Pharmaceutical composition for reducing local fat and use thereof

Abstract

The invention provides a pharmaceutical composition for reducing local fat. The pharmaceutical composition comprises a pharmaceutically acceptable aqueous solution, a plurality of drug-containing micelles formed by a surfactant, and resveratrol coated in the drug-containing micelles. The pharmaceutical composition for reducing local fat can reduce fat at an application site, and has the advantages of high stability, high adipose tissue living body availability, low side effect, slow release and the like.

Description

[0001] This application is a divisional application of an invention patent application with an application date of August 26, 2016, application number: 201680046431.9, and name: "Medical composition for reducing local fat and its use". Technical Field

[0002] The present invention relates to a pharmaceutical composition for reducing local fat, and in particular to a pharmaceutical composition comprising a plurality of drug-containing micelles and resveratrol encapsulated in the drug-containing micelles, and the pharmaceutical composition is used for reducing local fat. Background Art

[0003] In recent years, as more and more people's concept of beauty has changed, and the standards for self-health and body shape have been improved, people are no longer concerned with simply losing weight, but are more concerned with reducing local fat or sculpting curves to achieve a healthier and more beautiful body shape. Moreover, general weight loss methods, whether diet or exercise, cannot reduce fat in a specific single area. If you want to reduce fat in specific areas (such as the waist, abdomen, legs, arms, chin, and face, etc.), current technology can only be achieved through liposuction surgery.

[0004] At present, the main method to reduce local fat is liposuction. However, the liposuction process can cause serious damage to nerves, blood vessels, and other body tissues, and has the risk of infection, large amount of bleeding, long anesthesia time, and fatal risks of fat embolism and anesthesia allergy that cannot be prevented in advance. In addition, liposuction is also prone to severe bruising, redness, and swelling, severe pain, a recovery period of 3 to 6 months, and uneven liposuction sites. Therefore, statistics show that although most people want to use liposuction to improve local subcutaneous fat accumulation or body curves, the number of people who actually undergo liposuction is less than 40%, indicating that most consumers who want to improve body curves or reduce local fat will give up due to the side effects of liposuction and postoperative pain or risks.

[0005] Although some non-surgical local fat reduction pharmaceutical compositions or devices can reduce some of the side effects, most of them are ineffective and will produce other side effects, such as necrosis of surrounding normal cells, inflammation of surrounding tissues, and severe pain, and there are certain restrictions on the implementation site. Therefore, the market is still in great need of a local fat reduction pharmaceutical composition that can effectively reduce local fat, has lower side effects, better safety, and a shorter recovery period.

[0006] With high demands from both consumers and physicians, the development of localized fat-reducing pharmaceutical compositions that can break through current technological limitations will be an issue that urgently needs to be explored and resolved. Summary of the invention

[0007] In view of the defects of the prior art, the present invention provides a pharmaceutical composition for reducing local fat, comprising a plurality of drug-containing micelles formed by a surfactant, and resveratrol encapsulated in the drug-containing micelles. The pharmaceutical composition for reducing local fat can reduce fat at the site of application, and has the advantages of high stability, high bioavailability of adipose tissue, low side effects, and sustained release.

[0008] The present invention can promote the apoptosis reaction (apoptosis) of the fat cells at the application site, so as to achieve the purpose of reducing the local fat at the application site. The present invention can greatly improve the adverse reactions and side effects of the conventional technology that cause necrosis (necrosis) of the surrounding cells and inflammation, and the effect of local fat reduction is significantly better than other non-surgical medical compositions for reducing local fat. The present invention is suitable for application to the site where subcutaneous fat needs to be reduced by direct injection, subcutaneous implantation, implantation infusion, ointment or patch, etc., without the intervention or assistance of any surgical operation or instrument. Preferably, it is applied to the subcutaneous fat layer of the local site by subcutaneous fat injection. Preferably, the injection form of the medical composition of the present invention includes but is not limited to injection liquid or injection powder (powder for injection, or powder for solution for injection). The local fat referred to in the present invention includes but is not limited to the waist, abdomen, legs, arms, chin and face.

[0009] In the present invention, resveratrol refers to resveratrol extracted from natural plants or commercially available. Preferably, the purity of resveratrol is 90% to 100%.

[0010] In the present invention, the green tea extract refers to a mixture of green tea components extracted by any solvent and any extraction method, a commercially available green tea extract, any mixture containing at least 45% (weight percentage concentration) of epigallocatechin gallate (EGCG), or commercially available epigallocatechin gallate.

[0011] In the present invention, micelle refers to a microstructure formed by a surfactant, wherein the surfactant has a hydrophilic end and a lipophilic end (lipophilic end), and the surfactant forms the microstructure with the hydrophilic end facing outward and the lipophilic end (lipophilic end) facing inward. Preferably, the microstructure is spherical, spherical-like, or other microstructures.

[0012] In the present invention, drug-containing micelles refer to micelles containing resveratrol; that is, drug-containing micelles refer to micelles encapsulating or containing resveratrol.

[0013] In the present invention, the second lipophilic drug micelle refers to micelles containing other lipophilic drugs other than resveratrol. In other words, the second lipophilic drug micelle refers to micelles encapsulating or containing other lipophilic drugs.

[0014] Among them, other fat-soluble drugs refer to at least one or a combination of curcumin, quercetin, puerarin, and other fat-soluble drugs other than resveratrol; or, other fat-soluble drugs refer to fat-soluble drugs other than resveratrol.

[0015] In the present invention, the water-soluble drug refers to at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate, catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0016] In the present invention, the term "a state where no precipitate is generated" means a state where no precipitate is visible to the naked eye, that is, without the aid of artificial devices.

[0017] The present invention provides a pharmaceutical composition for reducing local fat, comprising:

[0018] A plurality of drug-containing micelles are uniformly distributed in the pharmaceutically acceptable aqueous solution; and

[0019] Resveratrol encapsulated in the drug-containing micelles;

[0020] In a preferred embodiment, the drug-containing micelle is a microstructure formed by a pharmaceutically acceptable surfactant, and the hydrophilic-lipophilic balance value (HLB value) of the surfactant is greater than 10.

[0021] In a preferred embodiment, the particle size of the drug-containing micelles is 3-250 nm.

[0022] In a preferred embodiment, the particle size of the drug-containing micelles is 5-50 nm.

[0023] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution, and the drug-containing micelles are uniformly distributed in the pharmaceutically acceptable aqueous solution. The pharmaceutically acceptable aqueous solution is water for injection, aqueous solution for injection, or physiological saline.

[0024] In a preferred embodiment, the surfactant is a nonionic surfactant.

[0025] In a preferred embodiment, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0026] In a preferred embodiment, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0027] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:4 to 1:500.

[0028] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:5 to 1:200.

[0029] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:8 to 1:80.

[0030] In a preferred embodiment, when the pharmaceutical composition is subjected to an accelerated stability test at a temperature of 25°C ± 2°C, a relative humidity of RH 60% ± 5%, and avoiding direct sunlight, the pharmaceutical composition remains in a state without precipitation for at least 24 hours.

[0031] In a preferred embodiment, when the pharmaceutical composition is subjected to an accelerated stability test at a temperature of 25°C ± 2°C, a relative humidity of RH 60% ± 5%, and avoiding direct sunlight, the pharmaceutical composition remains in a precipitate-free state for at least 6 months.

[0032] In a preferred embodiment, the concentration of resveratrol in the pharmaceutical composition is 0.2-166.7 mg / mL.

[0033] In a preferred embodiment, the concentration of resveratrol in the pharmaceutical composition is 2.5-60 mg / mL.

[0034] In a preferred embodiment, the pharmaceutical composition further comprises a green tea extract, and the green tea extract is dissolved in the pharmaceutically acceptable aqueous solution; wherein the green tea extract comprises:

[0035] A first green tea extract component, wherein the first green tea extract component is epigallocatechin gallate (EGCG).

[0036] In a preferred embodiment, the concentration of epigallocatechin gallate in the pharmaceutical composition is 0.25-300 mg / mL.

[0037] In a preferred embodiment, the concentration of epigallocatechin gallate in the pharmaceutical composition is 1-200 mg / mL.

[0038] In a preferred embodiment, the content of epigallocatechin gallate is 45-100% based on 100% by weight of the total weight of the green tea extract.

[0039] In a preferred embodiment, the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0040] In a preferred embodiment, the weight ratio of the resveratrol to the green tea extract is 20:1 to 1:20.

[0041] In a preferred embodiment, the weight ratio of the resveratrol to the green tea extract is 10:1 to 1:10.

[0042] In a preferred embodiment, the weight of the surfactant is 0.24-70 weight units based on the total weight of the resveratrol and the green tea extract as one weight unit; or, the weight ratio of the total weight of the resveratrol and the green tea extract to the surfactant is 4:1 to 1:70.

[0043] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution and a second lipid-soluble drug micelle, and the second lipid-soluble drug micelle is uniformly distributed in the pharmaceutically acceptable aqueous solution.

[0044] The second lipophilic drug micelle is another microstructure formed by the second surfactant, and another lipophilic drug (or a second lipophilic drug) is encapsulated in the second lipophilic drug micelle.

[0045] In a preferred embodiment, the hydrophilic-lipophilic balance value (HLB value) of the second surfactant is greater than 10.

[0046] In a preferred embodiment, the second surfactant is a nonionic surfactant.

[0047] In a preferred embodiment, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0048] In a preferred embodiment, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0049] In a preferred embodiment, the fat-soluble drug is at least one of quercetin, synephrine, puerarin, curcumin, and other fat-soluble drugs other than resveratrol, or a combination thereof.

[0050] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 30:1-1:20.

[0051] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 20:1-1:15.

[0052] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 15:1-1:10.

[0053] In a preferred embodiment, the pharmaceutical composition further includes a pharmaceutically acceptable aqueous solution and a water-soluble drug.

[0054] In a preferred embodiment, the water-soluble drug is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate (Catechingallate), catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0055] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 20:1-1:30.

[0056] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 15:1 to 1:20.

[0057] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 10:1 to 1:15.

[0058] In a preferred embodiment, the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the pharmaceutical composition is 0.25-300 mg / mL.

[0059] In a preferred embodiment, the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the pharmaceutical composition is 1-200 mg / mL.

[0060] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0061] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 20:1 to 1:20.

[0062] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 10:1 to 1:10.

[0063] In a preferred embodiment, the pharmaceutical composition further comprises a cosolvent to increase the solubility of the drug.

[0064] In a preferred embodiment, the co-solvent is at least one of polyethylene glycol, propylene glycol, ethanol, and other co-solvents, or a combination thereof.

[0065] In a preferred embodiment, the polyethylene glycol is at least one of polyethylene glycol 200 (PEG 200), polyethylene glycol 400 (PEG 400), polyethylene glycol 600 (PEG 600), and other polyethylene glycols, or a combination thereof.

[0066] In a preferred embodiment, the pharmaceutical composition further comprises a suspending agent (also called a suspending agent) for reducing the sedimentation rate of the drug or micelles.

[0067] In a preferred embodiment, the suspending agent is at least one of sodium alginate, glycerol, sodium carboxymethylcellulose, mannitol, and other suspending agents, or a combination thereof.

[0068] In a preferred embodiment, the pharmaceutical composition further comprises oil phase excipients to increase the stability of the pharmaceutical composition and the solubility of the drug.

[0069] In a preferred embodiment, the oil phase excipient is at least one of unsaturated fatty acids, glycerol, triglyceride, and other oil phase excipients, or a combination thereof.

[0070] In a preferred embodiment, the unsaturated fatty acid is at least one of oleic acid, castor oil, sesame oil, cottonseed oil, soybean oil, safflower oil, corn oil, and other unsaturated fatty acids, or a combination thereof.

[0071] In a preferred embodiment, the triglyceride is at least one of medium chain triglycerides and other triglycerides, or a combination thereof.

[0072] In a preferred embodiment, the pharmaceutically acceptable aqueous solution comprises a local anesthetic.

[0073] In a preferred embodiment, the local anesthetic is at least one of amides, p-aminobenzylamine lipids, and amino ethers, or a combination thereof.

[0074] In a preferred embodiment, the amide is at least one of dibucaine, lidocaine, mepivacaine HCl, bupivacaine HCl, pyrrocaine HCl, prilocaine HCl, digammacaine, and oxethazaine, or a combination thereof.

[0075] In a preferred embodiment, the p-aminobenzylamine is at least one of butacaine, dimethocaine, and tutocaine, or a combination thereof.

[0076] In a preferred embodiment, the amino ether is at least one of Quinisocaine and Pramocaine or a combination thereof.

[0077] In a preferred embodiment, the pharmaceutically acceptable aqueous solution comprises an antioxidant.

[0078] In a preferred embodiment, the antioxidant is at least one of beta-carotene, lutein, lycopene, bilirubin, vitamin A, vitamin C (also known as ascorbic acid), vitamin E, uric acid, nitric oxide, nitroxide, pyruvate, catalase, superoxide dismutase, glutathione peroxidases, N-acetyl cysteine, and naringenin, or a combination thereof.

[0079] The present invention further provides a subcutaneous fat layer injection injection or subcutaneous injection injection for reducing the amount of local subcutaneous fat, comprising:

[0080] A pharmaceutically acceptable aqueous solution;

[0081] A plurality of drug-containing micelles are uniformly distributed in the pharmaceutically acceptable aqueous solution; and

[0082] Resveratrol encapsulated in the drug-containing micelles;

[0083] The drug-containing micelle is a microstructure formed by a pharmaceutically acceptable surfactant, and the hydrophilic-lipophilic balance value (HLB value) of the surfactant is greater than 10.

[0084] In a preferred embodiment, the local subcutaneous fat mass is the subcutaneous fat mass at the application site.

[0085] In a preferred embodiment, the particle size of the drug-containing micelles is 3-250 nm.

[0086] In a preferred embodiment, the particle size of the drug-containing micelles is 5-50 nm.

[0087] In a preferred embodiment, the pharmaceutically acceptable aqueous solution is water for injection, aqueous solution for injection, or physiological saline.

[0088] In a preferred embodiment, the surfactant is a nonionic surfactant.

[0089] In a preferred embodiment, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0090] In a preferred embodiment, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0091] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:4 to 1:500.

[0092] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:5 to 1:200.

[0093] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:8 to 1:80.

[0094] In a preferred embodiment, the concentration of resveratrol in the subcutaneous fat layer injection or subcutaneous injection is 0.2-166.7 mg / mL.

[0095] In a preferred embodiment, the concentration of resveratrol in the subcutaneous fat layer injection or subcutaneous injection is 2.5-60 mg / mL.

[0096] In a preferred embodiment, the subcutaneous fat layer injection or subcutaneous injection further comprises a green tea extract, and the green tea extract is dissolved in the pharmaceutically acceptable aqueous solution; wherein the green tea extract comprises:

[0097] A first green tea extract component, wherein the first green tea extract component is epigallocatechin gallate (EGCG).

[0098] In a preferred embodiment, the concentration of epigallocatechin gallate in the subcutaneous fat layer injection or subcutaneous injection is 0.25-300 mg / mL.

[0099] In a preferred embodiment, the concentration of epigallocatechin gallate in the subcutaneous fat layer injection or subcutaneous injection is 1-200 mg / mL.

[0100] In a preferred embodiment, the content of epigallocatechin gallate is 45-100% based on 100% by weight of the total weight of the green tea extract.

[0101] In a preferred embodiment, the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0102] In a preferred embodiment, the weight ratio of the resveratrol to the green tea extract is 20:1 to 1:20.

