HK1259152B - Steroid derivative fxr agonist - Google Patents

Description

Steroid-derived FXR agonists

Technical Field

The present invention relates to a compound represented by formula (I), a tautomer or a pharmaceutically acceptable salt thereof, and to use thereof in preparing a drug for treating FXR-related diseases.

Background Art

The farnesoid X receptor (FXR) is an orphan nuclear receptor originally identified from a rat liver cDNA library (BM. Forman, et al., Cell 81: 687-693 (1995)). It is closely related to the insect ecdysone receptor. FXR is a member of the nuclear receptor family of ligand-activated transcription factors that includes steroid, retinoid, and thyroid hormone receptors (DJ. Mangelsdorf, et al., Cell 83: 841-850 (1995)). Northern and in situ analyses have shown that FXR is abundantly expressed in the liver, intestine, kidney, and adrenal gland (BM. Forman et al., Cell 81: 687-693 (1995) and W. Seol et al., Mol. Endocrinol. 9: 72-85 (1995)). FXR forms a heterodimer with the 9-cis retinoic acid receptor (RXR) to bind to DNA. FXR/RXR heterodimers preferentially bind to a component consisting of two nuclear receptor half-sites sharing the AG(G/T)TCA motif, which form inverted repeats and are separated by a single nucleotide (the IR-1 motif) (BM. Forman, et al., Cell 81: 687-693 (1995)). However, these compounds fail to activate mouse and human FXR, leaving the naturalness of the endogenous FXR ligand uncertain. Several naturally occurring bile acids bind and activate FXR at physiological concentrations (PCT WO 00/37077, published June 29, 2000). As described herein, bile acids that serve as FXR ligands include chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), and taurine and glycine conjugates of these bile acids.

WO-2005082925 discloses the use of INT747 in the preparation of a drug for treating FXR-related diseases.

Summary of the Invention

The purpose of the present invention is to provide a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

in,

Ring A is selected from a 5- to 12-membered aryl group, a 5- to 12-membered heteroaryl group containing 1-2 heteroatoms, a 5- to 6-membered non-aromatic heterocyclic group containing 1-2 heteroatoms, or a 5- to 6-membered cycloalkyl group, and the ring A is optionally substituted with 1, 2, or 3 _Ra groups or optionally oxoed with 1, 2, or 3 groups, wherein the heteroatoms are selected from N, O, or S;

L is selected from C _1-8 alkyl, C _1-8 heteroalkyl or C _2-8 alkenyl, said L is optionally substituted with 1, 2 or 3 R _b or optionally has 1, 2 or 3 oxo groups;

L ₁ is selected from O, N(R _d ), N(R _d )S(═O) ₂ or N(R _d )S(═O);

R ₁ is selected from H, C _1-6 alkyl, C _1-6 heteroalkyl, 5-6 membered aryl, 4-6 membered heteroaryl, C _3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, 5-6 membered aryl, 4-6 membered heteroaryl, C _3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _c or optionally has 1, 2 or 3 oxo groups;

_Ra , _Rb , _Rc and _Rd are each independently selected from H, F, Cl, Br, I, OH, CN, _NO2 , _NH2 , COOH, S(=O) _2OH , _C1-3 alkylamino, N,N-di( _C1-3 alkyl)amino, _C1-3 alkyl, _C1-3 alkyloxy or _C1-3 alkylthio, wherein the _C1-3 alkylamino, N,N-di( _C1-3 alkyl)amino, _C1-3 alkyl, _C1-3 alkyloxy or _C1-3 alkylthio is optionally substituted with 1, 2 or 3 R's;

R' is selected from F, Cl, Br, I, OH, _NH2 , _NO2 , CN, COOH, Me, Et, _CH2F , _CHF2 , _CF3 , _CH3O , _CH3S , NH( _CH3 ) or N( _CH3 ) ₂ .

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, _Ra , _Rb , _Rc and _Rd are each independently selected from H, F, Cl, Br, I, COOH, S(=O) _2OH , Me, _CF3 , _CHF2 , _CH2F , Et, OMe, NH( _CH3 ) or N( _CH3 ) ₂ .

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the ring A is selected from phenyl, pyridyl, pyridin-2(1H)-one, pyrimidinyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, isoxazolyl, isothiazolyl, bicyclo[1.1.1]pentanyl, benzoxazolyl, benzo[d]isoxazolyl, indazolyl, indolyl, quinolyl, isoquinolyl, quinazolinyl, 1H-pyrrolo[2,3-B]pyridinyl, indolizinyl, benzothiazolyl or benzothienyl, and the ring A is optionally substituted by 1, 2 or 3 _Ra .

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above-mentioned ring A is selected from the ring A optionally substituted by 1, 2 or 3 _Ra .

