Use of NLRP3 inhibitor for preventing and / or treating disease or condition related to weight gain

Abstract

Use of an NLRP3 inhibitor for preventing and / or treating a disease or condition related to weight gain. Specifically, the present disclosure relates to use of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a disease or condition related to weight gain, wherein each substituent is as defined in the description.

Description

Use of an NLRP3 inhibitor for the prevention and / or treatment of diseases or conditions associated with weight gain. Technical Field

[0001] This disclosure relates to the use of an NLRP3 inhibitor in a medicament for treating diseases or conditions associated with weight gain, and falls within the pharmaceutical field. Background Technology

[0002] While weight loss therapies can treat complications arising from weight gain, such as obesity-related complications, they can also affect body composition. Body composition includes free mass (FM), fat-free mass (FFM), lean body mass (LBM), skeletal muscle mass, bone mineral content, and total body water (TBW). Free mass refers to the mass of all adipose tissue; FFM is total weight minus total fat mass; LBM includes the mass of organs, skin, bones, total body water, and muscle mass minus total fat mass; skeletal muscle mass includes lean body mass minus connective tissue, skin, and other organs; and TBW is the sum of intracellular and extracellular water. GLP-1 receptor agonists (GLP-1RAs) used to induce weight loss in subjects requiring weight loss treatment can also induce LBM and / or skeletal muscle loss associated with GLP-1RA-induced weight loss.

[0003] In peripheral circulation or within macrophages, lipids / palmitic acid and other substances can activate NLRP3 to produce IL-1β, further leading to insulin resistance in organs such as muscle, fat, and liver, reducing glucose uptake, and causing diabetes and obesity (Nat Immunol.2021:22(5):550-559). Under normal circumstances, NLRP3 in muscle tissue is in an inactivated state, and muscles can take up glucose normally. In an inflammatory state, NLRP3 inflammasomes in muscle tissue are activated to produce inflammatory factors such as IL-1β and IL-18, triggering insulin resistance, leading to a reduction in GLUT-4 receptors on the cell membrane, reduced glucose uptake, insufficient energy supply, and muscle atrophy (Int J Mol Sci.2021 Mar 23;22(6):3254).

[0004] PCT / CN2024 / 088250 discloses a new class of NLRP3 inhibitors, and the applications of this class of inhibitors are currently being explored. Summary of the Invention

[0005] This disclosure provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of diseases or conditions associated with weight gain.

[0006] Among them, R 1 Selected from hydrogen, methyl, or difluoromethyl;

[0007] R 2 Each is independently selected from hydrogen, halogen, and C. 1-6 alkyl;

[0008] R 3 and R 4 Each is independently selected from hydrogen or halogen;

[0009] R 5 Each is independently selected from hydrogen, halogen, and C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0010] R 6 Selected from C 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0011] R 7 Each is independently selected from fluorine;

[0012] m and p are each independently selected from 0, 1, 2, and 3;

[0013] n is independently selected from 1, 2, and 3.

[0014] In another aspect, this disclosure provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of muscle loss caused by a GLP-1 receptor agonist, for example, in a medicament for the treatment or prevention of muscle loss caused by a disease or condition associated with weight gain treated with a GLP-1 receptor agonist.

[0015] Another aspect of this disclosure provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a GLP-1 receptor agonist in the preparation of a medicament for the prevention and / or treatment of diseases or conditions associated with weight gain.

[0016] In another aspect, this disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which, in combination with a GLP-1 receptor agonist, is used for the prevention and / or treatment of diseases or conditions associated with weight gain.

[0017] Another aspect of this disclosure provides a GLP-1 receptor agonist, which, in combination with a compound of formula (I) or a pharmaceutically acceptable salt thereof, is used for the prevention and / or treatment of diseases or conditions associated with weight gain.

[0018] This disclosure, in another aspect, provides a method for preventing and / or treating diseases or conditions associated with weight gain, comprising co-administering to a subject in need a compound of formula (I) or a pharmaceutically acceptable salt thereof and a GLP-1 receptor agonist.

[0019] Another aspect of this disclosure provides a method for inducing total weight loss and maintaining lean muscle mass in subjects who require weight loss, the method comprising co-administering to the subject in need a compound of formula (I) or a pharmaceutically acceptable salt thereof and a GLP-1 receptor agonist to maintain lean muscle mass while inducing fat and weight loss in the subject.

[0020] This disclosure, in another aspect, provides a method for increasing total weight loss induced by administering a GLP-1 receptor agonist to a subject in need, the method comprising co-administering to the subject in need a compound of formula (I) or a pharmaceutically acceptable salt thereof and a GLP-1 receptor agonist to increase the subject's total weight loss relative to weight loss induced by administering a predetermined amount of the GLP-1 receptor agonist alone.

[0021] This disclosure, in another aspect, provides a method for treating or preventing further muscle mass loss caused by administration of a GLP-1 receptor agonist to a subject in need, the method comprising: adding a compound of formula (I) or a pharmaceutically acceptable salt thereof to a GLP-1 receptor agonist treatment regimen of the subject in need to treat or prevent further loss of lean muscle mass in the subject.

[0022] In some implementations, the uses or methods provided in this disclosure, the R 1 Selected from hydrogen.

[0023] In some implementations, the uses or methods provided in this disclosure, the R 2 Each is independently selected from hydrogen.

[0024] In some implementations, the uses or methods provided in this disclosure, the R 3 and R 4 Each is independently selected from hydrogen.

[0025] In some implementations, the uses or methods provided in this disclosure, the R 5 Each is independently selected from hydrogen.

[0026] In some implementations, the uses or methods provided in this disclosure, the R 6 Selected from methyl or ethyl.

[0027] In some implementations, the uses or methods provided in this disclosure, the R 6 It is a methyl group.

[0028] In some embodiments, the uses or methods provided in this disclosure, wherein the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is compound 1 or a pharmaceutically acceptable salt thereof,

[0029] In some implementations, the uses or methods provided in this disclosure result in a reduction of the subject's total body weight while maintaining muscle mass and / or muscle strength.

[0030] In some implementations, the uses or methods provided in this disclosure, the diseases or conditions associated with weight gain are selected from overweight, obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, diabetes, overeating, fatty liver disease, non-alcoholic fatty liver disease, dyslipidemia, metabolic syndrome, insufficiency of satiety, hyperinsulinemia, and nocturnal hypoglycemia.

[0031] In some embodiments, the uses or methods provided in this disclosure refer to the diseases or conditions associated with weight gain selected from overweight or obesity.

