Preparation method of nano preparation drug based on polylipoic acid and nano preparation drug

Abstract

The method comprises the following steps: dissolving lipoic acid and a drug in an organic solvent to form a homogeneous solution, dropwise adding deionized water while stirring, then adding a reducing agent, initiating lipoic acid polymerization and drug co-assembly in situ at 20-40 DEG C to form a drug-loaded micelle solution, and carrying out freeze drying on the drug-loaded micelle solution to obtain the nano-preparation drug based on polylipoic acid. And dialyzing and purifying to obtain the nano preparation medicine based on polylipoic acid. The preparation method is characterized in that lipoic acid has dual characteristics of a drug carrier matrix and a dynamic crosslinking monomer, carrier synthesis and efficient drug encapsulation are synchronously realized through one-step in-situ polymerization-co-assembly, and the technical bottleneck that a traditional nano-carrier needs pre-synthesis and step-by-step drug loading is broken through. The method has the following practicability: (1) the process is simple and efficient, complex equipment is not needed, and reaction conditions are mild; (2) the drug loading performance is excellent, and the particle size is uniform and adjustable (50-300 nanometers); (3) a polylipoic acid skeleton endows the nano-drug preparation with environmental responsiveness, and intelligent drug release can be realized; and (4) the method is suitable for hydrophobic drugs and compound preparations. The method provides a high-load, high-stability and stimuli-responsive nanocrystallization solution for hydrophobic drug delivery, and has application value in the field of nano-medicines.

Description

Technical Field

[0001] This invention relates to the field of nanomedicine delivery technology, specifically to a method for preparing a nanomedicine formulation based on polythioctic acid and the nanomedicine formulation itself, which is used to improve the solubility, stability and delivery efficiency of the drug. Background Technology

[0002] Nanoparticle drug delivery systems are a key strategy for improving the bioavailability of poorly soluble drugs, achieving targeted therapy, and reducing toxic side effects. Among these, polymer micellar carrier technology is particularly noteworthy. However, existing technologies still face many challenges: traditional carrier materials (such as polylactic acid and liposomes) often require complex synthesis and purification steps; physical encapsulation methods have low drug loading efficiency, especially for poorly water-soluble natural active ingredients (such as curcumin and silymarin); preparation processes often rely on highly toxic organic solvents (such as chloroform) or high-energy emulsification methods, easily leading to drug inactivation and residual solvent risks; insufficient carrier structural stability may lead to drug burst release. Although lipoic acid possesses excellent biocompatibility and antioxidant properties, its inherent self-assembly characteristics and the potential for dynamic cross-linking of intermolecular disulfide bonds are still insufficient for systematic development in the field of drug co-delivery. Therefore, there is an urgent need to develop a novel nanoparticle formulation platform technology with simple and efficient processes, excellent drug loading performance, mild and controllable solvents, and stable and intelligent carriers. This invention is an innovative solution proposed to address this technological gap. Summary of the Invention

[0003] To address the aforementioned problems, this invention provides a method for preparing a nano-formulation drug based on polythioctic acid and the nano-formulation drug itself. The method involves dissolving thioctic acid and a drug in an organic solvent to form a homogeneous solution. Deionized water is then added dropwise under stirring, followed by the addition of a reducing agent. In-situ polymerization of thioctic acid is initiated, and the drug is co-assembled with the thioctic acid to form a drug-loaded micelle solution. After purification by dialysis, the polythioctic acid-based nano-formulation drug is obtained.

[0004] To achieve the above objectives, the first aspect of the present invention provides a method for preparing a nano-formulation drug based on polythioctic acid, comprising the following steps: (1) Dissolve thioctic acid and the drug in an organic solvent to form a homogeneous solution; (2) While stirring, add deionized water dropwise to the solution in step (1) to form an aqueous solution; (3) Add a reducing agent to the aqueous solution of step (2) while stirring, and stir to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 12 to 48 hours to obtain polythiooctanoic acid nanoparticles with a molecular weight cutoff of 500 to 3500 Da.

[0005] Furthermore, in step (1), the operating temperature is room temperature, and the mass ratio of thioctic acid, drug and organic solvent is 1:0.1 to 1:10 to 100.

