Drug carrier and micelle based on gallic acid-vitamin E polyethylene glycol succinate as well as preparation method and application of drug carrier and micelle

Abstract

The invention relates to a drug carrier, a micelle, an anti-inflammatory and / or anti-atherosclerosis drug preparation, and a preparation method and application thereof. The drug carrier is a conjugate of gallic acid and vitamin E polyethylene glycol succinate (GA-TPGS), the micelle is formed by self-assembly of GA-TPGS, and the anti-atherosclerosis pharmaceutical preparation comprises a GA-TPGS carrier and a carried drug. The anti-atherosclerosis pharmaceutical preparation can exert the functions of multiple components, and jointly interfere the functions of mononuclear / macrophages to exert the anti-inflammatory and anti-atherosclerosis effects. In addition, the anti-atherosclerosis pharmaceutical preparation is high in stability, the preparation method is simple and convenient, and industrial production is facilitated.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical preparations, and more particularly to pharmaceutical carriers, micelles, anti-inflammatory and / or anti-atherosclerotic pharmaceutical preparations based on gallic acid-vitamin E polyethylene glycol succinate, and preparation methods and applications thereof. Background Art

[0002] Atherosclerosis is a chronic vascular inflammatory disease, and anti-inflammatory therapy is one of the effective strategies to alleviate atherosclerosis. Atherosclerosis originates from vascular endothelial damage caused by lipid imbalance. Damaged vascular endothelial cells overexpress adhesion proteins to recruit monocytes in the blood, entering and triggering inflammation. Monocytes polarize into macrophages under the stimulation of the plaque microenvironment. Macrophages produce reactive oxygen species (ROS) and secrete inflammatory factors, further aggravating the plaque inflammatory response. Therefore, preventing monocyte recruitment and inhibiting macrophage inflammatory response can alleviate atherosclerosis.

[0003] In response to the inflammatory response of atherosclerosis, researchers have studied many targets of inflammation and designed a variety of drug delivery systems targeting macrophages. Due to the multiplicity and heterogeneity of plaques, it is difficult to completely inhibit plaque inflammation by targeting macrophages alone, and it is impossible to curb the pro-inflammatory response of macrophages from the source. Intervention of monocytes and macrophages at multiple links at the same time is conducive to the comprehensive inhibition of the progression of atherosclerotic inflammation. In addition, the toxic side effects and poor effects of drug delivery systems in actual applications have greatly limited their clinical use. The complex molecular design of carrier materials increases the uncontrollability of the chemical structure, which will lead to poor therapeutic effects. Therefore, it is not easy to select materials with therapeutic activity and high biosafety and design simple and feasible synthetic routes. Summary of the invention

[0004] One of the purposes of the present invention is to overcome the deficiencies of the prior art, and to provide a drug carrier and micelle with anti-inflammatory and anti-atherosclerotic properties, as well as their preparation methods and uses. The drug carrier and micelle contain functional target head gallic acid (GA) and vitamin E polyethylene glycol succinate (TPGS) skeleton. GA-TPGS can self-assemble to form nano-scale micelles, and GA binds to P-selectin on plaque endothelial cells, which on the one hand gives the micelles the ability to actively target plaque endothelial cells, and on the other hand inhibits the recruitment of monocytes by plaque endothelial cells, thereby hindering the adhesion of monocytes to plaque endothelium and inhibiting the initial link of plaque inflammation. TPGS, as the skeleton of the drug carrier, has strong anti-inflammatory and antioxidant capabilities, can reduce the level of plaque reactive oxygen, inhibit the secretion of inflammatory factors, and further alleviate atherosclerosis. Finally, the purpose of small side effects, high bioavailability, and good efficacy is achieved.

[0005] For the above and other purposes, the present invention provides a drug carrier, which comprises: a) a functional target head, wherein the functional target head is gallic acid (GA), and b) a skeleton, wherein the skeleton is vitamin E polyethylene glycol succinate (TPGS).

[0006] The present invention also provides a micelle, which comprises the drug carrier and water or a pharmaceutically acceptable aqueous solution.

[0007] As certain embodiments of the present invention, the aqueous solution is PBS or physiological saline.

[0008] As certain embodiments of the present invention, the particle size of the micelle is micrometer-scale or nanometer-scale.