[0103] In a preferred embodiment, the weight ratio of the resveratrol to the green tea extract is 10:1 to 1:10.

[0104] In a preferred embodiment, the weight of the surfactant is 0.24-70 weight units based on the total weight of the resveratrol and the green tea extract as one weight unit; or, the weight ratio of the total weight of the resveratrol and the green tea extract to the surfactant is 4:1 to 1:70.

[0105] In a preferred embodiment, the dosage of the administration site is 0.2-20 mg per square centimeter.

[0106] In a preferred embodiment, the dosage of the administration site is 0.4-12 mg per square centimeter.

[0107] In a preferred embodiment, the dosage at the administration site is 0.2-40 mg / kg.

[0108] In a preferred embodiment, the dosage at the administration site is 0.4-20 mg / kg.

[0109] In a preferred embodiment, the administration frequency is 1 to 12 times at intervals of 1 to 30 days.

[0110] In a preferred embodiment, the administration frequency is 1 to 8 times at intervals of 1 to 21 days.

[0111] In a preferred embodiment, the subcutaneous fat layer injection or subcutaneous injection further comprises a second lipid-soluble drug micelle, and the second lipid-soluble drug micelle is uniformly distributed in the pharmaceutically acceptable aqueous solution.

[0112] The second lipophilic drug micelle is a microstructure formed by another surfactant, and the other lipophilic drug is encapsulated in the second lipophilic drug micelle.

[0113] In a preferred embodiment, the hydrophilic-lipophilic balance value (HLB value) of the other surfactant is greater than 10.

[0114] In a preferred embodiment, the other surfactant is a nonionic surfactant.

[0115] In a preferred embodiment, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0116] In a preferred embodiment, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0117] Preferably, the lipophilic drug is at least one of curcumin, quercetin, puerarin, and other lipophilic drugs other than resveratrol, or a combination thereof.

[0118] In a preferred embodiment, the pharmaceutically acceptable aqueous solution further comprises a water-soluble drug.

[0119] In a preferred embodiment, the water-soluble drug is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate (Catechingallate), catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0120] In a preferred embodiment, the pharmaceutically acceptable aqueous solution comprises a local anesthetic.

[0121] In a preferred embodiment, the local anesthetic is at least one of amides, p-aminobenzylamine lipids, and amino ethers, or a combination thereof.

[0122] In a preferred embodiment, the amide is at least one of dibucaine, lidocaine, mepivacaine HCl, bupivacaine HCl, pyrrocaine HCl, prilocaine HCl, digammacaine, and oxethazaine, or a combination thereof.

[0123] In a preferred embodiment, the p-aminobenzylamine is at least one of butacaine, dimethocaine, and tutocaine, or a combination thereof.

[0124] In a preferred embodiment, the amino ether is at least one of Quinisocaine and Pramocaine or a combination thereof.

[0125] In a preferred embodiment, the pharmaceutically acceptable aqueous solution comprises an antioxidant.

[0126] In a preferred embodiment, the antioxidant is at least one of beta-carotene, lutein, lycopene, bilirubin, vitamin A, vitamin C (also known as ascorbic acid), vitamin E, uric acid, nitric oxide, nitroxide, pyruvate, catalase, superoxide dismutase, glutathione peroxidases, N-acetyl cysteine, and naringenin, or a combination thereof.

[0127] The present invention provides a method for reducing the amount of subcutaneous fat in a local area of ​​an individual, comprising applying a pharmaceutical composition to the local area of ​​the individual, wherein the pharmaceutical composition comprises:

[0128] a plurality of drug-containing micelles; and

[0129] Resveratrol encapsulated in the drug-containing micelles;

[0130] The drug-containing micelle is a microstructure formed by a pharmaceutically acceptable surfactant, and the hydrophilic-lipophilic balance value (HLB value) of the surfactant is greater than 10.

[0131] In a preferred embodiment, the particle size of the drug-containing micelles is 3-250 nm.

[0132] In a preferred embodiment, the particle size of the drug-containing micelles is 5-50 nm.

[0133] In a preferred embodiment, the surfactant is a nonionic surfactant.

[0134] In a preferred embodiment, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0135] In a preferred embodiment, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0136] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:4 to 1:500.

[0137] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:5 to 1:200.

[0138] In a preferred embodiment, the weight ratio of resveratrol to the surfactant is 1:8 to 1:80.

[0139] In a preferred embodiment, the concentration of resveratrol in the pharmaceutical composition is 0.2-166.7 mg / mL.

[0140] In a preferred embodiment, the concentration of resveratrol in the pharmaceutical composition is 2.5-60 mg / mL.

[0141] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution and a second lipid-soluble drug micelle, wherein the second lipid-soluble drug micelle is uniformly distributed in the pharmaceutically acceptable aqueous solution; wherein the second lipid-soluble drug micelle is another microstructure formed by a second nonionic surfactant, and a second lipid-soluble drug is encapsulated in the second lipid-soluble drug micelle.

[0142] In a preferred embodiment, the hydrophilic-lipophilic balance value (HLB value) of the second nonionic surfactant is greater than 10.

[0143] In a preferred embodiment, the second nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0144] In a preferred embodiment, the second nonionic surfactant is at least one of Cremophor ELP, Cremophor RH 40, and other Cremophor RH 40 derivatives or a combination thereof.

[0145] In a preferred embodiment, the second fat-soluble drug is at least one or a combination of quercetin, synephrine, puerarin, curcuminoid, curcumin, and other fat-soluble drugs other than resveratrol.

[0146] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 30:1-1:20.

[0147] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 20:1-1:15.

[0148] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 15:1-1:10.

[0149] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution and a water-soluble drug.

[0150] In a preferred embodiment, the water-soluble drug is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate (Catechingallate), catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0151] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 20:1-1:30.

[0152] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 15:1 to 1:20.

[0153] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 10:1 to 1:15.

[0154] In a preferred embodiment, the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the pharmaceutical composition is 0.25-300 mg / mL.

[0155] In a preferred embodiment, the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the pharmaceutical composition is 1-200 mg / mL.

[0156] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0157] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 20:1 to 1:20.

[0158] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 10:1 to 1:10.

[0159] In a preferred embodiment, the weight of the surfactant is 0.24-70 weight units based on the total weight of the resveratrol and the green tea extract as one weight unit; or, the weight ratio of the total weight of the resveratrol and the green tea extract to the surfactant is 4:1 to 1:70.

[0160] In a preferred embodiment, the dosage of the pharmaceutical composition applied to the local area is 0.2-20 mg per square centimeter.

[0161] In a preferred embodiment, the dosage of the pharmaceutical composition applied to the local area is 0.4-12 mg per square centimeter.

[0162] In a preferred embodiment, the dosage of the pharmaceutical composition applied to the local site is 0.2-40 mg / kg.

[0163] In a preferred embodiment, the dosage of the pharmaceutical composition applied to the local site is 0.4-20 mg / kg.

[0164] In a preferred embodiment, the frequency of administering the pharmaceutical composition is 1 to 12 times at intervals of 1 to 30 days.

[0165] In a preferred embodiment, the frequency of administering the pharmaceutical composition is 1 to 8 times at intervals of 1 to 21 days.

[0166] In a preferred embodiment, the individual is an animal or a human.

[0167] In a preferred embodiment, the pharmaceutical composition is injected or applied to the local area of ​​the individual.

[0168] In a preferred embodiment, the pharmaceutical composition further comprises at least one or a combination of a cosolvent, a suspending agent, and an oil phase excipient.

[0169] In a preferred embodiment, the oil phase excipient and / or cosolvent and the nonionic surfactant together form the microstructure.

[0170] The present invention provides a use of a pharmaceutical composition in preparing a drug or a subcutaneous injection formulation for reducing the amount of subcutaneous fat in a local area of ​​an individual; the pharmaceutical composition comprises:

[0171] a plurality of drug-containing micelles; and

[0172] Resveratrol encapsulated in the drug-containing micelles;

[0173] The drug-containing micelle is a microstructure formed by a pharmaceutically acceptable non-ionic surfactant, and the hydrophilic-lipophilic balance value (HLB value) of the non-ionic surfactant is greater than 10.

[0174] In a preferred embodiment, the particle size of the drug-containing micelles is 3-250 nm.

[0175] In a preferred embodiment, the particle size of the drug-containing micelles is 5-50 nm.

[0176] In a preferred embodiment, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0177] In a preferred embodiment, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0178] In a preferred embodiment, the weight ratio of resveratrol to the nonionic surfactant is 1:4 to 1:500.

[0179] In a preferred embodiment, the concentration of resveratrol in the pharmaceutical composition is 0.2-166.7 mg / mL.

[0180] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution and a second lipid-soluble drug micelle, wherein the second lipid-soluble drug micelle is uniformly distributed in the pharmaceutically acceptable aqueous solution; wherein the second lipid-soluble drug micelle is another microstructure formed by a second nonionic surfactant, and a second lipid-soluble drug is encapsulated in the second lipid-soluble drug micelle.

[0181] In a preferred embodiment, the hydrophilic-lipophilic balance value (HLB value) of the second nonionic surfactant is greater than 10.

[0182] In a preferred embodiment, the second nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0183] In a preferred embodiment, the second nonionic surfactant is at least one of Cremophor ELP, Cremophor RH 40, and other Cremophor RH 40 derivatives or a combination thereof.

[0184] In a preferred embodiment, the second fat-soluble drug is at least one of quercetin, synephrine, puerarin, curcuminoids, and other fat-soluble drugs other than resveratrol, or a combination thereof.

[0185] In a preferred embodiment, the weight ratio of the resveratrol to the second fat-soluble drug is 30:1-1:20.

[0186] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution and a water-soluble drug.

[0187] In a preferred embodiment, the water-soluble drug is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate (Catechingallate), catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0188] In a preferred embodiment, the weight ratio of the resveratrol to the water-soluble drug is 20:1-1:30.

[0189] In a preferred embodiment, the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the subcutaneous fat layer injection or subcutaneous injection is 0.25-300 mg / mL.

[0190] In a preferred embodiment, the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0191] In a preferred embodiment, the weight of the nonionic surfactant is 0.24 to 70 weight units based on the total weight of the resveratrol and the green tea extract as one weight unit; or, the weight ratio of the total weight of the resveratrol and the green tea extract to the nonionic surfactant is 4:1 to 1:70.

[0192] In a preferred embodiment, the medicine or subcutaneous injection contains a therapeutically effective amount of the pharmaceutical composition.

[0193] In a preferred embodiment, the therapeutically effective amount is 0.2-20 mg of the pharmaceutical composition per square centimeter of the local area.

[0194] In a preferred embodiment, the therapeutically effective amount is 0.2-40 mg of the pharmaceutical composition per kg body weight.

[0195] In a preferred embodiment, the drug or subcutaneous injection is administered to the administration site 1 to 12 times at intervals of 1 to 30 days.

[0196] In a preferred embodiment, the individual is an animal.

[0197] In a preferred embodiment, the drug is injected into the local area of ​​the individual or a subcutaneous injection is administered; or, the drug is applied to the local area of ​​the individual.

[0198] In a preferred embodiment, the pharmaceutical composition further comprises at least one or a combination of a cosolvent, a suspending agent, and an oil phase excipient.

[0199] In a preferred embodiment, the oil phase excipient and / or cosolvent and the nonionic surfactant together form the microstructure. BRIEF DESCRIPTION OF THE DRAWINGS

[0200] Figure 1A : Bar graph showing the effects of micellar-free resveratrol subcutaneous injection and micellar-free green tea extract subcutaneous injection on subcutaneous fat mass in rats.

[0201] Figure 1B : Bar graph showing the effects of subcutaneous injection of resveratrol without micelles and subcutaneous injection of green tea extract without micelles on relative total weight gain in rats.

[0202] Figure 2A : Bar graph showing the effects of resveratrol subcutaneous injections prepared with different excipients on the local subcutaneous fat mass in rats.

[0203] Figure 2B : Bar graph showing the effect of subcutaneous injection of resveratrol prepared with different excipients on the relative total weight gain of rats.

[0204] Figure 3 : Bar graph showing the effect of resveratrol pharmaceutical composition on subcutaneous fat mass in rats.

[0205] Figure 4A : Bar graph showing the effect of resveratrol compound pharmaceutical composition on subcutaneous fat mass in rats.

[0206] Figure 4B : Bar graph showing the effect of resveratrol compound pharmaceutical composition on relative total weight gain in rats.

[0207] Figure 5 : Effects of resveratrol-other fat-soluble drug compound pharmaceutical compositions on apoptosis of mature adipocytes.

[0208] Figure 6 : Effects of resveratrol-other water-soluble drug compound pharmaceutical compositions on apoptosis of mature adipocytes. DETAILED DESCRIPTION

[0209] Resveratrol is a polyphenol compound that is mainly found in red grape skin, Japanese knotweed, or red wine. Due to the fact that resveratrol is poorly soluble in aqueous solution, is easily metabolized into glucuronic acid and sulfate metabolites in the body, is quickly excreted through urine and feces, and has very poor body availability, it is quite difficult to develop a pharmaceutical composition for reducing local fat using resveratrol. The inventors faced this dilemma in the early stages of developing this pharmaceutical composition.

[0210] Experiment 1: Effects of subcutaneous injection of resveratrol and green tea extract on subcutaneous fat mass and body weight in rats

[0211] Preparation of green tea extract subcutaneous injection: Use water for injection to prepare green tea extract into a 5 mg / mL aqueous solution. Filter with a 0.2 μm filter membrane to obtain the 5 mg / mL green tea extract subcutaneous injection solution described in this experiment. It must be stored in a 4°C refrigerator away from light.

[0212] Preparation of resveratrol subcutaneous injection: Resveratrol was prepared into a 5 mg / mL solution using the commonly known excipient polysorbate 80 (Tween 80) for non-water-soluble drug injections, solvent ethanol, and water for injection. The detailed preparation method is as follows: 0.5 g of resveratrol was mixed with an appropriate amount of solvent ethanol to completely dissolve the resveratrol, and then 0.1 g of polysorbate 80 (Tween 80) was added to completely dissolve the polysorbate; the ethanol was evaporated by vacuum for 2 to 4 hours; after the ethanol was completely evaporated, water for injection was added to make the total volume 100 mL; after stirring evenly, it was filtered with a 0.2 μm filter membrane to obtain the 5 mg / mL resveratrol subcutaneous injection described in this experiment. It must be stored in a 4°C refrigerator away from light.

[0213] The experiment was conducted using 7-week-old male Sprague-Dawley rats. First, 18 rats were fed a high-fat diet (brand name: Research Diets, Inc.; model number: #D12492) to induce subcutaneous fat gain. After continuous feeding until the rats reached a body weight of 330±10g, the rats were randomly divided into 3 groups, namely a high-fat control group, a resveratrol group, and a green tea extract group, with 6 rats in each group, so that there was no statistical difference in the body weight of the rats in each group. The body weight of each rat was recorded and defined as the "pre-experimental body weight" of each rat. Then, the drug was administered in the following manner.

