Lipid-dissolving composition and application thereof

Abstract

The invention provides a fat-dissolving composition and application thereof. The fat-dissolving composition comprises an active pharmaceutical ingredient, an inclusion material, a pH regulator, one or more optional components selected from a buffering agent, a stabilizing agent, an osmotic pressure regulator and a complexing agent, and optional purified water, the active pharmaceutical ingredient is selected from one or more of deoxycholic acid, pharmaceutically acceptable salts of deoxycholic acid and deoxycholic acid derivatives, and the dosage range of the inclusion material is 4-35% based on the total weight of the fat-dissolving composition. Compared with a conventional preparation with the same dosage, the fat-dissolving composition disclosed by the invention has the advantages that under the condition that a good fat-dissolving effect is achieved, the adverse reaction of an administration part can be obviously relieved, and the fat-dissolving composition has higher safety.

Description

Technical Field

[0001] This invention relates to the field of injectable lipolysis drug formulation technology, and in particular to a lipolysis composition and its application, and an injectable formulation. Background Technology

[0002] Injectable lipolysis achieves fat reduction by injecting medication into subcutaneous fat tissue, directly destroying fat cells or inducing apoptosis. Compared to liposuction, it requires less skill from the operator, does not require general anesthesia or a sterile environment, and has not been found to cause skin laxity, unevenness, or other problems, significantly reducing treatment risks. Compared to lipolysis using medical devices such as cryolipolysis, it has advantages such as lower cost, more obvious effects, the ability to adjust the injection site according to the injection depth, and less likelihood of rebound, making it an important option in the field of localized fat reduction.

[0003] Deoxycholic acid is a cell-dissolving agent; when injected into body tissues, it can damage cell membranes, leading to cell lysis. Kybella, a currently marketed lipolysis drug, uses deoxycholic acid as its main ingredient and is a routine injectable solution. In clinical use, over 50% of subjects experienced injection site hematoma, pain, edema, and numbness; over 10% experienced injection site swelling, erythema, hardening, paresthesia, itching, and nodules. Furthermore, some subjects discontinued the drug or terminated the study due to adverse reactions, resulting in an overall adverse reaction rate exceeding 90%. Additionally, the adverse reactions observed clinically require at least four weeks to recover, necessitating a four-week dosing interval. A longer dosing interval inevitably weakens or even eliminates the drug's efficacy, requiring patients to wait at least two months to see results. These adverse reactions severely limit the product's clinical application.

[0004] CN110302082A discloses a deoxycholic acid body delivery formulation, belonging to the cosmetic field. It uses deoxycholic acid, lipid metabolism-related enzymes, and tricarboxylic acid cycle-related enzymes to prepare a related dosage form, requiring the use of multiple compounds for fat reduction. CN104971058B discloses a transdermal drug delivery formulation of deoxycholic acid, using hyaluronic acid, binders, moisturizers, and penetration enhancers as excipients. This formulation allows the sodium deoxycholate patch to maintain good mechanical and transdermal properties within its shelf life while effectively slowing down the deterioration of sodium deoxycholate. Both of these formulations are transdermal, requiring deoxycholic acid to penetrate the epidermis and dermis to reach the target tissue. During this process, some drug loss is inevitable, leading to waste. Furthermore, due to the mechanism of action of deoxycholic acid, it can damage the epidermis and dermis, resulting in unnecessary adverse reactions.

[0005] CN115721599A discloses a microneedle formulation of deoxycholic acid. Utilizing the ability of microneedles to directly penetrate the skin and reach the target site, this formulation can achieve targeted fat reduction and pain relief. However, the materials used in the microneedles pose biocompatibility risks; the thickness of the skin relative to fat varies depending on the degree of obesity or the location of the fat deposit, meaning that using the same type of microneedle cannot meet the needs of all populations and may lead to the risk of administering the drug to the wrong location, such as the dermis or nerve tissue. Using different types of microneedles can also cause inconvenience in clinical use.

[0006] CN116617160A discloses an aqueous suspension of deoxycholic acid, comprising deoxycholic acid, stabilizers, suspending agents, buffers, and wetting agents. In this patent application, the drug is undissolved and in a suspension state, which significantly reduces peak plasma concentration compared to ordinary injectable solutions. However, this patent application lacks specific practical safety and efficacy results. Because the drug acts locally, the pharmacokinetic-pharmacokinetic (PK-PD) correlation is unclear, and its safety and efficacy are uncertain.

[0007] CN115867321A discloses a deoxycholic acid-peptide conjugate, which is a composition for treating obesity. The composition contains a pharmaceutically acceptable carrier, excipients, and diluents, such as physiological saline, sterile water, Ringer's solution, buffered physiological saline, glucose solution, maltodextrin solution, glycerol, or ethanol. The deoxycholic acid-peptide conjugate is obtained by conjugating a synthesized peptide with deoxycholic acid. Compared to the single substance, it has high solubility in water and significantly enhanced anti-obesity activity. Although this invention has good lipolysis effects, the peptide is a newly synthesized substance, and its safety in vivo is unknown.

[0008] It is evident that developing a lipolysis formulation with high safety and efficacy has significant clinical value. Summary of the Invention

[0009] To address the aforementioned problems, this invention, through research, has yielded a highly safe and effective lipolysis composition. This composition utilizes conventional pharmaceutical excipients with known safety profiles commonly used in the field. It exhibits selective dissolution of adipocytes with minimal or no impact on other tissues. While achieving excellent lipolysis, it reduces adverse reactions at the administration site, improving site safety. Furthermore, this composition allows for shorter dosing intervals, which can be determined clinically based on efficacy to ensure optimal lipolysis. The composition is simple in composition and convenient for clinical application.

[0010] A first aspect of the present invention provides a fat-dissolving composition comprising:

[0011] Drug active ingredients,

[0012] Packaging materials,

[0013] pH adjuster

[0014] Optionally selected from one or more of buffers, stabilizers, osmotic pressure regulators, and complexing agents, and

[0015] Optional purified water,

[0016] The active pharmaceutical ingredient is selected from one or more of deoxycholic acid, pharmaceutically acceptable salts of deoxycholic acid, and deoxycholic acid derivatives.

[0017] The amount of inclusion material used ranges from 4% to 35%, based on the total weight of the fat-dissolving composition.

[0018] In some embodiments, the inclusion material is one or more selected from cyclodextrins and their derivatives; preferably one or more selected from hydroxypropyl-β-cyclodextrin, sulfonyl-β-cyclodextrin, methyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, glucosyl-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, α-cyclodextrin, and 2,6-dimethyl-β-cyclodextrin; more preferably one or two selected from hydroxypropyl-β-cyclodextrin and sulfonyl-β-cyclodextrin.

[0019] In some embodiments, the fat-dissolving composition has at least one of the following characteristics (1)-(10):

[0020] (1) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.1-5% of the active pharmaceutical ingredient, 4-35% of the inclusion material, 0.01-5% of the pH adjuster, 0-10% of the stabilizer, 0-5% of the buffer, 0-5% of the osmotic pressure adjuster, 0-1% of the complexing agent, and 0-500 ml of purified water / g of active pharmaceutical ingredient;

[0021] (2) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.2-4% of the active pharmaceutical ingredient, 5-30% of the inclusion material, 0.02-3% of the pH adjuster, 0-8% of the stabilizer, 0.01-3% of the buffer, 0-3% of the osmotic pressure adjuster, 0-0.8% of the complexing agent, and 0-400 ml of purified water / g of active pharmaceutical ingredient;

[0022] (3) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.3-3% of the active pharmaceutical ingredient, 8-30% of the inclusion material, 0.03-2% of the pH adjuster, 0-5% of the stabilizer, 0.03-2% of the buffer, 0-2% of the osmotic pressure adjuster, 0-0.5% of the complexing agent, and 0-300 ml of purified water / g of active pharmaceutical ingredient;