[0006] Furthermore, in step (2), the operating temperature is room temperature, and the mass ratio of deionized water to thioctic acid is 20 to 200:1.

[0007] Furthermore, in step (2), the dripping rate is 0.5 to 5 ml / min.

[0008] Furthermore, in step (3), the operating temperature is 20-40°C, the stirring time is 10-60 minutes, and the mass ratio of reducing agent to thioctic acid is 0.05-0.25:1.

[0009] Further, in step (1), the drug is any one or a combination of curcumin, doxorubicin, silymarin, quercetin, paeonol, baicalein, azathioprine, peony root extract, capecitabine, paclitaxel, irinotecan, and nilotinib.

[0010] Furthermore, in step (1), the organic solvent is selected from any one or a combination of N,N-dimethylformamide, dimethyl sulfoxide, and ethanol.

[0011] Furthermore, in step (3), the reducing agent is one of dithiothreitol, β-mercaptoethanol, and tris(2-carboxyethyl)phosphine.

[0012] A second aspect of the present invention provides a nanoparticle drug prepared by the above method.

[0013] Furthermore, the average particle size of the nano-formulation drug is 50–300 nanometers, and the drug loading efficiency is ≥9.5%.

[0014] The invention has a reasonable design and a simple preparation method, and has the following advantages: 1) It is the first to use thioctic acid as both a carrier backbone and an active polymerizing monomer. Under the action of a mild reducing agent, thioctic acid polymerization and drug encapsulation are achieved simultaneously with the drug in an aqueous environment, eliminating the cumbersome steps of pre-synthesizing the carrier and significantly improving process efficiency and drug loading efficiency (loading efficiency ≥ 9.5%).

[0015] 2) The unique co-assembly mechanism can efficiently load natural products or synthetic drugs with extremely poor water solubility but significant activity, such as curcumin and paclitaxel, thus solving key obstacles to their clinical application.

[0016] 3) Select low-toxicity water-soluble organic solvents (N,N-dimethylformamide, dimethyl sulfoxide, ethanol), and ensure uniform and stable micelle formation by precisely controlling the droplet acceleration rate of the aqueous phase (0.5-5 mL / min); combined with gentle dialysis purification, avoid destructive operations such as high temperature, ultrasound, and strong shearing, so as to protect the drug activity to the greatest extent and eliminate organic solvent residues.

[0017] 4) The polythioctic acid backbone is rich in dynamic disulfide bonds, which endows the nanocarrier with the potential for responsive drug release of glutathione, making it particularly suitable for targeted and controlled release in the tumor microenvironment.

[0018] 5) The average particle size of the obtained drug-loaded nanospheres is controlled in the range of 50 to 300 nanometers (verified by particle size analyzer), which is beneficial to the high permeability and retention effect of solid tumors and improves the therapeutic effect.

[0019] The polythioctic acid-based nanomedicine formulations prepared by this method provide an efficient, stable, and intelligent delivery solution for hydrophobic drugs, demonstrating translational value in the field of nanomedicine. Its simple process, controllable cost, and superior drug loading performance lay a solid technical foundation for the development of next-generation nanomedicines. Attached Figure Description

[0020] Figure 1 The ultraviolet-visible absorption spectrum and aqueous dispersion image of the polythiooctanoic acid nanoparticle drug loaded with curcumin prepared in Example 1 of the present invention. Figure 2 The particle size distribution curve of the polythioctic acid nanoparticle drug loaded with curcumin prepared in Example 1 of the present invention; Figure 3 Scanning electron microscope image of the polythioctic acid nanoparticle drug loaded with curcumin prepared in Example 1 of the present invention; Figure 4 The results show the cell viability of the polythioctic acid nanoparticles loaded with curcumin prepared in Example 1 of this invention, when co-cultured with L929 cells or B16 cells at different concentrations for 24 hours. Figure 5 Curves showing the release of curcumin from the polythioctic acid nanoparticle formulation loaded with curcumin prepared in Example 1 of this invention at different pH (a) and different concentrations of glutathione (b); Figure 6 The particle size distribution curve of the polythioctic acid nanoparticle formulation drug loaded with doxorubicin prepared in Example 2 of the present invention; Figure 7 The particle size distribution curve of the capecitabine-loaded polythioctic acid nanoparticle formulation prepared in Example 3 of the present invention; Figure 8 The particle size distribution curve of the polythioctic acid nanoparticle formulation prepared in Example 4 of the present invention, which is simultaneously loaded with paclitaxel and curcumin, is shown in water. Detailed Implementation