[0214] 5 mg / mL of resveratrol subcutaneous injection was injected into the subcutaneous fat layer of the lower groin of rats in the resveratrol group, and each injection amount was 8 mg (8 mg / kg) of resveratrol per kilogram of body weight; that is, 1.6 mL of the above 5 mg / mL resveratrol subcutaneous injection was injected per kilogram of body weight. 5 mg / mL of green tea extract subcutaneous injection was injected into the subcutaneous fat layer of the lower groin of rats in the green tea extract group, and each injection amount was 8 mg (8 mg / kg) of green tea extract per kilogram of body weight; that is, 1.6 mL of the above 5 mg / mL green tea extract subcutaneous injection was injected per kilogram of body weight. The high-fat control group was given the same volume of water for injection in the same injection method.

[0215] The injection site was the inguinal fat of the rats, and the injection was averaged on the left and right sides, and was injected once on the 1st, 3rd, and 5th days of the experiment. During the experiment, the rats were continuously fed a high-fat diet, and the body weight changes were recorded daily. The drinking water and food intake were recorded once a week. The rats were fasted on the 20th day of the experiment, and were sacrificed with carbon dioxide on the 21st day.

[0216] The body weight of each rat was recorded, which was defined as the "post-test body weight" of each rat. The "post-test body weight" of each rat was subtracted from the "pre-test body weight" to obtain the "total weight gain". The total weight gain of each group of rats was divided by the total weight gain of the rats in the high-fat control group to obtain the "relative total weight gain".

[0217] The fat of the left and right inguinal regions of the rats was weighed, and the amount of inguinal fat of each group was calculated. The data were presented as mean ± SD, and statistically analyzed by one-way ANOVA. Statistical results were expressed by symbols or letters. Different symbols or letters indicated statistical differences between the groups (p < 0.05), and the same symbols or letters indicated no statistical differences between the groups (p > 0.05).

[0218] See also Figure 1A as well as Figure 1B , Figure 1AThe bar graph shows the effects of micellar-free resveratrol subcutaneous injection and micellar-free green tea extract subcutaneous injection on subcutaneous fat mass in rats. Figure 1B The bar graph shows the effect of micellar-free resveratrol subcutaneous injection and micellar-free green tea extract subcutaneous injection on the relative total weight gain of rats. The inguinal fat mass is the sum of the inguinal fat mass on the left and right sides.

[0219] Figure 1A The results showed that after three subcutaneous fat injections, the amount of local fat at the injection site did not decrease significantly in the resveratrol group and the green tea extract group compared with the high-fat control group (p>0.05), indicating that directly injecting dissolved resveratrol or green tea extract into the subcutaneous fat layer cannot reduce the local fat at the injection site.

[0220] Figure 1B The results showed that after three subcutaneous fat injections, the body weight of rats in the resveratrol group and the green tea extract group did not decrease significantly compared with the high-fat control group (p>0.05), indicating that directly injecting dissolved resveratrol or green tea extract into the subcutaneous fat layer cannot reduce body weight.

[0221] Among them, green tea extract is an extremely water-soluble ingredient. Previous cell experiments have found that it can promote apoptosis of fat cells. However, when the green tea extract is dissolved and directly injected into the subcutaneous fat, it cannot reduce the local subcutaneous fat or body weight. From this experiment, it can be seen that directly dissolving resveratrol or green tea extract and injecting it into the subcutaneous fat layer is not enough to reduce local fat and body weight. In order to overcome this problem, the inventors further studied and developed the pharmaceutical composition containing resveratrol of the present invention.

[0222] Experiment 2: Effects of different types of resveratrol subcutaneous injections on subcutaneous fat mass and body weight in rats

[0223] Resveratrol physiological saline solution, resveratrol PEG solution, and resveratrol ELP solution were prepared in the following manner.

[0224] Preparation method of resveratrol physiological saline solution:

[0225] 500 mg of resveratrol was mixed with an appropriate amount of normal saline for injection to a final volume of 100 mL. The mixture was stirred evenly to completely dissolve the resveratrol to obtain a resveratrol normal saline solution, wherein the concentration of resveratrol in the resveratrol normal saline solution was 5 mg / mL.

[0226] Preparation method of resveratrol PEG solution:

[0227] Mix 15g of polyethylene glycol 400 (PEG400), 15g of glycerol, and an appropriate amount of physiological saline for injection to a final volume of 100mL. Stir evenly to completely dissolve polyethylene glycol 400 and glycerol to obtain a polyethylene glycol and glycerol mixed solution. Mix 450mg of resveratrol with an appropriate amount of polyethylene glycol and glycerol mixed solution to a final volume of 90ml. Stir evenly to completely dissolve resveratrol to obtain a resveratrol PEG solution. The resveratrol concentration in the resveratrol PEG solution is 5mg / mL.

[0228] Preparation method of resveratrol ELP solution:

[0229] Mix 500 mg of resveratrol with 100-160 mL of dichloromethane, and stir at 150-500 rpm at room temperature until resveratrol is completely dissolved. Add 20 g of polyoxyethylene 35 castor oil (Kolliphor ELP, referred to as ELP), stir evenly at a speed of 100-300 rpm to volatilize the dichloromethane. After the dichloromethane is completely volatilized, slowly add normal saline for injection to make the final volume reach 100 mL, stir evenly, and you can get a resveratrol ELP solution. The resveratrol concentration in the resveratrol ELP solution is 5 mg / mL, the concentration of polyoxyethylene 35 castor oil (ELP) is about 20% (weight percentage), and the weight ratio of resveratrol to polyoxyethylene 35 castor oil is 1:40.

[0230] The experiment was conducted using 6-week-old male Sprague-Dawley rats. First, 20 rats were fed a high-fat diet (brand name: Research Diets, Inc.; model number: #D12492) to induce subcutaneous fat gain. After continuous feeding until the rats reached a body weight of 330±10 g, the rats were randomly divided into 4 groups, namely, a control group, a saline group, a PEG group, and an ELP group, with 5 rats in each group, so that there was no statistical difference in the body weight of the rats in each group. The body weight of each rat was recorded, which was defined as the "pre-experimental body weight" of each rat. Then, the drug was administered in the following manner.

[0231] Resveratrol saline solution, resveratrol PEG solution, and resveratrol ELP solution were injected into the subcutaneous fat layer of the inguinal region of rats in the saline group, PEG group, and ELP group, respectively. The injection volume was 4 mL per kilogram of body weight (4 mL / kg), so that each injection dose was 20 mg of resveratrol per kilogram of body weight (20 mg / kg; calculated as 4 mL / kg×5 mg / mL=20 mg / kg). The control group was given the same volume of saline for injection in the same manner as above.

[0232] The injection site was the inguinal fat of the rats, and the injection was averaged on the left and right sides, and was injected once on the 1st, 2nd, 3rd, and 4th days of the experiment. During the experiment, the rats were continuously fed with high-fat diets, and the weight changes were recorded daily. The water intake and food intake were recorded once a week. The experiment lasted for 14 days, and the rats were sacrificed with carbon dioxide on the 15th day.

[0233] The weight of each rat was recorded, which was defined as the "post-test weight" of each rat. The "post-test weight" of each rat was deducted from the "pre-test weight" to obtain the "total weight gain". The total weight gain of each group of rats was divided by the total weight gain of the rats in the control group to obtain the "relative total weight gain".

[0234] The subcutaneous fat of the left and right inguinal regions of the rats was weighed, and the subcutaneous fat of the left and right inguinal regions was added up to calculate the inguinal subcutaneous fat mass. The inguinal subcutaneous fat mass of the rats in each group was divided by the inguinal subcutaneous fat mass of the rats in the control group to obtain the "relative weight of inguinal subcutaneous fat".

[0235] Data were presented as mean ± SD and analyzed by one-way ANOVA. Statistical results were expressed as symbols or letters. Different symbols or letters indicated statistical differences between the groups (p < 0.05), while the same symbols or letters indicated no statistical differences between the groups (p > 0.05).

[0236] See also Figure 2A and Figure 2B . Figure 2A This is a bar graph showing the effects of resveratrol subcutaneous injections prepared using different excipients on the local subcutaneous fat mass in rats. Figure 2B This is a bar graph showing the effects of subcutaneous resveratrol injections prepared with different excipients on the relative total weight gain of rats.

[0237] Figure 2AThe results showed that the relative weight of the subcutaneous fat in the lower groin of the rats in the control group was 100±27.6%, the relative weight of the subcutaneous fat in the lower groin of the rats in the saline group was 108.2±24.7%, the relative weight of the subcutaneous fat in the lower groin of the rats in the PEG group was 114.0±4.4%, and the relative weight of the subcutaneous fat in the lower groin of the rats in the ELP group was 72.5±0.0%. There was no significant difference in the relative weight of the subcutaneous fat in the lower groin of the rats in the saline group and the control group, indicating that directly injecting resveratrol into the subcutaneous fat layer at the application site could not reduce the fat (local fat) at the application site. There was no significant difference in the relative weight of the subcutaneous fat in the lower groin of the rats in the PEG group and the control group; there was a significant difference in the relative weight of the subcutaneous fat in the lower groin of the rats in the ELP group and the control group (p<0.05), and the relative weight of the subcutaneous fat in the lower groin of the rats in the ELP group was reduced by 27.5%.

[0238] Figure 2B The results showed that the relative total weight gain of rats in the control group was 100.0±30.8%, the relative total weight gain of rats in the normal saline group was 128.3±16.9%, the relative total weight gain of rats in the PEG group was 120.8±18.2%, and the relative total weight gain of rats in the ELP group was 101.3±22.0%. There was no significant difference among the four groups (p>0.05).

[0239] It can be seen from the above experiments that directly injecting resveratrol into the subcutaneous fat layer of the application site cannot reduce the fat (local fat) at the application site, nor can it reduce body weight. Injecting a resveratrol composition with the addition of excipient PEG (a common cosolvent) into the subcutaneous fat layer of the application site cannot reduce the fat (local fat) at the application site, nor can it reduce body weight. However, injecting a resveratrol composition with the addition of a nonionic surfactant ELP into the subcutaneous fat layer of the application site can significantly reduce the fat (local fat) at the application site. Therefore, it is necessary to further explore whether the resveratrol composition must contain a nonionic surfactant in order to reduce the subcutaneous fat (local fat) at the application site and reduce body weight.

[0240] Further analysis revealed that the resveratrol PEG solution used above did not contain micelles, but the resveratrol ELP solution contained micelles, and resveratrol was encapsulated in the micelles formed by ELP. Therefore, it is necessary to further explore the effect of micelles on reducing local fat and weight.

[0241] Experiment 3: Effects of subcutaneous injection of a single composition of resveratrol containing micelles on subcutaneous fat mass and body weight in rats

[0242] The resveratrol ELP fraction micellar formulation, the resveratrol HS-15 fraction micellar formulation, the resveratrol ELP micellar formulation, and the resveratrol HS-15 micellar formulation were formulated in the following manner.

[0243] Preparation method of resveratrol ELP partial micellar dosage form: Mix 20g of polyoxyethylene 35 castor oil (ie, ELP) with an appropriate amount of normal saline for injection to make the final weight reach 100g. Stir evenly to completely dissolve polyoxyethylene 35 castor oil (ie, ELP) to obtain a 20% ELP solution. Mix 400mg of resveratrol with an appropriate amount of 20% ELP solution to make the final weight reach 80g. Stir evenly to completely dissolve resveratrol to obtain a resveratrol ELP partial micellar dosage form. The resveratrol concentration in the resveratrol ELP partial micellar dosage form is about 5mg / mL, the concentration of polyoxyethylene 35 castor oil (ELP) is about 20% (weight percentage), and the weight ratio of resveratrol to polyoxyethylene 35 castor oil is about 1:40.

[0244] Preparation method of resveratrol HS-15 partial micellar dosage form: 20g of polyethylene glycol stearate 15 (Kolliphor HS 15, referred to as HS-15) is mixed with an appropriate amount of injection physiological saline to make the final weight reach 100g. Stir evenly to completely dissolve polyethylene glycol stearate 15 (i.e. HS-15) to obtain a 20% HS-15 solution. 400mg of resveratrol is mixed with an appropriate amount of 20% HS-15 solution to make the final weight reach 80g. Stir evenly to completely dissolve resveratrol to obtain a resveratrol HS-15 partial micellar dosage form. The resveratrol concentration in the resveratrol HS-15 partial micellar dosage form is about 5mg / mL, the concentration of polyethylene glycol stearate 15 (i.e. HS-15) is about 20% (weight percentage), and the weight ratio of resveratrol to polyethylene glycol stearate 15 (i.e. HS-15) is about 1:40.

[0245] The preparation method of the resveratrol ELP micelle dosage form is the same as the preparation method of the resveratrol ELP solution in Experiment 2.

[0246] Preparation method of resveratrol HS-15 micellar dosage form: 500 mg of resveratrol is mixed with 80-140 mL of dichloromethane, and stirred at 150-500 rpm at room temperature until resveratrol is completely dissolved. Add 20 g of polyethylene glycol stearate 15 (Kolliphor HS15, referred to as HS-15), stir evenly at a speed of 100-300 rpm to volatilize dichloromethane. After dichloromethane is completely volatilized, slowly add physiological saline for injection to make the final volume reach 100 g, stir evenly to form multiple drug-containing micelles, and then obtain resveratrol HS-15 micellar dosage form. The resveratrol concentration in the resveratrol HS-15 micellar dosage form is about 5 mg / g, the concentration of polyethylene glycol stearate 15 (HS-15) is 20% (weight percentage), and the weight ratio of resveratrol to polyethylene glycol stearate 15 (HS-15) is 1:40.

[0247] A particle size analyzer was used to determine whether the resveratrol ELP partial micelle formulation, the resveratrol HS-15 partial micelle formulation, the resveratrol ELP micelle formulation, and the resveratrol HS-15 micelle formulation contained micelles, and the particle size of the micelles was measured.

[0248] The results showed that some micelle formulations of resveratrol ELP and some micelle formulations of resveratrol HS-15 contained a large amount of precipitates, and the number of drug-containing micelles was small. Resveratrol ELP micelle formulations and resveratrol HS-15 micelle formulations were clear without stratification, and the number of drug-containing micelles was large.

[0249] It can be seen from this that although the resveratrol ELP partial micellar formulation and the resveratrol HS-15 partial micellar formulation contain a large amount of precipitates, the supernatant still contains micelles. Therefore, the resveratrol ELP partial micellar formulation, the resveratrol HS-15 partial micellar formulation, the resveratrol ELP micellar formulation, and the resveratrol HS-15 micellar formulation are all pharmaceutical compositions of the present invention.

[0250] The experiment was conducted using 6-week-old male Sprague-Dawley rats of the SD strain. First, 20 rats were fed a high-fat diet (brand name: Research Diets, Inc.; model number: #D12492) to induce subcutaneous fat increase. After continuous feeding until the rats reached a body weight of 330±10 g, the rats were randomly divided into 5 groups, namely, a control group, an ELP partial micelle group, a HS-15 partial micelle group, an ELP micelle group, and a HS-15 micelle group, with 4 rats in each group, so that there was no statistical difference in the body weight of the rats in each group. The body weight of each rat was recorded, which was defined as the "pre-experimental body weight" of each rat. Then, the drug was administered in the following manner.

[0251] The resveratrol ELP partial micelle formulation, resveratrol HS-15 partial micelle formulation, resveratrol ELP micelle formulation, and resveratrol HS-15 micelle formulation were mixed evenly (to make the precipitate in the partial micelle formulation evenly suspended), and then injected into the subcutaneous fat layer of the inguinal canal of rats in the ELP partial micelle group, HS-15 partial micelle group, ELP micelle group, and HS-15 micelle group, respectively. The injection volume for each time was 4 mL per kilogram of body weight (4 mL / kg), so that each injection dose was 20 mg of resveratrol per kilogram of body weight (20 mg / kg; calculated as 4 mL / kg×5 mg / mL=20 mg / kg). The control group was given the same volume of normal saline for injection in the same injection method as above.

