DEOXYCHOLIC ACID COMPOUNDS, PHARMACEUTICAL COMPOSITIONS AND USES THEREOF

Abstract

Disclosed are a deoxycholic acid compound and a pharmaceutical composition thereof, and use thereof in the preparation of drugs for treating liver diseases. The structure of the deoxycholic acid compound is as shown by Formula I, or pharmaceutically acceptable salts thereof. The compounds have significantly liver-targeting characteristics, and the compounds reduce the drug concentration in the circulatory system while improving the efficacy, thereby reducing the toxic side effects.

##STR00001##

Claims

1. A compound of Formula I: ##STR00031## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 and R.sup.2 each are independently selected from the group consisting of: H, —COR′, —CONHR′, —CONR′R″, or —COOR′; R.sup.3 is selected from the group consisting of H, methyl, or ethyl; R.sup.4 is selected from the group consisting of: H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heteroaryl or substituted heteroaryl, and heterocyclyl or substituted heterocyclyl; Linker is absent, or is selected from the group consisting of: alkylene or substituted alkylene, alkenylene or substituted alkenylene, alkynylene or substituted alkynylene, cycloalkylene or substituted cycloalkylene, arylene or substituted arylene, heteroarylene or substituted heteroarylene, and heterocyclylene or substituted heterocyclylene; R.sup.5 and R.sup.6 each are independently selected from the group consisting of: H, and alkyl or substituted alkyl; or R.sup.5 and R.sup.6 are linked to each other, and form a 3-7 membered ring with 0, 1, or 2 heteroatoms selected from the group consisting of O, S, or N, together with C atoms linked thereto; R is selected from the group consisting of: alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, alkylene-aryl, alkylene-substituted aryl, heteroaryl or substituted heteroaryl, and heterocyclyl or substituted heterocyclyl; or R is selected from the group consisting of: inorganic cations selected from the group consisting of sodium ions, potassium ions, magnesium ions, or calcium ions; or organic cations selected from the group consisting of ammonium, tetramethylammonium, tetraethylammonium, tetrapropylammonium, or tetrabutylammonium; R′ and R″ each are independently selected from the group consisting of: H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heteroaryl or substituted heteroaryl, and heterocyclyl or substituted heterocyclyl; and substituents of the substituted alkyl, the substituted alkylene, the substituted alkenyl, the substituted alkenylene, the substituted alkynyl, the substituted alkynylene, the substituted cycloalkyl, the substituted cycloalkylene, the substituted aryl, the substituted arylene, the alkylene-substituted aryl, the substituted heteroaryl, the substituted heteroarylene, the substituted heterocyclyl, or the substituted heterocyclylene each are independently selected from the group consisting of: halogen, cyano, amino, nitro, hydroxy, alkyl, alkoxyl, and modified alkyl.

2. The compound of Formula I or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of Formula I is not a compound: ##STR00032## optionally, R in the compound of Formula I is not methyl.

3. The compound of Formula I or the pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that: optionally, the “halogen” is selected from the group consisting of: F, Cl, Br, or I; optionally, “alkyl” in the “alkyl” and “alkoxyl” is C.sub.1-C.sub.20 linear or branched alkyl, optionally selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, and n-pentyl; optionally, “alkylene” in the “alkylene”, “alkylene-aryl”, and “alkylene-substituted aryl” is C.sub.1-C.sub.10 linear or branched alkylene, optionally selected from the group consisting of: methylene, ethylene, n-propylidene, isopropylidene, n-butylidene, isobutylidene, tert-butylidene, sec-butylidene, and n-pentylene; optionally, the “modified alkyl” is a group obtained by substituting one or more groups selected from the group consisting of —O—, —CO—, —NH.sub.2, —OH, halogen, —CN, and —NO.sub.2 for any carbon atom in the alkyl; optionally, the “alkenyl” is C.sub.2-C.sub.6 alkenyl; optionally, the “alkenylene” is C.sub.2-C.sub.6 alkenylene; optionally, the “alkynyl” is C.sub.2-C.sub.6 alkynyl; optionally, the “alkynylene” is C.sub.2-C.sub.6 alkynylene; optionally, the “cycloalkyl” is C.sub.3-C.sub.10 monocyclic or bicyclic cycloalkyl; optionally, the “cycloalkylene” is C.sub.3-C.sub.10 monocyclic or bicyclic cycloalkylene; optionally, “aryl” in the “aryl”, the “alkylene-aryl”, and the “alkylene-substituted aryl” is 6-10 membered aryl; and is optionally phenyl or naphthyl; optionally, the “arylene” is 6-10 membered arylene; and may be optionally phenylene or naphthylene; optionally, the “heteroaryl” is a 5-10 membered heteroaromatic ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S; optionally, the “heteroarylene” is a 5-10 membered heteroarylene ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S; optionally, the “heterocyclyl” is a 3-10 membered nonaromatic heterocyclic ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S; and optionally, the “heterocyclylene” is a 3-10 membered nonaromatic heterocyclylene ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S.

