Use of o-acyl-alpha-hydroxy acid compounds for weight loss and lipid reduction

Abstract

The application discloses application of O-acyl-alpha-hydroxy acid compounds in weight loss and lipid reduction, and belongs to the fields of biological medicine manufacturing, biological medicine use and functional food development. The O-acyl-alpha-hydroxy acid compounds can inhibit weight growth of high-fat diet-induced obese model mice and reduce body fat. Compared with GLP-1 receptor weight loss drugs which achieve the effect of reducing weight by inhibiting appetite, the O-acyl-alpha-hydroxy acid compounds are not dependent on appetite inhibition, and can reduce side effects such as gastrointestinal discomfort and muscle loss caused by inhibiting appetite. When combined or sequentially administered with GLP-1 drugs, the O-acyl-alpha-hydroxy acid compounds can maintain or enhance the weight loss effect while improving the side effects caused by GLP-1, and can prevent the risk of significant weight rebound after stopping GLP-1 by maintaining body weight. The application provides a safe and effective new strategy for weight loss and lipid reduction.

Description

Technical Field

[0001] This invention relates to the fields of biopharmaceutical manufacturing, biomedical applications, and the development of nutritional compositions, particularly to the application of O-acyl-α-hydroxy acid compounds in weight loss and lipid reduction. Background Technology

[0002] In recent years, with improved living standards and changes in dietary structure, the global incidence of overweight and obesity has continued to rise, and is showing a trend towards affecting younger people. Obesity not only significantly increases the risk of chronic diseases such as type 2 diabetes, fatty liver disease, and cardiovascular disease, but is also a major factor leading to preventable diseases and disability, becoming a serious public health and socioeconomic burden. Therefore, developing safe, effective, and long-term suitable nutritional intervention programs is of great significance for controlling weight and improving related metabolic disorders.

[0003] Currently, some drugs are approved for obesity management, such as orlistat and smegglutide, but they generally carry certain side effects, limiting their feasibility for long-term use. For example, orlistat may cause gastrointestinal discomfort and abnormal liver function, while GLP-1 receptor agonist peptide drugs are often accompanied by adverse reactions such as nausea, diarrhea, dizziness, and decreased lean body mass, and pose a risk of hypoglycemia in some individuals. Therefore, there is an increasing demand for functional ingredients with higher safety profiles suitable for pharmaceutical or functional food applications. Summary of the Invention

[0004] To address the adverse reactions caused by existing drugs, this invention proposes the application of O-acyl-α-hydroxy acid compounds in weight loss and lipid reduction. O-acyl-α-hydroxy acid compounds can be used to prepare drugs for treating or preventing obesity / overweight and / or hyperlipidemia, nutritional compositions or special medical foods for weight management and / or improving lipid indicators, and a dosing regimen for combining O-acyl-α-hydroxy acid compounds and their derivatives with GLP-1 drugs to reduce adverse reactions while achieving weight loss, and for weight maintenance after discontinuation of GLP-1.

[0005] The technical solution of the present invention is as follows: The first objective of this invention is to provide the use of an O-acyl-α-hydroxy acid compound in the preparation of a medicament for treating or preventing obesity / overweight, wherein the active ingredient of the medicament contains an O-acyl-α-hydroxy acid compound or an acceptable derivative thereof, thereby reducing body fat content and / or weight in obese / overweight individuals; the structure of the O-acyl-α-hydroxy acid compound is shown below: R 1 –C(O)O–CH(R)–COOH Among them, R 1For substitution or non-substitution: C 2-24 Straight-chain or branched, saturated or unsaturated aliphatic groups, C 3-12 Alicyclic groups or C 7-20 alkylaryl groups; R represents substituted or unsubstituted: C 1-22 Straight-chain or branched aliphatic groups; Substitution refers to the substitution of substituents such as halogens, hydroxyl groups, and C. 1-6 Alkyl, C 1-6 One or more of the following groups: alkoxy, amino, carboxyl, ester, amide, cyano, and nitro.

[0006] In one embodiment of the invention, R preferably comes from a branched-chain amino acid metabolite group: isobutyl–CH2CH(CH3)2 in α-hydroxyisocaproic acid (also known as α-hydroxy-4-methylvaleric acid, HICA), sec-butyl–CH(CH3)CH2CH3 in 2-hydroxy-3-methylvaleric acid (HMVA), or isopropyl–CH(CH3)2 in 2-hydroxy-3-methylbutyric acid (HIVA).

[0007] In one embodiment of the present invention, R 1 It is one of acetyl, propionyl, butyryl, hexanoyl, octanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, oleoyl, linoleoyl, linolenic acid, cyclopropyl, cyclohexyl, benzyl, phenethyl, and naphthylmethyl.

[0008] In one embodiment of the present invention, "unsaturated" refers to the presence of 1-6 carbon-carbon double bonds and / or carbon-carbon triple bonds (preferably 1-3 carbon-carbon double bonds) on the connecting aliphatic group; "substituted" refers to the presence of 1-3 substituents on the connecting group, the substituents being selected from: halogens, hydroxyl groups, C... 1-6 Alkyl, C 1-6 Alkoxy, amino, carboxyl, ester, amide, cyano, or nitro, or combinations thereof.

[0009] A second objective of this invention is to provide the use of an O-acyl-α-hydroxy acid compound in the preparation of a nutritional composition or food for weight management, wherein the active ingredient in the nutritional composition or food for weight management contains an O-acyl-α-hydroxy acid compound or an acceptable derivative thereof, and weight management includes reducing body fat percentage and / or reducing body weight.

[0010] In one embodiment of the present invention, the acceptable derivative is selected from food- or pharmaceutically acceptable salts, racemates, solvates, hydrates, crystal forms, prodrug derivatives, and derivatives that achieve stability or delivery optimization without significantly reducing safety / efficacy; the acceptable salt is an inorganic salt or an organic salt; the inorganic salt is selected from any one of sodium salts, calcium salts, potassium salts, and magnesium salts; the organic salt is selected from any one of meglumine salts, tromethamine salts, diethylamine salts, lysine salts, choline salts, arginine salts, tert-butylamine salts, and N,N-dibenzylethylenediamine salts.

[0011] In one embodiment of the present invention, an O-acyl-α-hydroxy acid compound is reacted with an alkaline inorganic salt or an organic amine in a solvent to form a salt, and the salt is obtained by filtration / concentration / drying. Alternatively, different crystal forms or solvates can be obtained by screening the solvent system and crystallization conditions to improve stability and manufacturability.

[0012] In one embodiment of the present invention, the medicine, nutritional composition or special medical food used to treat or prevent obesity / overweight further contains a second functional component, which is a GLP-1 drug, to achieve synergistic effect. The GLP-1 class of drugs are GLP-1 receptor agonists and / or drugs containing GLP-1 receptor agonist activity.

[0013] In one embodiment of the present invention, the GLP-1 drug is selected from: smegglutide, liraglutide, telpoglycinide, dulaglutide, exenatide, lixisenatide, abiglutide, or a pharmaceutically acceptable salt, prodrug, solvate, or crystal form thereof.

