Drugs and applications for removing fat or fat pads

Abstract

The invention belongs to the field of medicine and relates to a medicine for removing fat or fat pad. The medicine can be used to prepare medicine for treating adipose tissue diseases or preparing medicine for regressing adipose tissue tumors.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and relates to a drug for removing fat or fat pads and its application, and in particular to a drug for removing subcutaneous fat accumulation or fat pads resistant to diet and its application. Background Art

[0002] Currently, subcutaneous fat accumulations or fat cell proliferations such as lipomas or lipomas are treated surgically by liposuction or direct surgical removal. These types of treatments have known complications or risks associated with anesthesia, local reactions and possible infection, and often require hospitalization for monitoring.

[0003] Mesodermal lipolysis therapy is to inject drugs into the mesoderm, causing a large number of fat cells to necrotize and then dissolve fat. Mesodermal lipolysis therapy injection drugs mainly include cationic lipolysis injection drugs represented by phosphatidylcholine (as shown in Table 1) and anionic lipolysis injection drugs represented by sodium deoxycholate (as shown in Table 1). Figure 1 as well as Figure 2 As shown). The two types of drugs have different lipolysis mechanisms and their own strengths. The United States has approved the first sodium deoxycholate lipolysis injection product, but the product still has the above-mentioned side effects to a certain extent. A commercial product, which is an aqueous preparation containing phospholipids, bile acid, riboflavin, α-tocopherol, ethanol and water. However, the adverse reactions caused by these lipolysis drugs cannot be ignored. On the one hand, due to poor solubility, large volumes have to be injected. In order to increase solubility, a lot of solubilizers and pH regulators are added, resulting in severe pain and swelling after injection, which is difficult for patients to accept. On the other hand, due to the incompatibility of lipolysis ability and tissue compatibility of different lipolysis drugs, a series of inflammatory reactions will be caused in the peripheral tissues of the drug administration, which will cause inflammatory reactions and severe pain and swelling at the site of implementation, and even cause risks such as local tissue necrosis or infection. At the same time, the number of treatments and the scale of acupuncture need to be improved. In addition to the risks of severe postoperative pain, nerve paralysis, local tissue necrosis or infection, the dose of a single injection is also very limited. The RZL-012 for injection developed by Raziel Therapeutics of Israel (as shown in Table 1) is a new type of chemically synthesized small molecule compound, a cationic lipolytic drug, with the characteristics of long-term effect and good tissue compatibility. However, as a synthetic exogenous drug, its safety needs further investigation.

[0004] In general, the products currently on the market for reducing local fat require multiple local injections. However, the current products have problems such as large injection volume, complex dissolution-promoting excipients, poor fat dissolving effect, different fat dissolving mechanisms, and serious adverse reactions. Therefore, it is necessary to develop a product that can effectively reduce local fat with fewer side effects and better safety.

[0005] Table 1 Structural comparison of sodium deoxycholate, phosphatidylcholine and RZL-012

[0006] Summary of the invention

[0007] In order to solve the above technical problems existing in the background technology, the present invention provides a drug and application for removing fat or fat pads with high solubility, improved fat dissolving efficiency and small side effects.

[0008] In order to achieve the above object, the present invention adopts the following technical solution:

[0009] A drug for removing fat or fat pads, characterized in that the drug for removing fat or fat pads is a compound represented by formula I:

[0010]

[0011] in:

[0012] Said X is selected from O, S, NH or does not exist;

[0013] The R 4 Selected from H or OR 8 ;

[0014] The R 5 , R 6 , R 7 and R 8 Independently selected from H or -LR 9 ;

[0015] The L is a linking moiety or is absent, the L is a saturated or unsaturated hydrocarbon group, phosphorus, sulfur or a combination thereof, which may be optionally interrupted by one or more -O- and / or -NH- groups, and may be optionally replaced by one or more =O, -OH and / or -NH 2 Group substitution.

[0016] The above R 9 It is C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 a substituted or unsubstituted heteroaryl group;

[0017] or R 9 is a natural or non-natural amino acid residue or a peptide containing 2, 3 or more natural or non-natural amino acid residues;

[0018] or R 9 is a choline residue, a L-carnitine residue, or a betaine residue;

[0019] or R 9 is a phosphatidylcholine residue;

[0020] or R 9 yes

[0021] in:

[0022] R 1 , R 2 , and R 3 Independently selected from H, C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 a substituted or unsubstituted heteroaryl group;

[0023] A - Selected from Cl - Br - 、F - ,I - ,AcO - , citrate or any other negative ions;

[0024] R is selected from branched or straight chain -(CH 2 ) n -, where: n = 0, 1, 2, 3, ...;

[0025] All R or -(CH 2 ) n-group contains C, O, S, N, single, double or triple bonds, any CH 2 It may be substituted by O, S, or NH.

[0026] The amino acid residue is selected from valine, proline, lysine, leucine, methionine, D-methionine, serine, alanine, D-alanine, glycine, isoleucine, histidine, aminoisobutyric acid, phenylglycine, tryptophan, tyrosine, O-benzylserine, O-benzylglutamine, γ-aminobutyric acid or

[0027] in:

[0028] The R 11 and R 12 are each independently selected from the group consisting of H and a substituted or unsubstituted group selected from C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 heteroaryl;

[0029] Or the R 11 and R 12 Together with adjacent carbon atoms, they form substituted or unsubstituted C 3 -C 12 Heterocycloalkyl;

[0030] The R 13 is selected from the group consisting of H and a substituted or unsubstituted group selected from C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 heteroaryl; or when at least two amino acid residues are present, R 13It is the bond between two amino acid residues;

[0031] The R 14 is selected from the group consisting of H and a substituted or unsubstituted group selected from C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl; or R 13 and R 14 Together with the adjacent carbon and nitrogen atoms, they form C 3 -C 10 Heterocycloalkyl;

[0032] m is an integer selected from 0 to 10.

[0033] The above L represents absence, substituted or unsubstituted C 1 -C 9 alkyl,

[0034] or a combination thereof.

[0035] The above R 4 is H; said X is O, S or absent.

[0036] The above X is O.

[0037] The above-mentioned drug for removing fat or fat pad is a compound as shown below:

[0038]

[0039]

[0040]

[0041]

[0042] A use of a drug based on the removal of fat or fat pad as described above in the preparation of a drug for treating adipose tissue disorders or a drug for regressing adipose tissue tumors.

[0043] A use of a drug based on the removal of fat or fat pad as described above in the preparation of a drug for treating local fat distribution disorder.

