularized matrix layer 1, 1 layer of lipid-soluble drug layer, 1 layer of decellularized matrix layer 1, 1 layer of lipid-soluble drug layer 2, 4 layers of decellularized matrix layer 1, 1 layer of lipid-soluble drug layer 2, and 2 layers of decellularized matrix layer 1 are stacked sequentially from bottom to top, and then laminated to obtain the final product.

[0118] The system consists of three layers of decellularized matrix layer 1, one layer of lipid-soluble drug layer 1, one layer of lipid-soluble drug layer 2, and two layers of decellularized matrix layer 1. Comparative Example 1 This example provides a drug-loaded patch, which differs from Example 4 in that, Figure 2 (a) Drug-loaded patches have different A regions ( Figure 2 (a) Dark area) and area B ( Figure 2 (a) Light-colored area) distribution, the overlap rate between the edge of area A and the edge of the patch is 0. The lipid-soluble drug layer has two square-shaped spacer units with a side length of 1 cm.

[0119] Comparative Example 2 This example provides a drug-loaded patch, which differs from Example 4 in that, Figure 2 (b) Drug-loaded patches have different A regions ( Figure 2 (b) Dark areas) and Area B ( Figure 2 (b) Light-colored area distribution, the overlap rate between the edge of area A and the edge of the patch is 5%, such as Figure 2 (b).

[0120] Comparative Example 3 This example provides a drug-loaded patch, which differs from Example 1 in that the overlap rate between the edge of region A and the edge of the patch is 100% (the overlap rate between the outermost edge of the U-shaped strip and the edge of the patch is 100%). Figure 4 Drug-loaded patches have different A regions ( Figure 4 Dark area) and area B ( Figure 4 The light-colored area is distributed (equivalent to the rifampicin organic solution showing a zigzag pattern in the decellularized matrix layer 1 when preparing the lipid-soluble drug layer), wherein the width w of the strip is 0.5cm, the interval between each zigzag strip is 0.5cm, and the whole patch is a square with a side length of 5cm.

[0121] Comparative Example 4 This example provides a drug-loaded patch, which differs from Example 1 in that the decellularized matrix is ​​derived from the submucosa of the small intestine.

[0122] Specifically, the preparation method of the drug-loaded patch is as follows: (1) Preparation of the acellular matrix layer 1: use water to lay the acellular matrix derived from the small intestinal submucosa; (2) Preparation of the lipid-soluble drug layer 1 and the lipid-soluble drug layer 2: same as in Example 1.

[0123] (3) Forming: sequentially stack the 3 layers of the acellular matrix layer 1, the 1 layer of the lipid-soluble drug layer 1, the 1 layer of the lipid-soluble drug layer 2, and the 2 layers of the acellular matrix layer 1 from bottom to top, and then laminate, and the drug-loaded patch is obtained.

[0124] Comparative Example 5 This example provides a drug-loaded patch, which is different from Example 1 in that the concentration of the lipid-soluble drug in the A region of the drug-loaded patch is much greater than 1300 μg / cm 2 .

[0125] In this example, the rifampicin organic solution is prepared by adding rifampicin into 95 wt% ethanol aqueous solution, and the concentration of rifampicin in the rifampicin organic solution is 0.25 g / mL.

[0126] Comparative Example 6 This example provides a drug-loaded patch, which is different from Example 5 in that the concentration of the lipid-soluble drug in the A region of the drug-loaded patch is much lower than 400 μg / cm 2 .

[0127] In this example, the rifampicin organic solution is prepared by adding rifampicin into 90 wt% ethanol aqueous solution, and the concentration of rifampicin in the rifampicin organic solution is 0.005 g / mL.

[0128] Performance test 1. Overlapping rate and local drug concentration test: the overlapping rate of the A region edge and the patch edge can be obtained by measuring the length of the sample according to the formula "length of the overlapping part of the A region edge and the patch edge / patch circumference x 100%"; after the A region of the patch is cut, the total area of the A region of each patch sample is measured and calculated, and then the sample is immersed in methanol, the extraction liquid is taken every 24 h for sample testing by liquid chromatography, until the extraction liquid is colorless, and then the sample testing is stopped, the sample drug content is obtained by referring to the chromatogram of each drug standard, and the local drug concentration of the lipid-soluble drug in the A region = sample drug content / A region total area, and the results are shown in Table 1: Table 1

[0129] 2. Patch drug content test: The sample biological patch was immersed in methanol, and the extraction liquid was taken every 24 h for liquid chromatography sample test until the extraction liquid was colorless. The drug content of the sample was obtained by referring to the chromatogram of the standard drug, and the average deviation was recorded after repeating five times. The results are shown in Table 2.