[0009] As certain embodiments of the present invention, the particle size of the micelles ranges from 1 nm to 100 nm.

[0010] As certain embodiments of the present invention, the particle size of the micelle is 15 nm±10 nm.

[0011] The particle size range of the micelles is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 nm. The present invention also provides a method for preparing the drug carrier, which comprises: (1) activating gallic acid with a carboxyl activator to prepare activated gallic acid; (2) mixing a) the activated gallic acid obtained in step (1) and b) vitamin E polyethylene glycol succinate in a reaction solvent N,N-dimethylformamide, so that a) and b) react to form a bonded compound connected by an ester bond.

[0012] As certain embodiments of the present invention, in step (1), the carboxyl activator is selected from: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), hydroxysuccinimide (NHS), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide-4-dimethylaminopyridine (DMAP), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide-1-hydroxybenzotriazole (HOBT), dicyclohexylcarbodiimide (DCC)-1-hydroxybenzotriazole, and a mixed solvent of dicyclohexylcarbodiimide-hydroxysuccinimide and dicyclohexylcarbodiimide-4-dimethylaminopyridine.

[0013] As certain embodiments of the present invention, in step (1), the molar feed ratio is: gallic acid: activator = 1: (1 to 10); preferably, in step (1), the molar ratio of gallic acid to activator is 1:2.

[0014] As certain embodiments of the present invention, in step (1), the activation reaction is carried out in a reaction solvent, and the reaction solvent is selected from: water, N,N-dimethylformamide, formamide, a mixed solvent of N,N-dimethylformamide and formamide, dimethyl sulfoxide, a mixed solvent of dimethyl sulfoxide and N,N-dimethylformamide, a mixed solvent of dimethyl sulfoxide and formamide, and a mixed solvent of water and formamide.

[0015] As certain embodiments of the present invention, in step (1), the activation time is 30 min to 5 h, more preferably, the activation time is 3-4 h under the protection of nitrogen and away from light.

[0016] As certain embodiments of the present invention, in step (2), b) vitamin E polyethylene glycol succinate is first dissolved in a reaction solvent and then mixed with a).

[0017] As certain embodiments of the present invention, in step (2), the reaction temperature is 25-33°C, preferably, the reaction is carried out under the condition of light-proof and nitrogen protection, and the reaction time is 8-120h, more preferably, the reaction time is 72h.

[0018] As certain embodiments of the present invention, in step (2), the reaction solvent is selected from N,N-dimethylformamide.

[0019] As certain embodiments of the present invention, the feed ratio is: the mass ratio of a) activated gallic acid to b) vitamin E polyethylene glycol succinate is 1-10:1-10; preferably, the feed ratio is: the mass ratio of a) activated gallic acid to b) vitamin E polyethylene glycol succinate is 1:1.

[0020] As some embodiments of the present invention, step (3) is further included, wherein the product obtained in step (2) is purified, dialyzed with pure water, and freeze-dried to obtain a purified drug carrier.

[0021] The activation time for activating gallic acid by the carboxyl activator is selected from 30min, 40min, 50min, 1h, 1.1h, 1.2h, 1.3h, 1.4h, 1.5h, 1.6h, 1.7h, 1.8h, 1.9h, 2h, 2.1h, 2.2h, 2.3h, 2.4h, 2.5h, 2.6h, 2.7h, 2.8h, 2.9h, 3h, 3.1h, 3.2h, 3.3h, 3.4h, 3.5h, 3.6h, 3.7h, 3.8h, 3.9h, 4h, 4.1h, 4.2h, 4.3h, 4.4h, 4.5h, 4.6h, 4.7h, 4.8h, 4.9h, and 5h.

[0022] The molar ratio of gallic acid: activator is selected from 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, and 1:10.

[0023] b) Vitamin E polyethylene glycol succinate is dissolved in a reaction solvent and mixed with a) and the reaction temperature is selected from 25, 26, 27, 28, 29, 30, 31, 32, 33°C.

[0024] b) Vitamin E polyethylene glycol succinate is dissolved in a reaction solvent and mixed with a) and the reaction time is selected from 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 85, 90, 95, 100, 105, 110, 115, 120 hours.