[0014] In one embodiment of the present invention, the O-acyl-α-hydroxy acid compound and the GLP-1 class drug are administered in a co-formulation or in combination in separate formulations. The combination therapy involves first administering a GLP-1 class drug alone or simultaneously with the O-acyl-α-hydroxy acid compound to achieve weight loss; after the weight has been reduced to the expected target, the GLP-1 class drug is discontinued and the O-acyl-α-hydroxy acid compound is continued for weight maintenance, or the administration of the O-acyl-α-hydroxy acid compound is gradually reduced or even discontinued after the weight has been maintained.

[0015] In one embodiment of the present invention, at the end of the observation period of 2-12 consecutive weeks after the discontinuation of GLP-1 drugs, the increase in the weight of the test subject relative to the weight at the time of discontinuation of GLP-1 drugs shall not exceed 70% of the weight loss during the treatment period of GLP-1 drugs, preferably not more than 50%, and more preferably not more than 30%.

[0016] In one embodiment of the present invention, the combination therapy is used to reduce the incidence or severity of adverse reactions associated with the GLP-1 class of drugs while achieving weight loss, including one or both of lean body mass reduction or excessively reduced appetite.

[0017] In one embodiment of the present invention, the route of drug administration is selected from oral administration, intravenous injection, subcutaneous injection, transdermal absorption, or nasal spray, preferably oral administration or subcutaneous injection; the nutritional composition or special medical food is administered orally.

[0018] The dosage can be 0.1-300 mg / kg body weight per day, preferably 5-200 mg / kg body weight.

[0019] The application cycle can be continuous for no less than 4 weeks, preferably 8-16 weeks, and most preferably 12 weeks.

[0020] A third objective of this invention is to provide the use of an O-acyl-α-hydroxy acid compound in the preparation of a medicament, nutritional composition, or food for special medical purposes for the treatment or prevention of metabolic disorders caused by obesity / overweight.

[0021] In one embodiment of the present invention, a medicament, nutritional composition, or medical food for treating or preventing metabolic disorders caused by obesity / overweight is prepared, specifically including one or more of the following uses: a) Improves fatty liver disease; b) Improve liver function; c) Improve abnormal blood sugar levels; d) Improves dyslipidemia; O-acyl-α-hydroxy acid compounds or acceptable derivatives thereof, as active ingredients in medicaments for the purposes described herein, improve fatty liver disease; and / or improve liver function; and / or improve abnormal blood glucose levels; and / or improve abnormal blood lipid levels.

[0022] In one embodiment of the present invention, O-acyl-α-hydroxy acid compounds can reduce body fat content and / or reduce weight; and can exert weight management effects by enhancing the thermogenesis of adipose tissue and increasing energy consumption.

[0023] In one embodiment of the present invention, the drug contains an O-acyl-α-hydroxy acid compound as the active component and also includes pharmaceutical or food-grade excipients.

[0024] In one embodiment of the present invention, the pharmaceutical or food-grade excipient comprises a carrier system, which includes, but is not limited to, liposomes, nanoemulsions, solid lipid nanoparticles, microcapsules, hydrogels, micelles, exosomes, chylomicron mimics, or chemically modified forms.

[0025] In one embodiment of the present invention, the pharmaceutical or food-grade excipients include, but are not limited to, conventional excipients such as solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, and fragrances.

[0026] In one embodiment of the present invention, the dosage form of the drug includes, but is not limited to, tablets, capsules, elixirs, syrups, lozenges, inhalers, sprays, injections, films, patches, powders, granules, blocks, emulsions, suppositories, etc.

[0027] Nutritional compositions or special medical foods contain an effective amount of O-acyl-α-hydroxy acid compounds or their food-acceptable derivatives, and may contain food-grade excipients and nutritional matrices. The forms may include powders, granules, solid beverages, meal replacement powders, meal replacement bars, ready-to-drink nutritional beverages, etc., and may be formulated with nutritional systems such as proteins, lipids, carbohydrates, dietary fiber, vitamins, and minerals.

[0028] In one embodiment of the present invention, the excipient comprises a carrier system, which is preferably a liposome, emulsion, nanoparticle, microcapsule, hydrogel, micelle, exosome, or chylomicron mimic; the carrier system can be used to improve solubility, stability, sustained / controlled release, targeted delivery, or taste adaptation.

[0029] Beneficial effects: Compared with the prior art, the present invention has at least one of the following effects: (1) Reduce weight and / or body fat percentage; (2) Improve blood lipid indicators (e.g., TG, TC, LDL-C, HDL-C and their combinations); (3) Enhance the thermogenesis of adipose tissue to increase energy consumption; (4) When combined with GLP-1 drugs, it can reduce the incidence or severity of adverse reactions, increase the average daily food intake and / or energy intake and maintain the weight loss effect; after discontinuing GLP-1, the compound of the present invention can be continued to promote weight maintenance and reduce the rate of weight regain. Attached Figure Description

[0030] Figure 1a , Figure 1b The figure shows the effect of O-acyl-α-hydroxy acid compound on the intervention of high-fat diet-induced obesity (DIO) mice.

[0031] Figure 2a , Figure 2b The figure shows the effect of combining O-acyl-α-hydroxy acid compounds with GLP-1 drugs on the intervention of high-fat diet-induced obesity (DIO) mice.

[0032] Figure 3 The graph shows the results of sequential dosing (induction phase + maintenance phase) and weight maintenance after discontinuation of GLP-1.

[0033] Figure 4 The figure shows the results of different doses of HICA-PA intervention on high-fat diet-induced obesity (DIO) mice. Detailed Implementation

[0034] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0035] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that the upper and lower limits of the range and each intermediate value between them are specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0036] Unless otherwise stated, 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 is described. While only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail. Unless otherwise stated, “%” means percentage based on weight.

[0037] The term "heat production" as used in this article can be used interchangeably with the terms "thermogeneity," "heat generation," "thermogeneous effect," and "thermogenic effect." Similarly, "weight loss," "weight reduction," "lipid reduction," "fat reduction," "weight loss," and "resistance to weight gain" can be used interchangeably. In this article, thermogenic effect refers to the process by which mammals, including humans, regulate heat production.

[0038] "Effective dose" refers to the dosage or intake that produces the desired effects of weight management, weight loss, reduction of body fat percentage, and / or improvement of blood lipid levels. It can be adjusted according to the route of administration, dosage form, individual differences, and disease severity.

[0039] "Food or pharmaceutically acceptable" means that the salt, solvent, excipient, carrier, and derivative forms do not cause unacceptable toxicity, irritation, or allergic reactions in test subjects when used in appropriate doses and methods.

[0040] Unless otherwise stated, “derivatives” include at least: food / pharmaceutical acceptable salts, stereoisomers / racemates, solvates, and ester derivatives.

[0041] "GLP-1 class drugs" refers to GLP-1 receptor agonists and / or drugs containing GLP-1 receptor agonist activity, including but not limited to smegglutide, liraglutide, telpoxetine, dulaglutide, exenatide, lixisenatide, abiglutide and their pharmaceutically acceptable salts, prodrugs, solvates or crystal forms.

