LIVER-TARGETING COMPOUND HAVING THYROID HORMONE RECEPTOR AGONIST CHARACTERISTICS AND PHARMACEUTICAL COMPOSITION THEREOF

Abstract

The present invention belongs to the field of biomedicine, specifically relates to the field of targeting medicine. More specifically, the present invention relates to a liver-targeting compound having thyroid hormone receptor agonist characteristics and a pharmaceutical composition thereof. The compound is a compound represented by formula (1). The compound can be used for treating and/or preventing diseases caused by thyroid hormone dysregulation, and can also effectively reduce lipids in plasma and liver cells.

##STR00001##

Claims

1. A liver-targeting compound with the characteristics of thyroid hormone receptor agonist, which is a compound represented by formula (1): ##STR00014## wherein, n is an integer of 1 to 10; X is carbonyl; Y is NH or an oxygen atom.

2. The liver-targeting compound with the characteristics of thyroid hormone receptor agonist according to claim 1, which is compound GBL-0603: ##STR00015##

3. A method for preparing the liver-targeting compound with the characteristics of thyroid hormone receptor agonist according to claim 1, comprising a step of forming an ester or amide linkage between compound A and compound B: ##STR00016## wherein, n is an integer of 1 to 10; X.sub.1 is —COOH; Y.sub.1 is an amino group or a hydroxyl group.

4-6. (canceled)

7. A pharmaceutical composition for treatment and/or prevention of a disease caused by dysregulation of thyroid hormone, comprising a therapeutically effective amount of the liver-targeting compound with the characteristics of thyroid hormone receptor agonist according to claim 1, and optionally, a pharmaceutically acceptable excipient.

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutically acceptable excipient includes an intestinal absorption enhancer, wherein the intestinal absorption enhancer is sodium salts of medium-chain fatty acids, cholates, cyclodextrin, cationic polymers, anionic polymers or thiolated polymers.

9. The pharmaceutical composition according to claim 8, wherein the sodium salt of medium-chain fatty acid is sodium caprate.

10. The pharmaceutical composition according to claim 7, wherein the dosage form of the pharmaceutical composition is an injection, or an oral rapid or sustained release preparation.

11. A method for treatment and/or prevention of a disease caused by dysregulation of thyroid hormone comprising administering the thyroid hormone receptor agonist according to claim 1 to a mammal in need thereof.

12. The method according to claim 11, wherein the disease is obesity, hyperlipidemia, hypercholesterolemia, diabetes, non-alcoholic fatty liver disease, alcoholic fatty liver disease, atherosclerosis, cardiovascular diseases, hypothyroidism, or thyroid cancer.

13. The method according to claim 11, wherein the disease is non-alcoholic fatty liver disease.

14. The pharmaceutical composition according to claim 9, wherein the weight ratio of the liver-targeting compound with the characteristics of thyroid hormone receptor agonist to sodium caprate is 1:0.2 to 1:0.75.

15. The liver-targeting compound according to claim 1, wherein n is an integer of 1 to 3.

16. The method according to claim 3, wherein n is an integer of 1 to 3.

Description

DESCRIPTION OF THE DRAWINGS

[0029] In order to make the objects, technical solutions and beneficial effects of the present invention clearer, the following description of the drawings are provided:

[0030] FIG. 1 is a high-resolution mass spectrum of compound B;

[0031] FIG. 2 is a high-resolution mass spectrum of compound GBL-0603;

[0032] FIG. 3 is a diagram showing the effect of GBL-0603 on reducing CHO, TG and LDL-C in the serum of db/db obese model mice in Example 2;

[0033] FIG. 4 is a diagram showing the effect of GBL-0603 on reducing CHO and TG in liver tissue cells of db/db obese model mice in Example 2;

[0034] FIG. 5 is illustrative micrographs of liver sections stained with HE in Example 2, showing the effect of GBL-0603 on histopathological change;

[0035] FIG. 6 is a diagram showing the effect of GBL-0603 on the heart weight of db/db obese model mice in Example 2;

[0036] FIG. 7 is a diagram showing the effect of GBL-0603 on the liver weight of db/db obese model mice in Example 2;

[0037] FIG. 8 is a diagram showing the effect of GBL-0603 on CHO, TG and LDL-C in the serum of db/db obese model mice in Example 3;

[0038] FIG. 9 is a diagram showing the effect of GBL-0603 on CHO and TG in the liver of db/db obese model mice in Example 3;

[0039] FIG. 10 is a diagram showing the effect of GBL-0603 on the bone mineral density of normal mice in Example 4;

[0040] FIG. 11 is a diagram showing the effect of GBL-0603 on the bone mineral content of normal mice in Example 4;