[0025] The mass ratio of the activated gallic acid to vitamin E polyethylene glycol succinate is selected from 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 2:1, 2:3, 2:5, 2:7, 2:9, 3:1, 3:2, 3:4, 3:5, 3:7, 3:8, 3:10, 4:1, 4:3, 4:5, 4:7, 4:9, 5:10. :1, 5:2, 5:3, 5:4, 5:6, 5:7, 5:8, 5:9, 6:1, 6:5, 6:7, 7:1, 7:2, 7:3, 7:4, 7:5, 7:8, 7:9, 7:10, 8:1, 8:3, 8:5, 8:7, 8:9, 9:1, 9:2, 9:4, 9:5, 9:8, 9:10, 10:1, 10:3, 10:7, 10:9.

[0026] The present invention also provides a method for preparing micelles, wherein the drug carrier is prepared by adopting the method for preparing the drug carrier, and the drug carrier is dissolved in water or a pharmaceutically acceptable aqueous solution.

[0027] As certain embodiments of the present invention, the aqueous solution is PBS or physiological saline.

[0028] The present invention also provides an anti-inflammatory and / or anti-atherosclerotic drug preparation, wherein the anti-inflammatory and / or anti-atherosclerotic drug preparation is loaded physically or chemically on the hydrophobic part and / or hydrophilic part of the drug carrier or the micelle.

[0029] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug (A) is loaded on the hydrophobic part of the drug carrier or micelle, and the drug loading mode is physical action, and the drug loading method is selected from one or more of thin film dispersion method, emulsification ultrasonic rotary evaporation method, dialysis method, solvent injection method, direct dissolution method and solvent volatilization method.

[0030] As certain embodiments of the present invention, the loading method is emulsification ultrasonic rotary evaporation.

[0031] As certain embodiments of the present invention, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from dichloromethane, water, methanol, ethanol, chloroform, tetrahydrofuran, or a mixed solvent thereof.

[0032] As certain embodiments of the present invention, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from chloroform, dichloromethane, or tetrahydrofuran or a mixed solvent thereof.

[0033] As certain embodiments of the present invention, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from dichloromethane.

[0034] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug (B) is loaded on the hydrophilic part of the drug carrier or micelle, and the anti-inflammatory and / or anti-atherosclerotic drug is coupled to vitamin E polyethylene glycol succinate via a linker.

[0035] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug (A) and the anti-inflammatory and / or anti-atherosclerotic drug (B) may be the same or different.

[0036] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug comprises a cholesterol-lowering drug and / or a triglyceride-lowering drug.

[0037] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug is selected from one or more of statins, niacin drugs, fibrates, steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs.

[0038] The choice of lipid-lowering drugs should be based on the composition of blood lipids. If the cholesterol is high, cholesterol-lowering drugs should be used, such as statins, commonly used ones are atorvastatin and rosuvastatin. If the triglycerides are high, triglyceride-lowering drugs should be used, mainly fibrates.

[0039] As certain embodiments of the present invention, the statin is selected from one or more of fluvastatin, pitavastatin, atorvastatin, rosuvastatin, and simvastatin.

[0040] Statins: including fluvastatin, pitavastatin, atorvastatin, rosuvastatin, simvastatin, etc., suitable for patients with hypercholesterolemia, mixed hyperlipidemia with elevated cholesterol as the main feature, atherosclerosis and coronary heart disease.

[0041] As certain embodiments of the present invention, the nicotinic acid drug is selected from one or more of acipimox, inositol nicotinate, vitamin E nicotinate, mannitol nicotinate, and probucol.

[0042] Niacin drugs include acipimox, inositol nicotinate, vitamin E nicotinate, mannitol nicotinate, probucol, etc., which help lower blood cholesterol and triglyceride levels, dilate blood vessels, and maintain normal insulin function.

[0043] As certain embodiments of the present invention, the fibrate drug is selected from one or more of clofibrate, bezafibrate, and fenofibrate.

[0044] Fibrate drugs include clofibrate, bezafibrate, fenofibrate, etc., which are mainly used for patients with elevated triglycerides. They can also lower very low-density lipoprotein and low-density lipoprotein.

[0045] As certain embodiments of the present invention, the steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs are selected from one or more of aspirin, ibuprofen, indomethacin, rapamycin, curcumin, quercetin, and resveratrol.