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the ring A is selected from

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylthio, C _1-4 alkylamino, C _1-4 alkyl-C(＝O)NH-, C _2-4 alkylalkenyl or C _1-3 alkyl-OC _1-3 alkyl, and the L is optionally substituted by 1, 2 or 3 R _b .

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from the group consisting of L optionally substituted by 1, 2 or 3 R _b .

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from

In some embodiments of the present invention, the above L ₁ is selected from O, NH, NHS(═O) ₂ and NHS(═O).

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above-mentioned R ₁ is selected from H, C _1-3 alkyl, C _3-6 cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl, and the C _1-3 alkyl, C _3-6 cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl is optionally substituted with 1, 2 or 3 R _c , and the R _c is as defined above.

In some embodiments of the present invention, in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, the above R ₁ is selected from H, Me,

As a preferred embodiment of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, it is selected from the compound represented by formula (II) or a pharmaceutically acceptable salt thereof:

in,

_L2 is selected from _C1-6 alkyl, _C1-6 heteroalkyl or _C2-6 alkenyl;

Ring B is selected from a benzene ring, or a 5- to 6-membered heteroaromatic ring containing 1-2 heteroatoms, wherein the heteroatoms are selected from N, O or S;

The ring B is optionally substituted by 1, 2 or 3 R ₂ or optionally has 1 oxo; the R ₂ is selected from F, Cl, Br, I, OH, CN, NO ₂ , NH ₂ , COOH, S(═O) ₂ OH, C _1-3 alkylamino, N,N-di(C _1-3 alkyl)amino, C _1-3 alkyl, C _1-3 alkyloxy or C _1-3 alkylthio;

L ₁ is selected from O, NH, NHS(═O) ₂ or NHS(═O);

R ₁ is selected from H, C _1-6 alkyl, C _1-6 heteroalkyl, 5-6 membered aryl, 4-6 membered heteroaryl, C _3-6 cycloalkyl or 3-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _1-6 heteroalkyl, 5-6 membered aryl, 4-6 membered heteroaryl, C _3-6 cycloalkyl or 3-6 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R _c or optionally has 1, 2 or 3 oxo groups, wherein the R _c is selected from F, Cl, Br, I, OH, CN, NO ₂ , NH ₂ , COOH or S(═O) ₂ OH.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, L ₂ is selected from -(CH ₂ ) _1-6 -, -(CH ₂ ) _0-4 -X-(CH ₂ ) _0-4 - or -(CH ₂ ) _0-4 -Y-(CH ₂ ) _0-4 -, wherein X is selected from NH, O or S, and Y is selected from -CH=CH-, provided that the chain length of L ₂ is selected from 1-6, preferably 2-4.

In some embodiments of the present invention, _in the compound represented by formula (II) or a pharmaceutically acceptable salt _thereof , _L2 is selected from _-CH2CH2- , _{-CH2CH2CH2- , -CH} ＝CH-, _{-CH2CH ＝} CH- _, -CH＝CHCH2-, -CH2CH2O- _, -CH2CH2CH2O- _{, -CH2CH2NH- , -CH2CH2CH2NH- , -CH2CH2S- or -CH2CH2CH2S- ; preferably from -CH2CH2- , -CH2CH2CH2- , -CH} ＝ _{CH- , -CH2CH2O- , -CH2CH2CH2O- or -CH2CH2NH- .}

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, ring B is selected from a benzene ring, a 5-membered heteroaromatic ring containing 1 to 2 atoms selected from N, O or S, or a 6-membered heteroaromatic ring containing 1 to 2 atoms selected from N, and the ring B is optionally substituted with 1, 2 or 3 _R2 or optionally has 1 oxo substitution, and the _R2 is as defined above.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, ring B is selected from a benzene ring, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, pyridine, pyridazine, pyrimidine or pyrazine, and the ring B is optionally substituted with 1, 2 or 3 _R2 or optionally has 1 oxo substitution, and the _R2 is as defined above.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, the ring B is optionally substituted with one R ₂ or optionally oxoed, and the R ₂ is selected from F, Cl, Br, I, OH, CN, NO ₂ , NH ₂ , COOH or S(═O) ₂ OH, preferably F, Cl or Br.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, the position of the substitution or oxo substitution of R ₂ is at the ortho position (α position) relative to L ₂ on ring B.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, R ₁ is selected from H, C _1-3 alkyl, C _3-6 cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl, and the C _1-3 alkyl, C _3-6 cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl is optionally substituted with 1, 2 or 3 R _c , and R _c is as defined above.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, R ₁ is selected from H, Me,

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, the structural fragment L ₁ -R ₁ is selected from -OH, -NHSO ₂ CH ₃ , -NHSO ₂ C(CH ₃ ) ₃ , -NHCH ₂ CH ₂ SO ₂ OH, -NHCH ₂ COOH, -NHSO ₂ CH(CH ₃ ) ₂ , -NHCH ₃ , -OCH ₃ or

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, the structural fragment selected from L ₂ and R ₂ are as defined above.