[0130] Table 2

[0131] From Tables 1-2, it can be seen from Examples 1-8 and Comparative Examples 1-2 that when the overlap rate of the edge of the A area and the edge of the patch is within 90%, and the local drug concentration of the fat-soluble drug in the A area is 300-1300 μg / cm 2 , and more preferably 400-1200 μg / cm 2 , and especially 415.8-1194.3 μg / cm 2 , the average deviation of the drug loading of the fat-soluble drug in the patch is maintained at a low level (within 20%, and especially within 15%), and each antibiotic encapsulated by the antibacterial sustained-release patch has good drug loading stability, which is suitable for large-scale production.

[0132] Compared with Example 1 using only bladder basement membrane-derived decellularized matrix, the decellularized matrix of Example 7 includes not only bladder basement membrane but also small intestinal submucosa. From Tables 1-2, it can be seen that the drug loading and average deviation of the fat-soluble drug in the patch of Example 1 and Example 7 are similar. However, since the cost of bladder basement membrane is relatively high, the decellularized matrix derived from bladder basement membrane and small intestinal submucosa can be used, which is more cost-effective and more suitable for large-scale production.

[0133] Further, from Example 5 and Example 8, it can be seen that for the present technology, even if the number of layers is increased, the obtained patch product still has good drug content stability.

[0134] From Comparative Example 1 and Example 4, it can be seen that, when the acellular matrix is loaded with the liposoluble drug, if only the scheme of "spaced between the layers of biomaterials according to the radius of the inhibition zone" is implemented, without considering the local drug concentration range of the A zone liposoluble drug and the edge overlap rate, although the stability of the drug loading amount of the liposoluble drug is relatively low, it is also decreased by 34% compared to Example 4 which limits the local drug concentration range of the A zone liposoluble drug and the overlap rate of the A zone edge and the edge of the patch, which shows that when the acellular matrix is loaded with the liposoluble drug, the scheme of "spaced between the layers of biomaterials according to the radius of the inhibition zone" cannot produce beneficial effects on the stability of the drug loading amount without considering the local drug concentration of the liposoluble drug and its specific structural setting. It can be seen that the local drug concentration of the liposoluble drug and its specific structural setting are the key to the stability of the drug loading amount of the acellular matrix encapsulating the liposoluble drug.

[0135] 3. Animal experiment: After the rats were given gas anesthesia, the abdominal hair was shaved, a 5 cm long incision was made in the middle of the abdomen to fully expose the abdominal cavity, and the sample patch was placed on the abdominal wall of the rats with the same sample on both sides of the abdominal cavity and fixed with sutures. After the implantation was completed, the wound was closed. At 2h, 4h, 1 day, and 3 days after the operation, samples were taken, and after euthanasia, the abdominal cavity was opened, the morphology of the patch implantation site (including adhesion and granulation tissue growth) was observed, the sample was taken out, and the sample was extracted with PBS buffer solution for 24h. According to the drug standard curve, the drug loading amount was calculated and recorded as W (the standard of the drug was dissolved in PBS buffer solution to prepare a series of standard solutions of 1-500µg / mL, and the standard curve was obtained by liquid chromatography sample testing), the in vivo release rate = (the drug loading amount of each sample in performance test 1-W) / the drug loading amount of each sample in performance test 1 x 100%, and the results are shown in Tables 3-4.

[0136] Table 3

[0137] * " / " in Table 3 represents that the drug cannot be detected.

[0138] Table 4

[0139] Combining Tables 1-4, it can be seen from Examples 1-8 that when the overlap rate of the A zone edge and the edge of the patch is within 90%, and further within the range of 15%≤overlap rate≤86%, and the local drug concentration of the A zone liposoluble drug is 300-1300µg / cm 2 , and further, the local drug concentration is 400-1100µg / cm 2 , and especially 415.8-1055.2µg / cm 2The drug loading amount does not need to be considered, and the shape, pattern and interval distance of the A area do not need to be specially limited, so that the sustained release of the liposoluble drug loaded on the acellular matrix can be achieved, the release rate of the liposoluble drug in the initial release period (4h) is always maintained within 40%, and the drug is basically released at 72h (>90%), and the product has high stability, that is, the drug loading amount stability and release stability, and is especially suitable for batch large-scale production.

[0140] Compared with Example 1, the overlap rate of the A area edge of the patch of Comparative Example 3 and the patch edge is >90%, and the in vivo release rate of the liposoluble drug at 2h and 4h is significantly accelerated, which will increase the risk of bacterial infection during the temporary abdominal closure period (generally 3-5 days) of open abdominal surgery. If the local concentration of rifamycin in the A area is too high, such as Comparative Example 5, not only does the drug loading amount stability or release stability decrease significantly, but also the antibiotic release is slow, which will also cause a considerable amount of antibiotic to remain locally in the abdominal cavity during the definitive abdominal closure, which may have adverse effects on the human body and increase the risk of drug resistance. Further, granulation tissue growth has not been observed in Comparative Example 5 at all, which indicates that the function of the acellular matrix to promote tissue growth is inhibited.