[0044] The advantages of the present invention are:

[0045] The invention provides a drug for removing fat or fat pads. Compared with drugs for removing fat or fat pads such as sodium deoxycholate, the invention has the advantages of good solubility, high fat dissolving efficiency, good biocompatibility, low side effects, etc., and overcomes the shortcomings of drugs for removing fat or fat pads such as sodium deoxycholate or phosphatidylcholine, such as poor solubility, large injection volume, complex dissolution-promoting auxiliary materials leading to relatively serious adverse reactions, and difficulty in patients to accept, as well as poor fat dissolving effect, different fat dissolving mechanisms, and unpredictable safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 is the ball-and-stick model of the deoxycholate ion and its hydrophilic and hydrophobic sides;

[0047] Figure 2 is a scale model of the deoxycholate ion and its hydrophilic and hydrophobic sides;

[0048] Figure 3 MS ESI of compound LGR-008 + Spectrum;

[0049] Figure 4 For compound LGR-008 1 HNMR spectrum;

[0050] Figure 5 For compound LGR-008 13 C NMR spectrum;

[0051] Figure 6 This is a bar graph of the inhibitory effect of each group on the growth of pre-adipocytes detected by the cell survival rate test (MTT assay) of the present invention;

[0052] Figure 7 This is a bar graph of the inhibitory effect of each group on the growth of differentiated adipocytes detected by the cell survival rate test (MTT assay) of the present invention;

[0053] Figure 8 as well as Fig. 9 This is a bar graph of the inhibitory effect of each group on mature adipocyte growth detected by the cell survival rate test (MTT assay) of the present invention;

[0054] Fig.10 It is a representative comparative anatomical diagram of rats in the control group, sodium deoxycholate group and LGR-008 group of the present invention;

[0055] Fig.11 Schematic diagrams and representative comparative anatomical diagrams of fat sites obtained from the injected and uninjected groin of rats in the control group, sodium deoxycholate group and LGR-008 group of the present invention;

[0056] Fig.12It is a representative comparison diagram of fat obtained from the injected and non-injected groin of rats in the control group, sodium deoxycholate group and LGR-008 group of the present invention. DETAILED DESCRIPTION

[0057] The present invention provides a drug for removing fat or fat pads, which is a compound represented by formula I:

[0058]

[0059] in:

[0060] X is selected from O, S, NH or is absent;

[0061] R 4 Selected from H or OR 8 ;

[0062] R 5 , R 6 , R 7 and R 8 Independently selected from H or -LR 9 ;

[0063] L is a linking moiety or is absent, L is a saturated or unsaturated hydrocarbon group, phosphorus, sulfur or a combination thereof, which may be optionally interrupted by one or more -O- and / or -NH- groups, and may be optionally replaced by one or more =O, -OH and / or -NH 2 Group substitution.

[0064] R 9 It is C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 a substituted or unsubstituted heteroaryl group;

[0065] or R 9 is a natural or non-natural amino acid residue or a peptide containing 2, 3 or more natural or non-natural amino acid residues;

[0066] or R 9 is a choline residue, a L-carnitine residue, or a betaine residue;

[0067] or R 9is a phosphatidylcholine residue;

[0068] or R 9 yes

[0069] in:

[0070] R 1 , R 2 , and R 3 Independently selected from H, C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 a substituted or unsubstituted heteroaryl group;

[0071] A - Selected from Cl - Br - 、F - ,I - ,AcO - , citrate or any other negative ions;

[0072] R is selected from branched or straight chain -(CH 2 ) n -, where: n = 0, 1, 2, 3, ...;

[0073] All R or -(CH 2 ) n -group contains C, O, S, N, single, double or triple bonds, any CH 2 It may be substituted by O, S, or NH.

[0074] The amino acid residue is selected from valine, proline, lysine, leucine, methionine, D-methionine, serine, alanine, D-alanine, glycine, isoleucine, histidine, aminoisobutyric acid, phenylglycine, tryptophan, tyrosine, O-benzylserine, O-benzylglutamine, γ-aminobutyric acid or

[0075] in:

[0076] R 11 and R 12Each independently selected from the group consisting of H and substituted or unsubstituted groups, the substituted or unsubstituted groups selected from C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 heteroaryl;

[0077] or R 11 and R 12 Together with adjacent carbon atoms, they form substituted or unsubstituted C 3 -C 12 Heterocycloalkyl;

[0078] R 13 is selected from the group consisting of H and a substituted or unsubstituted group, the substituted or unsubstituted group being selected from C 1 -C 12 Alkyl, C 2 -C 12 Alkenyl, C 2 -C 12 Alkynyl, C 3 -C 15 Cycloalkyl, C 3 -C 15 Heterocycloalkyl, C 6 -C 15 Aryl or C 5 -C 15 heteroaryl; or when at least two amino acid residues are present, R 13 It is the bond between two amino acid residues;

[0079] R 14 is selected from the group consisting of H and a substituted or unsubstituted group, the substituted or unsubstituted group being selected from C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl; or R 13 and R 14 Together with the adjacent carbon and nitrogen atoms, they form C 3 -C 10 Heterocycloalkyl;

[0080] m is an integer selected from 0 to 10.

[0081] L represents absence, substituted or unsubstituted C 1 -C 9 alkyl,

[0082] or a combination thereof.

[0083] R 4 is H; X is O, S or absent.

[0084] X is O.

[0085] Exemplarily, the drug for removing fat or fat pad provided by the present invention can be one or a combination of the following compounds:

[0086]

[0087]

[0088]

[0089]

[0090] The present invention provides a drug for removing fat or fat pads, and also provides the use of the drug in preparing a drug for treating adipose tissue disorders or a drug for regressing adipose tissue tumors, or in preparing a drug for treating local fat distribution disorders.

[0091] Furthermore, the present invention provides the use of a drug for removing fat or fat pads in the preparation of a pharmaceutical product for reducing local fat and body weight.

[0092] The present invention further provides a medicine for reducing local fat and body weight, wherein the medicine comprises an effective dose of a composition for reducing local fat and body weight and a pharmaceutically acceptable carrier thereof.

[0093] The present invention encompasses both stereoisomers (eg, cis and trans isomers) and all currently known optical isomers of the compound (eg, R and S enantiomers), as well as racemic, diastereomeric and other similar isomeric mixtures.

[0094] The present invention relates to compounds, which, after preparation, separation and purification, have a content of not less than 95% (high purity) in the resulting components, which compounds will then be used in subsequent preparations of the present invention. In certain specific embodiments, the purity of the compounds of the present invention will be greater than 99%.

[0095] The present invention also relates to solvates and polymorphs of the relevant compounds. Solvates of the compounds of the present invention include, for example, hydrates.

[0096] The present invention also covers pharmaceutically acceptable acid addition salts, which are salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, and the like; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, decanoates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartates, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginate, ascorbates, salicylates, 4-aminosalicylates, naphthalene disulfonates, and the like. These salts can be prepared by methods known in the art.

[0097] The present invention also covers pharmaceutically acceptable base addition salts, which are salts formed with inorganic or organic bases that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, etc. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts and magnesium salts. The salt derived from organic base includes but is not limited to the following salt: primary amines, secondary amines and tertiary amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known in the profession.