[0032] In some implementation schemes, the subjects have or are susceptible to, or at risk of having, overweight, obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, diabetes, overeating, fatty liver disease, non-alcoholic fatty liver disease, dyslipidemia, metabolic syndrome, insufficiency of satiety, hyperinsulinemia, and nocturnal hypoglycemia, especially overweight or obese.

[0033] In some embodiments, patients with a BMI of 25 or greater are considered overweight.

[0034] In some embodiments, patients with a BMI of 30 or higher are considered obese.

[0035] In some implementations, GLP-1 agonists can also activate one or more other receptors or functions.

[0036] In some implementations, the GLP-1 agonist is also an agonist of the GIP receptor.

[0037] In some implementations, GLP-1 agonists are also glucagon receptor agonists.

[0038] In some implementations, GLP-1 agonists are also agonists of GIP receptors and glucagon receptors.

[0039] In some embodiments, the GLP-1 receptor agonist is a dual agonist of both the GLP-1 receptor and the GIP receptor.

[0040] In some embodiments, the GLP-1 receptor agonist comprises a GLP-1 analogue or a pharmaceutically acceptable salt thereof with the structure shown below.

[0041] In some implementations, the dosage of the GLP-1 receptor agonist is adjusted according to the patient's disease condition.

[0042] In various implementation schemes, the dose of the GLP-1 receptor agonist is at least 0.01 mg / kg to 20 mg / kg.

[0043] In some implementations, the GLP-1 receptor agonist is administered at a dose independent of the patient's body weight or surface area (fixed dose).

[0044] In some implementations, the dosage of the GLP-1 receptor agonist is selected from 0.01 mg to 1000 mg.

[0045] In some implementations, the dosage of the GLP-1 receptor agonist is selected from 0.1 mg to 50 mg.

[0046] In some implementations, the dosage of the GLP-1 receptor agonist is selected from 0.5 mg to 15 mg.

[0047] In some implementations, the dosage of the GLP-1 receptor agonist is selected from 1 mg to 10 mg.

[0048] In some implementations, the dosage of the GLP-1 receptor agonist is selected from 1 mg to 6 mg.

[0049] In various embodiments, the dosage is at least 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.7 mg, 0.6 mg, 0.8 mg, 0.9 mg, 1.0 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg, 3.2 mg, 3.3 mg, or 3.4 mg. 3.5mg, 3.6mg, 3.7mg, 3.8mg, 3.9mg, 4mg, 4.1mg, 4.2mg, 4.3mg, 4.4mg, 4.5mg, 4.6mg, 4.7mg, 4.8mg, 4.9mg, 5mg, 5.25mg, 5.5mg, 5.75mg, 6mg, 6.25mg, 6.5mg, 6.75mg, 7mg, 7.25mg, 7.5mg, 7.75mg, 8mg, 8.25mg, 8.5mg, 8.75mg, 9mg, 9.25mg, 9.5mg, 9.75mg, or 10mg.

[0050] In some implementation schemes, GLP-1 receptor agonists are administered once a month, twice a month, three times a month, every other week (qow), once a week (qw), twice a week (biw), three times a week (tiw), four times a week, five times a week, six times a week, every other day (qod), daily (qd), twice a day (Bid), or three times a day (tid).

[0051] In some implementations, the GLP-1 receptor agonist is administered orally or via parenteral administration, including but not limited to intravenous injection, subcutaneous injection, and intramuscular injection.

[0052] In some embodiments, the GLP-1 receptor agonist is administered via subcutaneous injection into the abdomen.

[0053] In some implementations, the GLP-1 receptor agonist is administered orally.

[0054] In some embodiments, the GLP-1 receptor agonist is formulated in an injectable form (e.g., an injection solution), and the specific GLP-1 receptor agonist injection solution further comprises at least one of a buffer, an osmotic pressure regulator, a pH regulator, and an antibacterial agent.

[0055] In some embodiments, the buffer is selected from one or more of acetate buffers, histidine buffers, phosphate buffers, succinate buffers, and citrate buffers. In some embodiments, the buffer is a phosphate buffer, such as disodium hydrogen phosphate.

[0056] In some embodiments, the pharmaceutical composition further comprises an osmotic pressure regulator. The osmotic pressure regulator includes, but is not limited to: salts (e.g., sodium chloride, phosphates, sodium citrate, boric acid, and sodium tartrate), sugars or sugar alcohols (lactose, trehalose, sucrose, glucose, mannitol, sorbitol, xylitol), amino acids (e.g., L-glycine, L-histidine, arginine, lysine, isoleucine, aspartic acid, tryptophan, threonine), polyhydroxy sugar alcohols [e.g., glycerol, 1,2-propanediol (also known as propylene glycol in this disclosure), 1,3-propanediol, 1,3-butanediol], polyethylene glycol (e.g., PEG 400), or mixtures thereof. In some embodiments, the osmotic pressure regulator is selected from one or more of propylene glycol, mannitol, sorbitol, xylitol, glycerol, lactose, trehalose, sucrose, glucose, sodium chloride, phosphates, sodium citrate, boric acid, and sodium tartrate. In some embodiments, the osmotic pressure regulator is sodium chloride. In some embodiments, the osmotic pressure regulator is propylene glycol or mannitol.

[0057] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable antimicrobial agent. In some embodiments, the pharmaceutically acceptable antimicrobial agent includes, but is not limited to: phenol, o-cresol, m-cresol, p-cresol, methylparaben, propylparaben, 2-phenoxyethanol, butylparaben, 2-phenylethanol, benzyl alcohol, ethanol, chlorobutanol, and thimerosal, bromonitrile, benzoic acid, imidacloprid, chlorhexidine, sodium dehydroacetate, chlorocresol, ethylparaben, benzyl chloride, or mixtures thereof. In some embodiments, the pharmaceutically acceptable antimicrobial agent is phenol.

[0058] In some embodiments, the pharmaceutical composition is selected from any component and concentration of pharmaceutical composition disclosed in WO2023083301. This disclosure incorporates the compositions, preparation methods, and other contents of the aforementioned patent by reference.