[0141] Comparative Examples 1-2 are only implemented according to the scheme of “placing rifamycin at a distance of the antibacterial circle radius between the layers of biological materials, and the surface of the patch is not missed”, without considering and having the specific overlap rate of the A area edge and the patch edge of the present technology. It can be seen that the drug release period is too long, and further, granulation tissue growth has not been observed in Comparative Examples 1-2 at all, which indicates that the function of the acellular matrix to promote tissue growth is inhibited. In contrast to Comparative Examples 1-2, the patch of Example 4 has the characteristics that the release rate of the liposoluble drug in the initial release period (4h) is always maintained within 40%, and the release period is maintained at 3-5 days, and granulation tissue growth is observed.

Claims

1. A drug-loaded patch, characterized in that, The drug-loaded patch is obtained by encapsulating a fat-soluble drug in a decellularized matrix; The drug-loaded patch comprises at least two regions, namely region A and region B, wherein the overlapping rate of the edge of region A with the edge of the patch is ≤ 90%; The A zone is loaded with at least a fat-soluble drug, and the concentration of any fat-soluble drug on the A zone is 300-1300 μg / cm 2 ; The drug-loaded patch is a multi-layer structure, and the multi-layer structure comprises a fat-soluble drug layer and at least two decellularized matrix layers.

2. The medicated patch of claim 2, wherein, The overlapping rate of the edge of region A with the edge of the patch is ≥ 0.

3. The medicated patch of claim 1 wherein, The geometric overlapping rate of region A with the edge of the patch is < 100% on any side length of the patch.

4. The medicated patch of claim 1 wherein, The fat-soluble drug is selected from one or more of the following: a fat-soluble vitamin drug, a hormone drug, an antibiotic drug, and an antitumor drug.

5. The medicated patch of claim 4 wherein, The vitamin drug is selected from one or more of the following: vitamin A, vitamin D, vitamin E, and vitamin K.

6. The medicated patch of claim 4, wherein the medicated patch is characterized by, The hormone drug is a glucocorticoid and / or a sex hormone, wherein the glucocorticoid is selected from one or more of the following: dexamethasone, methylprednisolone, betamethasone, and prednisone; and the sex hormone is selected from one or more of the following: testosterone, dihydrotestosterone, estradiol, and progesterone.

7. The medicated patch of claim 4 wherein, The antibiotic drug is selected from one or more of the following: rifamycin, tetracycline, a macrolide antibiotic, a quinolone antibiotic, and a nitroimidazole antibiotic.

8. The medicated patch of claim 4 wherein, The antitumor drug is selected from one or more of the following: paclitaxel, vincristine, lomustine, and cyclophosphamide.

9. The medicated patch of claim 1 wherein, The fat-soluble drug is encapsulated in the drug-loaded patch in the form of a suspension of a fat-soluble drug complex.

10. The medicated patch of claim 9, wherein, When the fat-soluble drug is encapsulated in the drug-loaded patch in the form of a suspension of a fat-soluble drug complex, the fat-soluble drug complex is obtained by reacting the fat-soluble drug with a metal cation or a liposome.

11. The medicated patch of claim 1 wherein, The B region of the drug-loaded patch is substantially free of the same fat-soluble drug loaded in the A region.

12. The medicated patch of claim 1 wherein, The decellularized matrix is derived from at least one of the following: small intestinal submucosa, peritoneum, dermis, bladder basement membrane, and pericardium.

13. The medicated patch of claim 12 wherein, The decellularized matrix comprises at least a bladder basement membrane-derived decellularized matrix.

14. The medicated patch of claim 13, wherein, The outermost layer in the multi-layer structure of the drug-loaded patch is a decellularized matrix layer derived from a bladder basement membrane decellularized matrix.

15. The medicated patch of claim 1 wherein, An organic solution of the fat-soluble drug is laid and / or sprayed onto a decellularized matrix layer to form a fat-soluble drug layer.

16. The medicated patch of claim 10 wherein, A suspension of a fat-soluble drug complex is laid and / or sprayed onto a decellularized matrix layer to form a fat-soluble drug layer.

17. The medicated patch of claim 1 wherein, The fat-soluble drug layer is continuously or discontinuously stacked.

18. The medicated patch of claim 1, wherein, The encapsulated drug amount of any liposoluble drug of the drug-loaded patch is 130-600 μg / cm 2 .

19. Use of the drug-loaded patch according to claim 1 in the preparation of an anti-pollution wound dressing material or an antitumor material.

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