[0041] FIG. 12 is a diagram showing the effect of GBL-0603 on the heart weight of normal mice in Example 4;

[0042] FIG. 13 is a diagram showing the effect of GBL-0603 on the liver weight of normal mice in Example 4;

[0043] FIG. 14 is a diagram showing the effect of GBL-0603 on the body weight of normal mice in Example 4:

[0044] FIG. 15 is a diagram showing the effect of GBL-0603 on the liver function of normal mice in Example 4:

[0045] FIG. 16 is a diagram showing the effect of Kylo-0101 on the liver function of normal mice in Example 4;

[0046] FIG. 17 is a diagram showing the effect of GBL-0603 on T3, fT3, T4, fT4 and TSH in the serum of normal mice in Example 4.

BEST MODE FOR CARRYING OUT THE INVENTION

[0047] The following examples illustrate some embodiments of the present disclosure, but the present invention is not limited thereto. In addition, while providing specific embodiments, the inventors anticipated application of these specific embodiments, for example, application of the compounds with specifically same or similar chemical structures in treatment of different liver-derived diseases.

Definitions

[0048] DMF refers to N,N-dimethylformamide;

[0049] HOBt refers to 1-hydroxybenzotriazole:

[0050] DIPEA refers to N,N-diisopropylethylamine;

[0051] Pd/C refers to palladium on activated carbon;

[0052] TBTU refers to O-benzotriazole-N,N,N′,N′-tetramethylurea tetrafluoroborate;

[0053] DCM refers to dichloromethane;

[0054] NBS refers to N-bromosuccinimide;

[0055] n-BuLi refers to n-butyl lithium;

[0056] TIPSCl refers to triisopropylchlorosilane;

[0057] THF refers to tetrahydrofuran;

[0058] MTBE refers to methyl tert-butyl ether;

[0059] TBAF refers to tetrabutylammonium fluoride.

[0060] Unless specified otherwise, the ratio of two substances involved in the present invention refers to the volume ratio.

[0061] Unless specified otherwise, the content involved in the present invention refers to volume percentage concentration.

Example 1: Preparation of Compound GBL-0603

1. Synthesis of Compound A

1.1 Synthesis of Compound A-c1

[0062] ##STR00005##

[0063] DMF (8 mL), cbz-6-aminocaproic acid (24 mg), HOBt (21.6 mg), dlSANC-c12 (84 mg) and DIPEA (53.5 mg) were added to a reaction flask in sequence. After the addition was completed, the reaction was stirred at room temperature overnight. TLC detection showed the reaction was qualified to be stopped and post-treated. The reaction was quenched with water and stood for phase separation. The aqueous phase was extracted with DCM three times, each of 20 mL. The combined organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatograph to obtain 72.8 mg of a white solid.

1.2 Synthesis of Compound A

[0064] ##STR00006##

[0065] Compound A-c1 (72.8 mg), methanol (15 mL) and Pd/C (3.4 mg) were added to a reaction flask in sequence, and vacuum/hydrogen replacement was performed. After the replacement. H.sub.2 was introduced, and the reaction was stirred for 1.0 h at 40° C. TLC showed the reaction is qualified. The reaction is stopped and filtered to remove Pd/C. The filtrate was concentrated to obtain 47 mg of a white solid.

2. Synthesis of Compound B

2.1 Synthesis of Compound B-c1

[0066] ##STR00007##

[0067] 2-isopropylphenol (30.3 mg) was weighed and dissolved in acetonitrile (10 mL), added with NBS (34.2 mg), and reacted at 35° C. for 6 h. The reaction solution was concentrated, dissolved again in petroleum ether (50 mL) and filtered to remove insolubles. The filtrate was washed with water (40 mL) and pyridine (40 mL) respectively, dried over anhydrous sodium sulfate, filtered, and rotary evaporated. The resulting residue (50.5 mg) was dissolved in acetonitrile (40 mL), added with anhydrous potassium carbonate (32.0 mg) and benzyl bromide (37.1 mg), and reacted at 40° C. for 5 h. The reaction mixture was cooled, filtered and purified by column chromatograph to give compound B-c1 (63.1 mg).

2.2 Synthesis of Compound B-c2

[0068] ##STR00008##

[0069] Compound B-c1 (63.1 mg) was weighed and dissolved in anhydrous THF (10 mL), cooled in an ice bath, added dropwise with a solution of 1.0 mol/L n-BuLi in n-hexane (5.0 mL), reacted for 3 h, added dropwise with DMF (1.5 mL), and reacted for another 3 h. The reaction was quenched with saturated ammonium chloride (5 mL), and extracted with ethyl acetate (10 mL). The organic phase was washed with pyridine (10 mL), dried over anhydrous sodium sulfate, filtered, and purified by column chromatograph to give compound B-c2 (33.4 mg).