[0023] (4) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.5-2% of the active pharmaceutical ingredient, 10-28% of the inclusion material, 0.05-1% of the pH adjuster, 0-3% of the stabilizer, 0.05-1% of the buffer, 0-1% of the osmotic pressure adjuster, 0-0.3% of the complexing agent, and 0-200 ml of purified water / g of active pharmaceutical ingredient;

[0024] (5) The pharmaceutically acceptable salt of the deoxycholic acid is sodium deoxycholate; the deoxycholic acid derivative is selected from one or more of the following: porcine deoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, sodium porcine deoxycholate, sodium ursodeoxycholate, sodium chenodeoxycholate, cholic acid, sodium cholate, glycocholic acid, sodium glycocholate, glycodeoxycholic acid, sodium glycodeoxycholate, taurocholic acid, sodium taurocholate, taurodeoxycholic acid, tauroursodeoxycholic acid, tauroporcine deoxycholic acid, taurochenodeoxycholic acid, glycotaurocholic acid, sodium glycotaurocholate, lithocholic acid, sodium lithocholic acid, sodium glycodeoxycholate, sodium phenylpropanecholate, sodium tyrosylcholate, sodium leucocholate, cholic acid dimer, cholic acid side chain amino acid conjugate, cholic acid side chain PEG conjugate, and cholic acid side chain glucose conjugate.

[0025] (6) The stabilizer is selected from one or more of injectable solvents and surfactants; the injectable solvent may be one or more of propylene glycol, ethanol, benzyl alcohol, dimethyl sulfoxide, N-methylpyrrolidone, liquid polyethylene glycol, dimethylacetamide, tetrahydrofuran polyethylene glycol ether, diethylene glycol monoethyl ether, and benzyl benzoate; the surfactant may be one or more of phosphatidylcholine, sorbitan fatty acid ester, polysorbate, polyoxyethylene fatty acid ester, polyoxyethylene fatty alcohol ether, and poloxamer.

[0026] (7) The pH adjuster is selected from one or more of pharmaceutically acceptable non-organic acids, organic acids, non-organic bases and organic bases; preferably selected from one or more of hydrochloric acid, phosphoric acid, acetic acid, sulfuric acid, citric acid, boric acid, lactic acid, methanesulfonic acid, maleic acid, tartaric acid, succinic acid, histidine, lysine, arginine, butyric acid, meglumine, ethylenediamine, ethanolamine, triethylamine, sodium hydroxide and potassium hydroxide;

[0027] (8) The buffer is selected from one or more of the following: disodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate, potassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium citrate, potassium citrate, sodium acetate, potassium acetate, tris(hydroxymethyl)aminomethane, potassium hydrogen phthalate, sodium hydrogen phthalate, sodium tetraborate, potassium tetraborate, sodium barbital, and glycine.

[0028] (9) The osmotic pressure regulator is selected from one or more of lactose, trehalose, xylitol, L-glucoside, sorbitol, maltose, glycine, sarcosine, sodium chloride, potassium chloride, glucose, sucrose and mannitol; preferably selected from one or more of trehalose, lactose, sodium chloride, potassium chloride, glucose, sucrose and mannitol;

[0029] (10) The complexing agent is one or more selected from ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid and calcium disodium ethylenediaminetetraacetic acid.

[0030] In the lipolysis composition of the present invention, a portion of the active pharmaceutical ingredient associates with the inclusion material to form an inclusion complex; this portion is referred to as the associated portion. The portion of the active pharmaceutical ingredient that does not associate with the inclusion material to form an inclusion complex is referred to as the free portion. In some embodiments, the free portion constitutes <30% of the total active pharmaceutical ingredient in the composition of the present invention by mass; optionally, it is <20%.

[0031] In some embodiments, the total daily dose of the lipolysis composition, based on the active pharmaceutical ingredient, is 0.1 mg to 1000 mg; preferably 1 mg to 500 mg.

[0032] A second aspect of the present invention provides the use of the lipolysis composition of the first aspect in the preparation of a medicament for the prevention or treatment of fat-related diseases.

[0033] In some embodiments, the fat-related diseases include, but are not limited to, one or more of the following: fat accumulation, lipoma, painful lipoma, cellulite, fat edema, fat hyperplasia, fat malnutrition, lipodystrophy, fat deposition after liposuction, and obstructive sleep apnea; optionally, the dosage form of the drug is an injectable preparation; more preferably, the application sites of the injectable preparation include, but are not limited to, one or more of the following: abdomen, under the eyes, under the chin, under the arms, buttocks, thighs, calves, back, bra line, armpits, stomach, and ankles.

[0034] A third aspect of the present invention provides an injectable formulation prepared from the lipolysis composition of the first aspect of the present invention.

[0035] Optionally, the injectable formulation may be administered via injection, including but not limited to one or more of subcutaneous injection, intramuscular injection, intravenous injection, and intralesional injection.

[0036] The association between the active pharmaceutical ingredient and the inclusion material in the lipolysis composition of the present invention has the following advantages:

[0037] ① It selectively dissolves adipocytes, which can reduce adverse reactions at the administration site and improve medication safety;

[0038] ② To exert a sustained fat-dissolving effect and ensure good fat-dissolving results;

[0039] ③ It can avoid the problem of weakened efficacy caused by prolonged dosing intervals due to adverse reactions when using Kybella. Clinically, the dosing interval can be determined according to the efficacy to ensure good fat-dissolving effect and improve patient comfort.

[0040] The lipolytic composition of the present invention, compared with conventional formulations of the same dosage (same dosage of active pharmaceutical ingredient), can significantly reduce adverse reactions at the administration site while maintaining good lipolytic effect. For example, it can reduce the incidence of adverse reactions at the administration site by more than 10%, preferably more than 20%, and more preferably more than 30%.

[0041] In summary, the fat-dissolving composition of the present invention, compared with conventional formulations of the same dosage (same dosage of active pharmaceutical ingredient), can significantly improve safety while maintaining good fat-dissolving effect, which has not been reported in the art. Attached Figure Description

[0042] To more clearly illustrate the technical solutions in the embodiments of this application and to more completely understand this application and its beneficial effects, the accompanying drawings used in the description of the embodiments will be briefly introduced below, but the present invention is not limited thereto.

[0043] Figure 1 Photographs showing the injection sites of mice at different time points in Comparative Example 1;

[0044] Figure 2 Photos showing the injection sites of mice at different time points in Comparative Example 2;

[0045] Figure 3 Photos showing the injection sites of mice at different time points in Comparative Example 3;

[0046] Figure 4 Photographs showing the injection sites of mouse mice at different time points after injection of composition 5;

[0047] Figure 5 Photographs showing the injection sites of mouse mice at different time points after injection of composition 6;

[0048] Figure 6 Photographs showing the injection sites of mouse mice at different time points after injection of composition 7;

[0049] Figure 7 A figure showing the results of tissue section observations in different tissues of mice 21 days after injection of different compositions;

[0050] Figure 8 A graph showing the survival percentage of each cell in different compositions. Detailed Implementation

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0052] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0053] In this invention, numerical ranges are involved. Unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe features or characteristics, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0054] This document only specifically discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form an unspecified range; and any lower limit can be combined with other lower limits to form an unspecified range, just as any upper limit can be combined with any other upper limit to form an unspecified range. Furthermore, each individually disclosed point or single value can itself serve as a lower or upper limit and be combined with any other point or single value or with other lower or upper limits to form an unspecified range.

[0055] Unless otherwise specified, the temperature parameters in this invention can be either constant temperature processing or processing within a certain temperature range. The constant temperature processing allows temperature fluctuations within the precision range controlled by the instrument.

[0056] In the description of the invention, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0057] Unless otherwise specified, all embodiments and optional embodiments of the present invention can be combined with each other to form new technical solutions. Unless otherwise specified, all technical features and optional technical features of the present invention can be combined with each other to form new technical solutions.

[0058] Unless otherwise specified, all steps of the present invention may be performed sequentially or randomly, but are preferably performed sequentially.

