Bone-targeted photo-thermal bionic nano-carrier and preparation method thereof

Abstract

The invention discloses a bone-targeted photo-thermal bionic nano-carrier and a preparation method thereof, bone-targeted ligand small molecules Aen in an anti-tumor drug carrier are introduced into an osteoporosis drug carrier to prepare the osteoporosis drug carrier with bone targeting, so that early release of drugs before reaching bone tissues is reduced, and the bioavailability of the drugs is improved. Black phosphorus (BPs) nanoparticles with good photo-thermal property, biocompatibility and biodegradability are used as raw materials, chitosan and other macromolecules are used for carrying out functional modification on the surfaces of the black phosphorus (BPs) nanoparticles, a bone promoting drug alendronate sodium (Aen) is loaded, drug-loaded black phosphorus nanoparticles (Aen at BPs) are formed, an erythrocyte membrane (RBM) is used for carrying out bionic camouflage on the Aen at BPs, a bone targeting ligand Aen is modified to the erythrocyte membrane, and the drug-loaded black phosphorus nanoparticles (Aen at BPs) are obtained. The prepared bone-targeted photo-thermal bionic nano-carrier is combined with photo-thermal and medicine synergistic treatment, so that the anti-osteoporosis treatment effect is improved.

Description

Technical Field

[0001] The present invention belongs to the field of nano drug carriers, and particularly relates to a bone-targeted photothermal biomimetic nano carrier and a preparation method thereof. Background Art

[0002] Osteoporosis is a degenerative disease characterized by a decrease in bone mass, deterioration of bone microstructure, and increased bone brittleness, leading to increased susceptibility to fractures. The risk of developing the disease increases with age. With the increasing life expectancy and the advent of an aging society, osteoporosis has become a significant health concern.

[0003] Currently, clinically used drugs for the treatment of osteoporosis primarily include hormonal drugs, estrogen receptor modulators, and aminobisphosphonates. The main drawbacks of hormonal drugs such as estradiol, androgens, and parathyroid hormone are the need for long-term administration, high drug costs, and the associated imbalances of other endocrine systems. Estrogen receptor modulators can strongly stimulate other tissues and organs and have the side effect of inducing thrombosis. Aminobisphosphonates, such as alendronate sodium (Alen), have numerous side effects, particularly gastrointestinal ulcers, esophageal irritation, diarrhea, or constipation, and their oral bioavailability is extremely low, often less than 1%. Therefore, the development of osteopromoting drug delivery systems is crucial for enhancing drug affinity for bone tissue and improving bioavailability. Summary of the Invention

[0004] The purpose of the present invention is to address the deficiencies of the existing technology and provide a bone-targeted photothermal biomimetic nanocarrier with good photothermal properties and high biocompatibility, which combines photothermal and drug synergistic treatment to improve the effect of anti-osteoporosis treatment.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] A method for preparing a bone-targeted photothermal biomimetic nanocarrier comprises the following steps:

[0007] Preparation of S1 drug-loaded black phosphorus nanoparticles (Alen@BPs)

[0008] BPs nanoparticles (50-200 nm) are prepared by mechanical or solvent exfoliation. Alkene is then loaded onto the BPs nanoparticle surface via electrostatic adsorption to create drug-loaded black phosphorus nanoparticles. The BPs particle surface can be modified with polymers such as PEG and chitosan to modulate its interaction with the Alkene drug molecules, thereby regulating drug loading and release.

[0009] S2 Synthesis of DSPE-Alen Distearoylphosphatidylethanolamine modified active ester (DSPE-NHS) was reacted with Alen to synthesize distearoylphosphatidylethanolamine (DSPE-Alen) with a terminal bone targeting ligand molecule Alen.

[0010] Preparation of S3 bone-targeted photothermal biomimetic nanocarriers A certain amount of red blood cell membrane vesicles were mixed with DSPE-Alen and sonicated to prepare Alen-modified red blood cell membrane vesicles, which were then sonicated with a certain amount of Alen@BPs dispersion. After ultrafiltration, washing, and freeze-drying, the bone-targeted photothermal biomimetic nanocarriers Alen@BPs@RBM-Alen were obtained.

[0011] Compared with the prior art, the present invention has the following beneficial effects:

[0012] (1) The present invention uses the diphosphonate drug Alen as a bone-targeting ligand to modify the carrier surface to enhance the affinity between the drug and bone tissue;

[0013] (2) The present invention uses red blood cell membranes to coat the nanocarriers, preventing premature drug release, prolonging the in vivo circulation time of the nanocarriers, and improving bone targeting;

[0014] (3) The present invention uses black phosphorus (BPs) as a nanocarrier and photothermal agent, combines photothermal and drug synergistic treatment, and enhances the therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The present invention will be further described below with reference to the accompanying drawings and examples.

[0016] Figure 1 The preparation process of bone-targeted photothermal biomimetic nanocarriers is shown. DETAILED DESCRIPTION

[0017] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments. These drawings are simplified schematic diagrams that only illustrate the basic concept of the present invention in a schematic manner, and therefore only show structures related to the present invention.

[0018] Unless otherwise specified, all reagents and materials used in the examples of the present invention were commercially available. 1. Specific embodiments

[0020] A method for preparing a bone-targeted photothermal biomimetic nanocarrier comprises the following steps:

[0021] A method for preparing a bone-targeted photothermal biomimetic nanocarrier comprises the following steps:

[0022] Preparation of S1 drug-loaded black phosphorus nanoparticles (Alen@BPs)

[0023] BPs nanoparticles (50-200 nm) are prepared by mechanical or solvent exfoliation. Alkene is then loaded onto the BPs nanoparticle surface via electrostatic adsorption to create drug-loaded black phosphorus nanoparticles. The BPs particle surface can be modified with polymers such as PEG and chitosan to modulate its interaction with the Alkene drug molecules, thereby regulating drug loading and release.