2.3 Synthesis of Compound B-c3

[0070] ##STR00009##

[0071] 4-bromo-3,5-dimethylphenol (30.8 mg) was weighed and dissolved in dichloromethane (50 mL), added with imidazole (18 mg), cooled, added dropwise with TIPSCl (25.5 mg) and reacted for 5 h. The reaction solution was diluted with dichloromethane (50 mL). The organic phase was washed with water (50 mL) and pyridine (50 mL) respectively, dried over anhydrous sodium sulfate, rotary evaporated, and purified by column chromatograph to give compound B-c3 (33.4 mg).

2.4 Synthesis of Compound B-c4

[0072] ##STR00010##

[0073] Compound B-c3 (36.1 mg) was weighed and dissolved in THF (60 mL), cooled in an ice bath, added dropwise with a solution of 1.0 mol/L n-BuLi in n-hexane (4 mL), reacted for 3 h. added dropwise with a solution of compound B-c2 (33.4 mg) in THF (5 mL), and reacted for another 3 h. The reaction was quenched with saturated ammonium chloride (20 mL), and extracted with ethyl acetate (20 mL). The organic phase was washed with pyridine (30 mL), dried over anhydrous sodium sulfate, filtered, and purified by column chromatograph to give compound B-c4 (54.3 mg).

2.5 Synthesis of Compound B

[0074] ##STR00011##

[0075] Compound B-c4 (54.3 mg) was weighed and dissolved in THF (50 mL), and added dropwise with a 1 mol/L TBAF solution (3 mL). TLC showed that the reaction is complete. The reaction was added with ethyl acetate (50 mL), washed with water (20 mL) and pyridine (20 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give a white solid (23.3 mg). The white solid was dissolved in DMF (5 mL), cooled, added with cesium carbonate (40.4 mg) and then with benzyl bromoacetate (15.9 mg), and reacted at 40° C. for 4 h. The reaction solution was diluted with MTBE (10 mL), filtered, and added with water (20 mL). The aqueous phase was extracted with MTBE (20 mL*2), and the combined organic phase was washed with pyridine (20 mL), dried over anhydrous sodium sulfate, filtered, rotary evaporated and purified by column chromatograph. The product was dissolved in acetic acid (5 mL), added with 10% Pd/C (0.2 g) as the catalyst, hydrogenated at room temperature overnight, filtered, rotary evaporated, and purified by column chromatograph to give compound B as a light yellow solid (15 mg). The high-resolution mass spectrum of compound B is shown in FIG. 1.

3. Synthesis of GBL-0603

3.1 Synthesis of GBL-0603-c1

[0076] ##STR00012##

[0077] DMF (3.0 mL), compound B (15 mg), TBTU (8.47 mg) and DIPEA (20.2 mg) were added to a reaction flask in sequence and reacted for 6 h. Then compound A (47 mg) was quickly added, and the mixture was stirred at room temperature for 2 h. HPLC detected that the reaction was qualified, and the reaction was terminated.

3.2 Synthesis of GBL-0603

[0078] ##STR00013##

[0079] After the GBL-0603-c1 reaction solution was tested to be qualified by HPLC in-process control, the pH value of the reaction solution was adjusted to 8-10 with a 1.0 mol/L ammonia solution under ice bath. After the pH value of the reaction solution was qualified, the ice bath was removed, and the reaction solution was stirred at room temperature for half an hour. HPLC in-process control analysis showed that the reaction was qualified. The pH value of the reaction solution was adjusted to 7.0 with glacial acetic acid. After the pH value of the reaction solution was qualified, it was concentrated to remove DMF in the reaction solution. The concentrated residue was dissolved in 35% acetonitrile/water, filtered, and lyophilized to give 29.47 mg of a lyophilized product. The high-resolution mass spectrum of GBL-0603 is shown in FIG. 2.

Example 2: Study on the Dose-Efficacy of GBL-0603 on Lipid Metabolism in db Mice

[0080] Laboratory Animals and Breeding:

[0081] 30 genetically obese model 7-week-old male mice (BKS-db) were selected. Mice need to adapt to the environment for a week before the experiment. Healthy mice were selected as test animals and reared in IVC cages at a density of 5 mice/cage, and the litter was changed twice a week. Requirements for laboratory animal room: room temperature 22-24° C., relative humidity 40 to 70%, automatic lighting. 12 h light-dark cycle (lights on at 08:00, lights off at 20:00). The laboratory animal room standards meet the national standard GB14925-2010 of the People's Republic of China.