[0098] The pharmaceutical products of the present invention can exist in a variety of forms. These forms include, but are not limited to, liquid, semisolid and solid dosage forms. Such as liquid solutions (e.g., injectable and infusible solutions), dispersions, suspensions, powders, lyophilized powders or liposomes, or through transdermal absorption of ointments and patches. The preferred form depends on the intended mode of administration and therapeutic application; preferably, the pharmaceutical products of the present invention are in the form of infusible solutions, and the preferred mode of administration is a non-intestinal mode, such as an injection solution. In an embodiment of the present invention, a pharmaceutical product containing a compound provided by the present invention that effectively reduces a dose of local fat is administered by subcutaneous injection; more preferably, a pharmaceutical product containing a compound provided by the present invention that effectively reduces a dose of local fat is administered by subcutaneous injection to the subcutaneous fat layer. In a specific embodiment, the ointment is formulated as known to those skilled in the art, including but not limited to active agents, waxes, water, vaseline, preservatives, higher alcohols, polyhydroxy alcohols, emulsifiers, solvents, thickeners, plasticizers, fragrances, buffers, antibiotics, tranquilizers or mixtures thereof.

[0099] The content of the drug for removing fat or fat pad provided by the present invention can be appropriately adjusted according to different parameters. The above parameters include but are not limited to the type of receptor, the weight of the receptor, the thickness and area of ​​the local fat of the receptor, etc. The drug for removing fat or fat pad provided by the present invention can be administered once, multiple times or continuously within 24 hours, or multiple times or continuously every week or month. The administration method is injection, subcutaneous implantation, implantable infusion, ointment or patch. Preferably, the injection method includes but is not limited to subcutaneous injection, subcutaneous implantation, intravenous drip, intravenous injection pump, and implantable injection pump.

[0100] The present invention further provides an application of a pharmaceutical product for reducing local fat and body weight, wherein an effective dose of the pharmaceutical product of the present invention is administered to a local part of a receptor, so that the local part of the receptor can achieve the effects of inhibiting fat cell growth, promoting fat cell apoptosis, reducing deposited fat and reducing body weight.

[0101] Synthesis method

[0102] The compounds and methods of the present invention will be better understood in conjunction with the following synthetic schemes, which illustrate methods for preparing the compounds of the present invention, which are intended to illustrate rather than limit the scope of the present invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications may be made without departing from the spirit of the present invention and the scope of the appended claims, including but not limited to changes and modifications related to the chemical structures, substituents, derivatives and / or methods of the present invention.

[0103] The following examples are intended to illustrate embodiments of the present invention and are not intended to limit the same in any way.

[0104] Scenario 1:

[0105] Solution 1-1:

[0106]

[0107] Solution 1-2:

[0108]

[0109] Reference patents CN101489985A, CN101500984A, CN103705496A and patent CN101687792A.

[0110] As shown in Scheme 1, compound LGR-001 was alternatively prepared from deoxycholic acid and 2-bromo-N,N-diethylethylamine and compound LGR-002 was prepared from deoxycholic acid and 3-bromo-N,N-diethylpropylamine:

[0111] The synthesis method of compound LGR-001 is:

[0112]

[0113] The synthesis method of compound LGR-004 is:

[0114]

[0115] The compound deoxycholeyl-L-carnitine is synthesized by:

[0116]

[0117] The synthesis method of the compound deoxycholeyl L-carnitine ethyl ester is:

[0118]

[0119] Scenario 2:

[0120] Solution 2-1:

[0121]

[0122] Solution 2-2:

[0123]

[0124] Reference patents CN108218945A, CN101489985A, CN101500984A, CN103705496A and patent CN101687792A.

[0125] As shown in Scheme 2, compound LGR-001 was alternatively prepared from deoxycholate and diethylaminoethanol and compound LGR-002 was prepared from deoxycholic acid and diethylaminopropanol hydrochloride:

[0126] The synthesis method of compound LGR-001 is:

[0127]

[0128] The synthesis method of compound LGR-004 is:

[0129]

[0130] Solution 3:

[0131] Solution 3-1:

[0132]

[0133] Reference patents WO2020260558A, CN101489985A, CN108218945A, CN101489985A, CN101500984A, CN103705496A and patent CN101687792A.

[0134] As shown in Scheme 3, the compound is prepared from deoxycholate and 2-bromo-N,N-dimethylethylamine, and further reacted with monoiodomethane to prepare compound LGR-003; the compound is prepared from deoxycholate and 3-bromo-N,N-dimethylpropylamine, and further reacted with monoiodomethane to prepare compound LGR-004; the compound is prepared from deoxycholate and 4-bromo-N,N-dimethylbutylamine, and further reacted with monoiodomethane to prepare compound LGR-005.

[0135] The synthesis method of compound LGR-003 is:

[0136]

[0137] The synthesis method of compound LGR-004 is:

[0138]

[0139] Solution 3-2:

[0140]

[0141] Reference patents WO2020260558A, CN108218945A, CN101489985A, CN101500984A, CN103705496A and patent CN101687792A.

[0142] As shown in Scheme 3, the compound is prepared from deoxycholic acid and dimethylaminoethanol hydrochloride, and further reacted with monoiodomethane to prepare compound LGR-003; the compound is prepared from deoxycholic acid and dimethylaminopropanol hydrochloride, and further reacted with monoiodomethane to prepare compound LGR-004; the compound is prepared from deoxycholic acid and diethylaminobutanol hydrochloride, and further reacted with monoiodomethane to prepare compound LGR-005.

[0143] The synthesis method of compound LGR-003 is:

[0144]

[0145] The synthesis method of compound LGR-004 is:

[0146]

[0147] Solution 4:

[0148]

[0149] Reference patents CN112135812A, CN101489985A2, CN101500984A, CN103705496A and patent CN101687792A.

[0150] As shown in Scheme 4, alternatively, compound 2-((1-chloroethoxy)carbonyl)oxy-N,N-dimethylethylammonium hydrochloride is prepared from 1-chloroethyl chloroformate and dimethylaminoethanol hydrochloride, deoxycholate and 2-((1-chloroethoxy)carbonyl)oxy-N,N-dimethylethylammonium hydrochloride are used to prepare compound LGR-006, which is further reacted with monoiodomethane.

[0151] The synthesis method of compound LGR-006 is:

[0152]

[0153] Solution 5:

[0154]

[0155] Reference patents CN112135812A, CN1135221A, CN101489985A, CN101500984A, CN103705496A and patent CN101687792A.

[0156] As shown in Scheme 4 and Scheme 5, the compound 2-((1-chloromethoxy)carbonyl)oxy-N,N-dimethylethylammonium hydrochloride is prepared from chloromethyl chloroformate and dimethylaminoethanol hydrochloride, and the other steps refer to the synthesis of LGR-006. The compound is prepared from deoxycholate and 2-((chloromethoxy)carbonyl)oxy-N,N-dimethylethylammonium hydrochloride, and further reacted with monoiodomethane to prepare compound LGR-007.