[0042] "Combined administration" includes administration of a common formulation or administration of separate formulations; "sequential administration" includes first administering a GLP-1 drug (or administering a GLP-1 drug and the compound of the present invention simultaneously) to achieve a weight loss effect, and then discontinuing GLP-1 while continuing to administer the compound of the present invention as a maintenance regimen.

[0043] "Average daily food intake / energy intake" can be calculated by recording the daily food intake (g / day) and taking the average value over a given period; the same recording method is used when comparing with the GLP-1 monotherapy group.

[0044] Example 1: Preparation of O-acyl-α-hydroxy acid compounds (R 1 =palmitoyl, capryloyl, DHA acyl) 1. Experimental Materials and Methods 1.1 Preparation of O-acyl-α-hydroxy acid compounds Compounds 1 (HICA), 2 (HMVA), 3 (HIVA), and 4 (α-hydroxyoleic acid, HLA) (1.0 mmol, 1.0 eq) were weighed as starting materials, and 4-dimethylaminopyridine (0.1 mmol, 0.1 eq) was dissolved in anhydrous dichloromethane. The reaction flask was placed in an ice-water bath at 0°C, and magnetic stirring was turned on to allow the system to cool completely. N,N-diisopropylethylamine (1.2 mmol, 1.2 eq) was slowly added dropwise to the cooled solution to pre-form the carboxylate and activate the system. Palmitoyl chloride / octanoyl chloride / DHA acyl chloride (1.05 mmol, 1.05 eq) was slowly added dropwise to the reaction mixture using a syringe or a constant-pressure dropping funnel, controlling the addition time to 15-20 minutes, and the reaction was stirred overnight. After the reaction was completed, an equal volume of water was added to the reaction solution to quench the reaction, and the organic phase was washed successively with dilute hydrochloric acid and saturated brine. After drying and concentration, the product was purified by silica gel column chromatography using n-hexane / ethyl acetate as the eluent to obtain the corresponding hydroxyl-terminated O-acyl derivative.

[0045] 1.2 Proton NMR spectrum The sample was dissolved in deuterated chloroform reagent and analyzed using a Bruker AVANCE II nuclear magnetic resonance spectrometer.1 HNMR analysis.

[0046] 2. Experimental Results 2.1 Compound 1 (HICA) derivatives (1) O-palmitoyl-HICA (R 1 =C16:0): Molecular formula C 22 H 42 O4; 1 H NMR (400 MHz, Chloroform-d) δ 5.04 (dd, J = 9.6, 3.9 Hz, 1H), 2.38 (t, J = 7.5 Hz, 2H), 1.81 (ddt, J = 17.9, 12.6, 5.6 Hz, 2H), 1.71 – 1.61 (m,3H), 1.25 (s, 24H), 0.97 (d, J = 6.3 Hz, 3H), 0.93 (d, J = 6.3 Hz, 3H), 0.88(t, J = 6.7 Hz, 3H). The molecular structural formula is: .

[0047] (2) O-octanoyl-HICA (R 1 =C8:0): Molecular formula C 14 H 26 O4; 1 H NMR (400 MHz, Chloroform-d) δ 4.95 (t, J = 7.3 Hz, 1H), 2.34 (td,J = 8.8, 1.8 Hz, 2H), 2.02 – 1.77 (m, 3H), 1.54 (ddd, J = 16.2, 8.8, 7.4 Hz,2H), 1.39 – 1.20 (m, 8H), 0.95 – 0.81 (m, 9H). The molecular structural formula is: .

[0048] (3) O-DHA acyl-HICA (R 1 =22:6): Molecular formula C 28 H 42 O4; 1H NMR (400 MHz, Chloroform-d) δ 5.66 – 5.32 (m, 12H), 4.98 – 4.90 (m, 1H), 2.82 – 2.65 (m, 10H), 2.45 – 2.30 (m, 2H), 2.20 – 1.90 (m, 4H), 1.95– 1.70 (m, 3H), 1.00 – 0.85 (m, 9H). The molecular structural formula is: .

[0049] 2.2 Compound 2 (HMVA) derivatives (1) O-palmitoyl-HMVA (R 1 =C16:0): Molecular formula C 22 H 42 O4; 1 H NMR (400 MHz, Chloroform- d ) δ 5.07 (d, J = 7.9 Hz, 1H), 2.35 (td, J = 8.8, 0.9 Hz, 2H), 2.15 (pt, J = 8.0, 6.4 Hz, 1H), 1.60 – 1.48 (m, 2H), 1.45– 1.20 (m, 26H), 1.01 (d, J = 7.9 Hz, 3H), 0.88 (td, J = 7.1, 4.1 Hz, 6H). The molecular structural formula is: .

[0050] (2) O-octanoyl-HMVA (R 1 =C8:0): Molecular formula C 14 H 26 O4; 1 H NMR (400 MHz, Chloroform- d ) δ 5.01 (d, J = 7.9 Hz, 1H), 2.35 (td, J = 8.8, 0.9 Hz, 2H), 2.15 (pt, J= 8.0, 6.3 Hz, 1H), 1.54 – 1.48 (m, 2H), 1.40– 1.22 (m, 10H), 1.01 (d, J = 7.9 Hz, 3H), 0.92 – 0.84 (m, 6H). The molecular structural formula is: .

[0051] (3) O-DHA acyl-HMVA (R 1 =22:6): Molecular formula C 28 H 42 O4; 1 H NMR (400 MHz, Chloroform- d ) δ 5.66 – 5.56 (m, 1H), 5.60 – 5.49 (m,1H), 5.52 – 5.34 (m, 10H), 5.07 (d, J = 7.9 Hz, 1H), 2.67 – 2.56 (m, 3H), 2.60 – 2.52 (m, 1H), 2.56 – 2.44 (m, 6H), 2.48 – 2.33 (m, 2H), 2.36 – 2.25 (m,2H), 2.15 (pt, J = 7.9, 6.3 Hz, 1H), 2.05 – 1.94 (m, 2H), 1.41 – 1.29 (m, 2H), 1.04 – 0.92 (m, 6H), 0.88 (t, J = 7.4 Hz, 3H). The molecular structural formula is: .

[0052] 2.3 Compound 3 (HIVA) derivatives (1) O-palmitoyl-HIVA (R 1 =C16:0): Molecular formula C 21 H 40 O4; 1H NMR (400 MHz, Chloroform-d) δ 4.91 (d, J = 4.3 Hz, 1H), 2.44 –2.38 (m, 2H), 2.28 (td, J = 6.8, 4.3 Hz, 1H), 1.70 – 1.63 (m, 2H), 1.03 (dd,J = 6.8, 5.8 Hz, 6H), 0.89 (d, J = 6.6 Hz, 4H). The molecular structural formula is: .

[0053] (2) O-octanoyl-HIVA (R 1 =C8:0): Molecular formula C 13 H 24 O4; 1 H NMR (400 MHz, Chloroform- d ) δ 4.81 (dq, J = 7.0, 1.5 Hz, 1H), 2.35(td, J = 8.8, 0.9 Hz, 2H), 2.16 (hept, J = 7.2 Hz, 1H), 1.54 (ddd, J = 16.2,8.7, 7.3 Hz, 2H), 1.37 – 1.23 (m, 8H), 1.01 (dd, J = 7.3, 1.4 Hz, 6H), 0.95 –0.84 (m, 3H). The molecular structural formula is: .