[0021] Before discussing the exemplary embodiments in more detail, it should be noted that some exemplary embodiments are described as processes or methods depicted as flowcharts. Although the flowcharts describe the operations as sequential processes, many of these operations can be performed in parallel, concurrently, or simultaneously. Furthermore, the order of the operations can be rearranged. The process can be terminated when its operation is completed, but may also have additional steps not included in the figures. The process can correspond to a method, function, procedure, subroutine, subroutine, etc.

[0022] It should be understood that although the terms "first," "second," etc., may be used herein to describe various units, these units should not be limited by these terms. These terms are used merely to distinguish one unit from another. For example, without departing from the scope of the exemplary embodiments, a first unit may be referred to as a second unit, and similarly, a second unit may be referred to as a first unit. The term "and / or" as used herein includes any and all combinations of one or more of the associated items listed.

[0023] A method for preparing a nano-formulation drug based on polythioctic acid includes the following steps: (1) Dissolve thioctic acid and the drug in an organic solvent to form a homogeneous solution; (2) While stirring, add deionized water dropwise to the solution in step (1) to form an aqueous solution; (3) Add a reducing agent to the aqueous solution of step (2) while stirring, and stir to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 12 to 48 hours to obtain a molecular weight cutoff of 500 to 3500 Da, thus preparing polythioctic acid nanoparticle formulations.

[0024] The present invention will be further described below with reference to specific embodiments, but the present invention is not limited to the following embodiments.

[0025] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0026] All chemical reagents used in this invention are of chemical purity or higher.

[0027] The room temperature range described in this invention is 20–35°C.

[0028] Unless otherwise specified, all reagents, materials, instruments, etc. used in the following examples are commercially available.

[0029] Example 1 (1) Dissolve 50 mg of lipoic acid and 10 mg of curcumin in 3 g of N,N-dimethylformamide and shake at room temperature for 30 minutes to form a homogeneous solution; (2) Add 6 grams of deionized water dropwise to the solution in step (1) while stirring at room temperature; (3) After the addition is complete, add 5 mg of dithiothreitol and stir at 30°C for 45 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 24 hours, and the molecular weight cutoff was 500 Da, thus obtaining polythioctic acid nanoparticles loaded with curcumin.

[0030] The polythioctic acid nanoparticles loaded with curcumin were characterized and tested as follows: Figure 1 The UV-Vis absorption spectrum and aqueous dispersion image of the polythioctic acid nanoparticle drug loaded with curcumin prepared in Example 1 are shown. The nanoparticle drug exhibits the characteristic absorption peak of curcumin in the spectrum, confirming the successful loading of curcumin. Simultaneously, its aqueous dispersion shows a homogeneous and stable state, indicating that the nanoparticle drug has good water dispersibility.

[0031] Figure 2 The particle size distribution curve of the curcumin-loaded polythioctic acid nanoparticle formulation prepared in Example 1 in water is shown. The results show that the nanoparticle formulation has a narrow particle size distribution in water, with an average particle size of 87 nanometers.

[0032] Figure 3 Scanning electron microscope (SEM) images of the curcumin-loaded polythioctic acid nanoparticle formulation prepared in Example 1. The images show that the nanoparticle formulation has a spherical morphology and a narrow particle size distribution.

[0033] Figure 4 The cell viability of the curcumin-loaded polythioctic acid nanoparticle formulation prepared in Example 1, at different concentrations, after co-culturing L929 or B16 cells in a 37°C, 5% CO2 incubator for 24 hours was measured. The results were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Figure 4 It can be seen that the nano-formulation drug formed by encapsulating curcumin exhibits low cytotoxicity and good biocompatibility in both cell types.