[0157] The synthesis method of compound LGR-007 is:

[0158]

[0159] Solution 6:

[0160] Scheme 6-1:

[0161]

[0162] Scheme 6-2:

[0163]

[0164] Scheme 6-3:

[0165]

[0166] Scheme 6-4:

[0167]

[0168] Scheme 6-5:

[0169]

[0170] Reference patents WO1995008559A, US5922703A, CN108218946A, CN101489985A, CN101500984A, CN103705496A, CN101687792A and Mohite AR, Phatake RS, Dubey P, Agbaria M, Shames AI, Lemcoff NG, Reany O. Thiourea-Mediated Halogenation of Alcohols. J Org Chem. 2020 Oct 16; 85(20): 12901-12911.

[0171] As shown in Scheme 6, the carboxyl group of deoxycholic acid is first protected, and then an esterification reaction is carried out. The product is separated by column chromatography, and the obtained compound is further reacted with iodomethane, and the reaction product is then decarboxylated to obtain the prepared compound LGR-008.

[0172] The synthesis method of compound LGR-008 is:

[0173]

[0174] Solution 7:

[0175] Scheme 7-1:

[0176]

[0177] Scheme 7-2:

[0178]

[0179] Scheme 7-3:

[0180]

[0181] Reference patents CN114716497A, US5922703A, CN108218946A, CN101489985A, CN101500984A, CN103705496A, CN101687792A and Mohite AR, Phatake RS, Dubey P, Agbaria M, Shames AI, Lemcoff NG, Reany O. Thiourea-Mediated Halogenation of Alcohols. J Org Chem. 2020 Oct 16; 85(20): 12901-12911.

[0182] As shown in Scheme 7, the carboxyl group of 12-ketolithocholic acid is first protected, and then an esterification reaction is carried out. The product is reduced with lithium tri-t-butoxyaluminohydride, and the obtained compound is further reacted with monoiodomethane. The reaction product is then decarboxylated to obtain the prepared compound LGR-008.

[0183] The synthesis method of compound LGR-008 is:

[0184]

[0185] Solution 8:

[0186] Scheme 8-1:

[0187]

[0188] Scheme 8-2:

[0189]

[0190] Scheme 8-3:

[0191]

[0192] Reference patents CN114716497A, US5922703A, CN108218946A, CN101489985A, CN101500984A, CN103705496A and patent CN101687792A.

[0193] As shown in Scheme 8, 12-ketolithocholic acid is first subjected to an esterification reaction with an acyl chloride, and the product is reduced with lithium tri-t-butoxyaluminohydride. The obtained compound is further reacted with iodomethane, and the reaction product is then decarboxylated to obtain the prepared compound LGR-008.

[0194] Some embodiments of the present invention relate to compounds of the chemical formula in Table 2:

[0195] Table 2 Synthesized Example Compounds

[0196]

[0197]

[0198]

[0199] Experimental Procedure for Synthesis of Example Compounds

[0200] Embodiment 1:

[0201] Synthesis of 2-(diethylamino)ethyl(R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoate (compound LGR-001).

[0202] Synthesis method 1: Add deoxycholic acid (0.39 g, 0.001 mol, 1 equivalent), 0.15 mL triethylamine and 100 mL acetonitrile to a single-mouth bottle at room temperature, stir to form a suspension, suspend 2-bromo-N, N-diethylethylamine hydrobromide (0.26 g, 0.001 mol, 1 equivalent) in 10 mL acetonitrile and add to the single-mouth bottle. Stir the mixture overnight at room temperature. Add 10 mL of 5% potassium carbonate 50% ethanol aqueous solution, and evaporate the solvent on a rotary evaporator. Add 20 mL of 10% hydrochloric acid aqueous solution to the evaporated mixture, fully rotate and mix on a rotary evaporator at 50°C, filter, and evaporate the solvent on a rotary evaporator at 50°C. The evaporated mixture is purified on a silica gel column (eluted with DCM / MeOH (10 / 1 to 1 / 2)), detected by TLC, developing agent: DCM / MeOH = 7 / 3, color developer: potassium bismuth iodide. After column chromatography, 2-(diethylamino)ethyl deoxycholate was obtained. 2-(diethylamino)ethyl deoxycholate was dissolved in 2 mL of 10% hydrochloric acid ethanol solution, heated to 50°C, and vacuum evaporated for 2 hours. After concentrating the 10% hydrochloric acid solution, the target compound LGR-001 (0.24 g, 46%) was obtained.

[0203] Synthesis method 2: Add 100mL DCM and deoxycholic acid (0.39g, 0.001mol, 1 equivalent) to a single-mouth bottle at room temperature, cool to 0°C, then add 2-bromo-N, N-diethylethylamine hydrobromide (0.26g, 0.001mol, 1 equivalent) to the single-mouth bottle, stir to form a suspension, and drop 0.15mL triethylamine. The mixed solution is stirred overnight at room temperature. Add 10mL of 5% potassium carbonate 50% ethanol aqueous solution to the reaction solution, and evaporate the solvent on a rotary evaporator. Add 20mL of 10% hydrochloric acid aqueous solution to the evaporated mixture, fully rotate and mix on a rotary evaporator at 50°C, filter, and evaporate the solvent on a rotary evaporator at 50°C. The evaporated mixture is purified on a silica gel column (eluted with DCM / MeOH (10 / 1 to 1 / 2)), and detected by TLC, with a developing agent: DCM / MeOH = 7 / 3, and a color developer: potassium bismuth iodide. After column chromatography, 2-(diethylamino)ethyl deoxycholate was obtained. 2-(diethylamino)ethyl deoxycholate was dissolved in 2 mL of 10% hydrochloric acid ethanol solution, and the mixture was evaporated under reduced pressure at 50°C for 2 hours. After the 10% hydrochloric acid ethanol solution was concentrated, the target compound LGR-001 (0.32 g, 60%) was obtained.

[0204] Synthesis method 3: Add 100mL DCM and deoxycholic acid (0.39g, 0.001mol, 1 equivalent) to a single-mouth bottle at room temperature, cool to 0°C, then add 2-bromo-N, N-diethylethylamine hydrobromide (0.26g, 0.001mol, 1 equivalent) to the single-mouth bottle, stir to form a suspension, and drop 0.15mL triethylamine. The mixed solution is stirred overnight at room temperature. The solvent is evaporated on a rotary evaporator. Add 10mL of 5% hydrochloric acid ethanol to the evaporated mixture, filter, wash the filter residue with 10mL of 5% hydrochloric acid ethanol, and evaporate the solvent on a rotary evaporator of the filtrate. Add 20mL of 10% hydrochloric acid aqueous solution to the evaporated mixture, fully rotate and mix on a rotary evaporator at 50°C, filter, and evaporate the solvent on a rotary evaporator of the filtrate at 50°C. The evaporated mixture was purified on a silica gel column (eluted with DCM / MeOH (10 / 1 to 1 / 2)), detected by TLC, developing solvent: DCM / MeOH = 7 / 3, color developing agent: potassium bismuth iodide. After column chromatography, 2-(diethylamino)ethyl deoxycholate was obtained. 2-(diethylamino)ethyl deoxycholate was dissolved in 2 mL of 10% hydrochloric acid ethanol solution, and vacuum evaporated at 50°C for 2 hours. After concentrating the 10% hydrochloric acid ethanol solution, the target compound LGR-001 (0.35 g, 67%) was obtained.