[0059] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0060] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this invention, solve the technical problem of this invention, and achieve the expected technical effect of this invention.

[0061] In this document, terms such as "preferred," "better," "more suitable," and "ideal" are merely descriptions of more effective implementation methods or embodiments and should be understood not to limit the scope of protection of this invention. If multiple "preferred" terms appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "preferred" term shall be independent.

[0062] In this invention, terms such as "further," "even more," and "particularly" are used for descriptive purposes and to indicate differences in content, but should not be construed as limiting the scope of protection of this invention.

[0063] In this invention, "optionally," "optionally," and "optional" mean that they are optional, that is, they are selected from either "with" or "without." If multiple "options" appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "option" is independent.

[0064] In this invention, "conventional formulation" refers to a formulation that does not contain inclusion materials or whose inclusion materials are outside the scope of this invention, such as the commercially available injectable Kybella or comparative examples 2 and 3.

[0065] A first aspect of the present invention provides a lipolysis composition comprising a pharmaceutically active ingredient, an inclusion material, a pH adjuster, optionally selected from one or more stabilizers, osmotic pressure regulators and complexing agents, and optionally purified water.

[0066] It is easy to understand that the active pharmaceutical ingredient mentioned in this invention refers to the component in the lipolysis composition that has a lipolysis effect.

[0067] It should be noted that "optional" indicates that the fat-dissolving composition may or may not contain this ingredient.

[0068] Not limited to any particular theory, the lipolysis composition of the present invention selectively dissolves adipocytes, ensuring a sustained lipolysis effect by maintaining the drug concentration in the local tissue within the therapeutic window at the administration site, thus guaranteeing a good lipolysis effect; it also reduces adverse reactions at the administration site and improves medication safety; at the same time, it avoids the problem of weakened efficacy caused by prolonged dosing intervals due to adverse reactions when using Kybella. Clinically, the dosing interval can be determined according to the efficacy to ensure a good lipolysis effect.

[0069] It should be noted that, compared with conventional lipolysis preparations of the same dosage (same dosage of active pharmaceutical ingredient), the lipolysis composition provided by the present invention reduces the incidence of adverse reactions by more than 10%, optionally by more than 20%, and more preferably by more than 30%, while still achieving good lipolysis effect.

[0070] In some embodiments, based on the total weight of the lipolysis composition, the lipolysis composition comprises 0.1-5% of the active pharmaceutical ingredient, 4-35% of the inclusion material, 0.01-5% of the pH adjuster, 0-10% of the stabilizer, 0-5% of the buffer, 0-5% of the osmotic pressure adjuster, 0-1% of the complexing agent, and 0-500 ml of purified water per g of active pharmaceutical ingredient.

[0071] It should be noted that the fat-dissolving composition provided by the present invention, after loading the drug, can reduce adverse reactions at the administration site.

[0072] Based on the total weight of the lipolysis composition, the amount of the active pharmaceutical ingredient contained in the lipolysis composition is 0.1-5%; for example, it can be, but is not limited to, 0.1%, 0.2%, 0.3%, 0.5%, 1%, 2%, 3%, 5% or any two of the above amounts.

[0073] Based on the total weight of the fat-dissolving composition, the amount of inclusion material contained in the fat-dissolving composition is 4-35%; for example, it can be, but is not limited to, 4%, 5%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, or any two of the above amounts.

[0074] Based on the total weight of the fat-dissolving composition, the amount of stabilizer contained in the fat-dissolving composition is 0-10%; for example, it can be, but is not limited to, 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or any range between two of the above amounts.

[0075] Based on the total weight of the lipolysis composition, the pH adjustment dose contained in the lipolysis formulation is 0.01-5%; for example, it can be, but is not limited to, 0.01%, 1%, 2%, 3%, 4%, 5% or any two of the above amounts.

[0076] Based on the total weight of the fat-dissolving composition, the buffer dosage contained in the fat-dissolving formulation is 0-5%; for example, it can be, but is not limited to, 0%, 0.1%, 0.2%, 0.3%, 0.5%, 1%, 2%, 3%, 4%, 5%, or any range between two of the above amounts.

[0077] Based on the total weight of the fat-dissolving composition, the weight percentage of the osmotic pressure regulator contained in the fat-dissolving composition is 0-5%; for example, it can be, but is not limited to, 0%, 0.1%, 0.2%, 0.3%, 0.5%, 0.8%, 1%, 2%, 3%, 5%, or any two of the above amounts.

[0078] Based on the total weight of the fat-dissolving composition, the weight percentage of the complexing agent contained in the fat-dissolving composition is 0-1%; for example, it can be, but is not limited to, 0%, 0.1%, 0.2%, 0.3%, 0.5%, 0.8%, 1.0%, or any two of the above amounts.

[0079] It should be noted that when the weight part is 0 parts, it indicates that the fat-dissolving composition does not contain this component.

[0080] When the weight ratios of the components in the lipolysis composition are within the above-mentioned ranges, the following beneficial effects are achieved: ① It selectively dissolves fat cells, reducing adverse reactions at the administration site and improving medication safety; ② It exerts a sustained lipolysis effect, ensuring good lipolysis results; ③ It avoids the problems of prolonged dosing intervals and long treatment cycles caused by adverse reactions when using commercially available injectable Kybella. Clinically, the dosing interval can be determined based on the efficacy to ensure good lipolysis results and improve patient comfort.

[0081] In some embodiments, based on the total weight of the lipolysis composition, the lipolysis composition comprises 0.2-4% of the active pharmaceutical ingredient, 5-30% of the inclusion material, 0.02-3% of the pH adjuster, 0-8% of the stabilizer, 0.01-3% of the buffer, 0-3% of the osmotic pressure adjuster, 0-0.8% of the complexing agent, and 0-400 ml of purified water per g of active pharmaceutical ingredient.

[0082] In some embodiments, based on the total weight of the lipolysis composition, the lipolysis composition comprises 0.3-3% of the active pharmaceutical ingredient, 8-30% of the inclusion material, 0.03-2% of the pH adjuster, 0-5% of the stabilizer, 0.03-2% of the buffer, 0-2% of the osmotic pressure adjuster, 0-0.5% of the complexing agent, and 0-300 ml of purified water per g of active pharmaceutical ingredient.

[0083] In some embodiments, based on the total weight of the lipolysis composition, the lipolysis composition comprises 0.5-2% of the active pharmaceutical ingredient, 10-28% of the inclusion material, 0.05-1% of the pH adjuster, 0-3% of the stabilizer, 0.05-1% of the buffer, 0-1% of the osmotic pressure adjuster, 0-0.3% of the complexing agent, and 0-200 ml of purified water per g of active pharmaceutical ingredient.

[0084] In some embodiments, the pharmaceutically active ingredient of the lipolytic composition includes one or more selected from deoxycholic acid, pharmaceutically acceptable salts of deoxycholic acid, and deoxycholic acid derivatives.

[0085] Deoxycholic acid, also known as 3α,12α-dihydroxy-5β-cholan-24-oic acid, has the following structure:

[0086]

[0087] Pharmaceutically acceptable salts of deoxycholic acid are preferably alkali metal salts, such as sodium or potassium salts, and more preferably sodium salts.

[0088] The deoxycholic acid derivative may be selected from one or more of the following: porcine deoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, sodium porcine deoxycholate, sodium ursodeoxycholate, sodium chenodeoxycholate, cholic acid, sodium cholate, glycocholic acid, sodium glycocholate, glycodeoxycholic acid, sodium glycodeoxycholate, taurocholic acid, sodium taurocholate, taurodeoxycholic acid, tauroursodeoxycholic acid, tauroporcine deoxycholic acid, taurochenodeoxycholic acid, glycotaurocholic acid, sodium glycotaurocholate, lithocholic acid, sodium lithochate, sodium glycodeoxycholate, sodium phenylpropanecholate, sodium tyrosocholate, sodium leucosterate, sodium leucosterate, cholic acid dimer, cholic acid side-chain amino acid conjugate, cholic acid side-chain PEG conjugate, and cholic acid side-chain glucose conjugate, but is not limited thereto.