[0024] S2 Synthesis of DSPE-Alen Distearoylphosphatidylethanolamine modified active ester (DSPE-NHS) was reacted with Alen to synthesize distearoylphosphatidylethanolamine (DSPE-Alen) with a terminal bone targeting ligand molecule Alen.

[0025] Preparation of S3 bone-targeted photothermal biomimetic nanocarriers A certain amount of red blood cell membrane vesicles were mixed with DSPE-Alen and sonicated to prepare Alen-modified red blood cell membrane vesicles, which were then sonicated with a certain amount of Alen@BPs dispersion. After ultrafiltration, washing, and freeze-drying, the bone-targeted photothermal biomimetic nanocarriers Alen@BPs@RBM-Alen were obtained.

[0026] 2. Structure and Performance Characterization of Bone-Targeted Photothermal Biomimetic Nanocarriers

[0027] The properties of nanocarriers primarily include their chemical composition and distribution, particle size, zeta potential, particle morphology and structure, drug loading capacity, drug release rate, photothermal performance, stability, and in vitro degradation. The chemical structure and interactions of the nanocarrier components were determined using FT-IR, 1H-NMR, XRD, and electron diffraction. The content and distribution of each component were determined using elemental analysis, thermogravimetric analysis, and transmission electron microscopy combined with ultrathin sectioning and staining techniques. The particle size distribution and zeta potential of the nanocarriers were determined using a Malvern Zetasizer. The morphology and size of the nanocarriers were characterized using scanning electron microscopy and transmission electron microscopy. The drug encapsulation and release behaviors of the nanocarriers were investigated using fluorescence analysis, ultraviolet spectrophotometry, and high-performance liquid chromatography. A certain mass of nanocarriers with different formulations was immersed in PBS, culture medium, or simulated body fluids. The mass change before and after immersion was measured at different time points (1 week, 2 weeks, 1 month, 2 months, and 4 months). The structural morphology of the nanocarriers was observed using scanning electron microscopy and transmission electron microscopy, and the stability and in vitro degradation rate of the nanocarriers were investigated.

[0028] 3. In vitro performance study of bone-targeted photothermal biomimetic nanocarriers

[0029] The biocompatibility, in vitro cellular uptake, and intracellular transport of the nanocarriers were primarily tested through in vitro cell experiments. The nanocarriers were co-cultured with hepatocytes, fibroblasts, and osteoblasts, and cell viability and proliferation were assessed using a CCK-8 assay to evaluate the biocompatibility of the nanocarriers. Laser confocal scanning microscopy (CLSM) and fluorescence spectroscopy were used to investigate the nanocarriers' in vitro cellular uptake, intracellular localization and distribution, bone tissue targeting, and inhibition of alkaline phosphatase (ALP) activity, to preliminarily evaluate the effectiveness of the nanosystem in treating osteoporosis.

[0030] IV. In vivo performance study of bone-targeted photothermal biomimetic nanocarriers

[0031] An osteoporosis animal model was established, and experimental and control groups were set up to receive medication respectively. The in vivo bone targeting ability of the nanocarrier and its in vivo distribution were investigated by fluorescence tracing. The tissue toxicity and biocompatibility of the nanocarrier were investigated by tissue sectioning, HE staining, and tissue immunoassay. The changes in bone density and urine calcium-creatinine ratio (U-Ca / Cr) were measured by micro-CT and kits to evaluate the safety and efficacy of the nanocarrier for the treatment of osteoporosis.

[0032] The above description is based on the ideal embodiment of the present invention. Based on the above description, relevant personnel can make various changes and modifications without departing from the technical scope of this invention. The technical scope of this invention is not limited to the content of the specification and must be determined according to the scope of the claims.

Claims

1. A method for preparing a bone-targeted photothermal biomimetic nanocarrier, comprising the following steps: Preparation of S1 drug-loaded black phosphorus nanoparticles (Alen@BPs) BPs nanoparticles with a size of 50 to 200 nm were prepared by mechanical or solvent exfoliation, and Alen was loaded on the surface of BPs nanoparticles by electrostatic adsorption to prepare drug-loaded black phosphorus nanoparticles. The surface of BPs particles can be modified by polymers such as PEG and chitosan to regulate their interaction with Alen drug molecules, thereby regulating their drug loading and release behaviors. Synthesis of S2 DSPE-Alen Distearoylphosphatidylethanolamine modified active ester (DSPE-NHS) was reacted with Alen to synthesize distearoylphosphatidylethanolamine with a terminal bone targeting ligand molecule Alen (DSPE-Alen). Preparation of S3 bone-targeted photothermal biomimetic nanocarrier A certain amount of red blood cell membrane vesicles were mixed with DSPE-Alen and ultrasonicated to prepare Alen-modified red blood cell membrane vesicles, which were then ultrasonicated with a certain amount of Alen@BPs dispersion. After ultrafiltration, washing and freeze-drying, the bone-targeted photothermal biomimetic nanocarrier Alen@BPs@RBM-Alen was obtained.

2. The method for preparing a bone-targeted photothermal biomimetic nanocarrier according to claim 1, characterized in that: Alen acts as both a drug and a targeting ligand, which can target bone tissue and provide treatment.

3. The method for preparing a bone-targeted photothermal biomimetic nanocarrier according to claim 1, characterized in that: BPs act as both nanocarriers and photothermal agents and can be naturally degraded into non-toxic PO4 in a physiological environment. 3- , which can serve as an essential bone component.

4. A nanocarrier prepared according to the method for preparing a bone-targeted photothermal biomimetic nanocarrier according to any one of claims 1 to 3.

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