4. The compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: optionally, R′ and R″ each are independently selected from the group consisting of: H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, benzyl, sec-butyl, n-pentyl, ##STR00033## optionally, R.sup.1 and R.sup.2 each are independently selected from the group consisting of H, acetyl, propionyl, isopropionyl, butyryl, and isobutyryl; optionally, R.sup.3 is selected from the group consisting of: H, methyl, or ethyl; optionally, R.sup.4 is selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, and n-pentyl; optionally, Linker is absent, or is selected from the group consisting of: —CH.sub.2—, —CH.sub.2CH.sub.2—, —CH.sub.2CH.sub.2CH.sub.2—, cyclopropylidene, ##STR00034## optionally, R.sup.5 and R.sup.6 each are independently selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, or R.sup.5 and R.sup.6 are linked to each other, and form, together with the C atoms linked thereto, any group selected from the following formulas: ##STR00035##  and optionally, R is selected from the group consisting of: ethyl, propyl, butyl, isopropyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, ##STR00036##  heterocyclyl or substituted heterocyclyl, sodium ions, potassium ions, magnesium ions, calcium ions, ammonium, tetramethylammonium, tetraethylammonium, tetrapropylammonium, or tetrabutylammonium; and Xs each are independently selected from the group consisting of: 0, 1, 2, or 3 groups selected from the group consisting of: F, Cl, Br, I, —NH.sub.2, —OH, -methoxyl, ethoxyl, propoxyl, isopropoxyl, —CN, methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl, or Xs each are independently a group obtained by substituting one or more groups selected from the group consisting of —O—, —CO—, and —NH.sub.2 for any carbon atom on C.sub.5-C.sub.10 linear or branched alkyl.

5. The compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that the compound of Formula I is selected from a compound of the following Formula II: ##STR00037## wherein: R.sup.3 and R are as defined in any one of claims 1 to 3.

6. The compound of Formula I or the pharmaceutically acceptable salt thereof according to claim 5, characterized in that R.sup.3 is selected from the group consisting of H, methyl, or ethyl; R is selected from the group consisting of ethyl, n-propyl, isopropyl, n-butyl, isobutyl, ##STR00038##  sodium ions, potassium ions, calcium ions, and Xs each are independently selected from the group consisting of: 0, 1, 2, or 3 groups selected from the group consisting of: F, Cl, Br, I, —NH.sub.2, —OH, -methoxyl, ethoxyl, propoxyl, isopropoxyl, —CN, methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl, or Xs each are independently a group obtained by substituting one or more groups selected from the group consisting of —O—, —CO—, and —NH.sub.2 for any carbon atom on C.sub.5-C.sub.10 linear or branched alkyl.

7. The compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, characterized in being selected from the following compounds: ##STR00039## ##STR00040## in Formula I-7, X is 1, 2, or 3 substituents, each independently selected from the group consisting of F, Cl, Br, I, —CN, —NH.sub.2, —NO.sub.2, or —OH, methyl, ethyl, propyl, isopropyl, methoxyl, ethoxyl, propoxyl, and isopropoxyl; and ##STR00041## in Formula I-8, X is 1, 2, or 3 substituents, each independently selected from the group consisting of F, Cl, Br, I, —CN, —NH.sub.2, —NO.sub.2, or —OH, methyl, ethyl, propyl, isopropyl, methoxyl, ethoxyl, propoxyl, and isopropoxyl.

8. The compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, characterized in that, the pharmaceutically acceptable salt includes a salt formed of a compound of Formula I and an acid; optionally, the acid includes an inorganic acid and an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid; and optionally, the organic acid includes formic acid, ascorbic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, gluconic acid, hydrogen tartaric acid, glucuronic acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzoic acid, benzenesulfonic acid, p-bromobenzenesulfonic acid, glutamic acid, salicylic acid, and pamoic acid.