[0059] In some embodiments, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof is administered at a dose independent of the patient's body weight or surface area (fixed dose), for example, 0.01 mg to 1000 mg, specifically 0.1 mg to 500 mg, 1 mg to 400 mg, 5 mg to 200 mg, 10 mg to 150 mg, for example, selected from: 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, 100mg, 105mg, 110mg, 115mg, 120mg, 125mg, 130mg, 135mg, 140mg, 145mg, 150mg, 155mg, 160mg, 165mg, 170mg, 175mg, 180mg, 185mg, 190mg, 195mg, 200mg, 205mg, 210mg, 215mg , 220mg, 225mg, 230mg, 235mg, 240mg, 245mg, 250mg, 255mg, 260mg, 265mg, 270mg, 275mg, 280mg, 285mg, 290 mg, 295mg, 300mg, 305mg, 310mg, 315mg, 320mg, 325mg, 330mg, 335mg, 340mg, 345mg, 350mg, 355mg, 360mg, 36 5mg, 370mg, 375mg, 380mg, 385mg, 390mg, 395mg, 400mg, 405mg, 410mg, 415mg, 420mg, 425mg, 430mg, 435mg, 440mg, 445mg, 450mg, 455mg, 460mg, 465mg, 470mg, 475mg, 480mg, 485mg, 490mg, 495mg, 500mg, or any value between two points.

[0060] In some implementations, the compound of formula (I) or its pharmaceutically acceptable salt is administered at the following frequencies: once a month, twice a month, three times a month, every other week (qow), once a week (qw), twice a week (biw), three times a week (tiw)), four times a week, five times a week, six times a week, every other day (qod), daily (qd), twice a day (qid), or three times a day (tid) over a period of time.

[0061] In some implementations, the duration of administration is from about one day to about one week, from about two weeks to about four weeks, from about one month to about two months, from about two months to about four months, from about four months to about six months, from about six months to about eight months, from about eight months to about one year, from about one year to about two years, or from about two years to about four years, or longer.

[0062] In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt is administered via parenteral administration or oral administration.

[0063] In some embodiments, the compound of formula (I) or its pharmaceutically acceptable salt is administered orally.

[0064] This disclosure also includes compounds identical to those described herein, but in which one or more atoms are labeled with isotopes whose atomic weights or mass numbers differ from those commonly found in nature. Examples of isotopes that can be incorporated into compounds of this disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as... 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl, etc.

[0065] Terminology Explanation

[0066] To facilitate understanding of this disclosure, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0067] Unless the context clearly requires otherwise, throughout the specification and claims, the words “comprising,” “having,” “including,” etc., should be understood as having an inclusive meaning, rather than an exclusive or exhaustive meaning; that is, the meaning of “including but not limited to.”

[0068] The terms “subject” and “patient” refer to mammals, especially primates, and particularly humans, especially patients who require relevant treatment.

[0069] When applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, "giving" and "treatment" refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. "Giving" and "treatment" can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cellular treatment includes contact between a reagent and cells, as well as contact between a reagent and a fluid, wherein the fluid is in contact with the cells. "Giving" and "treatment" also mean the treatment of, for example, cells, by means of a reagent, diagnostic agent, conjugate composition, or by means of another cell in vitro and ex vivo. When applied to humans, veterinary, or research subjects, "treatment" refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0070] "Treatment" means administering an oral or topical therapeutic agent, such as a composition comprising any of the compounds disclosed herein, to a patient who has symptoms of one or more diseases, and the therapeutic agent is known to have a therapeutic effect on these symptoms. Typically, a therapeutic agent is administered in a treated patient or population in an amount that effectively relieves symptoms of one or more diseases to induce the regression of such symptoms or inhibit their progression to any clinically measurable extent. The amount of a therapeutic agent that effectively relieves any specific disease symptom (also referred to as a "therapeuticly effective amount") can vary depending on a variety of factors, such as the patient's disease state, age, and weight, and the drug's ability to produce the desired therapeutic effect in the patient. Whether the disease symptoms have been relieved can be evaluated using any clinical testing methods commonly used by a physician or other healthcare professional to assess the severity or progression of the symptoms. Although the embodiments of this disclosure (e.g., treatment methods or products) may be ineffective in alleviating symptoms of each target disease, they should reduce symptoms of the target disease in a statistically significant number of patients, as determined by any statistical test known in the art, such as the Student t-test, chi-square test, U-test according to Mann and Whitney, Kruskal-Wallis test (H-test), Jonckheere-Terpstra test, and Wilcoxon test.

[0071] An "effective amount" includes the amount sufficient to improve or prevent the symptoms or condition of a medically diagnosed disease. An effective amount also means the amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the patient's overall health, the route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or administration regimen that avoids significant side effects or toxicity.

[0072] "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 20 carbon atoms. Alkyl groups containing 1 to 6 carbon atoms are also included. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, and their various branched isomers.

[0073] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0074] The terms “about” and “approximately” mean that a numerical value is within the acceptable error range of a specific value as determined by a person skilled in the art, the numerical value depending in part on how it is measured or determined (i.e., the limits of the measurement system). For example, “about” may mean within or above 1 standard deviation. Alternatively, “about” or “substantially includes” may mean a range of up to 20%, such as between 1% and 15%, between 1% and 10%, between 1% and 5%, between 0.5% and 5%, or between 0.5% and 1%. In this disclosure, each instance of a number or range of values ​​preceded by the term “about” also includes embodiments of a given number. Unless otherwise stated, when a specific value appears in this application and claims, the meaning of “about” or “substantially includes” should be assumed to be within the acceptable error range of that specific value.

[0075] The term “and / or”, such as “X and / or Y”, should be understood to mean “X and Y” or “X or Y” and should be used to provide clear support for both meanings or either meaning.

[0076] "Optional" or "optionally" means that the event or circumstances described below may, but do not have to, occur, including the circumstances in which the event or circumstances may or may not occur.

[0077] The term "natural GLP-1" refers to a naturally occurring molecule of the glucagon family of peptides or the venom exopeptide family, wherein: the glucagon family of peptides is encoded by the proglucagonogenamic gene and includes three highly homologous small peptides, namely glucagon (1-29), GLP-1 (1-37), and GLP-2 (1-33); and venom exopeptides are peptides expressed in lizards and, similar to GLP-1, are insulin-stimulating. In some embodiments, the term "natural GLP-1" also refers to human GLP-1 (7-37) and human GLP-1 (7-36). The term "natural GIP" refers to a peptide containing the human GIP sequence (1-42).

[0078] The term "GLP-1 analogue" refers to an amino acid modification or chemical modification having, compared to natural GLP-1 (particularly compared to human GLP-1 (7-37) and human GLP-1 (7-36)), up to 25, up to 24, up to 23, up to 22, up to 21, up to 20, up to 19, up to 18, up to 17, up to 16, up to 15, up to 14, up to 13, up to 12, up to 11, up to 10, up to 9, up to 8, up to 7, up to 6, up to 5, up to 4, up to 3, up to 2, or 1 amino acid modification or chemical modification, wherein the amino acid modification may be an amino acid substitution, addition, and / or deletion; and the chemical modification may be a chemical modification selected from the following groups: amide, carbohydrate, alkyl, acyl, ester, polyethylene glycol (PEG) group, sialylated group, glycosylated group, etc.