[0046] The present invention also provides a method for preparing the pharmaceutical preparation, wherein the anti-inflammatory and / or anti-atherosclerotic drug (A) is loaded on the hydrophobic part of the drug carrier or micelle, and the loading method is selected from one or more of a thin film dispersion method, an emulsification ultrasonic rotary evaporation method, a dialysis method, a solvent injection method, a direct dissolution method and a solvent volatilization method.

[0047] As some embodiments of the present invention, the loading method is emulsification ultrasonic rotary evaporation. As some embodiments of the present invention, the solvent used in the emulsification ultrasonic rotary evaporation is selected from dichloromethane, water, methanol, ethanol, chloroform, tetrahydrofuran, or a mixed solvent thereof.

[0048] As certain embodiments of the present invention, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from chloroform, dichloromethane, or tetrahydrofuran or a mixed solvent thereof.

[0049] As certain embodiments of the present invention, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from dichloromethane.

[0050] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug (B) is loaded on the hydrophilic part of the drug carrier or micelle and coupled to vitamin E polyethylene glycol succinate via a linker.

[0051] As certain embodiments of the present invention, the anti-inflammatory and / or anti-atherosclerotic drug (A) and the anti-inflammatory and anti-atherosclerotic drug (B) may be the same or different. The present invention also provides the use of the drug carrier, the micelle, or the pharmaceutical preparation in the preparation of anti-inflammatory and / or anti-atherosclerotic drugs.

[0052] Therefore, the drug carrier, micelle, anti-inflammatory and / or anti-atherosclerotic drug preparation based on vitamin E polyethylene glycol succinate, and the preparation method and application thereof of the present invention have achieved the following beneficial effects:

[0053] 1. Gallic acid and vitamin E polyethylene glycol succinate are directly connected through an ester bond, which makes the construction process easier, the structure simpler, the particles uniform, and the stability excellent.

[0054] 2. All components in gallic acid-vitamin E polyethylene glycol succinate are made of biocompatible and degradable materials. Compared with the existing technology, it eliminates the toxicity and metabolic excretion pressure caused by non-drug ingredients to the body to the greatest extent.

[0055] 3. Gallic acid binds to P-selectin on plaque endothelial cells, which, on the one hand, gives micelles the ability to actively target plaque endothelial cells, and on the other hand, inhibits the recruitment of monocytes by plaque endothelial cells, thereby hindering the adhesion of monocytes to plaque endothelial cells and inhibiting the initial stage of plaque inflammation. Vitamin E polyethylene glycol succinate, as the skeleton of the drug carrier, has strong anti-inflammatory and antioxidant capabilities, can reduce the level of plaque reactive oxygen, inhibit the secretion of inflammatory factors, and further alleviate atherosclerosis.

[0056] 4. The anti-inflammatory and / or anti-atherosclerotic drug preparation can interfere with the inflammatory response of monocytes / macrophages in atherosclerosis, producing a strong anti-atherosclerotic effect.

[0057] In short, the drug carrier and micelle of the present invention are simple in construction and highly stable, and after being loaded with anti-inflammatory and / or anti-atherosclerotic drugs, they can effectively inhibit the inflammatory cascade reaction and alleviate atherosclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] Figure 1 Shown is the H NMR spectrum of the product prepared in Example 1 of the present invention;

[0059] Figure 2 The particle size and potential of GA-TPGS / Cur NPs were measured by a dynamic light scattering particle size analyzer in Example 3 of the present invention, showing that the particle size of GA-TPGS / Cur NPs nanoparticles was about 20 nm, the PDI was 0.235, and the particle size was uniform;

[0060] Figure 3 The nanoparticle morphology is confirmed by transmission electron microscopy in Example 3 of the present invention, showing spherical nanoparticles;

[0061] Figure 4 The fluorescence image of the GA-TPGS NPs-treated group of human monocytic leukemia cells in Example 4 of the present invention is shown, and the fluorescence intensity is significantly reduced, indicating that GA-TPGS NPs can inhibit the adhesion of monocytes by competitively binding to P-selectin;

[0062] Figure 5 This shows the ability of Example 5 of the present invention to inhibit the secretion of reactive oxygen species (ROS). The fluorescence intensity of the curcumin, GA-TPGS NPs, and GA-TPGS / Cur NPs treatment groups decreased, and the decrease in the GA-TPGS / Cur NPs treatment group was more significant, indicating that GA-TPGS / Cur NPs can inhibit the secretion of ROS;