[0054] (3) O-DHA acyl-HIVA (R 1 =22:6): Molecular formula C 27 H 40 O4; 1H NMR (400 MHz, Chloroform-d) δ 5.66 – 5.34 (m, 12H), 2.67 – 2.61(m, 1H), 2.61 – 2.42 (m, 10H), 2.42 – 2.20 (m, 5H), 2.05 – 1.94 (m, 2H), 1.01(dd, J = 7.3, 1.5 Hz, 6H), 0.96 (td, J = 7.4, 0.8 Hz, 3H). The molecular structural formula is: .

[0055] 2.4 Compound 4 (HLA) derivatives (1) O-palmitoyl-HLA (R 1 =C16:0): Molecular formula C 34 H 62 O4; 1 H NMR (400 MHz, Chloroform- d ) δ 5.45 – 5.31 (m, 4H), 4.97 (t, J =6.5 Hz, 1H), 2.52–2.46 (m, 2H), 2.34 (td, J = 8.8, 1.7 Hz, 2H), 2.08 – 1.98(m, 4H), 1.95 – 1.78 (m, 2H), 1.60–1.48 (m, 2H), 1.48–1.20 (m, 38H), 0.89 (tt, J = 5.3, 1.7 Hz, 6H). The molecular structural formula is: .

[0056] (2) O-octanoyl-HLA(R) 1 =C8:0): Molecular formula C 26 H 46 O4; 1 H NMR (400 MHz, Chloroform- d) δ 5.45 – 5.31 (m, 4H), 4.97 (t, J =6.5 Hz, 1H), 2.52 (ddt, J = 5.1, 4.1, 1.1 Hz, 2H), 2.34 (td, J = 8.8, 1.7 Hz, 2H), 2.08 – 1.95 (m, 4H), 1.94 – 1.78 (m, 2H), 1.60 – 1.48 (m, 2H), 1.48 –1.21 (m, 22H), 0.95 – 0.84 (m, 6H). The molecular structural formula is: .

[0057] (3) O-DHA acyl-HLA (R 1 =22:6): Molecular formula C 40 H 62 O4; 1 H NMR (400 MHz, Chloroform- d ) δ 5.66 – 5.53 (m, 2H), 5.46 – 5.32 (m,14H), 4.97 (t, J = 6.5 Hz, 1H), 2.65 – 2.43 (m, 12H), 2.43 – 2.25 (m, 4H),2.08 – 1.94 (m, 6H), 1.93 – 1.80 (m, 2H), 1.49 – 1.21 (m, 14H), 1.00 – 0.94(m, 3H), 0.94 – 0.88 (m, 3H). The molecular structural formula is: .

[0058] Example 2: Effect of O-acyl-α-hydroxy acid compound administration on weight loss and lipid reduction in high-fat diet-induced obese mice 1. Experimental Materials and Methods 1.1 Mice Animal experiments were conducted in accordance with the ethical guidelines and protocols approved by Dalian University of Technology. All experiments used sex- and age-matched mice, which were randomly assigned to groups. Mice were housed in a specific, sterile animal facility with controlled temperature and humidity (25°C, 12:12 h light:dark cycle) and free access to food and water. The conditions for mice used in subsequent examples were the same.

[0059] Dietary-induced obesity (DIO) mouse modeling process: 4-6 week old C57BL / 6J mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. After one week of acclimatization, the mice were fed a high-fat diet (XTHF60, Synergistic Bio) for 4 weeks. Subsequently, an intervention was implemented, maintaining a high-fat diet throughout the intervention period.

[0060] 1.2 Intervention by O-acyl-α-hydroxy acid compounds Taking O-palmitoyl-α-hydroxy acid as an example, dietary obesity (DIO) mice were treated with intraperitoneal injection for 12 days according to the following grouping.

[0061] HFD group (high-fat feeding group): intraperitoneal injection of the same amount of solvent.

[0062] HICA group: 50 mg / kg HICA was administered intraperitoneally.

[0063] HICA-PA group (O-palmitoyl-HICA): intraperitoneal injection of 50 mg / kg O-palmitoyl-HICA.

[0064] HMVA group: intraperitoneal injection of 50 mg / kg HMVA.

[0065] HMVA-PA group (O-palmitoyl-HMVA): intraperitoneal injection of 50 mg / kg O-palmitoyl-HMVA.

[0066] HIVA group: Intraperitoneal injection of 50 mg / kg HIVA.

[0067] HIVA-PA group (O-palmitoyl-HIVA): Intraperitoneal injection of 50 mg / kg O-palmitoyl-HIVA.

[0068] HLA group: intraperitoneal injection of 50 mg / kg HLA.

[0069] HLA-PA group (O-palmitoyl-HLA): intraperitoneal injection of 50 mg / kg O-palmitoyl-HLA.

[0070] 1.3 Serum marker determination Triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (CRE) in mice were measured using a commercial kit (Nanjing Jiancheng) according to the instructions for use.

[0071] 1.4 Statistical Analysis Graphpad Prism (version 9.5) was used for plotting and statistical analysis. All statistical tests are fully described in the graph legends and conform to the criteria of a normal distribution with similar variances. No statistical methods were used to predetermine sample size. t-tests were used for comparisons between two groups. Repeated measures ANOVA was performed for assessment data of relevant samples. One-way ANOVA and multiple comparisons were used for assessments of more than two groups. Two-way ANOVA with multiple comparisons was used for assessments of two independent variables. Linear regression analysis was used for correlation analysis. Otherwise, data are expressed as mean ± sem unless otherwise stated. Statistical methods are the same in subsequent examples.

[0072] 2. Experimental Results Figure 1 shows the results of the intervention effect of O-palmitoyl-α-hydroxy acid on high-fat diet-induced obesity (DIO) mice. Figure 1a This includes Figures 1A-E; Figure 1b The experimental design flowchart shown in Figure 1F-M (Figure 1A) illustrates the following: After a one-week acclimatization period, mice were fed a high-fat diet for four weeks and then randomly divided into nine groups (n=5 / group) for intervention. These groups were: control group (HFD group), HICA intervention group (HICA group), O-palmitoyl-HICA intervention group (HICA-PA group), HMVA intervention group (HMVA group), O-palmitoyl-HMVA intervention group (HMVA-PA group), HIVA intervention group (HIVA group), O-palmitoyl-HIVA intervention group (HIVA-PA group), HLA intervention group (HLA group), and O-palmitoyl-HLA intervention group (HLA-PA group). During the intervention period, mice were continuously fed a high-fat diet (XTHF60, Synergy Biotech) for 12 days. The experimental groups received a daily intraperitoneal injection of 50 mg / kg, while the control group received an equal volume of solvent. Daily changes in body weight (Figure 1B) and food intake (Figure 1C) were monitored during the intervention period. The glucose tolerance test (oral administration of 1 g / kg glucose) and insulin tolerance test (intraperitoneal injection of 0.5 U / kg insulin) were performed 12 days after the intervention (Figure 1D). Serum parameters measured (Figure 1F-I) included total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL) levels. Alanine aminotransferase (ALT) (Figure 1J), aspartate aminotransferase (AST) (Figure 1K), blood urea nitrogen (BUN) (Figure 1L), and creatinine (CRE) levels were also measured (Figure 1M). Data are presented as mean ± standard error (mean ± sem) for independent biological samples.