[0252] The injection site was the inguinal fat of the rats, and the injection was averaged on the left and right sides, and was injected once on the 1st, 2nd, 3rd, 4th, 5th, and 6th days of the experiment. During the experiment, the rats were continuously fed with high-fat diets, and the weight changes were recorded daily. The water intake and food intake were recorded once a week. The experiment lasted for 14 days, and the rats were sacrificed with carbon dioxide on the 15th day.

[0253] The weight of each rat was recorded, which was defined as the "post-test weight" of each rat. The "post-test weight" of each rat was deducted from the "pre-test weight" to obtain the "total weight gain". The total weight gain of each group of rats was divided by the total weight gain of the rats in the control group to obtain the "relative total weight gain".

[0254] The subcutaneous fat of the left and right inguinal regions of the rats was weighed, and the subcutaneous fat of the left and right inguinal regions was added up to calculate the inguinal subcutaneous fat mass. The inguinal subcutaneous fat mass of the rats in each group was divided by the inguinal subcutaneous fat mass of the rats in the control group to obtain the "relative weight of inguinal subcutaneous fat".

[0255] Data were presented as mean ± SD and analyzed by one-way ANOVA. Statistical results were expressed as symbols or letters. Different symbols or letters indicated statistical differences between the groups (p < 0.05), while the same symbols or letters indicated no statistical differences between the groups (p > 0.05).

[0256] From the above dosage form preparation method and particle size analysis results, it can be seen that the ELP concentration and resveratrol concentration in the resveratrol ELP partial micelle dosage form and the resveratrol ELP micelle dosage form are the same, and only the number of drug-containing micelles is different. Therefore, compared with the resveratrol ELP partial micelle dosage form, if the resveratrol ELP micelle dosage form significantly reduces the local fat at the application site, it means that the formation of drug-containing micelles is the key factor for the resveratrol composition to significantly reduce the local fat at the application site; if the resveratrol ELP micelle dosage form significantly reduces body weight, it means that the formation of drug-containing micelles is the key factor for the resveratrol composition to significantly reduce body weight.

[0257] Similarly, the HS-15 concentration and the resveratrol HS-15 micellar dosage form are the same in the HS-15 partial micellar dosage form and the resveratrol HS-15 micellar dosage form, and only the number of drug-containing micelles is different. Therefore, if the resveratrol HS-15 micellar dosage form significantly reduces the local fat at the application site compared with the resveratrol HS-15 partial micellar dosage form, it means that the formation of drug-containing micelles is the key factor for the resveratrol composition to significantly reduce the local fat at the application site; if the resveratrol HS-15 micellar dosage form significantly reduces the body weight, it means that the formation of drug-containing micelles is the key factor for the resveratrol composition to significantly reduce the body weight.

[0258] The experimental results show that injecting the resveratrol ELP partial micelle formulation or the resveratrol HS-15 partial micelle formulation into the subcutaneous fat layer of the application site can reduce the fat (local fat) at the application site. On the other hand, injecting the resveratrol ELP micelle formulation or the resveratrol HS-15 micelle formulation into the subcutaneous fat layer of the application site can also reduce the fat (local fat) at the application site. Among the four formulations, the resveratrol ELP micelle formulation has the best local lipolysis effect.

[0259] Compared with the partial micellar formulation of resveratrol ELP, the micellar formulation of resveratrol ELP can significantly reduce local fat. Compared with the partial micellar formulation of resveratrol HS-15, the micellar formulation of resveratrol HS-15 can significantly reduce local fat.

[0260] It can be seen that the formation of micelles is the key factor for the resveratrol composition to significantly reduce local fat at the site of application, and the local fat-dissolving effect of drug-containing micelles formed by polyoxyethylene castor oil derivatives (such as ELP) is the best.

[0261] The experimental results show that injecting the resveratrol ELP partial micelle formulation or the resveratrol HS-15 partial micelle formulation into the subcutaneous fat layer of the application site can reduce body weight. On the other hand, injecting the resveratrol ELP micelle formulation or the resveratrol HS-15 micelle formulation into the subcutaneous fat layer of the application site can also reduce body weight. Among the four formulations, the resveratrol ELP micelle formulation has the best weight loss effect.

[0262] This experiment shows that the formation of micelles is the key factor for the resveratrol composition to significantly reduce body weight, and the drug-containing micelles formed by polyoxyethylene castor oil derivatives (such as ELP) have the best weight loss effect.

[0263] Experiment 4: Preparation of a pharmaceutical composition for reducing local fat

[0264] In this experiment, resveratrol was used to prepare a first pharmaceutical composition, and resveratrol and green tea extract were used to prepare a second pharmaceutical composition.

[0265] The steps for preparing the first pharmaceutical composition are as follows:

[0266] (a) mixing a first weight of resveratrol with a solvent, and stirring at 200-500 rpm at room temperature until the resveratrol is completely dissolved;

[0267] (b) adding a second weight of a pharmaceutically acceptable surfactant, stirring at a speed of 100 to 300 rpm to volatilize the solvent, wherein the surfactant has a hydrophilic-lipophilic balance value (HLB value) greater than 10;

[0268] (c) after the solvent is completely volatilized, slowly adding a third weight of a pharmaceutically acceptable aqueous solution and stirring evenly to obtain a plurality of drug-containing micelles; and

[0269] (d) After filtering with a 0.2 μm filter membrane, the filtrate containing drug-containing micelles is stored in a refrigerator away from light;

[0270] Wherein, in step (c), the drug-containing micelle is a microstructure formed by a surfactant, and resveratrol is encapsulated in the drug-containing micelle.

[0271] Preferably, the third weight is greater than or equal to 0 g.

[0272] Preferably, in step (a), the boiling point of the solvent is lower than the boiling point of pure water.

[0273] Preferably, in step (a), the solvent is a hydrophilic solvent.

[0274] Preferably, the hydrophilic solvent is at least one of methanol, ethanol, acetone and other hydrophilic solvents or a combination thereof.

[0275] Preferably, the solvent in step (a) is a lipophilic solvent.

[0276] Preferably, the lipophilic solvent is at least one of ether, benzene, chloroform, ethyl acetate, dichloromethane, hexane and other lipophilic solvents, or a combination thereof.

[0277] Preferably, in step (b), the surfactant is a nonionic surfactant.

[0278] Preferably, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0279] Preferably, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0280] Preferably, in steps (a) and (b), the weight ratio of the first weight of resveratrol to the second weight of surfactant is 1:4 to 1:500.

[0281] Preferably, in steps (a) and (c), the weight ratio of the first weight of resveratrol to the third weight of the pharmaceutically acceptable aqueous solution is 1:400 to 3:50.

[0282] Preferably, in step (c), the pharmaceutically acceptable aqueous solution is water for injection, aqueous solution for injection, or physiological saline.

[0283] Preferably, in step (c), the pharmaceutically acceptable aqueous solution comprises a local anesthetic.

[0284] Preferably, the local anesthetic is at least one of amides, p-aminobenzylamine lipids, and amino ethers, or a combination thereof.

[0285] Preferably, the amide is at least one of dibucaine, lidocaine, mepivacaine HCl, bupivacaine HCl, pyrrocaine HCl, prilocaine HCl, digammacaine, and oxethazaine, or a combination thereof.

[0286] Preferably, the p-aminobenzylamine is at least one of butacaine, dimethocaine, and tutocaine, or a combination thereof.

[0287] Preferably, the amino ether is at least one of Quinisocaine and Pramocaine or a combination thereof.

[0288] Preferably, in step (c), the pharmaceutically acceptable aqueous solution comprises an antioxidant.

[0289] Preferably, the antioxidant is at least one of beta-carotene, lutein, lycopene, bilirubin, vitamin A, vitamin C (also known as ascorbic acid), vitamin E, uric acid, nitric oxide, nitroxide, pyruvate, catalase, superoxide dismutase, glutathione peroxidases, N-acetyl cysteine, and naringenin, or a combination thereof.

[0290] The steps for preparing the second pharmaceutical composition are as follows:

[0291] (a1) mixing a fourth weight of resveratrol with a solvent, and stirring at 200-500 rpm at room temperature until the resveratrol is completely dissolved;

[0292] (b1) adding a fifth weight of a pharmaceutically acceptable surfactant, stirring at a speed of 100 to 300 rpm to volatilize the solvent, wherein the surfactant has a hydrophilic-lipophilic balance value (HLB value) greater than 10;

[0293] (c1) after the solvent is completely volatilized, slowly adding the sixth weight of the first pharmaceutically acceptable aqueous solution, and stirring evenly at a rotation speed of 100 to 300 rpm to form a plurality of drug-containing micelles; and

[0294] (d1) After filtering with a 0.2 μm filter membrane, the filtrate containing drug-containing micelles is stored in a refrigerator away from light;

[0295] The first pharmaceutically acceptable aqueous solution comprises a green tea extract in an amount of 70% by weight; the green tea extract comprises a first green tea extract component, and the first green tea extract component is epigallocatechin gallate.

[0296] Preferably, in step (a1), the boiling point of the solvent is lower than the boiling point of pure water.

[0297] Preferably, in step (a1), the solvent is a hydrophilic solvent.

[0298] Preferably, the hydrophilic solvent is at least one of methanol, ethanol, acetone and other hydrophilic solvents or a combination thereof.

[0299] Preferably, the solvent in step (a1) is a lipophilic solvent.

[0300] Preferably, the lipophilic solvent is at least one of ether, benzene, chloroform, ethyl acetate, dichloromethane, hexane and other lipophilic solvents, or a combination thereof.

[0301] Preferably, in step (b1), the surfactant is a nonionic surfactant.

[0302] Preferably, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0303] Preferably, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0304] Preferably, between step (c1) and step (d1), the method further comprises the following steps:

[0305] (c11) adding an eighth weight of the second pharmaceutically acceptable aqueous solution and stirring the mixture to allow the second pharmaceutically acceptable aqueous solution to completely dissolve.

[0306] Preferably, the green tea extract is dissolved in the first pharmaceutically acceptable aqueous solution, the drug-containing micelle is a microstructure formed by a surfactant, and resveratrol is encapsulated in the drug-containing micelle.

[0307] Preferably, the content of epigallocatechin gallate in the green tea extract in the first pharmaceutically acceptable aqueous solution is 45-100% based on 100% by weight of the total weight of the green tea extract.

[0308] Preferably, in steps (a1) and (c1), the weight ratio of the fourth weight of resveratrol to the seventh weight of green tea extract is 30:1 to 1:30.

[0309] Preferably, in steps (a1) to (c1), the weight of the fifth weight of the surfactant is 0.24 to 70 weight units, based on the total weight of the fourth weight of resveratrol and the seventh weight of the green tea extract as one weight unit; or, the weight ratio of the total weight of the resveratrol and the green tea extract to the surfactant is 4:1 to 1:70.

[0310] Preferably, in steps (a1), (c1), and (c11), based on the total weight of the fourth weight of resveratrol and the seventh weight of green tea extract as one weight unit, the total weight of the sixth weight of the first pharmaceutically acceptable aqueous solution and the eighth weight of the second pharmaceutically acceptable aqueous solution is 16 to 400 weight units; or, the sum of the fourth weight and the seventh weight: the sum of the sixth weight and the eighth weight is 1:400 to 3:50.

[0311] The total weight of the resveratrol and the green tea extract is one weight unit, and the total weight of the first pharmaceutically acceptable aqueous solution and the second pharmaceutically acceptable aqueous solution is 10 to 1000 weight units; or, the total weight of the resveratrol and the green tea extract: the total weight of the first pharmaceutically acceptable aqueous solution and the second pharmaceutically acceptable aqueous solution is 1:1000 to 1:10.

[0312] Preferably, in steps (c1) and (c11), the first pharmaceutically acceptable aqueous solution and the second pharmaceutically acceptable aqueous solution are water for injection, aqueous solution for injection, or physiological saline.

[0313] Preferably, in step (c1), the first pharmaceutically acceptable aqueous solution comprises a local anesthetic.

[0314] Preferably, the local anesthetic is at least one of amides, p-aminobenzylamine lipids, and amino ethers, or a combination thereof.

[0315] Preferably, the amide is at least one of dibucaine, lidocaine, mepivacaine HCl, bupivacaine HCl, pyrrocaine HCl, prilocaine HCl, digammacaine, and oxethazaine, or a combination thereof.

[0316] Preferably, the p-aminobenzylamine is at least one of butacaine, dimethocaine, and tutocaine, or a combination thereof.

[0317] Preferably, the amino ether is at least one of Quinisocaine and Pramocaine or a combination thereof.

[0318] Preferably, in step (c1), the pharmaceutically acceptable aqueous solution comprises an antioxidant.

[0319] Preferably, the antioxidant is at least one of beta-carotene, lutein, lycopene, bilirubin, vitamin A, vitamin C (also known as ascorbic acid), vitamin E, uric acid, nitric oxide, nitroxide, pyruvate, catalase, superoxide dismutase, glutathione peroxidases, N-acetyl cysteine, and naringenin, or a combination thereof.

[0320] In this experiment, resveratrol and a water-soluble drug are used to prepare a third pharmaceutical composition, and resveratrol and other fat-soluble drugs are used to prepare a fourth pharmaceutical composition and a fifth pharmaceutical composition.

[0321] The steps for preparing the third pharmaceutical composition are as follows:

[0322] (a2) mixing resveratrol and a solvent, and stirring at 200-500 rpm at room temperature until the resveratrol is completely dissolved;

[0323] (b2) adding a pharmaceutically acceptable surfactant, stirring at a speed of 100 to 300 rpm to volatilize the solvent, wherein the surfactant has a hydrophilic-lipophilic balance value (HLB value) greater than 10;

[0324] (c2) after the solvent is completely volatilized, slowly adding the first pharmaceutically acceptable aqueous solution and stirring evenly at a speed of 100 to 300 rpm to form a plurality of drug-containing micelles; and

[0325] (d2) After filtering with a 0.2 μm filter membrane, the filtrate containing drug-containing micelles is stored in a refrigerator away from light;

[0326] Wherein, the first pharmaceutically acceptable aqueous solution contains a water-soluble drug.

[0327] Preferably, the first pharmaceutically acceptable aqueous solution comprises a local anesthetic.

[0328] Preferably, the local anesthetic is at least one of amides, p-aminobenzylamine lipids, and amino ethers, or a combination thereof.

[0329] Preferably, the amide is at least one of dibucaine, lidocaine, mepivacaine HCl, bupivacaine HCl, pyrrocaine HCl, prilocaine HCl, digammacaine, and oxethazaine, or a combination thereof.

[0330] Preferably, the p-aminobenzylamine is at least one of butacaine, dimethocaine, and tutocaine, or a combination thereof.

[0331] Preferably, the amino ether is at least one of Quinisocaine and Pramocaine or a combination thereof.

[0332] Preferably, the pharmaceutically acceptable aqueous solution comprises an antioxidant.

[0333] Preferably, the antioxidant is at least one of beta-carotene, lutein, lycopene, bilirubin, vitamin A, vitamin C (also known as ascorbic acid), vitamin E, uric acid, nitric oxide, nitroxide, pyruvate, catalase, superoxide dismutase, glutathione peroxidases, N-acetyl cysteine, and naringenin, or a combination thereof.