[0063] Figure 6 Shown is the gross oil red staining of the aorta wall of mice in the PBS, curcumin, GA-TPGS NPs, and GA-TPGS / CurNPs groups in Example 6 of the present invention;

[0064] Figure 7 Shown is the aortic H&E staining of the aortic sinus of mice in the PBS, curcumin, GA-TPGS NPs, and GA-TPGS / CurNPs groups in Example 6 of the present invention, indicating that the preparation has a therapeutic effect on atherosclerosis. DETAILED DESCRIPTION

[0065] The present invention is further described below in conjunction with specific examples, and it should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of protection of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0066] It should be understood that the terms used in the examples of the present invention are intended to describe specific embodiments rather than to limit the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated, the singular forms "a", "an" and "the" include plural forms. When the examples give numerical ranges, it should be understood that unless otherwise specified in the present invention, both endpoints of each numerical range and any value between the two endpoints can be selected.

[0067] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, any methods, equipment, and materials of the prior art that are similar or equivalent to the methods, equipment, and materials in the embodiments of the present invention may be used to implement the present invention, based on the prior art mastery of those skilled in the art and the description of the present invention.

[0068] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in the field of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields. These techniques are well described in the literature, see Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL, Second edition, Cold Spring Harbor Laboratory Press, 1989 and Third edition, 2001; Ausubel et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987 and periodic updates; these series METHODS IN ENZYMOLOGY, Academic Press, San Diego; Wolffe, CHROMATINSTRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; METHODS IN ENZYMOLOGY, Vol. 304, Chromatin (PM Wassarman and AP Wolffe, eds.), Academic Press, San Diego, 1999; and METHODS IN MOLECULAR BIOLOGY, Vol. 304, Chromatin (PM Wassarman and AP Wolffe, eds.), Academic Press, San Diego, 1999. BIOLOGY, Vol. 119, Chromatin Protocols (PB Becker, ed.) Humana Press, Totowa, 1999, etc.

[0069] Example 1 Preparation of the pharmaceutical preparation of the present invention

[0070] 1. Synthesis of Gallic Acid-Vitamin E Polyethylene Glycol Succinate Drug Carrier:

[0071] Take 0.2g gallic acid, 450mg 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, 270mg hydroxysuccinimide, 72mg 4-dimethylaminopyridine and dissolve in 20mL N,N-dimethylformamide, activate at 35℃, 500rpm for 3h; weigh 3.55g vitamin E polyethylene glycol succinate and add it to the activated N,N-dimethylformamide solution, react at 35℃, 500rpm for 72h; after the reaction is completed, use a dialysis bag with a cutoff of 3500 to dialyze in DMF for 24h, and then dialyze in pure water for 48h; after freezing the dialysate at -40℃ for 8h, use a freeze dryer to freeze-dry, and the resulting product is a white solid. Nuclear magnetic hydrogen spectrum characterization of GA-TPGS, such as Figure 1 As shown, the H characteristic peak on the GA benzene ring appeared in the GA-TPGS spectrum, indicating that GA-TPGS was successfully synthesized.

[0072] 2. Preparation of Gallic Acid-Vitamin E Polyethylene Glycol Succinate Micelles GA-TPGS NPs:

[0073] Gallic acid-vitamin E polyethylene glycol succinate is dispersed in dichloromethane, pure water is added, and after ultrasonic emulsification, dichloromethane is removed by rotary evaporation to obtain self-assembled GA-TPGS NPs (gallic acid-vitamin E polyethylene glycol succinate nanoparticles). When used in animal experiments, the drug carrier powder is placed in a physiologically acceptable aqueous medium, such as PBS or saline.