[0073] 3. Conclusion The results showed that, compared with the control group, the treatment groups with free HICA, HMVA, HIVA, and HLA did not significantly inhibit weight gain within 12 days (p>0.05), while the corresponding O-acyl-α-hydroxy acid compounds (HICA-PA group, HMVA-PA group, HIVA-PA group, and HLA-PA group) showed a significant decreasing trend in body weight (p<0.05), and their food intake did not show a significant difference (Figure 1A-C). Metabolic function analysis showed that the treatment groups with HICA, HMVA, HIVA, HLA, and their O-acyl-α-hydroxy acid compounds all significantly improved glucose tolerance and insulin sensitivity in mice (Figure 1D-E). Regarding lipid metabolism, blood lipid indicators were significantly lower than those in the control group (TC, TG, and LDL, all p<0.05), but no statistically significant difference was observed in high-density lipoprotein cholesterol (HDL) (Figure 1F-I). Furthermore, regarding liver function, except for HMVA and HIVA-PA, AST levels in mice in all other intervention groups decreased significantly (p<0.05), while ALT levels decreased significantly only in HICA-PA and HIVA-PA (p<0.05), with no significant differences in the other groups (p>0.05, Figure 1J-K). Kidney function indicators BUN and CRE levels also decreased significantly (p<0.05, Figure 1L-M), indicating that O-acyl-α-hydroxy acid compounds significantly improved liver dysfunction caused by high-fat diets and improved liver and kidney function. In conclusion, O-acyl-α-hydroxy acid compounds can alleviate obesity symptoms induced by high-fat diets, reduce weight gain caused by high-fat diets, and improve liver and kidney function, providing data support for the development of potentially anti-obesity active substances.

[0074] Example 3: The effect of O-acyl-α-hydroxy acid compound in combination with GLP-1 drugs on weight loss and lipid reduction in high-fat diet-induced obese mice. 1. Experimental Materials and Methods 1.1 Mice Animal experiments were conducted in accordance with the ethical guidelines and protocols approved by Dalian University of Technology. All experiments used sex- and age-matched mice, which were randomly assigned to groups. Mice were housed in a specific, sterile animal facility with controlled temperature and humidity (25°C, 12:12h light:dark cycle) and free access to food and water. The conditions for mice used in subsequent examples were the same.

[0075] Dietary-induced obesity (DIO) mouse modeling process: 4-6 week old C57BL / 6J mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. After one week of acclimatization, the mice were fed a high-fat diet (XTHF60, Synergistic Bio) for 4 weeks. Subsequently, an intervention was implemented, maintaining a high-fat diet throughout the intervention period.

[0076] 1.2 GLP-1 class drug combination therapy Taking O-palmitoyl-α-hydroxy acid compound as an example, this study investigated the weight loss and lipid-lowering effects of GLP-1 class drugs in combination on obese mice. Diet-induced obese (DIO) mice were treated with intraperitoneal injection for 21 days according to the following group.

[0077] HFD group (high-fat feeding group): intraperitoneal injection of the same amount of solvent.

[0078] Liraglutide group (GLP-1): Intraperitoneal injection of 100 µg / kg Liraglutide.

[0079] HICA-PA group (O-palmitoyl-HICA): intraperitoneal injection of 50 mg / kg O-palmitoyl-HICA.

[0080] The HICA-PA+Liraglutide group (O-palmitoyl-HICA+GLP-1 combined administration): 50 mg / kg O-palmitoyl-HICA + 100 µg / kg Liraglutide were injected intraperitoneally.

[0081] HMVA-PA group (O-palmitoyl-HMVA): intraperitoneal injection of 50 mg / kg O-palmitoyl-HMVA.

[0082] HMVA-PA+Liraglutide group (O-palmitoyl-HMVA+GLP-1 combined administration): 50 mg / kg O-palmitoyl-HMVA + 100 µg / kg Liraglutide were injected intraperitoneally.

[0083] HIVA-PA group (O-palmitoyl-HIVA): Intraperitoneal injection of 50 mg / kg O-palmitoyl-HIVA.

[0084] HIVA-PA+Liraglutide group (O-palmitoyl-HIVA+GLP-1 combined administration): 50 mg / kg O-palmitoyl-HIVA + 100 µg / kg Liraglutide were injected intraperitoneally.

[0085] 1.3 Serum marker determination Triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (CRE) in mice were measured using a commercial kit (Nanjing Jiancheng) according to the instructions for use.

[0086] 1.4 Sucrose Preference Test Eight-week-old male C57BL / 6J mice were fed a standard diet and acclimatized for 48 hours before the experiment to reduce stress. The experiment assessed the preference of individually housed mice for a 1% sucrose solution (w / v) versus pure water. Two hours before the start of the dark cycle, mice were intraperitoneally injected with saline or a drug, and subsequently provided with a standard diet and two bottles of fluids: one water and one 1% sucrose solution. Intake was recorded. The bottles were randomly assigned among individuals. Sucrose solution preference was quantified by dividing the amount of sucrose ingested by the total fluid intake.

[0087] 1.5 Conditioned Taste Aversion Test Mice were housed individually and acclimatized to drinking from two bottles to confirm no preference for one side prior to habituation. Subsequently, mice were habituated to nighttime water restriction (days 1–3), followed by a 1-hour bottle presentation (two bottles) and intraperitoneal injection of saline. On day 4, at the start of conditioned reflex training, the two bottles were replaced with a novel 0.15% saccharin solution, given for 1 hour, followed by an intraperitoneal injection. Mice were allowed to drink the saccharin solution for another 30 minutes before resuming water restriction until the next restriction. Day 5 served as a washout period, using the same bottle regimen as days 1–3. Day 6 was the second conditioned reflex training period, with day 7 serving as the washout period. On day 8, conditioned taste aversion to saccharin solution was assessed using a standard two-bottle preference test (saccharin solution vs. water) (presented 1 hour after nighttime water restriction). Saccharin solution intake was calculated.

[0088] 1.6 Statistical Analysis Graphpad Prism (version 9.5) was used for plotting and statistical analysis. All statistical tests are fully described in the graph legends and conform to the criteria of a normal distribution with similar variances. No statistical methods were used to predetermine sample size. t-tests were used for comparisons between two groups. Repeated measures ANOVA was performed for assessment data of relevant samples. One-way ANOVA and multiple comparisons were used for assessments of more than two groups. Two-way ANOVA with multiple comparisons was used for assessments of two independent variables. Linear regression analysis was used for correlation analysis. Otherwise, data are expressed as mean ± sem unless otherwise stated. Statistical methods are the same in subsequent examples.