[0079] In some embodiments, the GLP-1 analog is a GLP-1 receptor agonist. In some embodiments, the GLP-1 analog is a dual agonist of both the GLP-1 receptor and the GIP receptor.

[0080] The term agonist is defined as a substance that activates the type of receptor under discussion. "Active," "activated," "activated," etc., refer to substances that are measured using detection methods known in the art.

[0081] The term "substitution" of an amino acid residue as used in this disclosure refers to the substitution of an amino acid residue by a different substance.

[0082] As used in this disclosure, the term "peptide" encompasses the category of peptides having modified amino and carboxyl terms. For example, amino acid chains containing terminal carboxylic acids with amide groups substituted are also included within the amino acid sequences named as natural amino acids.

[0083] All hydrogen atoms described in this disclosure can be replaced by their isotopes (protium, deuterium, tritium), and any hydrogen atom in the compounds disclosed herein can also be replaced by isotopic atoms.

[0084] "Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or absence of such event or environment. For example, "optionally alkyl-substituted heterocyclic group" means that the alkyl group may but does not have to be present, and the description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0085] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, that are independently substituted by a substituent. Substituents are only considered in their possible chemical positions, and those skilled in the art can determine (experimentally or theoretically) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0086] "Pharmaceutical composition" means containing one or more of the compounds described herein or their physiologically / medically acceptable salts or

[0087] A mixture of a prodrug and other chemical components; wherein said other components include, for example, physiological / pharmaceutical-grade carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertment of its biological activity.

[0088] "Buffer" refers to a buffer that is resistant to pH changes through the action of its acid-base conjugate components. Examples of buffers that maintain pH within an appropriate range include acetates, succinates, citrates, phosphates, gluconates, histidines, oxalates, lactates, phosphates, citrates, tartrates, fumarates, glycylglycine, and other organic acid buffers.

[0089] The three-letter and single-letter codes for amino acids used in this disclosure are as described in J. biol. chem, 243, p3558 (1968).

[0090] As used herein, "medicinal salt" is well known to those skilled in the art. In one embodiment, the GLP-1 analog is in a medicinal salt form. In another embodiment, the GLP-1 analog is in a non-salt form.

[0091] "Disorders associated with weight gain" (which may be referred to herein as "muscular disorders associated with weight gain" or "muscular disorders associated with fat gain") means a disease or condition associated with weight gain in mammalian subjects, such as obesity-related comorbidities. In some embodiments, weight gain includes fat gain. Attached Figure Description

[0092] Figure 1-A Average cumulative food intake (g) in high-fat diet-induced obese (DIO) mice.

[0093] Figure 1-B: Average body weight change (%) in high-fat diet-induced obese (DIO) mice.

[0094] Figure 2-A Average fat body ratio (%) in high-fat diet-induced obese (DIO) mice.

[0095] Figure 2-B Average lean body ratio (%) in high-fat diet-induced obese (DIO) mice. Detailed Implementation

[0096] The present disclosure will be explained in more detail below with reference to embodiments or experimental examples. The embodiments or experimental examples in the present disclosure are only used to illustrate the technical solutions in the present disclosure and are not intended to limit the substance and scope of the present disclosure.

[0097] Test conditions of the instruments used in the experiment:

[0098] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE NEO 500M NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. The spatial configurations of the optical isomers (isomers) of the compounds were further confirmed by measuring single-crystal parameters.

[0099] HPLC determinations were performed using a Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200LC high performance liquid chromatograph (ACQUITY UPLC BEH C18 1.7UM 2.1×50MM column, Ultimate XB-C18 3.0×150mm column, or Xtimate C18 2.1×30mm column).

[0100] MS measurements were performed using a Waters SQD2 mass spectrometer in positive / negative ion mode, with a mass scan range of 100–1200.

[0101] Chiral HPLC analysis was performed using Chiralpak IC-3 100×4.6mm ID, 3µm, Chiralpak AD-3 150×4.6mm ID, 3µm, Chiralpak AD-3 50×4.6mm ID, 3µm, Chiralpak AS-3 150×4.6mm ID, 3µm, Chiralpak AS-3 100×4.6mm ID, 3µm, ChiralCel OD-3 150×4.6mm ID, 3µm, Chiralcel OD-3 100×4.6mm ID, 3µm, ChiralCel OJ-H 150×4.6mm ID, 5µm, and Chiralcel OJ-3 150×4.6mm ID, 3µm columns.

[0102] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15 mm to 0.2 mm, and the diameter of the silica gel plates used for thin-layer chromatography separation and purification products is 0.4 mm to 0.5 mm.

[0103] Rapid column purification systems use either the Combiflash Rf150 (TELEDYNE ISCO) or Isolara One (Biotage).

[0104] Normal column chromatography generally uses Yantai Huanghai silica gel of 100-200 mesh, 200-300 mesh or 300-400 mesh as the carrier, or Changzhou Santai pre-filled ultrapure normal phase silica gel column (40-63μm, 60, 12g, 25g, 40g, 80g or other specifications).

[0105] Reversed-phase column chromatography typically uses Changzhou Sante pre-packed ultrapure C18 silica gel columns (20-45μm). 40g, 80g, 120g, 220g or other sizes).

[0106] The high-pressure column purification system uses Waters AutoP, in conjunction with the Waters XBridge BEH C18 OBD Prep Column. 5μm, 19mm×150mm or Atlantis T3 OBD Prep Column, 5μm, 19mm×150mm.

[0107] Chiral preparation columns used were DAICL CHIRALPAK IC (250 mm × 30 mm, 10 μm) or Phenomenex-Amylose-1 (250 mm × 30 mm, 5 μm).

[0108] XRPD (X-ray Powder Diffraction) was used for analysis: measurements were performed using a BRUKER D8 X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα1 rays. Kα2 rays Kβ rays Scanning mode: θ / 2θ, scanning range (2θ range): 3°~45°.

[0109] DSC stands for Differential Scanning Calorimetry: Measurements were performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10℃ / min. The specific temperature range was referenced from the corresponding spectra (mostly 25-250℃), and the nitrogen purging rate was 50mL / min.

[0110] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer, with a heating rate of 10℃ / min, and the specific temperature range was referenced from the corresponding spectrum (mostly 30-350℃). The nitrogen purging rate was 50mL / min.

[0111] DVS stands for Dynamic Moisture Adsorption: The detection method is SMSDVS Advantage, with humidity changing from 50% to 95% to 0% to 95% to 50% at 25℃, in 10% increments (the final step is 5%) (the specific humidity range is subject to the corresponding spectrum; the methods listed here are the most commonly used). The judgment criterion is Tmax 360min, and dm / dt not greater than 0.002%.