[0089] In some embodiments, the inclusion material is one or more selected from cyclodextrin derivatives; preferably one or more selected from hydroxypropyl-β-cyclodextrin, sulfonyl-β-cyclodextrin, methyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, glucosyl-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, α-cyclodextrin, and 2,6-dimethyl-β-cyclodextrin; more preferably one or two selected from hydroxypropyl-β-cyclodextrin and sulfonyl-β-cyclodextrin.

[0090] In some embodiments, the stabilizer is one or more selected from injectable solvents and surfactants; the injectable solvent may be one or more selected from propylene glycol, ethanol, benzyl alcohol, dimethyl sulfoxide, N-methylpyrrolidone, liquid polyethylene glycol, dimethylacetamide, tetrahydrofuran polyethylene glycol ether, diethylene glycol monoethyl ether, benzyl benzoate, and the surfactant may be one or more selected from phosphatidylcholine, sorbitan fatty acid ester, polysorbate, polyoxyethylene fatty acid ester, polyoxyethylene fatty alcohol ether, poloxamer, and sorbitol.

[0091] In some embodiments, the pH adjuster is selected from one or more of pharmaceutically acceptable non-organic acids, organic acids, non-organic bases, and organic bases; preferably, it is selected from one or more of hydrochloric acid, phosphoric acid, acetic acid, sulfuric acid, citric acid, boric acid, lactic acid, methanesulfonic acid, maleic acid, tartaric acid, succinic acid, histidine, lysine, arginine, butyric acid, meglumine, ethylenediamine, ethanolamine, triethylamine, sodium hydroxide, and potassium hydroxide.

[0092] In some embodiments, the buffer is selected from one or more of the following: disodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate, potassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium citrate, potassium citrate, sodium acetate, potassium acetate, tris(hydroxymethyl)aminomethane, potassium hydrogen phthalate, sodium hydrogen phthalate, sodium tetraborate, potassium tetraborate, sodium barbital, and glycine.

[0093] In some embodiments, the osmotic pressure regulator is one or more selected from lactose, trehalose, xylitol, L-glycoside, sorbitol, maltose, glycine, sarcosine, sodium chloride, potassium chloride, glucose, sucrose, and mannitol; preferably, it is one or more selected from trehalose, lactose, sodium chloride, potassium chloride, glucose, sucrose, and mannitol.

[0094] In some embodiments, the complexing agent is one or more selected from ethylenediaminetetraacetic acid (EDTA), disodium EDTA, and calcium disodium EDTA.

[0095] In the lipolysis composition of the present invention, a portion of the active pharmaceutical ingredient associates with the inclusion material to form an inclusion complex, referred to as the associated portion; further optionally, the portion of the active pharmaceutical ingredient that does not associate with the inclusion material to form an inclusion complex is referred to as the free portion. In some embodiments, the free portion accounts for <30% of the mass of the active pharmaceutical ingredient; optionally, it is <20%.

[0096] In some embodiments, the total daily dose of the fat-dissolving composition, based on the active pharmaceutical ingredient, is 0.1 mg to 1000 mg; optionally, it is 1 mg to 500 mg. The daily dose is primarily related to the area and amount of fat in the target fat-reducing region, and those skilled in the art can adjust the daily dose based on these factors. The dose for each treatment course is related to the area, amount of fat, and target fat reduction, and those skilled in the art can adjust the dose for each treatment course based on these factors.

[0097] A second aspect of the present invention provides the use of a lipolysis composition in the preparation of a medicament for the prevention or treatment of fat-related diseases.

[0098] In some embodiments, the fat-related diseases include, but are not limited to, one or more of the following: fat accumulation, lipoma, painful lipoma, cellulite, fat edema, fat hyperplasia, fat malnutrition, lipodystrophy, fat deposition after liposuction, and obstructive sleep apnea; optionally, the dosage form of the drug may be an injectable formulation; more preferably, the injection site may include one or more of the following: abdomen, below the eyes, below the chin, below the arm, buttocks, thighs, calves, back, bra line, armpits, stomach, and ankles.

[0099] The third and fourth aspects of the present invention each provide an injectable formulation prepared from the lipolysis composition of the first aspect of the present invention;

[0100] Optionally, the injectable formulation is administered via injection, including but not limited to one or more of subcutaneous injection, intramuscular injection, intravenous injection, and intralesional injection.

[0101] In some embodiments, the fat-dissolving composition can be prepared by the following method:

[0102] (1) Dissolve the buffer in purified water, adjust the pH to 10.0-11.0 using a pH adjuster, add the prescribed amount of active pharmaceutical ingredient, and dissolve completely; (2) Add the prescribed amount of inclusion material to the above solution and mix until clear and transparent; (3) Adjust the pH of the solution to 7.0-9.0 using a pH adjuster; (4) Filter using a 0.22μm microporous membrane; (5) Fill according to the drug content; (6) Optionally, freeze-dry the filled solution.

[0103] If the prescription contains stabilizers, osmotic pressure regulators and / or complexing agents, the above substances may be added in any of the steps (1), (2) and (3).

[0104] In some embodiments, the fat-dissolving composition can be prepared by the following method:

[0105] (1) Add the prescribed amount of active pharmaceutical ingredient and buffer to purified water, and adjust the pH with a pH adjuster until the active pharmaceutical ingredient is completely dissolved to obtain a clear and transparent solution; (2) Add the prescribed amount of inclusion material to the above solution and mix until clear and transparent; (3) Adjust the pH value of the solution to 7.0 to 9.0 with a pH adjuster; (4) Filter with a 0.22μm microporous membrane; (5) Fill according to the drug content; (6) Optionally, freeze-dry the filled solution.

[0106] If the prescription contains stabilizers, osmotic pressure regulators and / or complexing agents, the above substances may be added in any of the steps (1), (2) and (3).

[0107] In some embodiments, the fat-dissolving composition can be prepared by the following method:

[0108] (1) Weigh the prescribed amount of inclusion material into the prescribed amount of purified water, stir until completely dissolved, and obtain a clear and transparent solution; (2) Add buffer and pH adjuster, and dissolve to obtain a clear and transparent solution; (3) Add the prescribed amount of active pharmaceutical ingredient into the above solution, and dissolve completely; adjust the pH value to 7.0 to 9.0 using pH adjuster; (4) Filter the above solution using a 0.22μm microporous membrane; (5) Fill according to the drug content; (6) Optionally, freeze-dry the filled solution.

[0109] If the prescription contains stabilizers, osmotic pressure regulators and / or complexing agents, the above substances may be added in any of the steps (1), (2) and (3).

[0110] In the preparation method of the composition of the present invention, if the active pharmaceutical ingredient is dissolved first, it needs to be dissolved and clarified before adding cyclodextrin to the solution for inclusion; if the cyclodextrin is dissolved first, the cyclodextrin solution needs to be adjusted to alkaline before adding the active pharmaceutical ingredient for inclusion.

[0111] It should be noted that the composition prepared by this invention is a clear and transparent solution or a clear and transparent solution before lyophilization.

[0112] It should be noted that the pH value of the composition prepared by this invention is 7.0 to 9.0.

[0113] When used in vivo, the lipolysis composition of the present invention reduces adverse reactions at the application site and improves medication safety compared to conventional lipolysis preparations; it enables the drug to be released from the drug-inclusion material conjugate, ensuring a good lipolysis effect; it avoids the problem of weakened efficacy caused by prolonged dosing intervals due to adverse reactions when using commercially available injectable Kybella. Clinically, the dosing interval can be determined according to the efficacy to ensure a good lipolysis effect and improve patient comfort.

[0114] The technical solution of the present invention will be described in detail below with reference to specific embodiments. It should be understood that these embodiments are only for illustrating the present invention and are not intended to limit the scope of the present invention. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this invention, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0115] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0116] Example

[0117] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0118] Example 1: Preparation of the composition

[0119] 1. The formulations of compositions Z1 to Z14 are shown in Table 1.