[0334] Preferably, in step (a2), the boiling point of the solvent is lower than the boiling point of pure water.

[0335] Preferably, in step (a2), the solvent is a hydrophilic solvent.

[0336] Preferably, the hydrophilic solvent is at least one of methanol, ethanol, acetone and other hydrophilic solvents or a combination thereof.

[0337] Preferably, the solvent in step (a2) is a lipophilic solvent.

[0338] Preferably, the lipophilic solvent is at least one of ether, benzene, chloroform, ethyl acetate, dichloromethane, hexane and other lipophilic solvents, or a combination thereof.

[0339] Preferably, in step (b2), the surfactant is a nonionic surfactant.

[0340] Preferably, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0341] Preferably, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0342] Preferably, between step (c2) and step (d2), the method further comprises the following steps:

[0343] (c21) adding a second pharmaceutically acceptable aqueous solution and stirring evenly to allow the second pharmaceutically acceptable aqueous solution to completely dissolve.

[0344] Preferably, the water-soluble drug is dissolved in the first pharmaceutically acceptable aqueous solution, the drug-containing micelle is a microstructure formed by a surfactant, and resveratrol is encapsulated in the drug-containing micelle.

[0345] Preferably, the water-soluble drug in the first pharmaceutically acceptable aqueous solution is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate, catechin, epigallocatechingallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0346] Preferably, in steps (a2) and (c2), the weight ratio of the resveratrol to the water-soluble drug is 30:1 to 1:30.

[0347] Preferably, in steps (a2) to (c2), the weight of the surfactant is 0.24 to 70 weight units, based on the total weight of the resveratrol and the water-soluble drug as one weight unit; or, the weight ratio of the total weight of the resveratrol and the water-soluble drug to the surfactant is 4:1 to 1:70.

[0348] Preferably, in steps (a2), (c2), and (c21), the total weight of the first pharmaceutically acceptable aqueous solution and the second pharmaceutically acceptable aqueous solution is 16 to 400 weight units, based on the total weight of the resveratrol and the water-soluble drug as one weight unit; or, the sum of the fourth weight and the seventh weight: the sum of the sixth weight and the eighth weight is 1:400 to 3:50.

[0349] Preferably, in steps (c2) and (c21), the first pharmaceutically acceptable aqueous solution and the second pharmaceutically acceptable aqueous solution are water for injection, aqueous solution for injection, or physiological saline.

[0350] The steps for preparing the fourth pharmaceutical composition are as follows:

[0351] (A) a step of preparing a drug-containing micelle composition, for preparing a drug-containing micelle composition;

[0352] (B) a step of preparing a second lipid-soluble drug micelle subcomposition, for preparing a second lipid-soluble drug micelle subcomposition: and

[0353] (C) mixing the drug-containing micelle composition with the second lipid-soluble drug micelle composition to prepare the fourth pharmaceutical composition;

[0354] The step (A) of preparing the drug-containing micelle composition comprises the following steps (a3) ​​to (d3):

[0355] (a3) mixing resveratrol and the first solvent, and stirring at 200-500 rpm at room temperature until the resveratrol is completely dissolved;

[0356] (b3) adding a first pharmaceutically acceptable surfactant, stirring at a speed of 100 to 300 rpm to volatilize the first solvent, wherein the first surfactant has a hydrophilic-lipophilic balance value (HLB value) greater than 10;

[0357] (c3) after the first solvent is completely volatilized, slowly adding a pharmaceutically acceptable aqueous solution and stirring evenly to form a plurality of drug-containing micelles; and

[0358] (d3) After filtering with a 0.2 μm filter membrane, the filtrate is the drug-containing micelles secondary composition containing drug-containing micelles;

[0359] Furthermore, the step (B) of preparing the second lipid-soluble drug micelle composition comprises the following steps (a4) to (d4):

[0360] (a4) mixing another fat-soluble drug with the second solvent, and stirring at 200-500 rpm at room temperature until the other fat-soluble drug is completely dissolved;

[0361] (b4) adding a pharmaceutically acceptable second surfactant, stirring at a speed of 100 to 300 rpm to volatilize the second solvent, wherein the second surfactant has a hydrophilic-lipophilic balance value (HLB value) greater than 10;

[0362] (c4) after the second solvent is completely volatilized, slowly adding a pharmaceutically acceptable aqueous solution and stirring evenly to form a plurality of second lipid-soluble drug micelles; and

[0363] (d4) After filtering with a 0.2 μm filter membrane, the filtrate is the second lipid-soluble drug micelle sub-composition containing the second lipid-soluble drug micelle.

[0364] Wherein, in step (c3), the drug-containing micelle is a microstructure formed by a first surfactant, and resveratrol is encapsulated in the drug-containing micelle. In step (c4), the second lipid-soluble drug micelle is a microstructure formed by a second surfactant, and the other lipid-soluble drug is encapsulated in the second lipid-soluble drug micelle.

[0365] Preferably, the lipophilic drug is at least one of curcumin, quercetin, puerarin, and other lipophilic drugs other than resveratrol, or a combination thereof.

[0366] Preferably, in step (a3) ​​and / or step (a4), the boiling point of the first solvent and / or the second solvent is lower than the boiling point of pure water.

[0367] Preferably, in step (a3) ​​and / or step (a4), the first solvent and / or the second solvent is a hydrophilic solvent.

[0368] Preferably, the hydrophilic solvent is at least one of methanol, ethanol, acetone and other hydrophilic solvents or a combination thereof.

[0369] Preferably, the first solvent and / or the second solvent in step (a3) ​​and / or (a4) is a lipophilic solvent.

[0370] Preferably, the lipophilic solvent is at least one of ether, benzene, chloroform, ethyl acetate, dichloromethane, hexane and other lipophilic solvents, or a combination thereof.

[0371] Preferably, in step (b3) and / or (b4), the first surfactant and / or the second surfactant is a nonionic surfactant.

[0372] Preferably, the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0373] Preferably, the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0374] Preferably, in steps (a3) ​​and (b3), the weight ratio of the resveratrol to the first surfactant is 1:4 to 1:500.

[0375] Preferably, in steps (a4) and (b4), the weight ratio of the other fat-soluble drug to the second surfactant is 1:4 to 1:500.

[0376] Preferably, in steps (a3) ​​and (c3), the weight ratio of the resveratrol to the pharmaceutically acceptable aqueous solution is 1:400 to 3:50.

[0377] Preferably, in steps (a4) and (c4), the weight ratio of the other fat-soluble drug to the pharmaceutically acceptable aqueous solution is 1:400 to 3:50.

[0378] Preferably, in step (c3) and / or (c4), the pharmaceutically acceptable aqueous solution is water for injection, aqueous solution for injection, or physiological saline.

[0379] Preferably, in step (c3) and / or (c4), the pharmaceutically acceptable aqueous solution comprises a local anesthetic.

[0380] Preferably, the local anesthetic is at least one of amides, p-aminobenzylamine lipids, and amino ethers, or a combination thereof.

[0381] Preferably, the amide is at least one of dibucaine, lidocaine, mepivacaine HCl, bupivacaine HCl, pyrrocaine HCl, prilocaine HCl, digammacaine, and oxethazaine, or a combination thereof.

[0382] Preferably, the p-aminobenzylamine is at least one of butacaine, dimethocaine, and tutocaine, or a combination thereof.

[0383] Preferably, the amino ether is at least one of Quinisocaine and Pramocaine or a combination thereof.

[0384] Preferably, in step (c3) and / or (c4), the pharmaceutically acceptable aqueous solution comprises an antioxidant.

[0385] Preferably, the antioxidant is at least one of beta-carotene, lutein, lycopene, bilirubin, vitamin A, vitamin C (also known as ascorbic acid), vitamin E, uric acid, nitric oxide, nitroxide, pyruvate, catalase, superoxide dismutase, glutathione peroxidases, N-acetyl cysteine, and naringenin, or a combination thereof.

[0386] The steps for preparing the fifth pharmaceutical composition are as follows:

[0387] (a5) mixing resveratrol and other fat-soluble drugs with a solvent, and stirring at 200 to 500 rpm at room temperature until completely dissolved;

[0388] (b5) adding a pharmaceutically acceptable surfactant, stirring at a speed of 100 to 300 rpm to volatilize the solvent, wherein the surfactant has a hydrophilic-lipophilic balance value (HLB value) greater than 10;

[0389] (c5) after the solvent is completely volatilized, slowly adding a pharmaceutically acceptable aqueous solution and stirring evenly to form a plurality of drug-containing micelles and a plurality of second lipid-soluble drug micelles; and

[0390] (d5) After filtering with a 0.2 μm filter membrane, the filtrate containing the drug-containing micelles and the plurality of second lipid-soluble drug micelles is stored in a refrigerated state away from light.

[0391] The types and ranges of the solvent, surfactant, pharmaceutically acceptable aqueous solution, and other fat-soluble drugs used in the fifth pharmaceutical composition are the same as those of the fourth pharmaceutical composition. Moreover, the ratios and ranges of the components in the fifth pharmaceutical composition are also the same as those of the fourth pharmaceutical composition.

[0392] Preferably, the pharmaceutically acceptable aqueous solution comprises a local anesthetic and / or an antioxidant.

[0393] Preferably, the types and ranges of the local anesthetic and / or antioxidant used in the fifth pharmaceutical composition are the same as those in the fourth pharmaceutical composition.

[0394] Experiment 5: Determining the quality of pharmaceutical ingredients

[0395] Experiment 5-1 Group composition analysis

[0396] The pharmaceutical composition was allowed to stand for at least 20 minutes. If no stratification occurred, it was further tested using a particle size analyzer.

[0397] A particle size analyzer is used to determine whether the pharmaceutical composition contains micelles. If the particle size of the pharmaceutical composition is less than 250 nm, the PDI value is less than 0.4, the solution in the pharmaceutical composition is clear and transparent when observed with the naked eye, and the light path can be observed after irradiating the solution with a laser, it means that the pharmaceutical composition contains micelles.

[0398] If the pharmaceutical composition has micelles, the prepared composition is the pharmaceutical composition for reducing local fat of the present invention.

[0399] If the pharmaceutical composition does not separate into layers after standing and has micelles, the prepared composition is a preferred pharmaceutical composition for reducing local fat of the present invention.

[0400] Experiment 5-2 Analysis of the stability of pharmaceutical compositions using particle size distribution

[0401] The particle size distribution and polydispersity index (PDI) were measured using a particle size analyzer (purchased from Malvern). If the polydispersity index was less than 0.4, it indicated that the stability of the pharmaceutical composition was good, that is, the micelles in the pharmaceutical composition could exist stably.

[0402] Experiment 5-3 used accelerated stability testing to determine the stability of the pharmaceutical composition.

[0403] The storage condition of the pharmaceutical composition of the present invention is 2-8° C. In order to test the stability of the pharmaceutical composition, the inventor placed the pharmaceutical composition in a relatively high temperature and high humidity environment (temperature 25° C.±2° C., relative humidity RH60%±5%), and observed how long the micelles in the pharmaceutical composition could stably exist under the relatively high temperature state, so as to estimate how long the pharmaceutical composition could be stored under the condition of 2-8° C.

[0404] If a pharmaceutical composition can be stored at 25°C for n months, then the length of time that the pharmaceutical composition can be stored at 5°C is twice the length of n months. ((25-5) / 10) That is, the length of time that the pharmaceutical composition can be stored at 5°C is 2 times of n months.2 times, that is, 4 times.

[0405] For example, if a pharmaceutical composition can be stored at 25° C. for 6 months, then the pharmaceutical composition can be stored at 5° C. for 24 months (6 months×4 times=24 months).

[0406] Preferably, when the accelerated test is conducted at a temperature of 25±2 degrees Celsius (25±2° C.), a relative humidity of 60%±5%, and avoiding direct sunlight, the pharmaceutical composition remains in a precipitate-free state for at least 24 hours.

[0407] Preferably, when the accelerated test is conducted at a temperature of 25±2 degrees Celsius, a relative humidity of 60%±5%, and avoiding direct sunlight, the pharmaceutical composition remains in a precipitate-free state for at least 6 months.

[0408] Preferably, at a temperature of 2-8°C, the pharmaceutical composition remains free of precipitates for at least 24 months.

[0409] Experiment 6: Maximum drug loading capacity of drug-containing micelles formed by various nonionic surfactants

[0410] Since the maximum drug loading capacity of drug-containing micelles directly affects the injection volume, it has a great impact on the volume of drugs that the local subcutaneous fat layer (e.g., the subcutaneous fat layer of the face) must accommodate at one time, side effects, and burden. Therefore, this experiment will explore the maximum drug loading capacity of drug-containing micelles formed by various non-ionic surfactants to evaluate which non-ionic surfactant is the best excipient for preparing the pharmaceutical composition of the present invention.

[0411] Four nonionic surfactants were selected for this experiment. The four nonionic surfactants were polyoxyethylene 35 castor oil (ie, ELP), polyethylene glycol stearate 15 (ie, HS-15), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40, referred to as RH 40), and polysorbate 80 (ie, Tween 80).

[0412] The experiment was divided into 4 groups, namely ELP group, HS-15 group, RH40 group, and Tween 80 group.

[0413] Experimental steps:

[0414] (a') mixing 2.0 g (first weight example) of resveratrol with 300-500 mL of dichloromethane, and stirring at 150-500 rpm at room temperature until the resveratrol is completely dissolved;

[0415] (b') adding 18.0 g (second weight example) of a single nonionic surfactant described above, stirring at a rotation speed of 100 to 300 rpm to volatilize the dichloromethane; and

[0416] (c') After the solvent is completely volatilized, a composition of 20 g is obtained; 2 g of the composition is taken, 8 g (the third weight example) of physiological saline for injection is slowly added, and stirred evenly to obtain a composition to be tested. The resveratrol concentration in the composition to be tested is 20 mg / g, and the concentration of the nonionic surfactant is 18%.

[0417] The compositions to be tested in the ELP group, HS-15 group, RH40 group, and Tween 80 group were allowed to stand for at least 20 minutes to observe whether stratification occurred. If stratification occurred, it means that the resveratrol concentration was too high and the micelles in the first-stage composition were ruptured, that is, the pharmaceutical composition of the present invention with a resveratrol concentration of 20 mg / g could not be prepared using the nonionic surfactant.

[0418] The toxicity of various excipients (ELP, HS-15, RH40, and Tween 80) was evaluated, and the upper limit of the drug loading of drug-containing micelles formed by various excipients was estimated based on the injection concentration limit of excipients specified in the pharmacopoeia of various countries.

[0419] In order to find out the maximum drug loading limit of ELP, the inventors conducted subsequent experiment seven and found that the maximum drug loading of ELP is greater than or equal to 166.7 mg / g (when the ratio of resveratrol to ELP is 1:5, the prepared pharmaceutical composition contains 166.7 mg / g resveratrol).

[0420] The experimental results show that ELP is the best excipient for preparing the pharmaceutical composition of the present invention. In the pharmaceutical composition prepared by using ELP, the concentration of resveratrol can reach 166.7 mg / g.

[0421] Experiment 7: Preparation of pharmaceutical compositions using polyoxyethylene 35 castor oil (ELP)

[0422] In this experiment, a series of pharmaceutical compositions of the present invention were prepared by varying the ratio of resveratrol to polyoxyethylene 35 castor oil (ELP), and stability analysis was performed to determine the appropriate ratio of resveratrol to polyoxyethylene 35 castor oil (ELP) and the maximum drug loading when using ELP to prepare the pharmaceutical compositions of the present invention.