9. A pharmaceutical composition, characterized in comprising a therapeutically effective amount of a compound of the following Formula I: ##STR00042## or one or more of pharmaceutically acceptable salts thereof, and optionally a pharmaceutically acceptable carrier, in Formula I, R.sup.1 and R.sup.2 each are independently selected from the group consisting of: H, —COR′, —CONHR′, —CONR′R″, or —COOR′; R.sup.3 is selected from the group consisting of: H, methyl, or ethyl; R.sup.4 is selected from the group consisting of: H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heteroaryl or substituted heteroaryl, and heterocyclyl or substituted heterocyclyl; Linker is absent, or is selected from the group consisting of: alkylene or substituted alkylene, alkenylene or substituted alkenylene, alkynylene or substituted alkynylene, cycloalkylene or substituted cycloalkylene, arylene or substituted arylene, heteroarylene or substituted heteroarylene, and heterocyclylene or substituted heterocyclylene; R.sup.5 and R.sup.6 each are independently selected from the group consisting of: H, and alkyl or substituted alkyl; or R.sup.5 and R.sup.6 are linked to each other, and form a 3-7 membered ring with 0, 1, or 2 heteroatoms selected from the group consisting of O, S, or N, together with C atoms linked thereto; R is selected from the group consisting of: alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, alkylene-aryl, alkylene-substituted aryl, heteroaryl or substituted heteroaryl, and heterocyclyl or substituted heterocyclyl; or R is selected from the group consisting of: inorganic cations selected from the group consisting of sodium ions, potassium ions, magnesium ions, or calcium ions; or organic cations selected from the group consisting of ammonium, tetramethylammonium, tetraethylammonium, tetrapropylammonium, or tetrabutylammonium; R′ and R″ each are independently selected from the group consisting of: H, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heteroaryl or substituted heteroaryl, and heterocyclyl or substituted heterocyclyl; and substituents of the substituted alkyl, the substituted alkylene, the substituted alkenyl, the substituted alkenylene, the substituted alkynyl, the substituted alkynylene, the substituted cycloalkyl, the substituted cycloalkylene, the substituted aryl, the substituted arylene, the alkylene-substituted aryl, the substituted heteroaryl, the substituted heteroarylene, the substituted heterocyclyl, or the substituted heterocyclylene each are independently selected from the group consisting of: halogen, cyano, amino, nitro, hydroxy, alkyl, alkoxyl, and modified alkyl.

10. The pharmaceutical composition according to claim 9, characterized in that: optionally, the “halogen” is selected from the group consisting of: F, Cl, Br, or I; optionally, “alkyl” in the “alkyl” and “alkoxyl” is C.sub.1-C.sub.20 linear or branched alkyl, optionally selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, and n-pentyl; optionally, “alkylene” in the “alkylene”, “alkylene-aryl”, and “alkylene-substituted aryl” is C.sub.1-C.sub.10 linear or branched alkylene, optionally selected from the group consisting of: methylene, ethylene, n-propylidene, isopropylidene, n-butylidene, isobutylidene, tert-butylidene, sec-butylidene, and n-pentylene; optionally, the “modified alkyl” is a group obtained by substituting one or more groups selected from the group consisting of —O—, —CO—, —NH.sub.2, —OH, halogen, —CN, and —NO.sub.2 for any carbon atom in the alkyl; optionally, the “alkenyl” is C.sub.2-C.sub.6 alkenyl; optionally, the “alkenylene” is C.sub.2-C.sub.6 alkenylene; optionally, the “alkynyl” is C.sub.2-C.sub.6 alkynyl; optionally, the “alkynylene” is C.sub.2-C.sub.6 alkynylene; optionally, the “cycloalkyl” is C.sub.3-C.sub.10 monocyclic or bicyclic cycloalkyl; optionally, the “cycloalkylene” is C.sub.3-C.sub.10 monocyclic or bicyclic cycloalkylene; optionally, “aryl” in the “aryl”, the “alkylene-aryl”, and the “alkylene-substituted aryl” is 6-10 membered aryl; and is optionally phenyl or naphthyl; optionally, the “arylene” is 6-10 membered arylene; and may be optionally phenylene or naphthylene; optionally, the “heteroaryl” is a 5-10 membered heteroaromatic ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S; optionally, the “heteroarylene” is a 5-10 membered heteroarylene ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S; optionally, the “heterocyclyl” is a 3-10 membered nonaromatic heterocyclic ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S; and optionally, the “heterocyclylene” is a 3-10 membered nonaromatic heterocyclylene ring containing 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S.