In some embodiments of the present invention, in the compound represented by formula (II) or a pharmaceutically acceptable salt thereof, the structural fragment is selected from

The present invention also provides a compound represented by formula (III) or a pharmaceutically acceptable salt thereof:

in,

Ring A is a 5- to 12-membered aryl group, a 5- to 12-membered heteroaryl group, a 5- to 6-membered non-aromatic heterocyclic group, or a 5- to 6-membered cycloalkyl group optionally substituted with 1, 2, or 3 R groups;

L is C _1-6 alkyl, C _1-6 heteroalkyl, or C _2-6 alkenyl optionally substituted with 1, 2, or 3 R;

R is F, Cl, Br, I, OH, CN, NO ₂ , NH ₂ , or R is selected from C _1-3 alkylamino, N,N-di(C _1-3 alkyl)amino, C _1-3 alkyl, C _1-3 alkyloxy, C _1-3 alkylthio, optionally substituted with 1, 2 or 3 R′;

R' is selected from F, Cl, Br, I, OH, NH ₂ , NO ₂ , CN, COOH, Me, Et, CH ₂ F, CHF ₂ , CF ₃ , CH ₃ O, CH ₃ S, NH(CH ₃ ), N(CH ₃ ) ₂ ;

The "hetero" represents a heteroatom or heteroatom group selected from -C(=O)NH-, N, -NH-, -C(=NH)-, -S(=O) _2NH- , -S(=O)NH-, -O-, -S-, N, ＝O, ＝S, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) _2- , and -NHC(=O)NH-;

In any of the above cases, the number of heteroatoms or heteroatom groups is independently selected from 1, 2 or 3.

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, R is selected from F, Cl, Br, I, Me, CF ₃ , CHF ₂ , CH ₂ F, Et, OMe, NH(CH ₃ ), and N(CH ₃ ) ₂ .

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the ring A is phenyl, pyridyl, pyridin-2(1H)-one, pyrimidinyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, isoxazolyl, isothiazolyl, bicyclo[1.1.1]pentanyl, benzoxazolyl, benzo[d]isoxazolyl, indazolyl, indolyl, quinolyl, isoquinolyl, quinazolinyl, 1H-pyrrolo[2,3-B]pyridinyl, indolizinyl, benzothiazolyl, benzothienyl, etc., which are optionally substituted with 1, 2 or 3 R groups.

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the ring A is selected from the group consisting of

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the ring A is selected from

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylthio, C 1-4 alkylamino, C _1-4 alkyl-C(＝O)NH-, and C _2-4 alkylalkenyl, which are optionally substituted with 1, 2 or ₃ R.

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the above L is selected from the group consisting of

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from

In some embodiments of the present invention, in the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the above-mentioned L is selected from

The compound of the present invention is selected from:

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the above compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The present invention also provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating various diseases related to farnesoid X receptor, cholestatic liver disease, fibrotic disease, hypercholesterolemia, hypertriglyceridemia and cardiovascular disease.

The present invention also provides the use of the above-mentioned compound or its pharmaceutically acceptable salt or the above-mentioned pharmaceutical composition in the preparation of a drug for treating liver function impairment caused by non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), cholestatic liver disease, chronic liver disease, hepatitis C infection, alcoholic liver disease, liver fibrosis, primary sclerosing cholangitis (PSC), gallstones, biliary atresia, lower urinary tract symptoms and benign prostatic hyperplasia (BPH), ureteral stones, obesity, type 2 diabetes, atherosclerosis, arteriosclerosis, hypercholesterolemia and hyperlipidemia.

The compounds of the present invention are FXR agonists and can be used in a method for preventing or treating dyslipidemia or diseases associated with dyslipidemia, which comprises administering a therapeutically effective amount of the compounds of the present invention to a patient in need of such treatment.

The compounds of the present invention can be used to lower total cholesterol levels, lower LDL cholesterol levels, lower VLDL cholesterol levels, increase HDL cholesterol levels, and/or lower triglyceride levels. Lowering triglyceride levels as described herein refers to lowering triglycerides in a subject in need of treatment to a level below the initial triglyceride level of the subject prior to administration of the compounds of the present invention. For example, the compounds of the present invention can reduce hepatic triglyceride production or hepatic triglyceride secretion by reducing fat absorption. The compounds of the present invention can also lower serum triglycerides and hepatic triglycerides.

The compounds of the present invention can be used to prevent or treat cardiovascular diseases associated with hypertriglyceridemia and/or hypercholesterolemia in a subject (e.g., a mammal, particularly a human), such as but not limited to atherosclerosis, arteriosclerosis, hypercholesterolemia, hyperlipidemia, thrombosis, coronary artery disease, stroke or hypertension.