[0423] The experiment was divided into 9 groups, namely groups 1 to 9. The preparation method of the pharmaceutical composition of each group was roughly the same as the experimental steps of Experiment 6, except that the weight of resveratrol (the first weight in step (a')), the weight of ELP (the second weight in step (b')), and the weight of normal saline for injection (the third weight in step (c')) were different. In this experiment, the addition principle of the weight of resveratrol (the first weight), the weight of ELP (the second weight), and the weight of normal saline for injection (the third weight) is shown in Table 1.

[0424] In this experiment, the ratios (weight ratio) of resveratrol to ELP in groups 1 to 9 were 1:1, 1:2.5, 1:5, 1:8, 1:10, 1:40, 1:80, 1:200, and 1:500, respectively, and the final concentrations of resveratrol in the pharmaceutical compositions prepared in groups 1 to 9 were 1000 mg / g, 285.71 mg / g, 166.7 mg / g, 60 mg / g, 30 mg / g, 7.5 mg / g, 3.75 mg / g, 0.5 mg / g, and 0.2 mg / g, respectively. That is, in the preparation method of the pharmaceutical composition of groups 1 to 9, the weight ratio of resveratrol in step (a') to ELP in step (b') (ratio of the first weight to the second weight) is 1:1, 1:2.5, 1:5, 1:8, 1:10, 1:40, 1:80, 1:200, and 1:500, respectively, and after adding the third weight of normal saline for injection in step (c'), pharmaceutical compositions with final concentrations of resveratrol of 1000 mg / g, 285.71 mg / g, 166.7 mg / g, 60 mg / g, 30 mg / g, 7.5 mg / g, 3.75 mg / g, 0.5 mg / g, and 0.2 mg / g are prepared in sequence. Wherein, when the final drug concentration is expressed in mg / g, it represents the number of milligrams of resveratrol contained in each gram of the pharmaceutical composition.

[0425] A particle size analyzer is used to determine whether the pharmaceutical composition contains micelles and to measure the particle size of the micelles.

[0426] The particle size distribution and polydispersity index (PDI) are measured by a particle size analyzer to evaluate the stability of the pharmaceutical composition. The resveratrol content in the micelles is analyzed by high performance liquid chromatography (HPLC; such as HPLC-UV), which is defined as the "starting drug content".

[0427] The accelerated stability test is used to observe whether the pharmaceutical composition undergoes stratification under high temperature storage conditions (25 ± 2 degrees Celsius) for 3 months, and the drug content in the micelles is analyzed using high performance liquid chromatography (HPLC; e.g., HPLC-UV), which is defined as the "drug content after the accelerated test". The "drug content after the accelerated test" is divided by the "initial drug content" to obtain the "drug content percentage". If the drug content percentage is greater than or equal to 95%, it means that the stability of the pharmaceutical composition is excellent.

[0428] Table 1 Sample preparation table of pharmaceutical compositions prepared using ELP

[0429]

[0430]

[0431] Please refer to Table 2, which shows the stability analysis results of the pharmaceutical composition. Table 2 shows that when the weight ratio of resveratrol to ELP is 1:5 to 1:500, each group of pharmaceutical compositions has micelles, and the measured micelle particle size is 10 to 250 nm. Therefore, the pharmaceutical compositions prepared with a resveratrol to ELP ratio of 1:5 to 1:500 are all pharmaceutical compositions of the present invention that can be used to reduce local fat.

[0432] In terms of stability, when the ratio of resveratrol to ELP is 1:5 to 1:500, the PDI is less than 0.4. Therefore, if a pharmaceutical composition of the present invention that can be used to reduce local fat with better stability is to be prepared, the weight of ELP should be greater than or equal to 5 weight units, based on the weight of resveratrol as 1 weight unit. Preferably, the weight of ELP is 5 to 500 weight units, based on the weight of resveratrol as 1 weight unit. Preferably, the weight of ELP is 10 to 80 weight units, based on the weight of resveratrol as 1 weight unit. Preferably, the weight of ELP is 8 to 80 weight units, based on the weight of resveratrol as 1 weight unit.

[0433] Table 2 Stability analysis of pharmaceutical compositions

[0434]

[0435]

[0436] In the table above, blank columns represent that no analysis was performed.

[0437] From the data in Table 2, it can be seen that when the pharmaceutical compositions of groups 3, 5 to 7 were stored at 25 degrees Celsius for 3 months, the resveratrol drug content percentage of each group of samples was greater than 95%, and there was no obvious downward trend compared with the initial drug content. From this result, it can be seen that the pharmaceutical composition has good stability, and according to the empirical rule of the accelerated experiment, it is estimated that the pharmaceutical composition can be stored for at least 24 months under refrigeration at 2 to 8 degrees Celsius.

[0438] Experiment 8: Effects of preparing medical compositions using polyoxyethylene 35 castor oil on local lipolysis and weight loss

[0439] Example 1: Preparation of resveratrol pharmaceutical composition using polyoxyethylene 35 castor oil and resveratrol

[0440] (a') 0.4 g (first weight example) of resveratrol was mixed with 20-70 mL of ethanol, and stirred at 200-500 rpm at room temperature until the resveratrol was completely dissolved;

[0441] (b') adding 4 g (second weight example) of polyoxyethylene 35 castor oil and stirring at a speed of 100 to 300 rpm to evaporate the ethanol;

[0442] (c') after the ethanol is completely evaporated, slowly add physiological saline for injection to make the weight of the physiological saline for injection reach 80 g (the third weight example), and stir evenly to form a plurality of drug-containing micelles; and

[0443] (d') After filtering with a 0.2 μm filter membrane, the filtrate is the resveratrol pharmaceutical composition containing drug-containing micelles. Store in a dark place and refrigerate.

[0444] In this specific embodiment, the specific gravity of the injection saline solution is 1 g / mL. Therefore, the volume of 80 g of the injection saline solution is 80 mL. Accordingly, the concentration of resveratrol in the resveratrol pharmaceutical composition is about 5 mg / mL (0.4 g ÷ 80 mL = 0.005 g / mL = 5 mg / mL).

[0445] In this specific embodiment, the weight ratio of the first weight of resveratrol to the second weight of polyoxyethylene 35 castor oil (surfactant) is 0.4:4, that is, the weight ratio is 1:10.

[0446] In this specific embodiment, the weight ratio of the first weight of resveratrol to the third weight of normal saline for injection is 0.4:80, that is, the weight ratio is 1:200.

[0447] Example 2: Preparation of resveratrol compound pharmaceutical composition using polyoxyethylene 35 castor oil, resveratrol and green tea extract

[0448] (a1′) 0.36 g (the fourth weight example) of resveratrol is mixed with 20-70 mL of ethanol, and stirred at 200-500 rpm at room temperature until the resveratrol is completely dissolved;

[0449] (b1') adding 4 g (fifth weight example) of polyoxyethylene 35 castor oil, stirring evenly at a speed of 100 to 300 rpm to volatilize the ethanol;

[0450] (c1′) After the ethanol is completely volatilized, the sixth weight of the first injection physiological saline solution is slowly added and stirred at a rotation speed of 100 to 300 rpm to form a plurality of drug-containing micelles, wherein the first injection physiological saline solution contains 0.04 g (the seventh weight example) of green tea extract, and the composition of the green tea extract contains epigallocatechin gallate;

[0451] (c11') Add the eighth weight of the second injection saline solution to make the total volume of the injection saline solution 80 mL (that is, the total volume of the sixth weight of the first injection saline solution and the eighth weight of the second injection saline solution is 80 mL), and stir evenly to completely dissolve the second injection saline solution.

[0452] (d1') After filtering with a 0.2 μm filter membrane, the filtrate containing drug-containing micelles is the resveratrol compound pharmaceutical composition. Store in a dark and refrigerated place.

[0453] In this specific embodiment, the concentration of resveratrol and green tea extract in the resveratrol compound pharmaceutical composition is about 5 mg / mL [(0.36 g+0.04 g)÷80 mL=0.4 g÷80 mL=0.005 g / mL=5 mg / mL].

[0454] In this specific embodiment, if the total weight of the green tea extract (0.04 g) is 100 weight percent, the content of epigallocatechin gallate in the green tea extract is 95%.

[0455] In this specific embodiment, the weight ratio of the fourth weight of resveratrol to the seventh weight of green tea extract is 0.36:0.04, that is, the weight ratio is 9:1.

[0456] In this specific embodiment, the total weight of the fourth weight of resveratrol and the seventh weight of green tea extract (0.36 g + 0.04 g = 0.4 g) is taken as one weight unit, and the weight of the fifth weight of surfactant is 10 weight units (4 g ÷ 0.4 g = 10).

[0457] In this embodiment, the specific gravity of the injection saline solution is 1 g / mL. Therefore, in the example in which the total volume of the sixth weight of the first injection saline solution and the eighth weight of the second injection saline solution is 80 mL, the total weight of the saline solutions is 80 g.

[0458] Accordingly, taking the total weight of the fourth weight of resveratrol and the seventh weight of green tea extract (0.36 g + 0.04 g = 0.4 g) as one weight unit, the total weight of the sixth weight of the first injection of normal saline and the eighth weight of the second injection of normal saline (80 g) is 200 weight units (80 g ÷ 0.4 g = 200).

[0459] Example 3: Lipolysis effect of resveratrol pharmaceutical composition

[0460] The experiment was conducted using 6-week-old male Sprague-Dawley rats. First, 15 rats were fed a high-fat diet to induce subcutaneous fat gain. After continuous feeding until the rats reached a body weight of 330±10g, the rats were randomly divided into 3 groups, namely a high-fat control group, a low-dose resveratrol group, and a high-dose resveratrol group, with 5 rats in each group, so that there was no statistical difference in the body weight of the rats in each group.

[0461] The 5 mg / mL resveratrol pharmaceutical composition of Example 1 was injected into the subcutaneous fat layer of the lower groin of the rats in the low-dose resveratrol group, and the amount of each injection was 10 mg (10 mg / kg) of resveratrol per kilogram of body weight; that is, 2 mL of the 5 mg / mL resveratrol pharmaceutical composition of Example 1 was injected per kilogram of body weight. The 5 mg / mL resveratrol pharmaceutical composition of Example 1 was injected into the subcutaneous fat layer of the lower groin of the rats in the high-dose resveratrol group, and the amount of each injection was 20 mg (20 mg / kg) of resveratrol per kilogram of body weight; that is, 4 mL of the 5 mg / mL resveratrol pharmaceutical composition of Example 1 was injected per kilogram of body weight. The high-fat control group was given the same volume of normal saline for injection in the same injection method.

[0462] The injection site was the inguinal fat of the rats, and the injection was averaged on the left and right sides, and was injected once on the 1st, 2nd, 3rd, and 4th days of the experiment. During the experiment, the rats were continuously fed with a high-fat diet, and the weight changes were recorded daily. The drinking water and food intake were recorded once a week. The rats were fasted on the 14th day of the experiment, and sacrificed with carbon dioxide on the 15th day.

[0463] The fat of the left and right inguinal regions of the rats was weighed, and the amount of inguinal fat of each group was calculated. The data were presented as mean ± SD, and statistically analyzed by one-way ANOVA. Statistical results were expressed by symbols or letters. Different symbols or letters indicated statistical differences between the groups (p < 0.05), and the same symbols or letters indicated no statistical differences between the groups (p > 0.05).

[0464] See also Figure 3 And Table 3. Figure 3 The bar graph shows the effect of resveratrol pharmaceutical composition on the subcutaneous fat mass of rats. Table 3 shows the extent to which resveratrol pharmaceutical composition reduces the subcutaneous fat mass of rats. The subcutaneous fat mass of the inguinal region is the sum of the fat weights on the left and right sides.

[0465] The experimental results showed that compared with the high-fat control group, the amount of subcutaneous fat in the lower groin of rats in the low-dose resveratrol group (dosage was 10 mg / kg) had a decreasing trend, and the percentage of local fat reduction was 5.6%; while the amount of subcutaneous fat in the lower groin of rats in the high-dose resveratrol group (dosage was 20 mg / kg) showed a significant decrease compared with the high-fat control group (p<0.05), and the percentage of local fat reduction reached 18.1%.

[0466] Table 3. The extent to which resveratrol pharmaceutical composition reduces subcutaneous fat mass in rats

[0467]

[0468]

[0469] According to the inventor's experience, when the dosing frequency for rats is 4 times, the dosing frequency for humans is 1 to 12 times. Preferably, the dosing frequency for humans is 1 to 12 times every 1 to 30 days. Preferably, the dosing frequency for humans is 1 to 6 times every 1 to 30 days.

[0470] According to the inventor's experience, when the dosage for rats is 10 mg / kg to 20 mg / kg, the dosage for humans is 0.2 to 40 mg / kg.

[0471] Preferably, the dosage for human administration is 0.4 to 20 mg / kg.

[0472] Preferably, the dosage for human administration is 0.2 to 20 mg per square centimeter.

[0473] Preferably, the dosage for human administration is 0.4 to 12 mg per square centimeter.

[0474] Example 4: Fat-dissolving and weight-reducing effects of the resveratrol compound pharmaceutical composition

[0475] Preparation of green tea extract composition: The green tea extract was dissolved in physiological saline for injection to prepare a 5 mg / mL green tea extract composition.

[0476] The experiment was conducted using 6-week-old male Sprague-Dawley rats. First, 20 rats were fed a high-fat diet to induce subcutaneous fat increase. After continuous feeding until the rats reached a body weight of 330±10g, the rats were randomly divided into 4 groups, namely a high-fat control group, a green tea extract group, a resveratrol group, and a resveratrol-green tea extract compound group, with 5 rats in each group, so that there was no statistical difference in the body weight of the rats in each group. The body weight of each rat was recorded and defined as the "pre-experimental body weight" of each rat. Then, the drug was administered in the following manner.

[0477] The 5 mg / mL green tea extract composition of this example was injected into the subcutaneous fat layer of the lower groin of the rats in the green tea extract group, and the amount of each injection was 10 mg (10 mg / kg) of green tea extract per kilogram of body weight; that is, 2 mL of the 5 mg / mL green tea extract composition of this example was injected per kilogram of body weight. The 5 mg / mL resveratrol pharmaceutical composition of Example 1 was injected into the subcutaneous fat layer of the lower groin of the rats in the resveratrol group, and the amount of each injection was 10 mg (10 mg / kg) of resveratrol per kilogram of body weight; that is, 2 mL of the 5 mg / mL resveratrol pharmaceutical composition of Example 1 was injected per kilogram of body weight. The 5 mg / mL resveratrol compound pharmaceutical composition of Example 2 was injected into the subcutaneous fat layer of the lower groin of the rats in the resveratrol-green tea extract compound group, and the injection amount was 10 mg (10 mg / kg) of resveratrol and green tea per kilogram of body weight; that is, 2 mL of the 5 mg / mL resveratrol compound pharmaceutical composition of Example 2 was injected per kilogram of body weight. The high-fat control group was given the same volume of physiological saline for injection in the same injection method.

[0478] The injection site was the inguinal fat of the rats, and the injection was averaged on the left and right sides, and was injected once on the 1st, 2nd, 3rd, and 4th days of the experiment. During the experiment, the rats were continuously fed with a high-fat diet, and the weight changes were recorded daily. The drinking water and food intake were recorded once a week. The rats were fasted on the 14th day of the experiment, and sacrificed with carbon dioxide on the 15th day.