[0089] 2. Experimental Results Figure 2 shows the results of the intervention effect of O-acyl-α-hydroxy acid compound combined with GLP-1 drugs on high-fat diet-induced obesity (DIO) mice. Figure 2a Including Figures 2A-F, Figure 2bThe experimental design flowchart shown in Figure 2G-Q (Figure 2A) illustrates the following: After a one-week acclimatization period, mice were fed a high-fat diet for four weeks and then randomly divided into eight groups (n=5 / group) for intervention. These groups were: control group (HFD group), Liraglutide group (GLP-1 group), O-palmitoyl-HICA intervention group (HICA-PA group), O-palmitoyl-HICA and GLP-1 combined intervention group (HICA-PA+Liraglutide group), O-palmitoyl-HMVA intervention group (HMVA-PA group), O-palmitoyl-HMVA and GLP-1 combined intervention group (HMVA-PA+Liraglutide group), O-palmitoyl-HIVA intervention group (HIVA-PA group), and O-palmitoyl-HIVA and GLP-1 combined intervention group (HIVA-PA+Liraglutide group). During the intervention period, mice were continuously fed a high-fat diet (XTHF60, Synergy Biotech) for 21 days. The experimental groups received a daily dose of 50 mg / kg via intraperitoneal injection, while the control group received an equal volume of solvent. Daily body weight changes monitored during the intervention (Figure 2B), body weight changes of mice in each group on the last day (Figure 2C), and food intake (Figure 2D). Glucose tolerance test (oral administration of 1 g / kg glucose) and insulin tolerance test (intraperitoneal injection of 0.5 U / kg insulin) were performed 21 days after the intervention (Figure 2E). Serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL) were measured (Figure 2G-J). Alanine aminotransferase (ALT) (Figure 2K), aspartate aminotransferase (AST) (Figure 2L), blood urea nitrogen (BUN) (Figure 2M), and creatinine (CRE) levels were measured (Figure 2N). Lean body mass was analyzed by low-field nuclear magnetic resonance (Figure 2O). Sucrose intake by mice in the sucrose preference test (Figure 2P). Saccharin intake by mice in the conditioned taste aversion test (Figure 2Q). Data are presented as mean ± standard error (mean ± sem) of independent biological samples.

[0090] 3. Conclusion The experimental results showed that, compared with the control group, the body weight of mice in all groups decreased significantly after drug intervention (p<0.05), and the decrease in body weight was more pronounced in the combined drug group compared with GLP-1 alone (Figure 2A-B). After the drug administration, the difference in body weight loss between the HMVA-PA and HIVA-PA combined with GLP-1 groups was significantly higher than that in the GLP-1 alone group (p<0.05, Figure 2C), and the food intake of mice in all groups treated with GLP-1 increased significantly (p<0.05, Figure 2D). Metabolic function analysis showed that the combined drug intervention also significantly improved glucose tolerance and insulin sensitivity in mice (Figure 2E-F). In terms of lipid metabolism, the blood lipid indicators (TC, TG, and LDL) of mice in all groups were significantly lower than those in the control group after drug administration (p<0.05), but no statistically significant difference was found in high-density lipoprotein cholesterol (HDL) (p>0.05, Figure 2G-J). Furthermore, regarding liver function, the ALT level in mice treated with GLP-1 combined administration was significantly lower than that in the control group (p<0.05, Figure 2K), while no statistically significant difference was observed in AST levels (p>0.05, Figure 2L). Kidney function indicators BUN and CRE were significantly decreased (p<0.05, Figure 2M-N). However, the liver and kidney function indicators in mice treated with GLP-1 alone showed no significant difference compared to the control group (p>0.05, Figure 2K-N), indicating that the combined administration did not cause liver or kidney damage. In addition, low-field NMR analysis of the lean body mass of mice treated with the combined administration revealed that lean body mass significantly decreased after GLP-1 treatment alone (p<0.05), while lean body mass increased in all groups after combined administration, returning to the level of the control group, showing a significant difference compared to the GLP-1-only group (p<0.05, Figure 2O). Furthermore, based on the sucrose preference test and conditional taste aversion test to evaluate changes in appetite in mice after intervention, it was found that GLP-1 alone significantly decreased the intake of sucrose and saccharin in mice (p<0.05), while the intake of sucrose and saccharin increased in all groups after combined administration. Specifically, the combined administration of HIVA-PA and GLP-1 significantly increased the intake of sucrose in mice (p<0.05, Figure 2P), and the combined administration of HMVA-PA, HIVA-PA, and GLP-1 significantly increased the intake of saccharin in mice (p<0.05, Figure 2Q). In conclusion, the combined use of O-palmitoyl-α-hydroxy acid compounds and GLP-1 can not only effectively alleviate the obesity phenotype induced by a high-fat diet, but also alleviate the side effects of decreased appetite and lean body mass caused by GLP-1, providing experimental evidence for the development of potential anti-obesity active substances.

[0091] Example 4: Sequential drug administration (induction phase + maintenance phase) and weight maintenance after discontinuation of GLP-1 1.1 Mice Animal experiments were conducted in accordance with the ethical guidelines and protocols approved by Dalian University of Technology. All experiments used sex- and age-matched mice, which were randomly assigned to groups. Mice were housed in a specific, sterile animal facility with controlled temperature and humidity (25°C, 12:12h light:dark cycle) and free access to food and water. The conditions for mice used in subsequent examples were the same.

[0092] Dietary-induced obesity (DIO) mouse modeling process: 4-6 week old C57BL / 6J mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. After one week of acclimatization, the mice were fed a high-fat diet (XTHF60, Synergistic Bio) for 4 weeks. Subsequently, an intervention was implemented, maintaining a high-fat diet throughout the intervention period.

[0093] 1.2 Drug administration intervention Taking O-palmitoyl-α-hydroxyisocaproic acid as an example, this study investigated the weight loss and lipid-lowering effects of GLP-1 drugs after discontinuation in obese mice. Diet-induced obese (DIO) mice were treated with intraperitoneal injection according to the following group.

[0094] HFD group (high-fat feeding group): intraperitoneal injection of the same amount of solvent.

[0095] Treatment groups: Liraglutide group (GLP-1) and HICA-PA group.

[0096] Obese mice were intraperitoneally injected with 100 µg / kg Liraglutide for 15 days and then randomly divided into two groups (n=5 / group): the Liraglutide group (GLP-1 group): drug discontinuation; and the HICA-PA intervention group (HICA-PA group): intraperitoneal injection of 50 mg / kg HICA-PA every 3 days for 15 days.

[0097] 1.3 Statistical Analysis Graphpad Prism (version 9.5) was used for plotting and statistical analysis. All statistical tests are fully described in the graph legends and conform to the criteria of a normal distribution with similar variances. No statistical methods were used to predetermine sample size. t-tests were used for comparisons between two groups. Repeated measures ANOVA was performed for assessment data of relevant samples. One-way ANOVA and multiple comparisons were used for assessments of more than two groups. Two-way ANOVA with multiple comparisons was used for assessments of two independent variables. Linear regression analysis was used for correlation analysis. Otherwise, data are expressed as mean ± sem unless otherwise stated. Statistical methods are the same in subsequent examples.