[0112] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0113] Unless otherwise specified in the examples, all reactions can be carried out under an argon or nitrogen atmosphere.

[0114] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0115] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system used for column chromatography to purify the compounds, and the developing solvent system for TLC included: A: n-hexane / ethyl acetate system, B: dichloromethane / methanol system. The volume ratio of the solvent was adjusted according to the polarity of the compounds, and small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0116] Example 1

[0117] 5-(4-(((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)amino)-7,8-dihydro-5H-pyran[3,4-d]pyridazine-1-yl)-2,3-dihydrobenzofuran-4-ol

[0118] first step

[0119] tert-butyl((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)carbamate 1b

[0120] A solution of tert-butyl((3R,5R)-5-fluoropiperidin-3-yl)carbamate 1a (5.0 g, 22.91 mmol) in dichloroethane (120 mL) was added to an aqueous formaldehyde solution (30%, 3.34 mL, 34.36 mmol). The mixture was stirred at room temperature for 1 hour, followed by the addition of sodium triacetoxyborohydride (12.14 g, 57.27 mmol) and acetic acid (0.2 mL) in an ice bath. The reaction mixture was stirred overnight at room temperature. The solvent was removed by vacuum concentration. Water (30 mL) was added to the remaining solution, and the pH was adjusted to approximately 9 with ammonia. The mixture was extracted with dichloromethane (50 mL × 2). The organic phases were combined, washed with saturated brine (30 mL), and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to give compound 1b (5.1 g, 95% yield).

[0121] MS m / z (ESI): 233.4 [M+H]+.

[0122] Step 2

[0123] (3R,5R)-5-fluoro-1-methylpiperidin-3-amine 1c

[0124] Compound 1b (5.1 g, 21.95 mmol) was dissolved in dichloromethane (30 mL), and trifluoroacetic acid (15 mL) was added at room temperature. The mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure to obtain crude trifluoroacetate 1c (5.9 g, 110% yield), which was used directly in the next reaction.

[0125] MS m / z (ESI): 133.4 [M+H] + .

[0126] Step 3

[0127] 1-(3,6-dihydro-2H-pyran-4-yl)pyrrolidine 1bb

[0128] Tetrahydro-4H-pyran-4-one 1aa (1.0 g, 10 mmol) and potassium carbonate (0.14 g, 1 mmol) were mixed in tetrahydropyrrole (0.71 g, 10 mmol). The mixture was stirred at 0 °C until fully reacted. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated under vacuum to obtain crude compound 1bb (1.35 g), which could be used in the next step without further purification.

[0129] ES-MS m / z = 154.1 [M+H] + .

[0130] Step 4

[0131] 1,4-Dichloro-7,8-dihydro-5H-pyran[3,4-d]pyrazine 1cc

[0132] Compound 1bb (305 mg, 2 mmol) and 3,6-dichlorotetraazine (0.2 g, 1.32 mmol) were mixed in dichloromethane (5 mL). The mixture was stirred at 0 °C until fully reacted, diluted with water (5 mL), and extracted with dichloromethane (5 mL × 3). The liquid and liquid phases were separated, washed with brine (5 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to obtain the crude product, which was purified by rapid column chromatography (eluent: 0-20% ethyl acetate in petroleum ether) to give compound 1cc (24 mg, yield 18.3%).

[0133] MS m / z(ESI): 205.0 [M+H] + .

[0134] 1 H NMR (400MHz, CDCl3): δppm 4.70 (s, 2H), 4.05 (t, J = 5.5Hz, 2H), 2.53 (t, J = 6.0Hz, 2H).

[0135] Step 5

[0136] 1-Chloro-N-((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)-7,8-dihydro-5H-pyrano[3,4-d]pyrazine-4-amine1d-2

[0137] The crude trifluoroacetate of compound 1c (5.9 g, 21.94 mmol), compound 1cc (4.5 g, 21.94 mmol), tris(dibenzylacetone)dipalladium (0.8 g, 0.88 mmol), cesium carbonate (25.02 g, 76.79 mmol), and 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (2.73 g, 4.39 mmol) were mixed in dry toluene (120 mL) and stirred overnight at 110 °C under a nitrogen atmosphere. After cooling the reaction solution to room temperature, it was filtered, and the residue was washed with ethyl acetate (20 mL × 3). The filtrates were combined. The filtrate was concentrated under vacuum to obtain the crude product, which was then purified by rapid column chromatography. The purified product was further separated by SFC (column: ChiralPak AY, 250×20mm ID, 5μm, mobile phase: A: CO2; B: MeOH + 0.1% NH3H2O, gradient: B 40%; flow rate: 40mL / min; temperature: 35℃).

[0138] Compound 1d-2 (280 mg, yield 4.2%).

[0139] 1 H NMR (400MHz, DMSO) δ5.89(d,J=7.8Hz,1H),4.93(d,J=47.1Hz,1H),4.51-4.43(m,1H),4.41(s,2H),3.87(t,J=5.6Hz,2H ),2.96-2.83(m,2H),2.60(t,J=5.4Hz,2H),2.19(s,3H),2.18-2.04(m,2H),1.87(t,J=10.1Hz,1H),1.76-1.57(m,1H).

[0140] 19 F NMR (377MHz, DMSO) δ-180.76 (s).

[0141] MS m / z (ESI): 301.3 [M+H] + .

[0142] Step 6

[0143] 5-Bromo-4-(methoxymethoxy)-2,3-dihydrobenzofuran 1f

[0144] 5-Bromo-2,3-dihydrobenzofuran-4-ol 1e (5.9 g, 27.44 mmol) was dissolved in dichloromethane (60 mL). N,N-diisopropylethylamine (9.1 mL, 54.87 mmol) and chloromethyl ether (MOMCl) (2.87 g, 35.67 mmol) were added dropwise under ice bath conditions. The reaction was allowed to proceed to completion at room temperature. The solution was diluted with dichloromethane (200 mL), and the diluted solution was concentrated under vacuum to obtain a crude product. Purification was performed by rapid column chromatography to give compound 1f (6.05 g, 76.6% yield).

[0145] 1HNMR (400MHz, CDCl3), δ7.18(d,1H),6.38(d,1H),5.05(s,2H),4.48(t,2H),3.50(s,3H),3.24(t,2H).