[0074] Example 2 Particle size characterization of GA-TPGS NPs

[0075] The average particle size, PDI and Zeta potential of GA-TPGS NPs were measured by dynamic light scattering particle size analyzer (DLS). The results are shown in Table 1:

[0076] Table 1

[0077] Group Average particle size (nm) PDI Zeta(mV) GA-TPGS NPs 13.2±5.3 0.234±0.085 -8.2±1.6

[0078] Example 3 Preparation and Characterization of Curcumin (Cur)-loaded GA-TPGS Micelle (GA-TPGS / Cur)

[0079] 1) Example of carrying anti-tumor drugs by physical means

[0080] Taking curcumin as an example, GA-TPGS / Cur NPs were prepared by ultrasonic emulsification. Take hydrophobic curcumin, take GA-TPGS powder (GA-TPGS: curcumin = 9:1, w / w) and dissolve it in CH2Cl2, add 5 times the volume of deionized water, and ultrasonically form an emulsion (300W, 2s / 2s, 5min). Next, remove CH2Cl2 by vacuum rotary evaporation at 37°C, put the yellow clear preparation into a dialysis bag with a molecular weight cutoff of 1000, and then dialyze it in deionized water for 3h, and then filter it through a 0.22μm microporous filter membrane to obtain GA-TPGS / Cur NPs.

[0081] 2) Characterization of drug-loaded micelles

[0082] The particle size and potential of GA-TPGS / Cur NPs were determined by dynamic light scattering particle size analyzer, and the morphology of nanoparticles was confirmed by transmission electron microscopy (TEM). Figure 2 As shown in the figure, the particle size of GA-TPGS / Cur NPs nanoparticles is about 20 nm, the PDI is 0.235, and the particle size is uniform. The transmission electron microscopy image shows that the nanoparticles are spherical nanoparticles ( Figure 3 ).

[0083] Example 4 Investigation of the ability to inhibit the infiltration of monocytes

[0084] Human umbilical vein endothelial cells (HUVECs) were seeded into 24-well plates at a density of 40%. When the cells grew to about 80%, H2O2 was added to activate the cells. After 6 hours of stimulation, PBS and GA-TPGS NPs were added to the corresponding wells and incubated for 1 hour. Human monocytic leukemia cells (THP-1) labeled with BCEDF AM probe were added and incubated with the cells at 37°C and 30rpm for 30 minutes. After washing with PBS, DMSO was added to lyse the cells. The fluorescence intensity was detected at Ex=480nm and Em=520nm using a fluorescence spectrophotometer.

[0085] The results are as follows Figure 4 As shown in the figure, the fluorescence intensity of the GA-TPGS NPs-treated group was significantly reduced, indicating that GA-TPGS NPs could inhibit the adhesion of monocytes by competitively binding to P-selectin.

[0086] Example 5: Examination of the ability to inhibit the secretion of reactive oxygen species (ROS)

[0087] Mouse macrophage RAW264.7 cells were seeded into 6-well plates at a density of 40%. When the cells grew to about 80%, lipopolysaccharide (LPS) was added to activate the cells. After 6 hours of stimulation, PBS, curcumin, GA-TPGS NPs, and GA-TPGS / Cur NPs were added to the corresponding wells and incubated for 24 hours. DCFH-DA probe was added for staining and incubated with the cells at 37°C and 30rpm for 0.5 hours. After washing with PBS, the ROS content was analyzed by flow cytometry.

[0088] The results are as follows Figure 5 As shown in the figure, the fluorescence intensity of the curcumin, GA-TPGS NPs, and GA-TPGS / Cur NPs treatment groups decreased, and the decrease in the GA-TPGS / Cur NPs treatment group was more significant, indicating that GA-TPGS / Cur NPs can inhibit the secretion of ROS.

[0089] Example 6: Evaluation of therapeutic effects on atherosclerotic mice

[0090] 1. High-fat diet for Apoe- / - mice to establish AS model

[0091] 2. Dosage regimen The atherosclerotic mice were randomly divided into 4 groups (n=8). After the model was established, each group was injected with PBS, curcumin, GA-TPGS NPs, and GA-TPGS / Cur NPs through the tail vein twice a week for 2 months. Normal mice served as controls.

[0092] After the treatment, the mouse aorta was isolated and macroscopic oil red staining was performed. Macroscopic oil red staining showed that GA-TPGS / Cur NPs could significantly reduce the lipid content on the aortic wall. The mouse aortic sinus was sliced ​​and H&E stained to analyze the ratio of plaque to lumen and calculate the vascular stenosis rate. Aortic H&E staining confirmed that GA-TPGS / Cur NPs could reduce the area of ​​arterial plaque and reduce the lumen stenosis rate ( Figure 6 and Figure 7 ). This indicates that the preparation has a therapeutic effect on atherosclerosis.