11. The pharmaceutical composition according to claim 9 or 10, characterized in that: optionally, R′ and R″ each are independently selected from the group consisting of: H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, benzyl, sec-butyl, n-pentyl, ##STR00043## optionally, R.sup.1 and R.sup.2 each are independently selected from the group consisting of: H, acetyl, propionyl, isopropionyl, butyryl, and isobutyryl; optionally, R.sup.3 is selected from the group consisting of H, methyl, or ethyl; optionally, R.sup.4 is selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, and n-pentyl; optionally, Linker is absent, or is selected from the group consisting of: —CH.sub.2—, —CH.sub.2CH.sub.2—, —CH.sub.2CH.sub.2CH.sub.2—, cyclopropylidene, ##STR00044## optionally, R.sup.5 and R.sup.6 each are independently selected from the group consisting of: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, or R.sup.5 and R.sup.6 are linked to each other, and form, together with the C atoms linked thereto, any group selected from the following formulas: ##STR00045##  and optionally, R is selected from the group consisting of: ethyl, propyl, butyl, isopropyl, isobutyl, tert-butyl, sec-butyl, n-pentyl ##STR00046##  heterocyclyl or substituted heterocyclyl, sodium ions, potassium ions, magnesium ions, calcium ions, ammonium, tetramethylammonium, tetraethylammonium, tetrapropylammonium, or tetrabutylammonium; and Xs each are independently selected from the group consisting of: 0, 1, 2, or 3 groups selected from the group consisting of F, Cl, Br, I, —NH.sub.2, —OH, -methoxyl, ethoxyl, propoxyl, isopropoxyl, —CN, methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl, or Xs each are independently a group obtained by substituting one or more groups selected from the group consisting of —O—, —CO—, and —NH.sub.2 for any carbon atom on C.sub.5-C.sub.10 linear or branched alkyl.

12. The pharmaceutical composition according to any one of claims 9 to 11, characterized in that the compound of Formula I is selected from a compound of the following Formula II: ##STR00047## wherein: R.sup.3 and R are as defined in any one of claims 9 to 11.

13. The pharmaceutical composition according to claim 12, characterized in that R.sup.3 is selected from the group consisting of: H, methyl, or ethyl; R is selected from the group consisting of: ethyl, n-propyl, isopropyl, n-butyl, isobutyl, ##STR00048##  sodium ions, potassium ions, calcium ions, and Xs each are independently selected from the group consisting of 0, 1, 2, or 3 groups selected from the group consisting of F, Cl, Br, I, —NH.sub.2, —OH, -methoxyl, ethoxyl, propoxyl, isopropoxyl, —CN, methyl, ethyl, n-propyl, isopropyl, n-butyl, or isobutyl, or Xs each are independently a group obtained by substituting one or more groups selected from the group consisting of —O—, —CO—, and —NH.sub.2 for any carbon atom on C.sub.5-C.sub.10 linear or branched alkyl.

14. The pharmaceutical composition according to any one of claims 9 to 13, characterized in that the compound of the Formula I is selected from the following compounds: ##STR00049## ##STR00050## in Formula I-7, X is 1, 2, or 3 substituents, each independently selected from the group consisting of F, Cl, Br, I, —CN, —NH.sub.2, —NO.sub.2, or —OH, methyl, ethyl, propyl, isopropyl, methoxyl, ethoxyl, propoxyl, and isopropoxyl; and ##STR00051## in Formula I-8, X is 1, 2, or 3 substituents, each independently selected from the group consisting of F, Cl, Br, I, —CN, —NH.sub.2, —NO.sub.2, or —OH, methyl, ethyl, propyl, isopropyl, methoxyl, ethoxyl, propoxyl, and isopropoxyl.

15. The pharmaceutical composition according to any one of claims 9 to 14, characterized in that: the pharmaceutically acceptable salt of the compound of Formula I includes a salt formed of the compound of Formula I and an acid; optionally, the acid includes an inorganic acid and an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid; and optionally, the organic acid includes formic acid, ascorbic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, gluconic acid, hydrogen tartaric acid, glucuronic acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzoic acid, benzenesulfonic acid, p-bromobenzenesulfonic acid, glutamic acid, salicylic acid, and pamoic acid.

16. The pharmaceutical composition according to any one of claims 9 to 15, characterized in that: the dosage form of the pharmaceutical composition includes an oral preparation and an injection preparation; optionally, the oral preparation includes a solid preparation and a liquid preparation; optionally, the solid preparation includes tablets, powders, granules, and capsules; and optionally, the liquid preparation includes water or oil suspensions, and syrups.

17. Use of the compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the pharmaceutical composition according to any one of claims 9 to 16 in the preparation of a medicament for treating nonalcoholic steatohepatitis.