[0082] Drug Preparation:

[0083] 210 mg of GBL-0603 was accurately weighed and dissolved in 35 mL of a solvent to prepare a 6 mg/mL stock solution. Each of the dose groups was administered with the same volume by diluting the stock solution by 3, 10, 30 and 66.7 times respectively. The GBL-0603 stock solution was prepared every three days. After preparation, the stock solution was stored at 4° C. for later use.

[0084] Grouping and Dosing Schedule:

TABLE-US-00001 Group Experimental Mice type Quantity No. drugs Dosage Treatment manner db/db control 6 G1 Blank solvent — administered by gavage group (Vehicle) every day for two db/db 6 G2 GBL-0603 10 mg/kg consecutive weeks, administration 6 G3 3 mg/kg at 5 mL/kg group 6 G4 1 mg/kg 6 G5 0.45 mg/kg

[0085] Experimental Operation:

[0086] Before starting the experiment, blood was collected to test the total cholesterol (CHO) of the mice in each group, and the mice were weighed and randomly grouped according to body weight. The mice were weighed daily during the administration. After the last administration, the mice in each group were fasted for 6 h, and euthanized. Blood was collected from the heart, and the serum was separated to detect the levels of triglycerides (TG), total cholesterol (CHO), low-density lipoprotein cholesterol (LDL-C), ALT and AST in the serum. After blood collection, the liver was weighed, and a part of the middle lobe of the liver of the mice in each group was quick-frozen in liquid nitrogen and stored at −80° C. for later use. In addition, the middle lobe of the liver of the mice was fixed and embedded in paraffin. The heart was taken and weighed. The contents of CHO and TG in liver tissue were measured. Liver histopathological examination was performed by sectioning all mice and stained with HE, and comparing the steatosis, inflammation and ballooning of hepatocytes before and after treatment.

Example 3: Study on the Effect of Adding Sodium Caprate as an Accelerator

[0087] Animal breeding was the same as in Example 2. The grouping and dosing schedule are as follows:

TABLE-US-00002 Group Experimental Mice type Quantity No. drugs Dosage Treatment manner db/db mice 6 G1 Blank solvent — gavage (Vehicle) 6 G2 Drug A 10 mg/kg 6 G3 Drug B 10 mg/kg 5 G4 Drug C 10 mg/kg Remarks: Sodium caprate was not added in the prescription of Drug A, and was added at 20% and 75% by weight of the main drug in the prescriptions of Drugs B and C, respectively.

[0088] Experimental Operation:

[0089] For G2/G3/G4, the volume of the solvent was calculated according to the weight of the drug, and then the solvent was added and vortexed repeatedly until complete dissolution for use. After the preparation, the administration was finished within one hour.

[0090] Before starting the experiment, the mice were weighed and randomly grouped according to body weight. After the last administration, the mice in each group were fasted for 6 h, and euthanized. Blood was collected from the heart, and the serum was separated to detect the levels of triglycerides (TG), total cholesterol (CHO) and low-density lipoprotein cholesterol (LDL-C) in the serum, as well as the contents of CHO and TG in the liver tissue.

Example 4: Study on the Effects of GBL-0603 and Kylo-0101 on Thyroid and Liver Enzymes in Normal Mice

[0091] 66 C57BL/6J mice (half male and half female). The grouping and dosing schedule are shown in the table below:

TABLE-US-00003 Group Experimental Mice type Quantity No. drugs Dosage Treatment manner Blank control 6 1 Blank solvent — administered by gavage every (Vehicle) day for two consecutive weeks administration 6 2 GBL-0603 30 mg/kg group 6 3 10 mg/kg 6 4 3 mg/kg 6 5 1 mg/kg 6 6 0.3 mg/kg Blank control 6 7 Blank solvent — administered by subcutaneous (Control) injection every day for two administration 6 8 Kylo-0101 1 μg/kg consecutive weeks group 6 9 3 μg/kg 6 10 10 μg/kg 6 11 30 μg/kg

[0092] Experimental Operation:

[0093] Before starting the experiment, the mice were weighed and randomly grouped according to body weight. The mice were weighed weekly during the administration. After the last administration, the mice in each group were fasted for 6 h, and euthanized. Blood was collected from the heart, and the serum was separated to detect the contents of T3, fT3, T4, fT4, and TSH in the serum: bone mineral density, body weight, liver and heart weight; liver enzymes ALT, AST and GGT, in the blank group and the GBL-0603 administration group, as well as the liver enzymes ALT. AST and GGT in the blank group and Kylo-0101 administration group.