[0479] The body weight of each rat was recorded, which was defined as the "post-test body weight" of each rat. The "post-test body weight" of each rat was subtracted from the "pre-test body weight" to obtain the "total weight gain". The total weight gain of each group of rats was divided by the total weight gain of the rats in the high-fat control group to obtain the "relative total weight gain".

[0480] The fat of the left and right inguinal regions of the rats was weighed, and the amount of inguinal fat of each group was calculated. The data were presented as mean ± SD, and statistically analyzed by one-way ANOVA. Statistical results were expressed by symbols or letters. Different symbols or letters indicated statistical differences between the groups (p < 0.05), and the same symbols or letters indicated no statistical differences between the groups (p > 0.05).

[0481] Since each group was administered 10 mg / kg of the drug each time, the local lipolysis effect of the rats in the resveratrol-green tea extract compound group should be between the resveratrol group and the green tea extract group. If the local lipolysis effect of the rats in the resveratrol-green tea extract compound group is better than that of the resveratrol and green tea extract group, it means that the resveratrol and green tea extract in the resveratrol-compound pharmaceutical composition have a synergistic effect in the local lipolysis effect.

[0482] See also Figure 4A And Table 4. Figure 4A The bar graph of the effect of the resveratrol compound pharmaceutical composition on the subcutaneous fat mass of rats. Table 4 shows the extent to which the resveratrol compound pharmaceutical composition reduces the subcutaneous fat mass of rats. The subcutaneous fat mass of the inguinal region is the sum of the fat weights on the left and right sides.

[0483] Figure 4A The experimental results show that compared with the high-fat control group, the subcutaneous fat at the injection site of the rats in the green tea extract group did not decrease (the dosage was 10 mg of green tea extract per kilogram of body weight). Although there was no statistical difference in the amount of subcutaneous fat at the injection site of the rats in the resveratrol group compared with the high-fat control group, it showed a decreasing trend, and the percentage of local fat reduction was 5.6% (the dosage was 10 mg of resveratrol per kilogram of body weight). The amount of subcutaneous fat at the injection site of the rats in the resveratrol-green tea extract compound group showed a significant decrease (p<0.05), and the percentage of local fat reduction reached 18.9% (the dosage was 10 mg of resveratrol and green tea extract per kilogram of body weight). That is, the resveratrol compound pharmaceutical composition can more significantly achieve the effect of reducing local fat, and the effect is 3.4 times the effect of the resveratrol pharmaceutical composition.

[0484] This example shows that the composite pharmaceutical composition of resveratrol plus green tea extract of the present invention has a better local fat dissolving effect than resveratrol alone with the same dosage.

[0485] Comparison of the local lipolysis effects of rats in the resveratrol group, the green tea extract group, and the resveratrol-green tea extract compound group showed that the resveratrol and green tea extract in the resveratrol compound pharmaceutical composition have a synergistic effect in the local lipolysis effect.

[0486] Table 4. The extent to which the resveratrol compound pharmaceutical composition reduces the amount of subcutaneous fat in rats

[0487]

[0488]

[0489] See also Figure 4B and Table V. Figure 4B 1 is a bar graph showing the effect of the resveratrol compound pharmaceutical composition on the relative total weight gain of rats. Table 5 shows the extent to which the resveratrol compound pharmaceutical composition causes weight loss in rats.

[0490] Figure 4B The experimental results show that compared with the high-fat control group, the relative total weight gain of the rats in the green tea extract group did not decrease (the dosage was 10 mg of green tea extract per kilogram of body weight). Although there was no statistical difference in the relative total weight gain of the rats in the resveratrol group compared with the high-fat control group, it showed a decreasing trend, and the local fat reduction percentage was 7.4% (the dosage was 10 mg of resveratrol per kilogram of body weight). Compared with the high-fat control group, the relative total weight gain reduction percentage of the rats in the resveratrol-green tea extract compound group reached 15.9% (the dosage was 10 mg of resveratrol and green tea extract per kilogram of body weight). Although there was still no significant difference, the effect was better than that of the resveratrol group. According to the inventor's experience, as long as the dosage or frequency of administration is increased, the resveratrol single-prescription pharmaceutical composition and the resveratrol-green tea extract compound pharmaceutical group prepared by the present invention can achieve a significant weight loss effect.

[0491] Table 5. The extent to which the resveratrol compound pharmaceutical composition caused weight loss in rats

[0492]

[0493] Experiment 9: Effects of compound pharmaceutical compositions formed by resveratrol and other fat-soluble drugs on fat dissolution

[0494] In this experiment, resveratrol and other fat-soluble drugs are used to prepare compound pharmaceutical compositions to evaluate the lipolysis effects of various fat-soluble compound pharmaceutical compositions on mature adipocytes.

[0495] In this experiment, puerarin and quercetin were used to prepare various fat-soluble compound pharmaceutical compositions.

[0496] Experiment 9-1 Cytotoxicity Test

[0497] The cell viability test (MTT assay) was used to evaluate whether 50 ppm of resveratrol, puerarin, or quercetin was toxic to cells other than adipocytes. If it was not toxic, the lipolysis test was performed.

[0498] The experimental results show that 50 ppm of resveratrol, puerarin, and quercetin are not cytotoxic to somatic cells other than rat adipocytes, so this dose will not affect somatic cells.

[0499] Experiment 9-2 Lipolysis effect of mature adipocytes

[0500] The DMSO control cell culture medium, resveratrol cell culture medium, puerarin cell culture medium, quercetin cell culture medium, resveratrol-puerarin compound cell culture medium, and resveratrol-quercetin compound cell culture medium were prepared in the following manner.

[0501] DMSO control group cell culture medium: DMSO was mixed with an appropriate amount of sterile water to prepare a 0.5% DMSO solution. The 0.5% DMSO solution was mixed with a cell culture medium (product name Dulbecco's Modified Eagle Medium, purchased from Gibco) to prepare a DMSO control group cell culture medium, wherein the volume ratio of 0.5% DMSO solution to cell culture medium was 1:1000.

[0502] Resveratrol cell culture medium: Resveratrol was mixed with an appropriate amount of 0.5% DMSO solution to prepare a resveratrol solution. The resveratrol solution was mixed with a cell culture medium (product name Dulbecco's Modified Eagle Medium, purchased from Gibco) to prepare a resveratrol cell culture medium containing 50 ppm resveratrol, wherein the volume ratio of the resveratrol solution to the cell culture medium was 1:1000.

[0503] Puerarin cell culture medium: Puerarin (purchased from Sigma-Aldrich) was mixed with an appropriate amount of 0.5% DMSO solution to prepare a puerarin solution. The puerarin solution was mixed with the cell culture medium to prepare a puerarin cell culture medium containing 50 ppm puerarin, wherein the volume ratio of the puerarin solution to the cell culture medium was 1:1000.

[0504] Quercetin cell culture medium: Quercetin (purchased from Sigma-Aldrich) was mixed with an appropriate amount of 0.5% DMSO solution to prepare a quercetin solution. The quercetin solution was mixed with the cell culture medium to prepare a quercetin cell culture medium containing 50 ppm quercetin, wherein the volume ratio of the puerarin solution to the cell culture medium was 1:1000.

[0505] Resveratrol-puerarin compound cell culture solution: resveratrol, puerarin, and an appropriate amount of 0.5% DMSO solution are mixed to prepare a resveratrol-puerarin compound solution. The weight ratio of resveratrol to puerarin is 2:3. The resveratrol-puerarin compound solution is mixed with the cell culture medium to prepare a cell culture medium containing 50ppm of resveratrol-puerarin compound drugs, wherein the concentration of resveratrol is 20ppm, the concentration of puerarin is 30ppm, and the volume ratio of the resveratrol-puerarin compound solution to the cell culture medium is 1:1000.

[0506] Resveratrol-quercetin compound cell culture medium: resveratrol, quercetin, and an appropriate amount of 0.5% DMSO solution are mixed to prepare a resveratrol-quercetin compound solution. The weight ratio of resveratrol to quercetin is 2:3. The resveratrol-quercetin compound solution is mixed with the cell culture medium to prepare a cell culture medium containing 50ppm of the resveratrol-quercetin compound drug, wherein the concentration of resveratrol is 20ppm, the concentration of quercetin is 30ppm, and the volume ratio of the resveratrol-quercetin compound solution to the cell culture medium is 1:1000.

[0507] Experimental steps for the lipolysis effect of mature adipocytes:

[0508] 3T3-L1 preadipocytes (purchased from Taiwan Food Industry Development Research Institute, referred to as BCRC) were seeded in a 12-well plate so that each well contained 1×10 5 After 2 days of culture, the cells were cultured in a cell differentiation induction medium (DMI medium); which contained 0.5μM IBMX (purchased from Sigma-Aldrich), 0.1μM Dexamethasone (purchased from Sigma-Aldrich), and 5μg / ml Insulin (purchased from Humunlin R.)) for 2 days. Then, the cells were cultured in a culture medium containing 5μg / ml Insulin for 6 days. When the cell shape changed from spindle to spherical and many lipo droplets accumulated in the cells, it indicated that they had differentiated into mature adipocytes.

[0509] The mature adipocytes were divided into 6 groups, namely, DMSO control group, resveratrol group, puerarin group, quercetin group, resveratrol-puerarin compound group, and resveratrol-quercetin compound group.

[0510] The mature adipocytes in the DMSO control group, resveratrol group, puerarin group, quercetin group, resveratrol-puerarin compound group, and resveratrol-quercetin compound group were cultured with DMSO control group cell culture medium, resveratrol cell culture medium, puerarin cell culture medium, quercetin cell culture medium, resveratrol-puerarin compound cell culture medium, and resveratrol-quercetin compound cell culture medium, respectively, for 24 hours.

[0511] After the Annexin V protein (purchased from eBioscience) and Propidium iodide dye (abbreviated as PI; purchased from eBioscience) were mixed with each group of cells for a period of time, the proportion of cells in each group labeled by Annexin V protein and PI dye was analyzed by flow cytometry to evaluate the proportion of mature adipocytes undergoing apoptosis. Among them, mature adipocytes labeled by both Annexin V protein and PI dye represent that they have entered the apoptosis process; the more mature adipocytes undergo apoptosis, the better the lipolysis effect of the administered drug, and the lipolysis is through the apoptosis process rather than necrosis.

[0512] Data were presented as mean ± SD and analyzed by one-way ANOVA. Statistical results were expressed as symbols or letters. Different symbols or letters indicated statistical differences between the groups (p < 0.05), while the same symbols or letters indicated no statistical differences between the groups (p > 0.05).

[0513] Since the total dose of the drug administered in each group is 50ppm, the cell apoptosis effect of the resveratrol-puerarin compound group should be between the resveratrol group and the puerarin group. If the cell apoptosis effect of the resveratrol-puerarin compound group is better than that of the resveratrol group and the puerarin group, it means that the resveratrol and puerarin in the resveratrol-puerarin compound pharmaceutical composition have a synergistic effect on the fat-dissolving effect. Similarly, the cell apoptosis effect of the resveratrol-quercetin compound group should be between the resveratrol group and the quercetin group. If the cell apoptosis effect of the resveratrol-quercetin compound group is better than that of the resveratrol group and the quercetin group, it means that the resveratrol and quercetin in the resveratrol-quercetin compound pharmaceutical composition have a synergistic effect on the fat-dissolving effect.

[0514] See also Figure 5 . Figure 5 The effect of resveratrol-other lipophilic drugs combination on apoptosis of mature adipocytes.

[0515] Figure 5 The results showed that the percentage of cell apoptosis in the DMSO control group was 4.9±2.5%, the percentage of cell apoptosis in the resveratrol group was 19.0±1.1%, the percentage of cell apoptosis in the puerarin group was 7.2±3.7%, the percentage of cell apoptosis in the quercetin group was 5.9±2.6%, the percentage of cell apoptosis in the resveratrol-puerarin compound group was 50.6±3.8%, and the percentage of cell apoptosis in the resveratrol-quercetin compound group was 12.1±2.7%.

[0516] Comparison of the cell apoptosis effects of the resveratrol group, the puerarin group, and the resveratrol-puerarin compound group showed that the resveratrol-puerarin compound pharmaceutical composition had a synergistic effect in the fat-dissolving effect.

[0517] It can be seen that the compound pharmaceutical composition formed by resveratrol and various fat-soluble drugs can achieve the effect of dissolving fat, and resveratrol and various fat-soluble drugs have a synergistic effect in the effect of dissolving fat. Therefore, the present invention uses resveratrol and various fat-soluble drugs to prepare drug-containing micelles and second fat-soluble drug micelles, and then prepares resveratrol-other fat-soluble drug compound pharmaceutical compositions, which can be used as local fat dissolving and weight loss pharmaceutical compositions.

[0518] Experiment 10: Effects of the compound pharmaceutical composition formed by resveratrol and water-soluble drugs on fat dissolution

[0519] In this experiment, other water-soluble drugs other than green tea extract and resveratrol are used to prepare compound pharmaceutical compositions to evaluate the lipolysis effects of various resveratrol-water-soluble drug compound pharmaceutical compositions on mature adipocytes.

[0520] In this experiment, caffeine and L-carnitine were used to prepare various resveratrol-water-soluble drug compound pharmaceutical compositions.

[0521] Experiment 10-1 Cytotoxicity Test

[0522] The cell viability test (MTT assay) is used to evaluate whether 50ppm of caffeine and L-carnitine are toxic to cells other than fat cells. If they are not toxic, the lipolysis test is performed.

[0523] The experimental results show that 50 ppm of caffeine and L-carnitine are not cytotoxic to normal body cells other than rat fat cells, so this dose will not affect normal body cells.

[0524] Experiment 10-2 Lipolysis effect of mature adipocytes

[0525] Sterile water control cell culture medium, resveratrol cell culture medium, caffeine cell culture medium, L-carnitine cell culture medium, resveratrol-caffeine compound cell culture medium, and resveratrol-L-carnitine compound cell culture medium were prepared in the following manner.

[0526] Sterile water control group cell culture fluid: Sterile water was mixed with the cell culture fluid to prepare a sterile water control group cell culture fluid, wherein the volume ratio of sterile water to cell culture fluid was 1:1000.

[0527] Resveratrol cell culture medium: The preparation method is the same as that of the resveratrol cell culture medium in Experiment 9-2.

[0528] Caffeine cell culture medium: Caffeine (purchased from Sigma-Aldrich) was mixed with an appropriate amount of sterile water to prepare a caffeine solution. The caffeine solution was mixed with the cell culture medium to prepare a caffeine cell culture medium containing 50 ppm caffeine, wherein the volume ratio of the caffeine solution to the cell culture medium was 1:1000.

[0529] L-carnitine cell culture medium: L-carnitine (purchased from Sigma-Aldrich) was mixed with an appropriate amount of sterile water to prepare a L-carnitine solution. The L-carnitine solution was mixed with the cell culture medium to prepare a L-carnitine cell culture medium containing 50 ppm L-carnitine, wherein the volume ratio of the L-carnitine solution to the cell culture medium was 1:1000.

[0530] Resveratrol-caffeine compound cell culture medium: resveratrol, caffeine, and an appropriate amount of sterile water are mixed to prepare a resveratrol-caffeine compound solution, wherein the weight ratio of resveratrol to caffeine is 2:3. The resveratrol-caffeine compound solution is mixed with the cell culture medium to prepare a cell culture medium containing 50ppm of resveratrol-caffeine compound drugs, wherein the concentration of resveratrol is 20ppm, the concentration of caffeine is 30ppm, and the volume ratio of the resveratrol-caffeine compound solution to the cell culture medium is 1:1000.