[0146] Step 7

[0147] 1g of 2-(4-(methoxymethoxy)-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane

[0148] Under a nitrogen atmosphere, compound 1f (6.05 g, 23.35 mmol) was dissolved in acetonitrile (100 mL). Pinara-borane (5.08 mL, 35.03 mmol), triethylamine (9.74 mL, 70.05 mmol), and Pd(dppf)Cl2 (1.73 g, 2.34 mmol) were slowly added at room temperature, and the reaction temperature was raised to 80 °C. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated under vacuum to obtain the crude product. This crude product was purified by rapid column chromatography to give compound 1 g (5.7 g, yield 71.76%).

[0149] 1HNMR (400MHz, DMSO), δ7.42(d,1H),6.54(d,1H),5.03(s,2H),4.55(t,2H),3.45(s,3H),3.19(t,2H),1.26(s,12H).

[0150] Step 8

[0151] N-((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)-1-(4-(methoxymethoxy)-2,3-dihydrobenzofuran-5-yl)-7,8-dihydro-5H-pyran[3,4-d]pyridazine-4-amine1h

[0152] Under a nitrogen atmosphere, compound 1d-2 (270 mg, 0.90 mmol) was dissolved in 1,4-dioxane (10 mL) and water (1 mL). Cesium carbonate (731 mg, 2.24 mmol), (1,1'-bis(diphenylphosphine)ferrocene)palladium dichloride (65 mg, 0.09 mmol), and 1 g of compound (329 mg, 1.08 mmol) were added at room temperature. The reaction was heated to 95 °C and stirred. After the reaction was complete, the mixture was cooled to room temperature, filtered, and the filtrate was diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and filtered again. The filtrate was concentrated under vacuum to obtain the crude product, which was purified by rapid column chromatography to give compound 1h (260 mg, 65% yield).

[0153] MS m / z (ESI): 445.4 [M+H] + .

[0154] Step 9

[0155] 5-(4-(((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)amino)-7,8-dihydro-5H-pyran[3,4-d]pyridazine-1-yl)-2,3-dihydrobenzofuran-4-ol

[0156] At room temperature, a 1,4-dioxane solution (4M, 10 mL) of hydrogen chloride was added to a tetrahydrofuran solution (15 mL) containing 260 mg (0.59 mmol) of compound 1 h, and the mixture was stirred until the reaction was complete. After concentration, the residue was adjusted to pH ≈ 9 with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane (20 mL × 2), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by rapid column chromatography to give compound 1 (137 mg, yield 58%).

[0157] MS m / z (ESI): 401.4 [M+H] + .

[0158] 1HNMR(400MHz,DMSO)δ10.15(s,1H),7.00(d,J=8.4Hz,1H),6.35(d,J=8.4Hz,1H) ,5.66(d,J=7.6Hz,1H),4.95(d,J=47.4Hz,1H),4.57(t,J=8.8Hz,3H),4.48(s,2H ),3.76(t,J=5.6Hz,2H),3.14(t,J=8.8Hz,2H),2.99-2.86(m,2H),2.52-2.51(m ,2H),2.21(s,3H),2.17-2.08(m,2H),1.91(t,J=10.0Hz,1H),1.79-1.62(m,1H).

[0159] 19 F NMR (377MHz, DMSO) δ-180.65 (s).

[0160] Test Example 1: Determination of NLRP3 inflammasome inhibitory activity in THP1-Null cells

[0161] 1. Experimental instruments and reagents

[0162] 1.1 Experimental Apparatus

[0163] Plate reader:PerkinElmer 2104 EnVision Multilabel Plate Readers

[0164] 1.2 Experimental Reagents

[0165] 2. Experimental Design

[0166] Day 1: THP1-Null cells were seeded in 96-well plates at 10 × 10⁶ cells / well. 4 Cells / 180 μL of hygromycin B- and Normocin-free medium / well. Add 20 μL of PMA (final concentration 100 nM) to each well and incubate at 37°C in 5% CO2 for 24 hours.

[0167] Day 2: Dilute the compound with phenol red-free RPMI 1640 medium containing 25 mM HEPES and 0.5% DMSO. Remove the medium and pretreat cells with 160 μL of medium containing the compound, incubating at 37°C for 1 hour in 5% CO2. Then prepare LPS in phenol red-free RPMI 1640 medium containing 25 mM HEPES. Add 20 μL of 9 μg / mL LPS (final concentration 1 μg / mL) to each well and incubate at 37°C for 3.5 hours in 5% CO2. Add 20 μL of 50 mM ATP (final concentration 5 mM) to the compound-treated wells and high control wells, and add 20 μL of medium to the low control wells, incubating at 37°C for 0.5 hours in 5% CO2. Transfer 160 μL of the supernatant to a new plate and store at -80°C.

[0168] Day 3: The supernatant was used for IL-1β release assay in THP1-Null cells according to the manufacturer's instructions.

[0169] (1) Take 16 μL of IL1β standard (Std 0-Std 7) and add it to each standard well, or take 16 μL of the sample to be tested and add it to each sample well.

[0170] (2) Add 4 μL of premixed IL1β antibody working solution to all wells.

[0171] (3) Seal the plate and incubate overnight at room temperature.

[0172] (4) Read the ratio of fluorescence wavelengths 665 / 620 using the Envision instrument.

[0173] 3. Experimental Results

[0174] Activation of the NLRP3 inflammasome leads to the release of the inflammatory cytokine IL-1β, while dysregulation of NLRP3 inflammasome activation drives the development of many diseases. The data in the table show that the compounds disclosed in this paper have NLRP3 inhibitory activity.

[0175] Table 1. Tests on the inhibitory activity of the compounds disclosed in this paper against NLRP3.

[0176] Test Example 2. Toxicity assay of NLRP3 inhibitor in HepG2 cells

[0177] 1. Experimental instruments and reagents

[0178] 1.1 Experimental Apparatus

[0179] PerkinElmer 2104 EnVision Multilabel Plate Readers

[0180] Multidrop TM Pico 8 Digital Dispenser

[0181] 1.2 Experimental Reagents

[0182] Compound A: Compound 50 prepared according to the method of Example 45 of WO2022135567A.

[0183] 2. Experimental Design

[0184] Day 1: HepG2 cells were seeded in 384-well plates at a density of 2 × 10⁶ cells / well. 3 Cells / 50 μL DMEM medium containing 10% FBS / well, cultured overnight at 37°C with 5% CO2.

[0185] Day 2: via Multidrop TM The Pico 8 Digital Dispenser was used to add serially diluted compounds to 50 μL of cell culture medium, resulting in final compound concentrations of 150, 50, 16.67, 5.56, 1.85, 0.62, 0.21, 0.069, and 0.023 μM. The DMSO concentration was less than 0.3%. Cells were incubated at 37°C in a 5% CO2 incubator for 2 days.