[0093] Example 7 Safety Investigation

[0094] After the treatment, blood was collected from each group of mice; the blood was centrifuged (3000rpm, 5min) to obtain plasma samples, and the expression levels of various enzymes in the plasma were detected using a specific plasma enzyme kit on an automatic biochemical analyzer, and a number of biochemical index parameters related to liver and kidney function were measured.

[0095] The indicators of each treatment group were normal, indicating that GA-TPGS / Cur NPs had no liver and kidney toxicity within the investigated indicators.

[0096] The above is only a preferred embodiment of the present invention, and is not any formal or substantial limitation of the present invention. It should be pointed out that ordinary technicians in this technical field can make several improvements and supplements without departing from the method of the present invention, and these improvements and supplements should also be regarded as the protection scope of the present invention. Any technician familiar with this profession, without departing from the spirit and scope of the present invention, can make some changes, modifications and evolutions of the technical content disclosed above, which are equivalent embodiments of the present invention; at the same time, any changes, modifications and evolutions of any equivalent changes made to the above embodiments based on the essential technology of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A drug carrier, comprising: a) a functional target head, wherein the functional target head is gallic acid (GA), and b) a backbone, wherein the backbone is vitamin E polyethylene glycol succinate (TPGS).

2. A micelle comprising the drug carrier according to claim 1 and water or a pharmaceutically acceptable aqueous solution.

3. The micelle according to claim 2, wherein Also includes any one or more of the following features: A1) the aqueous solution is selected from PBS or physiological saline; A2) The particle size of the micelle is micrometer or nanometer; Preferably, the particle size of the micelles ranges from 1 nm to 100 nm; More preferably, the particle size of the micelle is 15 nm±10 nm.

4. The method for preparing a drug carrier according to claim 1, wherein: The method comprises: (1) activating gallic acid with a carboxyl activator to prepare activated gallic acid; (2) Mixing a) the activated gallic acid obtained in step (1) and b) vitamin E polyethylene glycol succinate in a reaction solvent, N,N-dimethylformamide, to allow a) and b) to react to form a bonded compound connected by an ester bond.

5. The method for preparing a drug carrier according to claim 4, wherein: Also includes any one or more of the following features: B1) In step (1), the carboxyl activating agent is selected from: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), hydroxysuccinimide (NHS), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide-4-dimethylaminopyridine (DMAP), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide-1-hydroxybenzotriazole (HOBT), dicyclohexylcarbodiimide (DCC)-1-hydroxybenzotriazole, and a mixed solvent of dicyclohexylcarbodiimide-hydroxysuccinimide and dicyclohexylcarbodiimide-4-dimethylaminopyridine, B2) In step (1), the molar feed ratio is: gallic acid: activator = 1: (1-10); preferably, in step (1), the molar ratio of gallic acid to activator is 1:2; B3) in step (1), the activation reaction is carried out in a reaction solvent selected from the group consisting of water, N,N-dimethylformamide, formamide, a mixed solvent of N,N-dimethylformamide and formamide, dimethyl sulfoxide, a mixed solvent of dimethyl sulfoxide and N,N-dimethylformamide, a mixed solvent of dimethyl sulfoxide and formamide, and a mixed solvent of water and formamide; B4) In step (1), the activation time is 30 min to 5 h, more preferably 3-4 h under the protection of nitrogen and away from light; B5) In step (2), b) vitamin E polyethylene glycol succinate is first dissolved in a reaction solvent and then mixed with a), The reaction temperature is 25-33°C. Preferably, the reaction is carried out under the condition of light-proof and nitrogen protection. The reaction time is 8-120h. More preferably, the reaction time is 72h. B6) In step (2), the reaction solvent is selected from N,N-dimethylformamide; B7) the feed ratio is: the mass ratio of a) activated gallic acid to b) vitamin E polyethylene glycol succinate is 1-10:1-10; preferably, the feed ratio is: the mass ratio of a) activated gallic acid to b) vitamin E polyethylene glycol succinate is 1:1; B8) In step (3), the product obtained in step (2) is purified, dialyzed with pure water, and freeze-dried to obtain a purified drug carrier.

6. A method for preparing micelles, wherein: The drug carrier is prepared by the preparation method of the drug carrier according to any one of claims 4 to 5, and the drug carrier is dissolved in water or a pharmaceutically acceptable aqueous solution; preferably, the aqueous solution is PBS or physiological saline.