[0205] LCMS (ESI) m / z: 492.2 [M+1] + ; 1 H NMR (400 MHz, CDCl 3 )δ4.31-4.30(m, 2H), 3.73-3.59(m, 2H), 3.35-3.37(m, 2H), 3.26(q, J=7.2Hz, 4H), 2.29-2.13(m, 1H) , 2.07-2.01 (m, 1H), 1.81-1.23 (m, 24H), 1.13-0.91 (d, J=6.3Hz, 6H), 0.90-0.80 (m, 6H), 0.54 (s, 3H).

[0206] Embodiment 2:

[0207] Synthesis of 3-(diethylamino)propyl(R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoate (Compound LGR-002). The method described in Example 1 was followed, except that 2-bromo-N,N-diethylethylamine hydrobromide was replaced with 3-bromo-N,N-diethylpropylamine hydrobromide to obtain the target compound. LCMS (ESI) m / z: 506.2 [M+1] + ; 1 H NMR (400 MHz, CDCl 3 )δ4.41-4.36(m, 2H), 3.66-3.53(m, 2H), 3.31-3.30(m, 2H), 3.21(q, J=7.4Hz, 4H), 2.21-2.13(m, 1H) , 2.11-2.05 (m, 1H), 1.81-1.23 (m, 26H), 1.13-0.91 (d, J=6.3Hz, 6H), 0.90-0.80 (m, 6H), 0.55 (s, 3H).

[0208] Embodiment 3:

[0209] Synthesis of 2-(((R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)oxy)-N,N,N-trimethylethan-1-aminium (Compound LGR-003). The method described in Example 1 was followed, except that 2-bromo-N,N-diethylethylamine hydrobromide was replaced with 2-bromo-N,N,N-trimethylethylammonium hydrobromide to obtain the target compound. LCMS (ESI) m / z: 478.2 [M] + ; 1 H NMR (400 MHz, CDCl 3)δ4.39-4.37(m, 2H), 3.69-3.57(m, 2H), 3.38-3.35(m, 2H), 3.00(s, 9H), 2.20-2. 13(m, 1H), 2.11-2.05(m, 1H), 1.85-1.21(m, 24H), 0.93-0.85(m, 6H), 0.60(s, 3H).

[0210] Embodiment 4:

[0211] Synthesis of 3-(((R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)oxy)-N,N,N-trimethylpropan-1-aminium (Compound LGR-004). The method described in Example 1 was followed, except that 2-bromo-N,N-diethylethylamine hydrobromide was replaced with 3-bromo-N,N,N-trimethylpropylammonium hydrobromide to obtain the target compound. LCMS (ESI) m / z: 492.2 [M] + ; 1 HNMR (400 MHz, CDCl 3 )δ4.35-4.32(m, 2H), 3.68-3.58(m, 2H), 3.35-3.32(m, 2H), 3.02(s, 9H), 2.21-2. 16 (m, 1H), 2.10-2.06 (m, 1H), 1.83-1.25 (m, 26H), 0.90-0.83 (m, 6H), 0.58 (s, 3H).

[0212] Embodiment 5:

[0213] Synthesis of 4-(((R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)oxy)-N,N,N-trimethylbutan-1-aminium (Compound LGR-005). The method described in Example 1 was followed, except that 2-bromo-N,N-diethylethylamine hydrobromide was replaced with 4-bromo-N,N,N-trimethylbutylammonium hydrobromide to obtain the target compound. LCMS (ESI) m / z: 506.2 [M] + ;1 HNMR (400 MHz, CDCl 3 )δ4.30-4.25(m, 2H), 3.61-3.53(m, 2H), 3.31-3.27(m, 2H), 3.01(s, 9H), 2.20-2. 13(m, 1H), 2.10-2.05(m, 1H), 1.85-1.21(m, 28H), 0.90-0.82(m, 6H), 0.54(s, 3H).

[0214] Embodiment 6:

[0215] Synthesis of 2-(((1-(((R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)oxy)ethoxy)carbonyl)oxy)-N,N,N-trimethylethan-1-aminium (Compound LGR-006). 2-((1-Chloroethoxy)carbonyl)oxy-N,N-dimethylethanaminium hydrochloride (0.23 g, 0.001 mol), 10 mg of NaI and sodium deoxycholate (0.42 g, 0.001 mol) were added to acetone (30 mL) and 2.0 mL of triethylamine was added. The reaction was heated to 50 °C for 12 h. The solvent was evaporated on a rotary evaporator. Add 10 mL of 5% hydrochloric acid ethanol to the evaporated mixture, filter, wash the filter residue with 10 mL of 5% hydrochloric acid ethanol, and evaporate the solvent on a rotary evaporator. Add 20 mL of 10% hydrochloric acid aqueous solution to the evaporated mixture, fully rotate and mix on a rotary evaporator at 50°C, filter, and evaporate the solvent on a rotary evaporator at 50°C. The evaporated mixture is redissolved in THF (5 mL) solution, 0.3 mL of MeI (5 mmol) is added, and the mixture is stirred at room temperature in the dark overnight, and the solvent is evaporated on a rotary evaporator at 50°C. The reactants are mixed with silica gel and purified on a silica gel column (eluted with DCM / MeOH (10 / 1 to 1 / 2)), and detected by TLC, with a developing agent of DCM / MeOH = 7 / 3 and a color developer of potassium bismuth iodide. The compound obtained after column chromatography was dissolved in 2 mL of 10% hydrochloric acid ethanol solution, heated to 50°C, and vacuum evaporated for 1 hour. After concentrating the 10% hydrochloric acid solution, the target compound LGR-006 (270 mg, 45%) was obtained. LCMS (ESI) m / z: 566.2 [M] + ; 1 HNMR (400 MHz, CDCl 3)δ7.85-7.75(m, 1H), 4.35-4.31(m, 2H), 3.64-3.53(m, 2H), 3.37-3.33(m, 2H), 3.01(s, 9H) , 2.23-2.15(m, 1H), 2.11-2.08(m, 1H), 1.85-1.25(m, 27H), 0.90-0.83(m, 6H), 0.55(s, 3H).

[0216] Embodiment 7:

[0217] Synthesis of 2-((((((R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-3,12-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoyl)oxy)methoxy)carbonyl)oxy)-N,N,N-trimethylethan-1-aminium (Compound LGR-007). The method described in Example 6 was followed, except that 1-chloroethyl chloroformate was replaced with chloromethyl chloroformate to obtain the target compound. LCMS (ESI) m / z: 552.2 [M] + ; 1 HNMR (400 MHz, CDCl 3 )δ5.88(s, 2H), 4.33-4.30(m, 2H), 3.66-3.56(m, 2H), 3.36-3.33(m, 2H), 3.01(s, 9H), 2. 20-2.13 (m, 1H), 2.11-2.08 (m, 1H), 1.81-1.23 (m, 24H), 0.90-0.81 (m, 6H), 0.57 (s, 3H).