[0098] 2. Experimental Results Figure 3 This graph shows how O-palmitoyl-HICA inhibits obesity and improves glucose homeostasis after discontinuation of GLP-1 inhibitors. Figure 3 A) Experimental Design Flowchart: After a 1-week acclimatization period, mice were fed a high-fat diet for 4 weeks, followed by intraperitoneal injection of GLP-1 for 15 consecutive days. After discontinuation of the injection, mice were randomly divided into two groups (n=5 / group): the Liraglutide group (GLP-1 group): drug discontinuation; and the O-palmitoyl-HICA intervention group (HICA-PA group): intraperitoneal injection of 50 mg / kg HICA-PA every three days for 15 days, with a continuous high-fat diet (XTHF60, Synergy Biotech) during the intervention period. After the drug administration ended, mice were continued to be fed a high-fat diet for 30 days, and their body weight and food intake were observed. Daily changes in body weight during the intervention period were monitored (…). Figure 3 B) and food intake ( Figure 3 C). Data are presented as mean ± standard error (mean ± sem) of independent biological samples.

[0099] 3. Conclusion The experimental results showed that, compared with the control group, the body weight of mice in all groups significantly decreased after GLP-1 intervention (p<0.05). After GLP-1 administration was stopped after 15 days, the body weight of mice in the Liraglutide group rebounded gradually. After continuous administration of HICA-PA, the body weight remained stable without significant rebound (p>0.05). After administration ended on day 30, mice were continuously fed a high-fat diet. The results showed that on day 60, the body weight of mice in the HICA-PA group decreased by 1.54±0.42 g compared to their initial body weight on day 0. The increase in body weight after drug withdrawal was 43.38% of the decrease at the end of administration, with no significant rebound. Figure 3 B). Furthermore, on day 60, there was a significant difference in body weight between the HICA-PA group and the Liraglutide group (p<0.05). However, on day 60 after the end of administration, there was no significant difference in food intake between the HICA-PA and GLP-1 drug groups (p>0.05). Figure 3 C). In summary, O-palmitoyl-HICA can improve weight rebound after discontinuation of GLP-1 drugs and effectively alleviate the obesity phenotype induced by a high-fat diet, providing experimental evidence for the development of potential anti-obesity drugs.

[0100] Example 5: Effects of different doses of HICA-PA on weight loss and lipid reduction in high-fat diet-induced obese mice 1. Experimental Materials and Methods 1.1 Mice Animal experiments were conducted in accordance with the ethical guidelines and protocols approved by Dalian University of Technology. All experiments used sex- and age-matched mice, which were randomly assigned to groups. Mice were housed in a specific, sterile animal facility with controlled temperature and humidity (25°C, 12:12 h light:dark cycle) and free access to food and water. The conditions for mice used in subsequent examples were the same.

[0101] Dietary-induced obesity (DIO) mouse modeling process: 4-6 week old C57BL / 6J mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. After one week of acclimatization, the mice were fed a high-fat diet (XTHF60, Synergistic Bio) for 4 weeks. Subsequently, an intervention was implemented, maintaining a high-fat diet throughout the intervention period.

[0102] 1.2 Intervention with different doses of HICA-PA Diet-induced obesity (DIO) mice were treated by gavage for 4 weeks in the following groups.

[0103] HFD group (high-fat feeding group): administered the same amount of solvent via gavage.

[0104] Low-dose HICA-PA group (HICA-PA 25): 25 mg / kg HICA-PA administered by gavage.

[0105] HICA-PA medium-dose group (HICA-PA 50): 50 mg / kg HICA-PA administered by gavage.

[0106] High-dose HICA-PA group (HICA-PA 100): 100 mg / kg HICA-PA administered by gavage.

[0107] 1.3 Statistical Analysis Graphpad Prism (version 9.5) was used for plotting and statistical analysis. All statistical tests are fully described in the graph legends and conform to the criteria of a normal distribution with similar variances. No statistical methods were used to predetermine sample size. For assessments of more than two groups, one-way ANOVA and multiple comparisons were used. Correlation analysis was performed using linear regression. Otherwise, data are expressed as mean ± sem unless otherwise stated. The statistical methods used in subsequent examples are the same.

[0108] 2. Experimental Results Figure 2 shows the effects of different doses of HICA-PA on DIO mice. Figure 4A) Experimental Design Flowchart: After a 1-week acclimatization period, mice were fed a high-fat diet for 4 weeks and then randomly divided into 4 groups (n=6 / group) for intervention: a control group (HFD group), a 25 mg / kg HICA-PA intervention group (HICA-PA 25), a 50 mg / kg HICA-PA intervention group (HICA-PA 50), and a 100 mg / kg HICA-PA intervention group (HICA-PA 100). During the intervention period, mice were continuously fed a high-fat diet (XTHF60, Synergistic Bio) for 4 weeks. Weekly weight changes were monitored during the intervention period. Figure 4 B) and food intake ( Figure 4 C). Data are presented as mean ± standard error (mean ± sem) of independent biological samples.

[0109] 3. Conclusion The experimental results showed that, compared with the HFD control group, the body weight of obese mice induced by a high-fat diet after 4 weeks of continuous gavage administration of HICA-PA was significantly reduced (p<0.05), and this reduction was concentration-dependent. At week 4, the average body weight of the 25 mg / kg HICA-PA intervention group, the 50 mg / kg HICA-PA intervention group, and the 100 mg / kg HICA-PA intervention group decreased by 1.38 g, 2.20 g, and 2.80 g respectively compared with the control group. Figure 4 B). However, HICA-PA gavage had no significant effect on food intake in mice (p>0.05, Figure 4 C). In summary, HICA-PA can effectively alleviate the obesity phenotype induced by a high-fat diet, and the weight loss effect is dose-dependent, providing experimental evidence for the development of potential anti-obesity drugs.

[0110] Comparative Example 1 Compare the effects of compound compositions, such as α-hydroxyisohexanoic acid (HICA) and a physical mixture of palmitic acid, 2-hydroxy-3-methylbutyric acid (HIVA) and a physical mixture of palmitic acid, 2-hydroxy-3-methylvaleric acid (HMVA) and a physical mixture of palmitic acid, and palmitic acid-9-hydroxystearic acid (9-PAHSA), on reducing body weight in high-fat diet-induced obese mice.

[0111] Referring to Example 1, after a one-week adaptation period, mice were randomly divided into 5 groups (n=6 / group): control group (HFD group), 50 mg / kg physical mixture of α-hydroxyisocaproic acid and palmitic acid (HICA+PA group), 50 mg / kg physical mixture of 2-hydroxy-3-methylbutyric acid and palmitic acid (HIVA+PA group), 50 mg / kg physical mixture of 2-hydroxy-3-methylvaleric acid and palmitic acid (HMVA+PA group), and 50 mg / kg palmitic acid-9-hydroxystearic acid (9-PAHSA group). Other parameters remained unchanged. After 12 days of intervention, the indicators were compared with those before intervention, and changes in body weight, epididymal fat, subcutaneous fat, and brown fat were detected. The results are shown in Table 1.

[0112] Table 1

[0113] Note: Tukey's HSD multiple comparison analysis showed that different letters indicated significant differences between groups (p<0.05).