18. Use of the compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the pharmaceutical composition according to any one of claims 9 to 16 in the preparation of a medicament for treating primary biliary cirrhosis.

19. Use of the compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the pharmaceutical composition according to any one of claims 9 to 16 in the preparation of a medicament for treating a biliation-associated disease; optionally, the biliation-associated disease includes portal hypertension, bile acid diarrhea, alcoholic hepatitis, and primary sclerotic cholangitis.

20. A method for treating nonalcoholic steatohepatitis, characterized in administering, to a patient, a therapeutically effective amount of the compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the pharmaceutical composition according to any one of claims 9 to 16.

21. A method for treating primary biliary cirrhosis, characterized in administering, to a patient, a therapeutically effective amount of the compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the pharmaceutical composition according to any one of claims 9 to 16.

22. A method for treating a biliation-associated disease, characterized in administering, to a patient, a therapeutically effective amount of the compound of Formula I or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the pharmaceutical composition according to any one of claims 9 to 16; optionally, the biliation-associated disease includes portal hypertension, bile acid diarrhea, alcoholic hepatitis, and primary sclerotic cholangitis.

Description

DETAILED DESCRIPTION

[0104] The specific embodiments of the present disclosure will be elaborated below. It should be appreciated that the specific embodiments described herein are merely intended to exemplarily illustrate, and not limit the present disclosure.

Example 1: Preparation of Compounds

[0105] 1. Preparation of Compound 4:

##STR00025##

[0106] A. Raw Material 1 (0.8 g, 1.90 mmol, 1.0 eq), glycine methyl ester hydrochloride (2,356 mg, 2.85 mmol), EDCI (546 mg, 2.85 mmol), and DMAP (696 mg, 5.70 mmol) were dissolved in DCM (10 mL), and reacted overnight at room temperature under nitrogen protection. The reaction was detected by TLC to be completed. The reactant was rotary-evaporated to remove DCM, and EtOAC was added for dissolution. The solution was washed with 1N HCl 5 mL x 2 to remove excess DMAP. The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound 3 (900 mg, 96%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.94 (s, 1H), 4.0 (d, J=5.1 Hz, 2H), 3.76 (s, 3H), 3.70 (s, 1H), 3.44-3.36 (m, 1H), 2.34-2.27 (m, 1H), 2.18-2.10 (m, 1H), 1.97-0.89 (m, 36H), 0.65 (s, 3H).

##STR00026##

[0107] B. Compound 3 (900 mg, 1.83 mmol), and NaOH (111 mg, 2.78 mmol) were added to CH.sub.3OH—H.sub.2O (4 mL, V:V=1:1), and subjected to a reflux reaction for 3 h. The reaction was detected by TLC to be completed. The reactant was rotary-evaporated to remove methanol, and extracted by adding EtOAC to remove organic impurities. The aqueous phase was adjusted with 1N HCl till the pH was about 5. A white solid was precipitated, and dissolved and extracted by adding EtOAC. The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound 4 (795 mg, 91%). .sup.1H NMR (400 MHz, DMSO) δ 12.4 (s, 1H), 8.1 (t, J=5.9 Hz, 1H), 4.3 (s, 1H), 4.05 (d, J=4.9 Hz, 1H), 3.7 (d, J=5.9 Hz, 2H), 3.5 (s, 1H), 3.16-3.10 (m, 1H), 2.18-2.11 (m, 1H), 2.07-2.01 (m, 1H), 1.97-0.89 (m, 34H), 0.61 (s, 3H); m/z(ESI) [M-H].sup.−=476.4.

[0108] 2. Preparation of Compound I-4:

##STR00027##

[0109] Raw Material 4 (1.3 g, 2.72 mmol), isopropanol (245 mg, 4.08 mmol), EDCI (782 mg, 4.08 mmol), and DMAP (498 mg, 4.08 mmol) were dissolved in DCM (10 mL), and reacted overnight at room temperature under nitrogen protection. The reaction was detected by TLC to be completed. The reactant was rotary-evaporated to remove DCM, and EtOAC was added for dissolution. The solution was washed with 1N HCl 10 mL x 2 to remove excess DMAP. The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (DCM:MeOH=20:1) to give Compound I-4 (653 mg, 46%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 5.94 (s, 1H), 5.10-5.04 (m, 1H), 4.0 (d, J=5.0 Hz, 2H), 3.7 (s, 1H), 3.44-3.36 (m, 1H), 2.34-2.27 (m, 1H), 2.17-2.07 (m, 1H), 1.97-0.89 (m, 42H), 0.65 (s, 3H).