[0531] Resveratrol-L-carnitine compound cell culture fluid: resveratrol, L-carnitine, and an appropriate amount of sterile water are mixed to prepare a resveratrol-L-carnitine compound solution, wherein the weight ratio of resveratrol to L-carnitine is 2:3. The resveratrol-L-carnitine compound solution is mixed with the cell culture fluid to prepare a cell culture fluid containing 50ppm of resveratrol-L-carnitine compound drugs, wherein the concentration of resveratrol is 20ppm, the concentration of L-carnitine is 30ppm, and the volume ratio of the resveratrol-L-carnitine compound solution to the cell culture fluid is 1:1000.

[0532] The method for preparing mature adipocytes was the same as that in Experiment 9-2.

[0533] The mature adipocytes were divided into 6 groups, namely, sterile water control group, resveratrol group, caffeine group, L-carnitine group, resveratrol-caffeine compound group, and resveratrol-L-carnitine compound group.

[0534] The mature adipocytes in the sterile water control group, resveratrol group, caffeine group, L-carnitine group, resveratrol-caffeine compound group, and resveratrol-L-carnitine compound group were cultured with sterile water control group cell culture medium, resveratrol cell culture medium, caffeine cell culture medium, L-carnitine cell culture medium, resveratrol-caffeine compound cell culture medium, and resveratrol-L-carnitine compound cell culture medium, respectively, for 24 hours.

[0535] After the Annexin V protein (purchased from eBioscience) and Propidium iodide dye (PI; purchased from eBioscience) were mixed with each group of cells for a period of time, the proportion of cells in each group labeled by Annexin V protein and PI dye was analyzed by flow cytometry to evaluate the proportion of mature adipocytes undergoing apoptosis. Among them, mature adipocytes labeled by both Annexin V protein and PI dye represent that they have entered the apoptosis process; the more mature adipocytes undergo apoptosis, the better the lipolysis effect of the administered drug, and the lipolysis is through the apoptosis process rather than necrosis.

[0536] Since the total dose of the drug administered to each group is 50ppm, and the proportion of resveratrol accounts for 40% and caffeine accounts for 60%, the cell apoptosis effect of the resveratrol-caffeine compound group should be close to the average of the resveratrol group and the caffeine group. If the cell apoptosis effect of the resveratrol-caffeine compound group is significantly better than the average of the resveratrol group and the caffeine group, it means that the resveratrol and caffeine in the resveratrol-caffeine compound pharmaceutical composition have a synergistic effect on the fat dissolving effect. Similarly, since the total dose of the drug administered to each group is 50ppm, and the proportion of resveratrol accounts for 40% and L-carnitine accounts for 60%, the cell apoptosis effect of the resveratrol-L-carnitine compound group should be close to the average of the resveratrol group and the L-carnitine group. If the cell apoptosis effect of the resveratrol-L-carnitine compound group is significantly better than the average values ​​of the resveratrol group and the L-carnitine group, it means that the resveratrol and L-carnitine in the resveratrol-L-carnitine compound pharmaceutical composition have a synergistic effect in the fat dissolving effect.

[0537] See also Figure 6 . Figure 6 It is the effect of resveratrol-other water-soluble drug compound pharmaceutical composition on the apoptosis of mature adipocytes.

[0538] Figure 6 The results showed that the percentage of cell apoptosis in the sterile water control group was 9.6±1.5%, the percentage of cell apoptosis in the resveratrol group was 19.0±1.1%, the percentage of cell apoptosis in the caffeine group was 6.9±1.1%, the percentage of cell apoptosis in the L-carnitine group was 5.2±1.2%, the percentage of cell apoptosis in the resveratrol-caffeine compound group was 43.1±4.5%, and the percentage of cell apoptosis in the resveratrol-L-carnitine compound group was 19.3±0.5%.

[0539] Comparison of the cell apoptosis effects among the resveratrol group, the caffeine group, and the resveratrol-caffeine compound group revealed that the resveratrol and caffeine in the resveratrol-caffeine compound pharmaceutical composition have a synergistic effect in the fat-dissolving effect.

[0540] Comparison of the cell apoptosis effects of the resveratrol group, the L-carnitine group, and the resveratrol-L-carnitine compound group revealed that the resveratrol and L-carnitine in the resveratrol-L-carnitine compound pharmaceutical composition have a synergistic effect in the fat-dissolving effect.

[0541] It can be seen that the compound pharmaceutical composition formed by resveratrol and various water-soluble drugs can achieve the effect of dissolving fat, and resveratrol and various water-soluble drugs have a synergistic effect in the effect of dissolving fat. Therefore, the present invention uses resveratrol and various water-soluble drugs to prepare a resveratrol-water-soluble drug compound pharmaceutical composition containing drug-containing micelles, which can be used as a local fat dissolving and weight loss pharmaceutical composition.

[0542] It can be seen from the embodiments of the present invention that the first pharmaceutical composition, the second pharmaceutical composition, the third pharmaceutical composition, the fourth pharmaceutical composition, the fifth pharmaceutical composition, and other pharmaceutical compositions provided by the present invention can all reduce the amount of local fat. Therefore, the first pharmaceutical composition, the second pharmaceutical composition, the third pharmaceutical composition, the fourth pharmaceutical composition, the fifth pharmaceutical composition, and other pharmaceutical compositions provided by the present invention can be used to prepare subcutaneous implant devices, subcutaneous implants, implantable infusions, ointments or salve, or patches, and can be applied to the part where subcutaneous fat needs to be reduced through percutaneous absorption methods such as subcutaneous implants, implantable infusions, smears or patches.

[0543] Preferably, the first pharmaceutical composition, the second pharmaceutical composition, the third pharmaceutical composition, the fourth pharmaceutical composition, the fifth pharmaceutical composition, and other pharmaceutical compositions provided by the present invention can reduce fat at the application site through subcutaneous fat injection. Therefore, the first pharmaceutical composition, the second pharmaceutical composition, the third pharmaceutical composition, the fourth pharmaceutical composition, the fifth pharmaceutical composition, and other pharmaceutical compositions provided by the present invention can be used to prepare subcutaneous fat layer injection injections or subcutaneous injection injections for reducing local subcutaneous fat.

[0544] Therefore, the invention content of this application includes the following items:

[0545] 1. A pharmaceutical composition for reducing local fat, comprising:

[0546] a plurality of drug-containing micelles; and

[0547] Resveratrol encapsulated in the drug-containing micelles;

[0548] The drug-containing micelle is a microstructure formed by a pharmaceutically acceptable polyoxyethylene castor oil derivative, and the hydrophilic-lipophilic balance value (HLB value) of the polyoxyethylene castor oil derivative is greater than 10.

[0549] 2. The pharmaceutical composition as described in item 1, wherein the particle size of the drug-containing micelles is 3 to 250 nm.

[0550] 3. The pharmaceutical composition according to item 1, wherein the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0551] 4. The pharmaceutical composition according to item 1, wherein the weight ratio of the resveratrol to the surfactant is 1:4 to 1:500.

[0552] 5. The pharmaceutical composition according to item 4, wherein the weight ratio of the resveratrol to the surfactant is 1:5 to 1:80.

[0553] 6. The pharmaceutical composition according to item 1, wherein the concentration of resveratrol in the pharmaceutical composition is 0.2 to 166.7 mg / mL.

[0554] 7. The pharmaceutical composition according to item 6, wherein the concentration of resveratrol in the pharmaceutical composition is 2.5 to 60 mg / mL.

[0555] 8. The pharmaceutical composition as described in item 1, further comprising a pharmaceutically acceptable aqueous solution.

[0556] 9. The pharmaceutical composition according to item 1 or 8, further comprising a second lipid-soluble drug micelle; the second lipid-soluble drug micelle is another microstructure formed by a second polyoxyethylene castor oil derivative, and a second lipid-soluble drug is encapsulated in the second lipid-soluble drug micelle.

[0557] 10. The pharmaceutical composition according to item 9, wherein the second polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0558] 11. The pharmaceutical composition according to item 9, wherein the second fat-soluble drug is at least one of quercetin, synephrine, puerarin, curcumin, and other fat-soluble drugs other than resveratrol, or a combination thereof.

[0559] 12. The pharmaceutical composition according to item 11, wherein the weight ratio of the resveratrol to the second fat-soluble drug is 30:1 to 1:20.

[0560] 13. The pharmaceutical composition according to item 1 or 8, further comprising a water-soluble drug.

[0561] 14. The pharmaceutical composition according to item 13, wherein the water-soluble drug is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate, catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0562] 15. The pharmaceutical composition according to item 14, wherein the weight ratio of the resveratrol to the water-soluble drug is 20:1 to 1:30.

[0563] 16. The pharmaceutical composition according to item 14, wherein the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the pharmaceutical composition is 0.25 to 300 mg / mL.

[0564] 17. The pharmaceutical composition as described in item 14, wherein the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0565] 18. A method for reducing the amount of subcutaneous fat in a local area of ​​a subject, comprising applying a pharmaceutical composition to the local area of ​​the subject, wherein the pharmaceutical composition comprises:

[0566] a plurality of drug-containing micelles; and

[0567] Resveratrol encapsulated in the drug-containing micelles;

[0568] The drug-containing micelle is a microstructure formed by a pharmaceutically acceptable non-ionic surfactant, and the hydrophilic-lipophilic balance value (HLB value) of the non-ionic surfactant is greater than 10.

[0569] 19. The method as described in item 18, wherein the particle size of the drug-containing micelles is 3 to 250 nm.

[0570] 20. The method as described in item 18, wherein the nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0571] 21. The method according to item 20, wherein the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0572] 22. The method according to item 18, wherein the weight ratio of the resveratrol to the nonionic surfactant is 1:4 to 1:500.

[0573] 23. The method according to item 18, wherein the concentration of resveratrol in the pharmaceutical composition is 0.2 to 166.7 mg / mL.

[0574] 24. The method according to item 18, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable aqueous solution.

[0575] 25. The method according to item 18 or 24, wherein the pharmaceutical composition further comprises a second lipid-soluble drug micelle; wherein the second lipid-soluble drug micelle is another microstructure formed by a second nonionic surfactant, and a second lipid-soluble drug is encapsulated in the second lipid-soluble drug micelle.

[0576] 26. The method according to item 25, wherein the hydrophilic-lipophilic balance value (HLB value) of the second nonionic surfactant is greater than 10.

[0577] 27. The method as described in item 26, wherein the second nonionic surfactant is at least one of polysorbate 80 (Tween 80), 2-hydroxyethyl 12-hydroxyoctadecanoate (solutol HS15), polyoxyethylene castor oil derivatives, and other nonionic surfactants, or a combination thereof.

[0578] 28. The method as described in item 27, wherein the second nonionic surfactant is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

[0579] 29. The method according to item 25, wherein the second fat-soluble drug is at least one or a combination of fat-soluble drugs other than quercetin, synephrine, puerarin, curcuminoids and other resveratrol.

[0580] 30. The method as described in item 29, wherein the weight ratio of the resveratrol to the second fat-soluble drug is 30:1 to 1:20.

[0581] 31. The method according to item 18 or 24, wherein the pharmaceutical composition further comprises a water-soluble drug.

[0582] 32. The method according to item 31, wherein the water-soluble drug is at least one of green tea extract, epigallocatechin gallate, epicatechin, epicatechin gallate, epigallocatechin, gallocatechin gallate, gallocatechin, catechin gallate, catechin, epigallocatechin gallate (EGCG), caffeine, carnitine (also known as carnitine or carnitine), L-carnitine, synephrine, chlorogenic acid, and other water-soluble drugs, or a combination thereof.

[0583] 33. The method as described in item 32, wherein the weight ratio of the resveratrol to the water-soluble drug is 20:1 to 1:30.

[0584] 34. The method according to item 32, wherein the water-soluble drug is epigallocatechin gallate, and the concentration of epigallocatechin gallate in the pharmaceutical composition is 0.25 to 300 mg / mL.

[0585] 35. The method as described in item 32, wherein the water-soluble drug is green tea extract, and the weight ratio of the resveratrol to the green tea extract is 30:1 to 1:30.

[0586] 36. The method of item 35, wherein the weight of the nonionic surfactant is 0.24 to 70 weight units, based on the total weight of the resveratrol and the green tea extract as one weight unit; or, the weight ratio of the total weight of the resveratrol and the green tea extract to the nonionic surfactant is 4:1 to 1:70.

[0587] 37. The method according to item 18, wherein the dosage of the pharmaceutical composition applied to the local area is 0.2 to 20 mg per square centimeter.

[0588] 38. The method according to item 18, wherein the dosage of the pharmaceutical composition applied to the local site is 0.2 to 40 mg per kg injected.

[0589] 39. The method according to item 18, wherein the frequency of administering the pharmaceutical composition is 1 to 12 times at intervals of 1 to 30 days.

[0590] 40. The method according to item 18, wherein the individual is an animal or a human.

[0591] 41. The method according to item 18, wherein the pharmaceutical composition is injected or applied to the local area of ​​the individual.

[0592] 42. The method according to item 18, wherein the pharmaceutical composition further comprises at least one of a cosolvent, a suspending agent, and an oil phase excipient or a combination thereof.

[0593] 43. According to the method described in item 42, at least one of the oil phase excipient and the co-solvent forms the microstructure together with the non-ionic surfactant.

[0594] The above description is only a preferred embodiment of the present invention and is not intended to limit the claims of the present invention. Therefore, all other changes or modifications that do not deviate from the spirit disclosed by the present invention should be included in the claims of the present invention.

Claims

1. A pharmaceutical composition for reducing local fat, comprising: a plurality of drug-containing micelles; and Resveratrol encapsulated in the drug-containing micelles; in, The drug-containing micelle is a microstructure formed by a pharmaceutically acceptable polyoxyethylene castor oil derivative, and the hydrophilic-lipophilic balance value (HLB value) of the polyoxyethylene castor oil derivative is greater than 10.

2. The pharmaceutical composition according to claim 1, wherein the particle size of the drug-containing micelles is 3-250 nm.

3. The pharmaceutical composition according to claim 1, wherein the polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives, or a combination thereof.

4. The pharmaceutical composition according to claim 1, wherein The weight ratio of the resveratrol to the surfactant is 1:4 to 1:

500.

5. The pharmaceutical composition according to claim 4, wherein The weight ratio of the resveratrol to the surfactant is 1:5 to 1:

80.

6. The pharmaceutical composition according to claim 1, wherein The concentration of resveratrol in the pharmaceutical composition is 0.2-166.7 mg / mL.

7. The pharmaceutical composition according to claim 6, wherein The concentration of resveratrol in the pharmaceutical composition is 2.5-60 mg / mL.

8. The pharmaceutical composition of claim 1, further comprising a pharmaceutically acceptable aqueous solution.

9. The pharmaceutical composition according to claim 1 or 8, further comprising a second lipid-soluble drug micelle; the second lipid-soluble drug micelle is another microstructure formed by a second polyoxyethylene castor oil derivative, and a second lipid-soluble drug is encapsulated in the second lipid-soluble drug micelle.

10. The pharmaceutical composition according to claim 9, wherein the second polyoxyethylene castor oil derivative is at least one of polyoxyethylene 35 castor oil (Cremophor ELP), polyoxyethylene 40 hydrogenated castor oil (Cremophor RH 40), and other polyoxyethylene castor oil derivatives or a combination thereof.

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