[0034] Figure 5 The curcumin release curves of the polythioctic acid nanoparticle formulation loaded with curcumin prepared in Example 1 under different pH (a) and different concentrations of glutathione (GSH) (b) conditions are shown. The results show that the release behavior of this nanoparticle formulation is pH-responsive and redox-responsive, which is beneficial for achieving targeted drug release, improving efficacy and enhancing adaptability.

[0035] Example 2 (1) Dissolve 50 mg of thioctic acid and 5 mg of doxorubicin in 3 g of dimethyl sulfoxide and shake at room temperature for 30 minutes to form a homogeneous solution; (2) Add 1.5 g of deionized water dropwise to the solution from step (1) while stirring at room temperature; (3) After the addition is complete, add 12.5 mg of dithiothreitol and stir at 30°C for 60 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 36 hours, and the molecular weight cutoff was 1800 Da, thus obtaining polythioctic acid nanoparticles loaded with doxorubicin.

[0036] Figure 6 The particle size distribution curve of the polythioctic acid nanoparticle formulation loaded with doxorubicin prepared in Example 2 of this invention in water. Figure 6 It can be seen that the nano-formulation drug has a narrow particle size distribution in water, with an average particle size of 142 nanometers.

[0037] Example 3 (1) Dissolve 50 mg of lipoic acid and 50 mg of capecitabine in 5 g of ethanol and stir at room temperature for 30 minutes to form a homogeneous solution; (2) Add 10 grams of deionized water dropwise to the solution in step (1) while stirring at room temperature; (3) After the addition is complete, add 5 mg of β-mercaptoethanol and stir at 20°C for 60 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 36 hours, and the molecular weight cutoff was 1800 Da, thus obtaining a polythioctic acid nanoparticle drug loaded with capecitabine.

[0038] Figure 7 The particle size distribution curve of the capecitabine-loaded polythioctic acid nanoparticle formulation prepared in Example 3 of this invention in water. Figure 7 It can be seen that the nano-formulation drug has a narrow particle size distribution in water, with an average particle size of 158 nanometers.

[0039] Example 4 (1) Dissolve 100 mg of lipoic acid, 10 mg of paclitaxel and 10 mg of curcumin together in 6 g of N,N-dimethylformamide and stir at room temperature for 30 minutes to form a homogeneous solution; (2) Add 15 grams of deionized water dropwise to the solution in step (1) while stirring at room temperature; (3) After the addition is complete, add 5 mg of β-mercaptoethanol and stir at 40°C for 30 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 24 hours, and the molecular weight cutoff was 3500 Da, thus obtaining a polythioctic acid nanoparticle drug loaded with both paclitaxel and curcumin.

[0040] Figure 8 The particle size distribution curve of the polythioctic acid nanoparticle formulation prepared in Example 4 of this invention, simultaneously loaded with paclitaxel and curcumin, in water. Figure 8 It can be seen that the particle size distribution of this nano-formulation drug in water is relatively narrow, with an average particle size of 137 nanometers.

[0041] Example 5 (1) Dissolve 20 mg of lipoic acid and 5 mg of silymarin in 1 g of ethanol and shake at room temperature for 30 minutes to form a homogeneous solution; (2) Add 2.5 g of deionized water dropwise to the solution in step (1) while stirring at room temperature; (3) After the addition is complete, add 2 mg of tris(2-carboxyethyl)phosphine and stir at 40 °C for 10 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 12 hours, and the molecular weight cutoff was 3500 Da, thus obtaining polythioctic acid nanoparticles loaded with silymarin.

[0042] Example 6 (1) Dissolve 200 mg of lipoic acid and 120 mg of baicalein in 2 g of dimethyl sulfoxide and shake at room temperature for 30 minutes to form a homogeneous solution; (2) Add 18 g of deionized water dropwise to the solution from step (1) while stirring at room temperature; (3) After the addition is complete, add 20 mg of tris(2-carboxyethyl)phosphine and stir at 30°C for 60 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 48 hours, and the molecular weight cutoff was 1800 Da, thus obtaining polythioctic acid nanoparticles loaded with baicalin.