[0218] Embodiment 8:

[0219] Synthesis of (R)-4-((3R,5R,8R,9S,10S,12S,13R,14S,17R)-12-hydroxy-10,13-dimethyl-3-(2-(trimethylammonio)acetoxy)hexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoate (compound LGR-008).

[0220] Deoxycholic acid (19.6 g, 0.05 mol, 1 equivalent) and 20 mL of triethylamine were added to 200 mL of DMF, and the mixture was thoroughly stirred to dissolve. Benzyl chloride (6.3 g, 0.05 mol, 1 equivalent) was added, and the mixture was stirred at room temperature overnight. TLC showed that the conversion of the raw material was complete. Water was added dropwise to quench the mixture, and the mixture was extracted with ethyl acetate three times. The organic layers were combined and concentrated to obtain a crude product. The product was purified by silica gel column chromatography (PE / EA=20 / 1) to obtain 17.50 g of benzyldeoxycholic acid LGR-008-A as a brown solid.

[0221] Add LGR-008-A (12.0 g, 0.025 mol, 1 equivalent) and 20 mL of triethylamine to 400 mL of toluene, stir thoroughly to dissolve, then cool to 0°C, gradually add N, N, N-trimethylacetyl chloride hydrochloride (4.7 g, 0.025 mol, 1 equivalent) in batches while stirring, and then stir at room temperature overnight. Add 250 mL of 5% potassium carbonate 50% ethanol aqueous solution to the reaction solution, and evaporate the solvent on a rotary evaporator. Add 1000 mL of 10% hydrochloric acid aqueous solution to the evaporated mixture, rotate and mix thoroughly on a rotary evaporator at 50°C, filter, and evaporate the solvent on a rotary evaporator at 50°C. The evaporated mixture was purified on a silica gel column (eluted with DCM / MeOH (10 / 1 to 1 / 1)), detected by TLC, developing solvent: DCM / MeOH = 7 / 3, color developer: potassium bismuth iodide. After column chromatography, 10.7 g of compound LGR-008-B was obtained,

[0222] Dissolve 5.0 g of compound LGR-008-B in 50 ml of methanol, add Pd / C (0.050 g, 10%), stir overnight at room temperature under hydrogen atmosphere, filter, and evaporate the solvent on a rotary evaporator at 50°C. The evaporated mixture was dissolved in 20 mL of 10% hydrochloric acid ethanol solution, heated to 50°C, and evaporated under reduced pressure for 2 hours. After concentrating the 10% hydrochloric acid ethanol solution, the target compound LGR-008 (2.0 g, 54%) was obtained. The mass spectrum, hydrogen spectrum and carbon spectrum of compound LGR-008 are shown as follows: Figure 3 , Figure 4 as well as Figure 5 As shown, LCMS (ESI) m / z: 492.2 [M+H] + ; 1 HNMR (400 MHz, CD 3 OD) δ5.11 (s, 1H), 4.37 (s, 2H), 3.97 (s, 1H), 3.35 (s, 9H), 2.43-2.17 (m, 2H), 1.81-1.20 (m, 24H), 0.90-0.80 (m, 6H), 0.71 (s, 3H).

[0223] Embodiment 9:

[0224] Synthesis of (R)-4-((3R, 5R, 8R, 9S, 10S, 12S, 13R, 14S, 17R)-3-(((R)-3-acetoxy-4-(trimethylammonio)butanoyl)oxy)-12-hydroxy-10, 13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoate (compound LGR-009). The method described in Example 8 was followed, except that N, N, N-trimethylacetyl chloride hydrochloride was replaced with 2-acetoxy-4-chloro-N, N, N-trimethyl-4-oxobutane-1-amine hydrochloride to first prepare LGR-009-B, and then the benzyl group was removed according to the method described in Example 8 to obtain the target compound. LCMS (ESI) m / z: 578.4 [M] + ; 1 HNMR (400 MHz, CD 3 OD) δ5.61 (s, 1H), 4.74 (s, 1H), 3.94-3.82 (m, 3H), 3.19 (s, 9H), 2.75-2.69 (m, 2H), 2 .43-2.15(m, 2H), 2.09(S, 3H), 1.81-1.20(m, 24H), 0.90-0.80(m, 6H), 0.70(s, 3H).

[0225] Embodiment 10:

[0226] Synthesis of (R)-4-((3R, 5R, 8R, 9S, 10S, 12S, 13R, 14S, 17R)-3-((dimethylglycyl)oxy)-12-hydroxy-10, 13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanoic acid (compound LGR-010). The method described in Example 8 was followed, except that N, N, N-trimethylacetyl chloride hydrochloride was replaced with N, N-dimethylacetyl chloride hydrochloride to first prepare LGR-010-B, and then the benzyl group was removed according to the method described in Example 8 to obtain the target compound. LCMS (ESI) m / z: 478.4 [M] + ; 1 HNMR (400 MHz, CD 3OD) δ5.17 (s, 1H), 4.33 (s, 2H), 3.91 (s, 1H), 3.35 (s, 6H), 2.45-2.13 (m, 2H), 1.80-1.20 (m, 24H), 0.91-0.82m, 6H), 0.69 (s, 3H).

[0227] Example 11: Solubility Experiment

[0228] Since bile acid drugs have poor solubility, one of the purposes of the present invention is to improve the solubility of bile acid drugs. Therefore, the solubility of the deoxycholic acid derivative or a pharmaceutically acceptable salt thereof, deoxycholic acid and sodium deoxycholate in water was measured.

[0229] The solubility of the compound was determined by the saturated shake flask method: 50 mL of PBS solution with a pH of 7.4 was added to a 250 mL conical flask, and deoxycholic acid, sodium deoxycholate and solid test substances LGR-001 to LGR-008 were crushed with a mortar and sieved. Excess deoxycholic acid, sodium deoxycholate and solid test substances LGR-001 to LGR-008 powder were divided into 3 parts and added to the PBS dissolution medium respectively, and ultrasonicated for one hour, with continuous vortexing and shaking during the period. Solution balance: The suspension was stirred for 24 hours. The temperature of the suspension was controlled at 37 ± 0.5 ° C throughout the dissolution stage. After stirring, it was allowed to stand for 2 hours and the solid was completely precipitated. Take 0.5 mL of the supernatant, add it to an EP tube containing 0.5 mL of methanol, and use HPLC-UV analysis to test the solubility.

[0230] Table 3 Comparison of solubility of deoxycholic acid, sodium deoxycholate and the present invention

[0231]

[0232] The solubility of the salt-formed compounds (Examples 1-10) of the present invention is greater than 4*100 mg / mL at a pH of 7.4, while the solubility of deoxycholic acid and sodium deoxycholate is 0.007*100 mg / mL and 0.13*100 mg / mL, respectively. Therefore, the compounds of the present invention are beneficial to the preparation, reduce the injection volume, and thus reduce side effects such as pain.