[0120] Table 1. Composition of the composition

[0121]

[0122]

[0123] 2. Preparation process of Z1-Z5:

[0124] (1) Accurately weigh the prescribed amount of buffer and stabilizer into the prescribed amount of purified water and stir until completely dissolved;

[0125] (2) Slowly add the prescribed amount of the active pharmaceutical ingredient to the above solution and stir;

[0126] (3) Add the prescribed amount of sodium hydroxide and stir until the solution is clear;

[0127] (4) Add the prescribed amount of inclusion material, stir at 60°C for 2 hours until the solution is clear; add the osmotic pressure regulator and stir until the solution is clear, then adjust the pH of the solution to 7.0-9.0 with hydrochloric acid;

[0128] (5) The above solution was filtered through a 0.22 μm microporous membrane to obtain a clear and transparent solution;

[0129] (6) Dispense and sterilize compositions Z3 and Z4 to obtain the final product;

[0130] (7) The compositions Z1, Z2 and Z5 are packaged and freeze-dried to obtain the freeze-dried formulation.

[0131] 3. Preparation process of Z6-Z14:

[0132] (1) Accurately weigh the prescribed amount of inclusion material and stabilizer, add them to the prescribed amount of purified water, stir at 60°C until dissolved, and obtain a clear and transparent solution;

[0133] (2) Weigh the prescribed amount of buffer and sodium hydroxide into the solution obtained in (1), sonicate until dissolved, and obtain a clear and transparent solution;

[0134] (3) Weigh the prescribed amount of active pharmaceutical ingredient into the solution obtained in (2), stir at 60°C for 2 hours until the solution is clear; add osmotic pressure regulator and stir until the solution is clear, and adjust the pH of the solution to 7.0-9.0 with hydrochloric acid;

[0135] (4) The above solution was filtered through a 0.22 μm microporous membrane to obtain a clear and transparent solution;

[0136] (5) Dispense and sterilize the compositions Z6, Z7, Z8, and Z10 to obtain the final product;

[0137] (6) The compositions Z9, Z11, Z12, Z13 and Z14 are packaged and freeze-dried to obtain the freeze-dried preparation.

[0138] Example 2, Preparation of the Composition

[0139] 1. The formulations of compositions Z15 to Z28 are shown in Table 2.

[0140] Table 2. Composition of the composition

[0141]

[0142]

[0143] 2. Preparation process of Z15-Z21:

[0144] (1) Accurately weigh the prescribed amount of buffer and stabilizer into the prescribed amount of purified water and stir until completely dissolved;

[0145] (2) Slowly add the prescribed amount of the active pharmaceutical ingredient to the above solution and stir;

[0146] (3) Add the prescribed amount of sodium hydroxide and stir until the solution is clear;

[0147] (4) Add the prescribed amount of inclusion material, stir at 60°C for 2 hours until the solution is clear; adjust the pH of the solution to 7.0-9.0 with hydrochloric acid;

[0148] (5) The above solution was filtered through a 0.22 μm microporous membrane to obtain a clear and transparent solution;

[0149] (6) Dispense and sterilize the compositions Z17, Z18, Z19, Z20, and Z21 to obtain the final product;

[0150] (7) The compositions Z15 and Z16 are packaged and freeze-dried to obtain the freeze-dried formulation.

[0151] 3. Preparation process of Z22-Z28:

[0152] (1) Accurately weigh the prescribed amount of buffer and stabilizer, add them to the prescribed amount of purified water, and adjust the pH value to 10.0-11.0 using a pH adjuster;

[0153] (2) Add the prescribed amount of active ingredient and stir until completely dissolved;

[0154] (3) Add the prescribed amount of inclusion material, stir at 60°C for 2 hours until the solution is clear; adjust the pH of the solution to 7.0-9.0 with hydrochloric acid;

[0155] (4) The above solution was filtered through a 0.22 μm microporous membrane to obtain a clear and transparent solution;

[0156] (5) Dispense and sterilize the compositions Z22, Z23, and Z24 to obtain the final product;

[0157] (6) The compositions Z25, Z26, Z27 and Z28 are packaged and freeze-dried to obtain the freeze-dried formulation.

[0158] Comparative Example 1: Preparation of Commercially Available Injectable Solution

[0159] 1. Comparative Example 1 was prepared by referring to the commercially available injectable Kybella, and its formulation composition is shown in Table 3.

[0160] Table 3. Formulation of Kybella Injection

[0161]

[0162] 2. Preparation process

[0163] Weigh the prescribed amounts of benzyl alcohol, disodium hydrogen phosphate, sodium chloride, and sodium hydroxide into water for injection. Stir and sonicate until completely dissolved. Add the prescribed amount of deoxycholic acid, and stir and sonicate until completely dissolved. Slowly adjust the pH to 8.3 with hydrochloric acid / sodium hydroxide to obtain a drug-containing solution. Filter, dispense, seal, and then sterilize by moist heat to obtain a clear and transparent liquid. Comparative Examples 2-3: Preparation of cyclodextrin formulations with different dosages.

[0164] 1. By varying the proportions of the inclusion materials, a formulation was prepared, and the composition of the formulation is shown in the table below.

[0165] Table 4. Prescription composition of Comparative Examples 2-3

[0166]

[0167] 2. Preparation process

[0168] (1) Accurately weigh the prescribed amounts of dipotassium hydrogen phosphate and potassium dihydrogen phosphate into the prescribed amount of purified water, and stir until completely dissolved;

[0169] (2) Slowly add the prescribed amount of the active pharmaceutical ingredient to the above solution and stir;

[0170] (3) Add the prescribed amount of sodium hydroxide and stir until the solution is clear;

[0171] (4) Add the prescribed amount of inclusion material, stir at 60°C for 2 hours until the solution is clear; adjust the pH of the solution to 7.0-9.0 with hydrochloric acid;

[0172] (5) The above solution was filtered through a 0.22 μm microporous membrane to obtain a clear and transparent solution;

[0173] (6) Dispense and sterilize Comparative Examples 2 and 3 to obtain the final product;

[0174] Preparation of different types of cyclodextrin preparations (Comparative Examples 4-5)

[0175] 1. By changing the type of inclusion material, a formulation was prepared, and the formulation composition is shown in the table below.

[0176] Table 5. Prescription composition of Comparative Example 4 and Comparative Example 5

[0177]

[0178] 2. Preparation process

[0179] (1) Accurately weigh the prescribed amounts of dipotassium hydrogen phosphate and potassium dihydrogen phosphate into the prescribed amount of purified water, and stir until completely dissolved;

[0180] (2) Slowly add the prescribed amount of the active pharmaceutical ingredient to the above solution and stir;

[0181] (3) Add the prescribed amount of sodium hydroxide and stir until the solution is clear;

[0182] (4) Add the prescribed amount of inclusion material and stir at 60°C for 2 hours; adjust the pH of the solution to 7.0–9.0 using hydrochloric acid;

[0183] (5) Filter the above solution using a 0.22 μm microporous membrane;

[0184] The preparation process revealed that the solutions of Comparative Example 4 and Comparative Example 5 were both milky white liquids. This demonstrates that using γ-cyclodextrin or β-cyclodextrin as inclusion materials cannot yield a clear and transparent composition.

[0185] Experimental Example 1: Inclusion Rate Determination

[0186] Take 0.5 ml of the drug solution prepared in Example 1 or the drug solution obtained by reconstituted the lyophilized drug solution with an appropriate amount of water into an ultrafiltration tube (molecular weight 0.3 kDa). Set the centrifugation speed to 4000 rpm and the centrifugation time to 15 min. Measure the total active pharmaceutical ingredient content and the free active pharmaceutical ingredient content of the drug solution respectively, and calculate the inclusion rate of the composition.

[0187] The inclusion rate is calculated as follows:

[0188] Inclusion rate % = (Total active pharmaceutical ingredient content - Free active pharmaceutical ingredient content) / Total active pharmaceutical ingredient content.