[0110] 3. Preparation of Compound I-5:

##STR00028##

[0111] Raw Material 4 (2.27 g, 4.75 mmol), phenylmethanol (770 mg, 7.12 mmol), EDCI (1.36 g, 7.12 mmol), and DMAP (870 mg, 7.12 mmol) were dissolved in DCM (30 mL), and reacted overnight at room temperature under nitrogen protection. The reaction was detected by TLC to be completed. The reactant was rotary-evaporated to remove DCM, and EtOAC was added for dissolution. The solution was washed with 1N HCl 10 mL x 2 to remove excess DMAP. The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (DCM:MeOH=20:1) to give Compound I-5 (1.72 g, 64%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.39-7.32 (m, 5H), 5.94 (t, J=4.8 Hz, 1H), 5.18 (s, 2H), 4.08 (d, J=5.2 Hz, 2H), 3.7 (s, 1H), 3.44-3.36 (m, 1H), 2.34-2.27 (m, 1H), 2.17-2.07 (m, 1H), 1.97-0.89 (m, 36H), 0.65 (s, 3H); m/z(ESI) [M+Na].sup.+=590.5.

[0112] 4. Preparation of Compound I-6:

##STR00029##

[0113] A. Raw Material 4 (2.27 g, 4.75 mmol), Compound 5 (2.20 g, 7.12 mmol), EDCI (1.36 g, 7.12 mmol), and DMAP (870 mg, 7.12 mmol) were dissolved in DCM (30 mL), and reacted overnight at room temperature under nitrogen protection. The reaction was detected by LC-MS to be completed. The reaction was quenched by adding water. The reactant was extracted. The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (DCM:MeOH=20:1) to give Compound 6 (750 mg, 21%). .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.15 (d, J=8.5 Hz, 2H), 7.03 (d, J=8.5 Hz, 2H), 5.98 (t, J=5.2 Hz, 1H), 5.0 (d, J=8.0 Hz, 1H), 4.54 (dd, J=6.3 Hz, J=13.8 Hz, 1H), 4.28 (d, J=5.2 Hz, 2H), 4.15 (t, J=7.2 Hz, 2H), 3.70 (s, 1H), 3.40 (s, 1H), 3.13-3.02 (m, 2H), 2.37-2.29 (m, 1H), 2.21-2.13 (m, 1H), 1.97-0.89 (m, 48H), 0.65 (s, 3H).

##STR00030##

[0114] B. Compound 6 (750 mg, 0.98 mmol) was dissolved in DCM (10 mL), into which HCl gas was charged, and the solution was changed from clear to turbid. The reaction was detected by TLC to be completed. The reactant was rotary-evaporated to remove the solvent, and extracted twice with ether, and the reactant was turned into a foamed solid, thereby giving Compound I-6 (550 mg, 84%). .sup.1H NMR (400 MHz, DMSO) δ 8.67 (d, J=4.2 Hz, 3H), 8.45 (t, J=5.7 Hz, 1H), 7.30 (d, J=8.5 Hz, 2H), 7.07 (d, J=8.5 Hz, 2H), 4.24 (dd, J=6.8 Hz, J=12.4 Hz, 1H), 4.14-4.08 (m, 4H), 3.49 (s, 1H), 3.24-3.03 (m, 3H), 2.22-2.16 (m, 1H), 2.12-2.05 (m, 1H), 1.97-0.89 (m, 39H), 0.59 (s, 3H); m/z(ESI) [M-HCl+H].sup.+=669.5.

[0115] 5. Preparation of Compound I-1:

[0116] Compound 3 was hydrolyzed with NaOH, or Compound 4 was treated with an aqueous NaOH solution to give compound I-1.

Test Example 1: Pharmacokinetic Test

[0117] 1. Formulation of preparation and dosing: An appropriate amount of the test sample was precisely weighed, and mixed with an appropriate volume of a solvent to obtain a clear solution or a homogeneous suspension. The preparation was administered to animals within 4 hours after formulation. The dosage formulation would be administered by oral gavage in accordance with facility standard operating procedures. The dosage volume would be determined by the body weights of the animals collected on the morning of administering.

[0118] 2. Blood collection: Blood (approx. 0.2 ml per point in time) would be acupunctured each time from the jugular vein of every animal into a polypropylene tube. All blood samples would be transferred to pre-cooled commercial EDTA-K2 test tubes, and kept on wet ice until they were centrifuged.