[0233] Example 12: Adipocyte growth inhibition experiment

[0234] 3T3-L1 (mouse embryonic fibroblasts) are continuous substrain cells of 3T3 (Swiss mice) obtained by cloning and separation. They are morphologically similar to fibroblasts and are long spindle-shaped. Studies have found that it is a precursor cell that can proliferate and differentiate into adipocytes under certain conditions. When 3T3-L1 cells go from rapid division to full growth and contact inhibition, the cells reverse from pre-fat to fat-like. In the culture medium, high serum content can promote the accumulation of fat in 3T3-L1 cells. The 3T3-L1 cell line can not only be stably propagated but also has good specificity for differentiation into adipocytes. It has now become the most commonly used cell line for inducing differentiation into adipocytes. 3T3-L1 cells can be used to study diabetes, obesity, etc., and have become an internationally recognized cell model for studying fat metabolism. It has great scientific research value for the study of adipocyte proliferation and differentiation.

[0235] Example 12-1 Preadipocyte growth inhibition experiment.

[0236] In this example, 3T3-L1 preadipocytes were cultured at 1×10 4 Cells (cells / well) were cultured in 96-well plates. In addition to the DMSO solvent control group (control group), 50ppm of deoxycholic acid (DA), 100ppm of choline, 100ppm of L-carnitine, 100ppm of betaine and 50ppm of the compounds of the present invention: LGR-001, LGR-003, LGR-005 and LGR-008 were added to different wells, with a total of 8 groups and 3 replicates for each group. After treatment and culture for 48 hours, the cell growth status was recorded by taking photos, and the cell survival rate test (MTT assay) was used to analyze the growth inhibition effect of each group on the precursor adipocyte 3T3-L1. The data of each group are expressed as mean ± SD.

[0237] The results are as follows Figure 6 As shown, compared with the DMSO solvent control group, the compounds of the present invention LGR-001, LGR-003, LGR-005 and LGR-008 can effectively inhibit the growth of preadipocytes (p < 0.05), and compared with the deoxycholic acid (DA) group, choline group, L-carnitine group and betaine group, they also have better inhibitory effects and reach significant differences (p < 0.05).

[0238] Example 12-2 Experiment on inhibition of growth of differentiated adipocytes.

[0239] In this embodiment, 3T3-L1 is converted into 1×10 5 Cells (cells / well) were cultured in a 12-well plate. On the fourth day, the culture medium containing 5 μg / mL differentiation agent insulin, 1 μM dexamethasone, and 0.5 mM 3-isobutyl-1-methylxanthine was used. DMSO solvent was added to different wells as a control group, 100 ppm deoxycholic acid (DA), 100 ppm choline, 100 ppm L-carnitine, 100 ppm betaine, and 100 ppm compounds of the present invention: LGR-001, LGR-003, LGR-005, and LGR-008 were added to the wells for experiments. There were 8 groups in total and each group was repeated 3 times. After drug treatment and culture for 48 hours, the cell growth status was recorded by taking photos, and the inhibitory effect of each experimental substance on differentiated adipocytes was analyzed by cell survival test. The data of each group were expressed as mean±SD.

[0240] The results are as follows Figure 7 As shown, compared with the DMSO solvent control group, the compounds LGR-001, LGR-003, LGR-005 and LGR-008 of the present invention can significantly inhibit the growth of differentiated adipocytes (p < 0.05), among which the compound LGR-008 has the best inhibitory effect on the growth of differentiated adipocytes, and the compound LGR-008 also has a better inhibitory effect on the growth of differentiated adipocytes compared with other groups (p < 0.05). Moreover, the effects of the drugs LGR-001, LGR-003, LGR-005 and LGR-008 for removing fat or fat pads provided by the present invention on inhibiting the growth of differentiated adipocytes are significantly better than those of the deoxycholic acid (DA) group, choline group, L-carnitine group and betaine group (p < 0.05).

[0241] Example 12-3 Mature adipocyte growth inhibition experiment.

[0242] This example compares the inhibitory effect of the composition of the present invention and the known local lipolysis ingredients deoxycholic acid (DA), choline and L-carnitine on mature adipocytes. In this example, 3T3-L1 cells were cultured in a 12-well plate at 3×104 cells / well. On the fourth day, the culture medium containing 5μg / mL insulin, 1μM dexamethasone and 0.5mM 3-isobutyl-1-methylxanthine was used. After two days, the culture medium containing 5μg / mL insulin was used and cultured for four days. After the 3T3-L1 cells were differentiated and matured, 100ppm of deoxycholic acid (DA) was added to different wells, except for the two solvent control groups of PBS and DMSO. 100 ppm of deoxycholic acid (DA), 100 ppm of choline, 100 ppm of L-carnitine, 100 ppm of betaine, and 100 ppm of the compounds of the present invention: LGR-001, LGR-003, LGR-005, and LGR-008 were added to the wells for the experiment, and there were 8 groups in total, and each group of experiments was repeated 3 times; except for the two solvent control groups of PBS and DMSO, 100 ppm of deoxycholic acid (DA), 100 ppm of choline, 100 ppm of L-carnitine, 100 ppm of betaine, and 100 ppm of the compounds of the present invention: LGR-009-B, LGR-009, LGR-010-B, and LGR-010 were added to different wells for the experiment, and there were 8 groups in total, and each group of experiments was repeated 3 times. After treatment with drugs and culture for 48 hours, the cell growth status was recorded by photographing, and the inhibitory effect of each experimental substance on differentiated adipocytes was analyzed by cell survival test. The data of each group are expressed as mean±SD.

[0243] The results are as follows Figure 8 as well as Fig. 9 As shown, the drugs for removing fat or fat pads provided by the present invention can effectively inhibit the growth of mature adipocytes, and the effects of the drugs for removing fat or fat pads provided by the present invention, LGR-001, LGR-003, LGR-005, LGR-008, LGR-009-B, LGR-009, LGR-010-B and LGR-010 on inhibiting the growth of mature adipocytes are significantly better than those of the deoxycholic acid (DA) group, choline group, L-carnitine group and betaine group (p < 0.05).

[0244] Example 13: Rat Animal Experiment

[0245] Test system: Male SD rats were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. at 6 weeks of age and were adaptively raised for one week.