[0189] The results are shown in Table 6.

[0190] Table 6. Inclusion rate results of different compositions

[0191]

[0192]

[0193] As a result, the inclusion rate of the composition of the present invention is as high as 80% or more, that is, the active pharmaceutical ingredient that associates with the inclusion material to form an assemblage can account for more than 80% of the total active pharmaceutical ingredient.

[0194] Experiment Example 2, Stability Considerations

[0195] The lipolysis compositions prepared in Example 1 were subjected to long-term testing at 25℃±2℃ and 60%±5%RH, and accelerated testing at 40℃±2℃ and 75%±5%RH. The content of the active pharmaceutical ingredient, the content of related substances, and the pH value of each lipolysis composition were determined. The methods for determining the content of the active pharmaceutical ingredient and the methods for determining related substances are as follows:

[0196] 1. Content of active pharmaceutical ingredient

[0197] Mobile phase: Mobile phase A: 0.1% formic acid-water solution, Mobile phase B: 0.1% formic acid-acetonitrile solution; Gradient elution was performed according to Table 7.

[0198] Table 7. Gradient elution program

[0199]

[0200] Preparation of test solution: First, take the sample and prepare a drug solution containing about 10 μg / ml with solvent (methanol:water = 80:20, v / v) as the test solution.

[0201] Preparation of reference solution: Accurately weigh an appropriate amount of the drug active ingredient reference standard, and dilute it with a solvent (methanol:water = 80:20, v / v) to prepare a drug solution containing approximately 10 μg / ml, which is used as the reference solution.

[0202] Chromatographic conditions: Method: Determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512); Detector: CAD detector; Column: YMC Pack Pro C18 (4.6 mm × 150 mm, 3 μm); Injection volume: 25 μL; Column temperature: 30 ℃; Flow rate: 1 mL / min; Run time: 43 min.

[0203] Analytical method: The content was calculated based on peak area using the external standard method.

[0204] 2. Related substances

[0205] The determination method and chromatographic conditions are the same as those for content determination, and will not be repeated here.

[0206] The results of the determination of the contents of related substances and active pharmaceutical ingredients are shown in Table 8.

[0207] Table 8. Results of determination of active pharmaceutical ingredient and related substances content during the stability storage of the composition

[0208]

[0209] As shown in the table above, after six months of accelerated and long-term exposure conditions, the content of active pharmaceutical ingredients and related substances in each of the above compositions did not change significantly, indicating that the compositions provided by the present invention have good stability.

[0210] 3. The pH value of the composition is shown in the table below.

[0211] Table 8. Results of pH value determination of the composition during the stability storage period.

[0212]

[0213] As shown in the table above, after 6 months of accelerated and long-term exposure conditions, the pH values ​​of all the compositions did not change significantly, with changes not exceeding 0.5. This indicates that the pH values ​​of the compositions of the present invention remain stable during stable storage, consistently between 7.0 and 9.0.

[0214] Experiment 3: Safety and efficacy study in high-fat mice

[0215] Experimental animals: 12-week-old male C57BL / 6 mice were fed a high-fat diet for 12 weeks.

[0216] Laboratory animal source: Laboratory Animal Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences

[0217] Laboratory animal IACUC number: 2023-10-GY-71

[0218] Laboratory animal housing conditions: 25±5℃, 60±5%RH

[0219] Experimental grouping and dosage: High-fat mice were randomly divided into groups of 6. The abdominal area of ​​the mice was shaved and disinfected with alcohol. The composition shown in the table below was injected subcutaneously into the inguinal fat pad on one side of the mouse, and an equal volume of saline was injected into the other side in the same manner. Injections were repeated on day 1 and day 14. The experimental groups and dosages are shown in Table 9.

[0220] Table 9. Dosage information for in vivo studies of lipolysis formulations

[0221] Group Dosage volume Dosage (mg) Comparative Example 1 0.1ml*2 dots 2 Comparative Example 2 0.1ml*2 dots 2 Comparative Example 3 0.1ml*2 dots 2 Z5 0.1ml*2 dots 2 Z6 0.1ml*2 dots 2 Z7 0.1ml*2 dots 2

[0222] The experimental mice were treated as follows:

[0223] 1. Incidence of adverse reactions at the injection site:

[0224] The injection sites of the experimental mice were photographed and recorded at different time points after injection; the results are shown in Table 10 and... Figure 1-6 As shown.

[0225] Table 10. Injection sites in high-fat mice at different time points after drug administration

[0226]

[0227] From the above table and Figure 1-6The results showed that adverse reactions occurred at the injection sites of Comparative Examples 1 and 2 at different time points after a single injection, with an incidence rate of 100%, and the reactions did not fully recover after 14 days, indicating that the time interval between repeated injections of Comparative Examples 1 and 2 should be at least 14 days. No obvious abnormalities were observed at the injection sites of the same dose of composition Z5, Z6, and Z7, with an incidence rate of 0%, indicating that the safety of the composition of this application after a single injection is significantly better than that of Comparative Examples 1 and 2, and the incidence of adverse reactions at the injection site can be reduced by at least 60% compared to Comparative Examples 1 and 2.

[0228] After repeated injections, adverse reactions occurred at the injection sites of Comparative Examples 1 and 2, with an incidence rate of 67%; while no obvious abnormalities were observed at the injection sites of the same dosage compositions Z5, Z6, and Z7, with an incidence rate of 0%. This shows that the safety of the composition of this application after repeated injections is significantly better than that of Comparative Examples 1 and 2. The time interval between repeated injections can be set according to requirements, and the incidence of adverse reactions at the injection site can be reduced by 67% compared to Comparative Examples 1 and 2.

[0229] 2. Measurement of fat reduction effect at the injection site:

[0230] Animals were euthanized 7 days after the first injection and 21 days after the first injection (7 days after the repeat injection), the inguinal fat pad was separated, weighed, and the fat loss was quantitatively evaluated. The results are shown in Table 11.

[0231] Table 11. Results of quantitative evaluation of fat at different time points after drug administration

[0232]

[0233] As shown in the table above, after a single injection, the fat weight reduction of compositions Z5, Z6, and Z7 was 22.54-24.98%, while the fat weight reduction of Comparative Example 1 was 16.53%, indicating that the compositions of the present invention have a good fat-dissolving effect after a single injection. After repeated injections, the fat weight reduction of compositions Z5, Z6, and Z7 was 26.26-33.53%, while the fat weight reduction of Comparative Example 1 was 21.98%, indicating that the compositions of the present invention have a good fat-dissolving effect after repeated injections.

[0234] Commercially available injectable Kybella (i.e., Comparative Example 1 of this invention) has severe adverse reactions in clinical practice, with a recovery time of at least 4 weeks. Therefore, the clinical dosing interval is 4 weeks. However, Kybella is a common injectable solution, and excessively long dosing intervals can lead to reduced efficacy. The aqueous lipolysis formulation of this invention significantly reduces adverse reactions compared to Comparative Example 1. Therefore, the aqueous lipolysis formulation of this invention does not have the dosing interval problem caused by adverse reactions. Clinically, the dosing interval can be determined based on efficacy to ensure good lipolysis effect.

[0235] Comparing Comparative Examples 2 and 3 with Z5, Z6, and Z7, it can be seen that Comparative Example 2, containing 3% inclusion material, showed severe adverse reactions at the injection site in mouse studies; Comparative Example 3, containing 40% inclusion material, did not show significant adverse reactions in mouse studies, but its efficacy was significantly lower than that of the composition of the present invention; while the composition of the present invention, while ensuring good lipolysis effect, can significantly reduce the incidence of adverse reactions at the administration site, indicating that the inclusion material in the composition of the present invention has a dosage range requirement, which is higher than 3% and lower than 40%.