[0043] Example 7 (1) Dissolve 80 mg of lipoic acid and 24 mg of irinotecan in 4 g of N,N-dimethylformamide and shake at room temperature for 30 minutes to form a homogeneous solution; (2) Add 16 grams of deionized water dropwise to the solution from step (1) while stirring at room temperature; (3) After the addition is complete, add 10 mg of dithiothreitol and stir at 35°C for 60 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 30 hours, and the molecular weight cutoff was 500 Da, thus obtaining polythioctic acid nanoparticles loaded with irinotecan.

[0044] Example 8 A method for preparing a nano-formulation drug based on polythioctic acid, comprising the following steps: (1) Dissolve 150 mg of lipoic acid, 30 mg of nilotinib, 30 mg of doxorubicin and 30 mg of curcumin together in 12 g of N,N-dimethylformamide and stir at room temperature for 30 minutes to form a homogeneous solution; (2) While stirring at room temperature, add 30 grams of deionized water dropwise to the solution from step (1); (3) After the addition is complete, add 25 mg of dithiothreitol and stir at 30°C for 45 minutes to obtain a co-assembled micelle solution; (4) The co-assembled micelle solution was dialyzed in deionized water for 24 hours, and the molecular weight cutoff was 3500 Da, thus obtaining a polythioctic acid nanoparticle drug loaded with nilotinib, doxorubicin and curcumin.

[0045] The above examples demonstrate that the polythioctic acid-based nanomedicine formulations prepared by this method provide an efficient, stable, and intelligent delivery solution for hydrophobic drugs, demonstrating translational value in the field of nanomedicine. Its simple process, controllable cost, and superior drug loading performance lay a solid technical foundation for the development of next-generation nanomedicines.

[0046] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A method for preparing a nano-preparation drug based on lipoic acid, comprising the following steps: (1) dissolving lipoic acid and a drug in an organic solvent to form a homogeneous solution; (2) adding deionized water dropwise to the solution of step (1) under stirring to form an aqueous solution; (3) adding a reducing agent to the aqueous solution of step (2) under stirring, and stirring to obtain a co-assembled micelle solution; (4) dialyzing the co-assembled micelle solution in deionized water for 12-48 hours with a molecular weight cut-off of 500-3500 Da to obtain a lipoic acid nano-preparation drug.

2. The method of claim 1, wherein the polythioic acid-based nanofomulation drug is prepared by the following steps of: In step (1), the operation temperature is room temperature, and the mass ratio of lipoic acid, drug and organic solvent is 1:0.1-1:10-100.

3. The method for preparing a polythioctic acid-based nanoparticle drug according to claim 1, characterized in that: In step (2), the operation temperature is room temperature, and the mass ratio of deionized water to lipoic acid is 20-200:

1.

4. The method for preparing a nano-formulation drug based on polythioctic acid according to claim 1, characterized in that: In step (2), the dropwise adding speed is 0.5-5 mL / min.

5. The method for preparing a polythioctic acid-based nanoparticle drug according to claim 1, characterized in that: In step (3), the operation temperature is 20-40℃, the stirring time is 10-60 minutes, and the mass ratio of reducing agent to lipoic acid is 0.05-0.25:

1.

6. The method for preparing a polythioctic acid-based nanoparticle drug according to claim 1, characterized in that: In step (1), the drug is any one or a combination of curcumin, doxorubicin, silymarin, quercetin, paeonol, baicalein, azathioprine, plumbagin, capecitabine, paclitaxel, irinotecan and nilotinib.

7. The method for preparing a polythioctic acid-based nanoparticle drug according to claim 1, characterized in that: In step (1), the organic solvent is selected from any one or a combination of N, N-dimethylformamide, dimethyl sulfoxide and ethanol.

8. The method for preparing a polythioctic acid-based nanoparticle drug according to claim 1, characterized in that: In step (3), the reducing agent is one of dithiothreitol, β-mercaptoethanol and tris (2-carboxyethyl) phosphine. 9.A nano-preparation drug based on lipoic acid prepared by the method according to any one of claims 1-8.

10. The nanoformulation drug of claim 9, wherein: The average particle size of the nano-preparation drug is 50-300 nm, and the drug loading efficiency is ≥9.5%.

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