[0119] 3. Plasma processing: Blood samples would be centrifuged (3200 rpm, 10 min) at approximately 4° C. Plasma was collected separately and transferred to pre-labeled PP tubes in wet ice at each point in time, and then immediately precipitated by ACN (6 IS) (the ratio of plasma: ACN was 1:4). Centrifugation (10 min, 12,000 rpm) was carried out again to obtain a supernatant. The supernatant was quickly frozen on dry ice and kept at −70±10° C. before LC/MS/MS analysis.

[0120] 4. Liver processing: Liver tissues were collected at each point in time, washed twice with pre-cooled deionized water, and dried with filter paper. Liver tissues were immediately homogenized with 10-times volume of a methanol-water solution (1:2, v/v). After homogenization, a portion of liver tissue homogenate (e.g., 200 uL of homogenate) was immediately measured and delivered to an analyst. After precipitated on wet ice, the samples were centrifuged, and a supernatant was collected and stored in a refrigerator at −70±10° C. before LC-MS/MS analysis. From the remaining liver homogenate, 800 uL of the homogenate was measured as a spare.

[0121] Compounds (1-4, 1-5, 1-6 and OCA, 0.0576 mmol/kg for each) were administered to rats by gavage, respectively. Rat plasma and liver samples were collected at points in time of 0.25, 0.5, 1, 2, 4, 8, 12, and 24 h, respectively, and tested by LC-MS/MS according to the above-mentioned method to determine the concentrations of the in vivo active metabolites of the above-mentioned compounds in plasma and liver (see Tables 1-4).

[0122] After Compound I-4 was administered to rats by gavage, the in vivo active compounds in the rats were Compound 4. The concentrations of Compound 4 in plasma and liver were as shown in Table 1.

TABLE-US-00002 TABLE 1 Compound 4 PK Parameters Plasma Liver Rsq_adj 0.930 0.868 No. points used for T.sub.1/2 5.00 5.00 C.sub.max (ng/mL of ng/g) 889 10494 T.sub.max (h) 1.00 1.00 T.sub.1/2 (h) 4.79 5.61 T.sub.last (h) 24.0 24.0 AUC.sub.0-last (ng .Math. h/mL or ng .Math. h/g) 4845 105664 AUC.sub.0-inf (ng .Math. h/mL or ng .Math. h/g) 4997 111955 MRT.sub.0-last (h) 5.91 6.80 MRT.sub.0-inf (h) 6.67 8.22 AUC.sub.Extra (%) 3.04 5.62 AUMC.sub.Extra (%) 14.1 21.9 AUC Ratio 21.8 Total AUC in plasma and liver = 110506, AUC Ratio = liver/plasma = 105664/4845 = 21.8.

[0123] After Compound I-5 was administered to rats by gavage, the in vivo active compounds in the rats were Compound 4. The concentrations of Compound 4 in plasma and liver were as shown in Table 2.

TABLE-US-00003 TABLE 2 Compound 4 PK Parameters Plasma Liver Rsq_adj 0.961 0.994 No. points used for T.sub.1/2 7.00 4.00 C.sub.max (ng/mL of ng/g) 2047 31790 T.sub.max (h) 0.250 0.250 T.sub.1/2 (h) 4.24 4.58 T.sub.last (h) 24.0 24.0 AUC.sub.0-last (ng .Math. h/mL or ng .Math. h/g) 7634 142808 AUC.sub.0-inf (ng .Math. h/mL or ng .Math. h/g) 7831 147182 MRT.sub.0-last (h) 5.19 6.31 MRT.sub.0-inf (h) 5.81 7.03 AUC.sub.Extra (%) 2.51 2.97 AUMC.sub.Extra (%) 13.0 12.9 AUC Ratio 18.7 Total AUC in plasma and liver = 150442, AUC Ratio = liver/plasma = 142808/7634 = 18.7.

[0124] After Compound I-6 was administered to rats by gavage, the in vivo active compounds in the rats were Compound 4. The concentrations of Compound 4 in plasma and liver were as shown in Table 3.

TABLE-US-00004 TABLE 3 Compound 4 PK Parameters Plasma Liver Rsq_adj 0.747 0.754 No. points used for T.sub.1/2 4.00 4.00 C.sub.max (ng/mL of ng/g) 775 18040 T.sub.max (h) 2.00 2.00 T.sub.1/2 (h) 6.04 6.34 T.sub.last (h) 24.0 24.0 AUC.sub.0-last (ng .Math. h/mL or ng .Math. h/g) 3858 83872 AUC.sub.0-inf (ng .Math. h/mL or ng .Math. h/g) 4131 90846 MRT.sub.0-last (h) 5.67 6.02 MRT.sub.0-inf (h) 7.46 8.10 AUC.sub.Extra (%) 6.62 7.68 AUMC.sub.Extra (%) 29.0 31.4 AUC Ratio 21.7 Total AUC in plasma and liver = 87730, AUC Ratio = liver/plasma = 83872/3858 = 21.7.