[0236] 3. Observation results of tissue sections at the injection site:

[0237] Animals were euthanized 21 days after the first injection (7 days after the repeat injection). Adipose tissue, intramuscular tissue, and epidermal tissue were separated from the injection site, sectioned in paraffin, stained with hematoxylin and eosin (HE), and their morphology was observed. The results are shown in Table 12. Figure 7 As shown (no obvious abnormalities were observed in Comparative Example 3 by the naked eye, so no tissue section was examined).

[0238] Table 12. Pathological section results of different tissues at different injection sites for different compositions

[0239] Composition Fat muscle skin Comparative Example 1 Moderate lipolysis Mild fibrosis Mild bleeding, epidermal hyperplasia, mild to moderate inflammation, moderate fibrosis Comparative Example 2 Moderate lipolysis Mild fibrosis Mild bleeding, epidermal hyperplasia, mild to moderate inflammation, moderate fibrosis Z5 Severe lipolysis No abnormalities Mild inflammation, mild fibrosis Z6 Moderate lipolysis No abnormalities Mild inflammation, mild fibrosis Z7 Moderate lipolysis No abnormalities No obvious abnormalities

[0240] Depend on Figure 7 The tissue section results in Table 12 show that the degree of fat dissolution in compositions Z5, Z6, and Z7 is moderate to severe, while it is moderate in comparative examples 1 and 2. This indicates that the fat dissolution degree of the compositions of the present invention is better than that of comparative example 1. The muscle tissue showed no obvious abnormalities in compositions Z5, Z6, and Z7, while mild fibrosis was observed in comparative examples 1 and 2. This indicates that the irritant effect of the compositions of the present invention on muscles is less than that of comparative examples 1 and 2. The skin tissue showed mild inflammation and mild fibrosis in compositions Z5 and Z6, while no obvious abnormalities were observed in composition Z7. However, the degree of inflammation and fibrosis observed in comparative examples 1 and 2 was more severe than that in compositions Z5, Z6, and Z7. In addition, hemorrhage and epidermal hyperplasia were also observed, indicating that the irritant effect of compositions Z5, Z6, and Z7 on the skin is much less than that of comparative examples 1 and 2.

[0241] Tissue section studies showed that the compositions Z5, Z6, and Z7 of the present invention had a greater degree of fat solubility than Comparative Examples 1 and 2, but less irritation to muscles and skin than Comparative Examples 1 and 2.

[0242] In summary, based on the observation results of adverse reactions at the injection site and the results of tissue section studies, the safety of the composition of the present invention is significantly improved compared with Comparative Example 1 and Comparative Example 2. Based on the results of fat reduction effect measurements at the injection site, the efficacy of the composition of the present invention is significantly improved compared with Comparative Example 1 and Comparative Example 2.

[0243] Experiment Example 4: Safety and efficacy study in pigs

[0244] Laboratory animals: Bama pigs, ~30kg.

[0245] Laboratory animal source: Shanghai University of Traditional Chinese Medicine

[0246] Laboratory animal IACUC number: PZSHUTCM2211070010

[0247] Laboratory animal housing conditions: 25±5℃, 60±5%RH

[0248] Experimental grouping and drug dosage: Four 2*3cm areas were selected on the back of the piglets for hair removal and disinfection. Each area was divided into 6 grids at 1cm intervals.

[0249] The same composition was injected into the fat layer at the center point of each grid, once every week for four consecutive weeks. The injection sites were dissected 28 days after the last injection. The experimental groups and dosages are shown in Table 13.

[0250] Table 13. Dosage information for in vivo studies in experimental pigs

[0251] Composition Single-point drug volume Dosage per region: mg Comparative Example 1 0.2ml 12 Z3 0.4ml 12 Z9 0.16ml 12 Z13 0.2ml 12

[0252] The experimental pigs were treated as follows:

[0253] 1. Incidence of adverse reactions at the injection site:

[0254] The injection sites of piglets were photographed and recorded at different time points after injection. The results are shown in Table 14 below.

[0255] Table 14. Gross observations of injection sites in experimental pigs

[0256]

[0257] The results showed that Comparative Example 1 caused hardening at the injection site, with an adverse reaction rate of 100%. In particular, the injection site of Comparative Example 1 also showed slight bulging. However, compositions 3, 9, and 13 of the present invention did not show any abnormalities in pigs, indicating that the compositions of the present invention have better safety than Comparative Example 1 and can reduce the incidence of adverse reactions at the injection site by 100%.

[0258] 1. Determination of fat reduction effect and drug residue at the injection site: Animals were sacrificed 28 days after the last injection, and tissues at the injection site were separated. The fat dissolution at the injection site was observed, and the fat thickness at the injection site and the surrounding fat thickness were measured. The changes in fat thickness were calculated. The results of fat thickness measurement at the administration sites for different groups are shown in Table 15.

[0259] Table 15. Fat reduction results at different injection sites in different groups

[0260] Composition visual observation fat thickness reduction Comparative Example 1 No dents 24.56％ Z3 There is a dent 32.56％ Z9 There is a dent 31.85％ Z13 There is a dent 33.74％

[0261] As shown in the table above, the fat thickness reduction rate at the administration site of the composition of the present invention is 31.85-33.74%, indicating that the composition of the present invention has a good fat-dissolving effect.

[0262] Experiment Example 5: Cellular Experiment Investigation

[0263] 1. Cell type

[0264] (1) Adipocytes (3T3-L1) were derived from the Shanghai Institute of Materia Medica, Chinese Academy of Sciences;

[0265] (2) Skeletal muscle cells (C2C12; mouse muscle cells, based on the second generation), derived from the Shanghai Institute of Materia Medica, Chinese Academy of Sciences;

[0266] (3) Vascular endothelial cells (HUVEC: human endothelial cells, based on the 3rd generation) were obtained from the Shanghai Institute of Materia Medica, Chinese Academy of Sciences;

[0267] (4) Human epidermal keratinocytes (HACAT) were obtained from the Shanghai Institute of Materia Medica, Chinese Academy of Sciences.

[0268] 2. Research Process

[0269] Comparative Example 1 and compositions Z6, Z9 and Z10 were diluted to a drug active ingredient concentration of 125 μg / ml, and then 100 μl of each was added to each of the above-mentioned cells in good growth condition. CCK-8 reagent was added at the same time, and the cells were placed in a cell culture incubator for routine culture for 1 day, and cell viability was detected.

[0270] 3. Test Results

[0271] The survival percentages of each cell in different compositions are shown in Table 16 below. Figure 8 As shown in the figure. The survival percentages of different cells and adipocytes were compared and their ratios were calculated; the results are shown in Tables 16-18.

[0272] Table 16. Percentage of cell viability in different compositions

[0273] Cell types Comparative Example 1 Z6 Z9 Z10 fat cells 80.87％ 51.23％ 43.99％ 39.12％ muscle cells 57.97％ 78.14％ 77.85％ 75.24％ Epidermal keratinocytes 39.38％ 89.36％ 88.79％ 86.97％ venous endothelial cells 57.20％ 75.63％ 76.34％ 78.85％

[0274] Table 17. Differences in cell survival percentage in different compositions relative to the survival percentage in Comparative Example 1

[0275] Cell types Z6 Z9 Z10 muscle cells 20.17％ 19.88％ 17.27％ Epidermal keratinocytes 49.98％ 49.41％ 47.59％ venous endothelial cells 18.43％ 19.14％ 21.65％

[0276] Table 18. Ratio of survival percentage of different cells to survival percentage of adipocytes in different compositions

[0277] Cell types Comparative Example 1 Z6 Z9 Z10 Muscle cells / fat cells 0.72 1.53 1.77 1.92 Epidermal keratinocytes / adipocytes 0.49 1.74 2.02 2.22 Venous endothelium / adipocytes 0.71 1.48 1.74 2.02

[0278] As can be seen from the results in Tables 17 and 18, Comparative Example 1, in addition to its destructive effect on adipocytes, also exhibits destructive effects on muscle cells, epidermal cells, and endothelial cells. At the same concentration, the survival percentage of cells other than adipocytes (such as muscle cells, epidermal keratinocytes, and venous endothelial cells) in the composition of the present invention is at least 17% higher than that in Comparative Example 1.