[0246] Test method and drug administration: After 24 rats were continuously fed with high-fat diet for 2 weeks, the subcutaneous fat of the rats thickened. 18 rats weighing 310±10g were taken and randomly divided into 3 groups, 6 rats in each group, namely control group, sodium deoxycholate group and LGR-008 group. The 6 rats in the control group were numbered 1-1 to 1-6, the 6 rats in the sodium deoxycholate group were numbered 2-1 to 2-6, and the 6 rats in the LGR-008 group were numbered 3-1 to 3-6. Drug administration began, and the drug was administered by subcutaneous injection into the subcutaneous fat: the sodium deoxycholate group was administered with sodium deoxycholate solution, the LGR-008 group was administered with a solution of the compound LGR-008 of Example 8 of the present invention, and the control group was administered with normal saline. The injection site was the lower inguinal fat on the left side of the rat, with four injection points (2.5 mg / kg / point) on the upper, lower, left and right sides. The injection dose for each rat was 10 mg / kg / time per kg of the recipient, while the control group was injected with an equal volume of sterile saline for injection, once every 2 days, for a total of 3 injections. The day of the start of drug injection was set as the first day, and the drug was repeated in the above manner on the 3rd and 5th days. During the experiment, the body weight change and average daily feed intake were recorded daily. The last body weight was measured on the 15th day. The rats were fasted for 24 hours, and then the rats were killed and the fat in the same area of ​​the abdomen, upper inguinal and lower inguinal areas including the injection site was weighed to calculate the subcutaneous fat mass.

[0247] Experimental results: The fat weight of the treated side and the fat weight of the untreated side, the fat weight change ratio, and the number of rats with fat necrosis on the treated side of each group of rats on the 15th day are shown in Table 4.

[0248] Table 4 Fat weight on the drug-treated side and the non-drug-treated side of each group of rats on the 15th day and their change ratio

[0249]

[0250] * Fat weight is expressed as mean±SD.

[0251] ** Weight change ratio = (fat weight on the side without drug administration - fat weight on the side with drug administration) / fat weight on the side without drug administration * 100%.

[0252] Experimental conclusion: There was no significant difference in the weight of fat on the non-medicated side among the control group, sodium deoxycholate group and LGR-008 group (p>0.05), and there was a significant difference in the average weight of fat on the medicated side among the control group, sodium deoxycholate group and LGR-008 group, and the average weight of fat on the medicated side of the LGR-008 group was significantly lower than that of the control group (p<0.05). The weight of fat on the medicated side of the rats in the LGR-008 group injected with the compound LGR-008 of the present invention and the group injected with sodium deoxycholate, including the same area of ​​the injection site, was reduced compared with the weight of fat on the non-medicated side, which was reduced by 44.4% and 26.3% respectively, and the average weight of fat on the medicated side of the LGR-008 group was significantly lower than that of the sodium deoxycholate group (p<0.05).

[0253] like Fig.10 The representative comparative anatomical diagram of rats in the control group, sodium deoxycholate group and LGR-008 group of the present invention shows that after the rats were injected with the compound LGR-008 of the present invention three times, the fat on the injection side was significantly reduced. Fig.11 The diagram is a schematic diagram and representative comparative anatomical diagram of the fat sites obtained from the injected and uninjected sides of the groin of rats in the control group, sodium deoxycholate group and LGR-008 group of the present invention. After the rats were injected with the compound LGR-008 of the present invention three times, the fat on the injected side was significantly reduced, and there was no necrosis in the fat and its surrounding tissues. Fig.12 The figure is a representative comparison of fat obtained from the injected and non-injected groin of rats in the control group, sodium deoxycholate group and LGR-008 group of the present invention. After the rats were injected with the compound LGR-008 of the present invention three times, the fat on the injected side was significantly reduced, and there was no fat necrosis.

Claims

1. A drug for removing fat or fat pads, characterized in that: The drug for removing fat or fat pads is a compound represented by Formula I: in: Said X is selected from O, S, NH or does not exist; Said R4 is selected from H or OR8; Said R5, R6, R7 and R8 are independently selected from H or -L-R9; The L is a linking moiety or is absent, the L is a saturated or unsaturated hydrocarbon group, phosphorus, sulfur or a combination thereof, which may be optionally interrupted by one or more -O- and / or -NH- groups, and may be optionally substituted by one or more =O, -OH and / or -NH2 groups.

2. The drug for removing fat or fat pad according to claim 1, characterized in that: The R9 is C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C3-C 15 Cycloalkyl, C3-C 15 Heterocycloalkyl, C6-C 15 Aryl or C5-C 15 a substituted or unsubstituted heteroaryl group; or R9 is a natural or unnatural amino acid residue or a peptide containing 2, 3 or more natural or unnatural amino acid residues; or R9 is a choline residue, a L-carnitine residue or a betaine residue; or R9 is a phosphatidylcholine residue; Or R9 is in: R1, R2, and R3 are independently selected from H, C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C3-C 15 Cycloalkyl, C3-C 15 Heterocycloalkyl, C6-C 15 Aryl or C5-C 15 a substituted or unsubstituted heteroaryl group; A - Selected from Cl - Br - 、F - ,I - ,AcO - , citrate or any other negative ions; R is selected from branched or straight chain -(CH2) n -, wherein: n is an integer selected from 0 to 10; All R or -(CH2) n The -group contains C, O, S, N, a single bond, a double bond, or a triple bond, and any CH2 can be replaced by O, S, or NH.

3. The drug for removing fat or fat pad according to claim 2, characterized in that: The amino acid residue is selected from valine, proline, lysine, leucine, methionine, D-methionine, serine, alanine, D-alanine, glycine, isoleucine, histidine, aminoisobutyric acid, phenylglycine, tryptophan, tyrosine, O-benzylserine, O-benzylglutamine, γ-aminobutyric acid or in: The R 11 and R 12 Each is independently selected from the group consisting of H and a substituted or unsubstituted group selected from C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C3-C 15 Cycloalkyl, C3-C 15 Heterocycloalkyl, C6-C 15 Aryl or C5-C 15 heteroaryl; Or the R 11 and R 12 Together with adjacent carbon atoms, it forms a substituted or unsubstituted C3-C 12 Heterocycloalkyl; The R 13 Selected from the group consisting of H and substituted or unsubstituted groups, the substituted or unsubstituted groups are selected from C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C3-C 15 Cycloalkyl, C3-C 15 Heterocycloalkyl, C6-C 15 Aryl, C5-C 15 heteroaryl; or when at least two amino acid residues are present, R 13 It is the bond between two amino acid residues; The R 14 is selected from the group consisting of H and a substituted or unsubstituted group selected from C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl; or R 13 and R 14 Together with the adjacent carbon and nitrogen atoms, they form C3-C 10 Heterocycloalkyl; m is an integer selected from 0 to 10.

4. The drug for removing fat or fat pad according to claim 1, 2 or 3, characterized in that: The L represents absence, substituted or unsubstituted C1-C9 alkyl, or a combination thereof.

5. The drug for removing fat or fat pad according to claim 4, characterized in that: The R4 is H; the X is O, S or absent.

6. The drug for removing fat or fat pad according to claim 5, characterized in that: Said X is O.

7. The drug for removing fat or fat pad according to claim 6, characterized in that: The drug for removing fat or fat pads is a compound as shown below:

8. Use of the drug for removing fat or fat pad according to claim 7 in the preparation of a drug for treating adipose tissue disorders or a drug for regressing adipose tissue tumors.

9. Use of the drug for removing fat or fat pad according to claim 8 in the preparation of a drug for treating local fat distribution disorder.

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