[0125] After the obeticholic acid was administered to rats by gavage, the in vivo active compounds in the rats were obeticholic acid (OCA) and Compound 4. Their concentrations in plasma and liver were as shown in Table 4.

TABLE-US-00005 TABLE 4 OCA Compound 4 PK Parameters Plasma Liver Plasma Liver Rsq_adj 0.287 0.629 0.963 0.998 No. points used for T.sub.1/2 6.00 6.00 4.00 3.00 C.sub.max (ng/mL) 24333 17930 35.6 2859 T.sub.max (h) 0.500 0.500 1.00 1.00 T.sub.1/2 (h) 4.35 3.33 28.7 14.5 T.sub.last (h) 24.0 24.0 24.0 24.0 AUC.sub.0-last (ng .Math. h/mL) 48093 31894 263 11136 AUC.sub.0-inf (ng .Math. h/mL) 50289 32264 575 15344 AUC Ratio 0.663 42.3 Total AUC in plasma and liver AUC = 91386; AUC Ratio = liver/plasma = 43030/48356 = 0.89.

[0126] After Compound 4 was administered to rats by gavage, the in vivo active compounds in the rats were Compound 4. The concentrations of Compound 4 in plasma and liver were as shown in Table 5.

TABLE-US-00006 TABLE 5 Compound 4 PK Parameters Plasma Liver Rsq_adj 0.997 0.994 No. points used for T.sub.1/2 3.00 3.00 C.sub.max (ng/mL of ng/g) 788 16830 T.sub.max (h) 4.00 4.00 T.sub.1/2 (h) 4.79 4.30 T.sub.last (h) 24.0 24.0 AUC.sub.0-last (ng .Math. h/mL or ng .Math. h/g) 7309 107187 AUC.sub.0-inf (ng .Math. h/mL or ng .Math. h/g) 7463 110040 MRT.sub.0-last (h) 5.56 6.25 MRT.sub.0-inf (h) 6.10 6.77 AUC.sub.Extra (%) 2.14 2.19 AUMC.sub.Extra (%) 10.8 9.78 AUC Ratio 14.7 Total AUC in plasma and liver AUC = 117503, AUC Ratio = liver/plasma = 14.7.

[0127] From the data listed in Tables 1, 2, 3 and 4, it could be seen that after gavage administration of the compound OCA to rats, lots of drug OCA (a sum of AUC of OCA in plasma+AUC of OCA-Gly was 48360) circulated in the blood, which might lead to side effects on skin caused by OCA, and the total effective drug amount (a sum of AUC of OCA in liver+AUC of OCA-Gly was 43030) in the liver was reduced accordingly. However, after Compounds I-4, I-5 and I-6 of the present disclosure were administered to rats by gavage in the same molar dosage respectively, the total effective drug amount in the liver was significantly increased. In particular, after administration of Compound I-5, the AUC of the effective drug in the liver reached 142808, which was 3.3 times that of the effective drug in the liver in case of direct administration of OCA, but the AUC of the effective drug in the blood decreased from 48360 in case of direct administration of OCA to 7634 by 84%. After obeticholic acid-glycine conjugate 4 and benzyl ester I-5 thereof were administered to rats by gavage in the same molar dosage, the comparison showed that the AUC ratio of the obeticholic acid-glycine conjugate 4 generated by the benzyl ester I-5 in liver/plasma reached 18.7, while the corresponding AUC ratio in case of direct administration of the obeticholic acid-glycine conjugate 4 was only 14.7. Enrichment of esterase in the liver might be one of the reasons why the benzyl ester I-5 exhibited a higher liver-targeting characteristic. The total AUC, i.e., 150442 of the Compound 4 generated by the benzyl ester I-5 in the liver and plasma was also significantly increased by 28% in comparison to the total AUC, i.e., 117503 of the Compound 4 when directly administered by gavage to rats.

[0128] Therefore, when the compounds of the present disclosure are used to treat the corresponding liver diseases, it is expected to significantly reduce the side effects on skin, while improving the efficacy of treating the liver diseases, and the dosage required to achieve the same efficacy is significantly reduced.