[0279] As can be seen from Table 18, the ratio of the viability of other cells such as adipocytes, epidermal cells, and venous endothelial cells to that of adipocytes is less than 1, indicating that Comparative Example 1 has a greater destructive effect on other cells than on adipocytes; while the composition of the present invention has a less destructive effect on other cells than on adipocytes, indicating that the composition of the present invention is more selective in dissolving adipocytes than Comparative Example 1, which can reduce the attack or damage to other cells, and further verifies that the composition of the present invention has better safety.

[0280] At the same drug concentration, the percentage of fat cell survival in Comparative Example 1 was higher than that in the composition of the present invention, indicating that the fat-dissolving effect of the composition of the present invention is higher than that in Comparative Example 1.

[0281] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0282] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A fat-dissolving composition, said fat-dissolving composition comprising: Drug active ingredients, Packaging materials, pH adjuster Optionally selected from one or more of buffers, stabilizers, osmotic pressure regulators, and complexing agents, and Optional purified water, in, The active pharmaceutical ingredient is selected from one or more of deoxycholic acid, pharmaceutically acceptable salts of deoxycholic acid, and deoxycholic acid derivatives. The amount of inclusion material used ranges from 4% to 35%, based on the total weight of the fat-dissolving composition.

2. The fat-dissolving composition according to claim 1, wherein, The inclusion material is selected from one or more of cyclodextrins and their derivatives; preferably selected from one or more of hydroxypropyl-β-cyclodextrin, sulfonyl-β-cyclodextrin, methyl-β-cyclodextrin, carboxymethyl-β-cyclodextrin, hydroxyethyl-β-cyclodextrin, glucosyl-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin, α-cyclodextrin, and 2,6-dimethyl-β-cyclodextrin; more preferably selected from one or two of hydroxypropyl-β-cyclodextrin and sulfonyl-β-cyclodextrin.

3. The fat-dissolving composition according to claim 1 or 2, wherein, The fat-dissolving composition has at least one of the following characteristics (1) to (10): (1) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.1-5% of the active pharmaceutical ingredient, 4-35% of the inclusion material, 0.01-5% of the pH adjuster, 0-10% of the stabilizer, 0-5% of the buffer, 0-5% of the osmotic pressure adjuster, 0-1% of the complexing agent, and 0-500 ml of purified water / g of active pharmaceutical ingredient; (2) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.2-4% of the active pharmaceutical ingredient, 5-30% of the inclusion material, 0.02-3% of the pH adjuster, 0-8% of the stabilizer, 0.01-3% of the buffer, 0-3% of the osmotic pressure adjuster, 0-0.8% of the complexing agent, and 0-400 ml of purified water / g of active pharmaceutical ingredient; (3) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.3-3% of the active pharmaceutical ingredient, 8-30% of the inclusion material, 0.03-2% of the pH adjuster, 0-5% of the stabilizer, 0.03-2% of the buffer, 0-2% of the osmotic pressure adjuster, 0-0.5% of the complexing agent, and 0-300 ml of purified water / g of active pharmaceutical ingredient; (4) Based on the total weight of the lipolysis composition, the lipolysis composition contains 0.5-2% of the active pharmaceutical ingredient, 10-28% of the inclusion material, 0.05-1% of the pH adjuster, 0-3% of the stabilizer, 0.05-1% of the buffer, 0-1% of the osmotic pressure adjuster, 0-0.3% of the complexing agent, and 0-200 ml of purified water / g of active pharmaceutical ingredient; (5) The pharmaceutically acceptable salt of the deoxycholic acid is sodium deoxycholate; the deoxycholic acid derivative is selected from one or more of the following: porcine deoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, sodium porcine deoxycholate, sodium ursodeoxycholate, sodium chenodeoxycholate, cholic acid, sodium cholate, glycocholic acid, sodium glycocholate, glycodeoxycholic acid, sodium glycodeoxycholate, taurocholic acid, sodium taurocholate, taurodeoxycholic acid, tauroursodeoxycholic acid, tauroporcine deoxycholic acid, taurochenodeoxycholic acid, glycotaurocholic acid, sodium glycotaurocholate, lithocholic acid, sodium lithocholic acid, sodium glycodeoxycholate, sodium phenylpropanecholate, sodium tyrosylcholate, sodium leucocholate, cholic acid dimer, cholic acid side chain amino acid conjugate, cholic acid side chain PEG conjugate, and cholic acid side chain glucose conjugate. (6) The stabilizer is selected from one or more of injectable solvents and surfactants; the injectable solvent may be one or more of propylene glycol, ethanol, benzyl alcohol, dimethyl sulfoxide, N-methylpyrrolidone, liquid polyethylene glycol, dimethylacetamide, tetrahydrofuran polyethylene glycol ether, diethylene glycol monoethyl ether, and benzyl benzoate; the surfactant may be one or more of phosphatidylcholine, sorbitan fatty acid ester, polysorbate, polyoxyethylene fatty acid ester, polyoxyethylene fatty alcohol ether, and poloxamer. (7) The pH adjuster is selected from one or more of pharmaceutically acceptable non-organic acids, organic acids, non-organic bases and organic bases; preferably selected from one or more of hydrochloric acid, phosphoric acid, acetic acid, sulfuric acid, citric acid, boric acid, lactic acid, methanesulfonic acid, maleic acid, tartaric acid, succinic acid, histidine, lysine, arginine, butyric acid, meglumine, ethylenediamine, ethanolamine, triethylamine, sodium hydroxide and potassium hydroxide; (8) The buffer is selected from one or more of the following: disodium hydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate, potassium phosphate, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium citrate, potassium citrate, sodium acetate, potassium acetate, tris(hydroxymethyl)aminomethane, potassium hydrogen phthalate, sodium hydrogen phthalate, sodium tetraborate, potassium tetraborate, sodium barbital, and glycine. (9) The osmotic pressure regulator is selected from one or more of lactose, trehalose, xylitol, levonorgestrel, sorbitol, maltose, glycine, sarcosine, sodium chloride, potassium chloride, glucose, sucrose and mannitol; preferably selected from one or more of trehalose, lactose, sodium chloride, potassium chloride, glucose, sucrose and mannitol; (10) The complexing agent is one or more selected from ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid and calcium disodium ethylenediaminetetraacetic acid.

4. The fat-dissolving composition according to claim 1, wherein, The active pharmaceutical ingredient comprises an associated portion that associates with the inclusion material to form an inclusion complex, and optionally, a free portion that does not associate with the inclusion material to form an inclusion complex. Preferably, the free portion accounts for <30% by mass of the active pharmaceutical ingredient; optionally, it accounts for <20%.

5. The fat-dissolving composition according to claim 1, wherein, The total daily dose of the lipolysis composition, based on the active pharmaceutical ingredient, is 0.1 mg to 1000 mg; preferably 1 mg to 500 mg.

6. Use of the lipolysis composition according to any one of claims 1-5 in the preparation of a medicament for the prevention or treatment of fat-related diseases.

7. The application according to claim 6, wherein, The fat-related diseases include, but are not limited to, one or more of the following: fat accumulation, lipoma, painful lipoma, cellulite, fat edema, fat hyperplasia, fat malnutrition, lipodystrophy, fat deposition after liposuction, and obstructive sleep apnea; optionally, the dosage form of the drug is an injectable preparation; more preferably, the injection site includes, but is not limited to, one or more of the following: abdomen, below the eyes, below the chin, below the arm, buttocks, thighs, calves, back, bra line, armpits, stomach, and ankles.

8. An injectable formulation prepared from the lipolysis composition as described in any one of claims 1-5.

9. The injectable formulation according to claim 8, wherein, The injectable preparation is administered via injection, including but not limited to one or more of the following: subcutaneous injection, intramuscular injection, intravenous injection, and intralesional injection.

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