[0186] Day 4: Add 25 μL of the assay kit (Celltiter Glo assay kit) to each well and shake (in the dark) for 3 min. Incubate at room temperature in the dark for 10 min, then read the plate using the Envision instrument.

[0187] 3. Experimental Results

[0188] Table 2. Cytotoxicity test of the disclosed compounds against HepG2 cells.

[0189] Compared to compound A, compound 1 exhibits weaker cytotoxicity against HepG2.

[0190] Test Example 3: Inhibition of hERG potassium ion channels by compound

[0191] 1. Experimental materials and instruments

[0192] Compound A: Compound 50 prepared according to the method of Example 45 of WO2022135567A.

[0193] 2. Cell lines and cell culture

[0194] HEK293 cell line (catalog number: K1236) stably expressing the hERG ion channel was purchased from Invitrogen. This cell line was cultured in a medium containing 85% DMEM, 10% dialyzed fetal bovine serum, 0.1 mM non-essential amino acid solution, 100 U / mL penicillin-streptomycin solution, 25 mM HEPES, 5 μg / mL blastomycin, and 400 μg / mL genimycin. When the cell density reached 40%–80% of the culture dish bottom area, the cells were digested with trypsin and passaged three times per week. Before experiments, cells were cultured at a density of 5 × 10⁵ cells in 6 cm culture dishes, induced with 1 μg / mL doxycycline for 48 hours, then digested and seeded on slides for subsequent manual patch-clamp experiments.

[0195] 3. Solution preparation

[0196] 1) Extracellular fluid (in mM): 132 sodium chloride, 4 potassium chloride, 3 calcium chloride, 0.5 magnesium chloride, 11.1 glucose and 10 HEPES (pH adjusted to 7.35 with sodium hydroxide).

[0197] 2) Intracellular fluid (in mM): 140 potassium chloride, 2 magnesium chloride, 10 EGTA, 5 magnesium ATP and 10 HEPES (pH adjusted to 7.35 using potassium hydroxide).

[0198] 4. Preparation of the solution of the compound to be tested

[0199] 1) The test compound was dissolved in DMSO and prepared into a stock solution with a final concentration of 10 mM.

[0200] 2) The stock solution was serially diluted with DMSO at a ratio of 1:3 to prepare three other intermediate concentration solutions with concentrations (mM) of 3.33, 1.11 and 0.37.

[0201] 3) Before the experiment, the intermediate solution of the test compound gradient was diluted again with extracellular fluid at a ratio of 1:1000 to prepare a series of working solutions with final concentrations (μM): 10, 3.33, 1.11, and 0.37. The 30 μM working solution was prepared by diluting the 10 mM stock solution 333.33 times. The content of DMSO in the working solutions was 0.1-0.3% (volume ratio).

[0202] 4) Five working solutions at different concentration gradients (30, 10, 3.33, 1.11, and 0.37 μM) were used to determine the potential inhibitory effect of the compound on hERG channels, and to fit dose-response curves and calculate IC50. 50 .

[0203] 5. Experimental Design

[0204] 1) Place the small glass slide containing HEK293 cells in the culture dish into the perfusion tank of the micromanipulation table.

[0205] 2) Using an Olympus IX51, IX71, or IX73 inverted microscope, position the appropriate cells in the center of the field of view. Use a ×10 objective lens to locate the tip of the glass electrode and center it in the field of view. Then, use the micromanipulator to move the electrode down while adjusting the coarse focus knob to slowly bring the electrode closer to the cells.

[0206] 3) When you get close to the cell, switch to a 40x objective lens for observation. Use the micromanipulator to fine-tune the setting so that the electrode gradually gets closer to the cell surface.

[0207] 4) Apply negative pressure to form a seal with a resistance higher than 1G between the electrode tip and the cell membrane.

[0208] 5) In voltage clamping mode, the instantaneous capacitive current C fast Compensation is then performed. Short bursts of negative pressure are then repeatedly applied to rupture the membrane, eventually forming a whole-cell recording pattern.

[0209] 6) Under the condition that the membrane potential is clamped at -60mV, the slow capacitive current C slow The cell membrane capacitance (Cm) and input membrane resistance (Ra) are compensated separately.

[0210] 7) After the cells stabilize, change the clamping voltage to -90mV, set the sampling frequency to 20kHz, and the filtering frequency to 10kHz. The leakage current detection conditions are: clamping voltage changed to -80mV, time duration 500ms.

[0211] 8) The hERG current testing method is as follows: A depolarization command voltage of 4.8 seconds is applied to depolarize the membrane potential from -80mV to +30mV. Then, a repolarization voltage of 5.2 seconds is applied instantaneously to reduce the membrane potential to -50mV to remove channel inactivation, thereby allowing the hERG tail current to be observed. The peak value of the tail current is the magnitude of the hERG current.

[0212] 9) The hERG currents used to detect the test compounds were continuously recorded for 120 seconds before drug administration to assess the stability of hERG current generation in the test cells. Only stable cells within the acceptable range of the evaluation criteria were allowed to proceed to the subsequent compound detection.

[0213] Testing the inhibitory effect of the test compound on hERG current: First, the hERG current measured in extracellular fluid containing 0.1% DMSO was used as the baseline. After the hERG current stabilized for at least 5 minutes, the solution containing the test compound was sequentially perfused around the cells from low to high concentration. After each perfusion, approximately 5 minutes were allowed for the compound to fully act on the cells while simultaneously recording the hERG current. Once the recorded current stabilized, the last 5 hERG current values ​​were recorded, and their average was taken as the final current value at the specific concentration. After testing the compound, 150 nM of dofilad (positive control) was added to the same cell to completely inhibit its current, serving as a positive control for that cell. Simultaneously, the positive control compound dofilad was detected synchronously before and after the test drug experiment using the same patch-clamp system to ensure the reliability and sensitivity of the entire detection system.

[0214] 6. Data Analysis

[0215] 1) After injecting blank solvent or compound gradient solution, calculate the average of the five consecutive current values ​​obtained after stabilization, and use these averages as the "tail current magnitude". 空白 "and tail current magnitude" 化合物 ".

[0216] 2) The current suppression percentage is calculated using the following formula.

[0217] 3) The dose-response curve was fitted using Graphpad Prism 8.0 software and the IC was calculated. 50 value.

[0218] 7. Experimental Results

[0219] Table 3. Effects of the compounds disclosed herein on hERG potassium ion channels IC50 50 The result of the value

[0220] Compared to compound A, compound 1 disclosed herein has a weaker inhibitory effect on hERG potassium ion channels.