[0114] The comparison revealed that although the physical mixture of 50 mg / kg HICA, HIVA, and HMVA with palmitic acid could reduce the body weight of mice, there was no significant difference compared with the control group (p>0.05), and there was no significant reduction in subcutaneous fat, epididymal fat, and brown fat (p>0.05).

[0115] Some commercially available hydroxy fatty acid esters, such as palmitic acid-9-hydroxystearic acid (9-PAHSA), are believed to have anti-diabetic and anti-inflammatory effects (Mark M, et al., Cell, 2014;159(2):318-332) and can regulate glucose metabolism. However, combined with the comparative experiment in Example 1, it can be seen that in the diet-induced obesity (DIO) mouse model, 9-PAHSA did not significantly reduce the weight of mice, and its weight loss and lipid-lowering abilities were significantly lower than those of O-acyl-α-hydroxy acid compounds (p<0.05).

[0116] Comparative Example 2 Comparing carboxyl-terminated esters of compounds, taking α-hydroxyisohexanoic acid as an example, we synthesized α-hydroxyisohexanoic acid-O-palmitoyl ester, with the following chemical structural formula: The effects of O-palmitoyl-α-hydroxyisohexanoic acid (HICA-PA) and α-hydroxyisohexanoic acid-O-palmitoyl ester (HICA-O-PA) on reducing body weight in high-fat diet-induced obese mice were compared.

[0117] Referring to Example 1, after a one-week adaptation period, mice were randomly divided into three groups (n=6 / group): the control group (HFD group), the 50 mg / kg O-palmitoyl-α-hydroxyisohexanoic acid (HICA-PA group), and the 50 mg / kg α-hydroxyisohexanoic acid-O-palmitoyl ester (HICA-O-PA group). Other parameters remained unchanged. After 12 days of intervention, the body weight, epididymal fat, subcutaneous fat, and brown fat were compared with the pre-intervention indicators. The results are shown in Table 2.

[0118] Table 2

[0119] Note: Tukey's HSD multiple comparison analysis showed that different letters indicated significant differences between groups (p<0.05).

[0120] The comparison revealed that the weight gain in mice treated with 50 mg / kg carboxyl-terminated HICA-O-PA was not significantly different from that in the control group (p>0.05), and there was no significant reduction in epididymal fat, subcutaneous fat, or brown fat (p>0.05). However, 50 mg / kg hydroxyl-terminated HICA-PA not only significantly reduced mouse weight (p<0.05) but also significantly reduced epididymal and subcutaneous fat (p<0.05).

[0121] Therefore, the core innovation of this invention lies in the fact that it is the first time that O-acyl-α-hydroxy acid compounds have been proposed and verified mechanistically to effectively improve body fat and reduce weight, providing a new prevention / treatment method with effectiveness and higher safety for obese or overweight patients.

[0122] The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.

Claims

1. The use of O-acyl-α-hydroxy acid compounds in the preparation of medicaments for treating or preventing obesity / overweight, characterized in that, The active ingredient in the drug contains an O-acyl-α-hydroxy acid compound or an acceptable derivative thereof, which reduces body fat content and / or weight in obese / overweight individuals; the structure of the O-acyl-α-hydroxy acid compound is shown below: R¹–C(O)O–CH(R)–COOH Among them, R 1 For substitution or non-substitution: C 2-24 Straight-chain or branched, saturated or unsaturated aliphatic groups, C 3-12 Alicyclic groups or C 7-20 alkylaryl groups; R represents substituted or unsubstituted: C 1-22 Straight-chain or branched aliphatic groups; Substituents used are halogens, hydroxyl groups, and C. 1-6 Alkyl, C 1-6 One or more of the following groups: alkoxy, amino, carboxyl, ester, amide, cyano, and nitro.

2. The application according to claim 1, characterized in that, R is isobutyl–CH2CH(CH3)2, sec-butyl–CH(CH3)CH2CH3 or isopropyl–CH(CH3)2.

3. The application of O-acyl-α-hydroxy acid compounds in the preparation of nutritional compositions or special medical foods for weight management, characterized in that, The active ingredient in the nutritional composition or food for special medical use contains an O-acyl-α-hydroxy acid compound or an acceptable derivative thereof, which enables weight management including reducing body fat content and / or reducing weight.

4. The application according to any one of claims 1-3, characterized in that, Acceptable derivatives are selected from food or pharmaceutically acceptable salts, racemic mixtures, and solvates; acceptable salts are inorganic or organic salts; inorganic salts are selected from any one of sodium, calcium, potassium, and magnesium salts; organic salts are selected from any one of meglumine salts, tromethamine salts, diethylamine salts, lysine salts, choline salts, arginine salts, tert-butylamine salts, and N,N-dibenzylethylenediamine salts.

5. The application according to any one of claims 1-3, characterized in that, It also contains a second functional component, which is a GLP-1 drug, to achieve synergistic effects; The GLP-1 class of drugs are GLP-1 receptor agonists and / or drugs containing GLP-1 receptor agonist activity.

6. The application according to claim 5, characterized in that, GLP-1 drugs are selected from: smegglutide, liraglutide, telpoxetine, dulaglutide, exenatide, lixisenatide, abiglutide, or their pharmaceutically acceptable salts, prodrugs, solvates, or crystal forms.

7. The application according to claim 5, characterized in that, O-acyl-α-hydroxy acid compounds can be administered in co-formulation or in combination with the GLP-1 class drugs in separate formulations; The combination therapy involves first administering a GLP-1 class drug alone or simultaneously with the O-acyl-α-hydroxy acid compound to achieve weight loss; after the weight has been reduced to the expected target, the GLP-1 class drug is discontinued and the O-acyl-α-hydroxy acid compound is continued for weight maintenance, or the administration of the O-acyl-α-hydroxy acid compound is gradually reduced or even discontinued after the weight has been maintained.

8. The application according to claim 7, characterized in that, At the end of the observation period of 2-12 weeks after discontinuation of GLP-1 drugs, the increase in the subject's weight relative to the weight at the time of discontinuation of GLP-1 drugs shall not exceed 70% of the weight loss during the GLP-1 drug treatment period, preferably not more than 50%, and more preferably not more than 30%.

9. The application according to any one of claims 1-3, characterized in that, The route of drug administration is selected from oral, intravenous, subcutaneous, transdermal absorption or nasal spray, with oral and subcutaneous injection being preferred; the route of administration for nutritional compositions or special medical foods is oral.

10. The use of O-acyl-α-hydroxy acid compounds in the preparation of pharmaceuticals, nutritional compositions, or special medical foods for the treatment or prevention of metabolic disorders caused by obesity / overweight, specifically including one or more of the following uses: a) improving fatty liver disease; b) improving liver function; c) improving abnormal blood glucose; d) improving abnormal blood lipids, characterized in that, O-acyl-α-hydroxy acid compounds or acceptable derivatives thereof, as active ingredients in medicaments for the purposes described herein, improve fatty liver disease; and / or improve liver function; and / or improve abnormal blood glucose levels; and / or improve abnormal blood lipid levels.

Images