7. An anti-inflammatory and / or anti-atherosclerotic pharmaceutical preparation, wherein: The anti-inflammatory and / or anti-atherosclerotic drug preparation is loaded physically or chemically on the hydrophobic part and / or the hydrophilic part of the drug carrier according to claim 1 or the micelle according to claim 2.

8. The pharmaceutical preparation according to claim 7, wherein The anti-inflammatory and / or anti-atherosclerotic drug (A) is loaded in the hydrophobic part of the drug carrier or micelle, The drug loading method is physical loading, and the drug loading method is selected from one or more of a thin film dispersion method, an emulsified ultrasonic rotary evaporation method, a dialysis method, a solvent injection method, a direct dissolution method, and a solvent volatilization method; preferably, the drug loading method is selected from an emulsified ultrasonic rotary evaporation method; wherein the solvent used in the emulsified ultrasonic rotary evaporation method is selected from dichloromethane, water, methanol, ethanol, chloroform, tetrahydrofuran, or a mixed solvent thereof; more preferably, the solvent used in the emulsified ultrasonic rotary evaporation method is selected from chloroform, dichloromethane, or tetrahydrofuran, or a mixed solvent thereof; More preferably, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from dichloromethane.

9. The pharmaceutical preparation according to claim 7 or 8, wherein The anti-inflammatory and / or anti-atherosclerotic drug (B) is loaded in the hydrophilic part of the drug carrier or micelle, The anti-inflammatory and / or anti-atherosclerotic drug is coupled to vitamin E polyethylene glycol succinate via a linker; The anti-inflammatory and / or anti-atherosclerotic drug (A) and the anti-inflammatory and / or anti-atherosclerotic drug (B) may be the same or different.

10. The pharmaceutical preparation according to any one of claims 7 to 9, wherein The anti-inflammatory and / or anti-atherosclerotic drugs include one or both of cholesterol-lowering drugs and / or triglyceride-lowering drugs; Preferably, the anti-inflammatory and / or anti-atherosclerotic drug is selected from one or more of statins, niacin drugs, fibrates, steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs; More preferably, the statin is selected from one or more of fluvastatin, pitavastatin, atorvastatin, rosuvastatin, and simvastatin; The nicotinic acid drug is selected from one or more of acipimox, inositol nicotinate, vitamin E nicotinate, mannitol nicotinate, and probucol; The fibrate drug is selected from one or more of clofibrate, bezafibrate, and fenofibrate; The steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs are selected from one or more of aspirin, ibuprofen, indomethacin, rapamycin, curcumin, quercetin, and resveratrol.

11. A method for preparing the pharmaceutical preparation according to any one of claims 7 to 10, wherein: The anti-inflammatory and / or anti-atherosclerotic drug (A) is loaded on the hydrophobic part of the drug carrier or micelle, and the loading method is selected from one or more of a thin film dispersion method, an emulsification ultrasonic rotary evaporation method, a dialysis method, a solvent injection method, a direct dissolution method and a solvent volatilization method; Preferably, the loading method is emulsification ultrasonic rotary evaporation; More preferably, the solvent used in the emulsification ultrasonic rotary evaporation method is selected from dichloromethane, water, methanol, ethanol, chloroform, tetrahydrofuran, or a mixed solvent thereof; Further preferably, the solvent used in the emulsification ultrasonic rotary evaporation method is chloroform, dichloromethane, or tetrahydrofuran or a mixed solvent thereof; Most preferably, the solvent used in the emulsification ultrasonic rotary evaporation method is dichloromethane.

12. A method for preparing a pharmaceutical preparation as claimed in any one of claims 11, wherein: The anti-inflammatory and / or anti-atherosclerotic drug (B) is loaded on the hydrophilic part of the drug carrier or micelle and coupled to the vitamin E polyethylene glycol succinate via a linker; The anti-inflammatory and / or anti-atherosclerotic drug (A) and the anti-inflammatory and / or anti-atherosclerotic drug (B) may be the same or different.

13. Use of the drug carrier according to claim 1, the micelle according to claim 2, or the pharmaceutical preparation according to any one of claims 5 to 7 in the preparation of anti-inflammatory and / or anti-atherosclerotic products.

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