[0221] Example 2. Preparation of GLP-1 analogs

[0222] The GLP-1 analogue is compound 18# prepared in Example 1 of WO2023083301 (which is incorporated herein by reference in its entirety).

[0223] The molecular structure of compound 18# is: H-YAibEGTFTSDYSIYK(OEG-OEG-yGlu-C20-OH)EKIAAQEFVNWLLAGGPSSGAPPPS-NH2

[0224] The structure is as follows:

[0225] Test Example 4. Efficacy of drugs in high-fat diet-induced obese (DIO) mice

[0226] 4.1 Information on experimental animals and materials

[0227] Male C57BL / 6J mice (Jiangsu Jicui Yaokang Biotechnology Co., Ltd.), 20-21 weeks old, weighing 27-31g.

[0228] DIO male mice (Jiangsu Jicui Yaokang Biotechnology Co., Ltd.), 20-21 weeks old, weighing 44-55g.

[0229] The manufacturer of 60% HFD (Cat#D12492) is Research Diets.

[0230] Compound 1, The preparation method is described in Example 1.

[0231] 4.2 Experimental Design

[0232] The dosage and administration regimen for this independent experiment are shown in Table 4, with 10 animals in each group.

[0233] Table 4. Experimental design for the efficacy study of the test drug in DIO mice " / " indicates not applicable; "sc" indicates subcutaneous administration; "BID" indicates twice daily; "Q3D" indicates once every three days; the dosage unit for compound 1 is mg / kg; the dosage unit for compound 18 is nmol / kg.

[0234] 4.3 Experimental Procedure

[0235] Five-week-old C57BL / 6J mice were fed 60% HFD for 15 weeks to induce a DIO (digestive iodine disorder) obesity model. Mice were weighed and randomly assigned to groups: a normal control group (C57BL / 6J mice fed a standard diet) and all other groups were DIO mice. The mice were administered the drug for 28 days while maintaining a high-fat diet. Body weight was monitored daily, and food intake was monitored every three days. At the endpoint, fat mass and lean body mass were measured.

[0236] 4.4 Data Analysis

[0237] Statistical analysis was performed using Graphpad Prism 9 software. The analysis was based on the raw data, and the results are expressed as mean ± SEM. One-way ANOVA and Dunnett's test were used for statistical analysis, with p < 0.05 considered statistically significant.

[0238] 4.5 Experimental Results

[0239] As shown in the figure, in the DIO mouse model, compound 1 alone was effective at 10 mg / kg. When compound 1 was used in combination with compound 18, it demonstrated a better weight loss effect (approximately 10% further weight reduction compared to compound 18 alone), and there was no significant change in cumulative food intake between the combination group and the compound 18-only group. The combination group showed better fat reduction and muscle preservation effects compared to compound 18 alone (****p<0.0001, ***p<0.001, **p<0.01 vs Model). ## p < 0.01 vs compound 18-10 nmol / kg).

[0240] Table 5. Results of drug efficacy and body weight assay in DIO mice

Claims

1. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prevention and / or treatment of a disease or disorder associated with weight gain, wherein, R 1 selected from hydrogen, methyl or difluoromethyl; R 2 each independently is selected from hydrogen, halogen, C 1-6 alkyl; R 3 and R 4 are each independently selected from hydrogen or halogen; R 5 each independently is selected from hydrogen, halogen, C 1-6 alkyl or C 1-6 haloalkyl; R 6 selected from C 1-6 alkyl or C 1-6 haloalkyl; R 7 each independently is selected from fluorine; m and p are each independently selected from 0, 1, 2, and 3; n is independently selected from 1, 2, and 3.

2. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of muscle loss caused by a GLP-1 receptor agonist; preferably, use in a medicament for the treatment and / or prevention of muscle loss caused by a disease or condition related to weight gain treated with a GLP-1 receptor agonist.

3. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a GLP-1 receptor agonist in the preparation of a medicament for the prevention and / or treatment of diseases or conditions associated with weight gain.

4. A method for preventing and / or treating a disease or condition associated with weight gain, comprising co-administering to a subject in need a compound of formula (I) or a pharmaceutically acceptable salt thereof and a GLP-1 receptor agonist.

5. The use or method according to any one of claims 1 to 4, said R 1 is selected from hydrogen.

6. The use or method according to any one of claims 1 or 5, said R 2 each independently is selected from hydrogen.

7. The use or method according to any one of claims 1 to 6, said R 3 and R 4 are each independently selected from hydrogen.

8. Use or method according to any one of claims 1 to 7, said R 5 each independently is selected from hydrogen.

9. Use or method according to any one of claims 1 to 8, said R 6 is selected from methyl or ethyl, preferably methyl.

10. The use or method according to any one of claims 1 to 9, wherein the compound of Formula (I) or a pharmaceutically acceptable salt thereof is Compound 1 or a pharmaceutically acceptable salt thereof, 11. The use or method according to any one of claims 1 to 10, wherein the subject's total weight is reduced while maintaining muscle mass and / or muscle strength.

12. The use or method according to any one of claims 1 to 11, wherein the disease or condition related to weight gain is selected from overweight, obesity, obesity-related gallbladder disease, obesity-induced sleep apnea, diabetes, overeating, fatty liver disease, non-alcoholic fatty liver disease, dyslipidemia, metabolic syndrome, insufficiency of satiety, hyperinsulinemia and nocturnal hypoglycemia; preferably overweight or obesity.

13. The use or method according to any one of claims 1 to 12, wherein the GLP-1 receptor agonist is a dual agonist of the GLP-1 receptor and the GIP receptor, preferably a GLP-1 analogue comprising the structure shown below, or a pharmaceutically acceptable salt thereof, 14. The use or method according to any one of claims 1 to 13, wherein the dosage of the compound of formula (I) or its pharmaceutically acceptable salt is selected from 0.01 mg to 1000 mg, and the frequency of administration is selected from once a month, twice a month, three times a month, every other week, once a week, twice a week, three times a week, four times a week, five times a week, six times a week, every other day, once a day, twice a day, or three times a day.

15. The use or method according to claim 14, wherein the compound of formula (I) or its pharmaceutically acceptable salt is administered by parenteral administration or oral administration, preferably oral administration.

16. The use or method according to any one of claims 1 to 15, wherein the dosage of the GLP-1 receptor agonist is selected from 0.01 mg to 1000 mg, and the frequency of administration is selected from once a month, twice a month, three times a month, every other week, once a week, twice a week, three times a week, four times a week, five times a week, six times a week, every other day, once a day, twice a day, or three times a day; preferably once a week or every other week.

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