The compounds of the present invention can be used to prevent or treat liver/biliary system diseases in a subject (e.g., a mammal, particularly a human), such as, but not limited to, cholestatic liver diseases, hypercholesterolemia, hypertriglyceridemia, or fibrotic diseases, specifically, such as, but not limited to, non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), gallstones, non-alcoholic cirrhosis, biliary atresia, cholestatic liver disease, chronic liver disease, hepatitis infection (type B or C), alcoholic liver disease, or liver fibrosis.

The compounds of the present invention can be used to prevent or treat obesity in a subject (eg, a mammal, particularly a human).

The compounds of the present invention can be used to prevent or treat diabetes, or diseases related to insulin resistance and glucose intolerance in a subject (eg, mammal, particularly human).

The compounds of the present invention can be used to prevent or treat lower urinary tract symptoms (nearby areas) and benign prostatic hyperplasia (BPH) or ureteral stones in a subject (eg, a mammal, particularly a human).

On the other hand, the present invention provides the use of a compound as shown in Formula I, Formula II or Formula III, a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a method for preventing or treating diseases that benefit from FXR stimulation, wherein the diseases that benefit from FXR stimulation include cardiovascular disease, liver/biliary system disease, obesity, diabetes, lower urinary tract symptoms (nearby area) and benign prostatic hyperplasia (BPH) or ureteral stones.

Another aspect of the present invention provides a method for preventing or treating diseases that benefit from FXR agonism, comprising administering to a patient a therapeutically effective amount of Formula I, Formula II or Formula III, a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition. The diseases that benefit from FXR agonism include cardiovascular disease, liver/biliary system disease, obesity, diabetes, lower urinary tract symptoms (nearby area) and benign prostatic hyperplasia (BPH) or ureteral stones.

The cardiovascular disease includes the cardiovascular disease related to hypertriglyceridemia and/or hypercholesterolemia. The cardiovascular disease further includes atherosclerosis, arteriosclerosis, hypercholesterolemia, hyperlipidemia, thrombosis, coronary artery disease, stroke or hypertension. The liver/biliary system disease includes cholestatic liver disease, high cholesterol disease, high triglyceride disease or fibrotic disease. The liver/biliary system disease further includes non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), gallstones, non-alcoholic cirrhosis, biliary atresia, cholestatic liver disease, chronic liver disease, hepatitis infection (type B or type C), alcoholic liver disease or liver fibrosis.

Definition and Explanation

Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, as long as the valence of the particular atom is normal and the compound after the substitution is stable.

The term "oxo" refers to the replacement of two hydrogen atoms on a specific atom by =O. When an aromatic ring undergoes oxo, the ring may retain or lose its aromaticity. It is understood that the group after oxo is stable. For example, when a benzene ring undergoes oxo, it can be selected from:

The terms "optionally" or "optionally" mean that the subsequently described event or situation may or may not occur, and the description includes both the occurrence of the event or situation and the non-occurrence of the event or situation. For example, an ethyl group is "optionally" substituted with a halogen, which means that the ethyl _group may be unsubstituted ( _CH2CH3 ), _{monosubstituted} (such as _CH2CH2F ), polysubstituted (such as _CHFCH2F , _{CH2CHF2 ,} etc.), or fully substituted ( _CF2CF3 ); for another example, "optionally, the ring B is substituted with 1, 2, or 3 _R2 or has 1 oxo" _means that the ring B is substituted with 1, 2, or 3 _R2 or has 1 oxo, or that the ring B is not substituted with _R2 or has no oxo. It will be understood by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern that is sterically impossible and/or cannot be synthesized will be introduced.

Herein, C _mn means that the moiety has an integer number of carbon atoms in a given range. For example, "C _1-6 " means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms.

Numerical ranges herein refer to each integer within the given range. For example, " _C1-3 alkyl" means that the group can be selected from _C1 , _C2 , and _C3 alkyl groups; " _C3-10 cycloalkyl" means that the group can be selected from _C3 , _C4 , _C5 , _C6 , _C7 , _C8 , _C9 , and _C10 cycloalkyl groups; and "5-6 membered cycloalkyl" means that the group can be selected from 5- and 6-membered heterocycloalkyl groups.

When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and/or their variants are permissible only if such combinations result in stable compounds.

Herein, when some groups can be substituted or oxoed, the substitution or oxo is independent of each other, including substitution without oxo, oxo without substitution, or both substitution and oxo. For example, when Ring A is optionally substituted with 1, 2, or 3 _Ras or oxoed, the following include: 1) Ring A has no substituents and no oxo; 2) Ring A is substituted with only 1, 2, or 3 _Ras ; 3) Ring A has only 1, 2, or 3 oxoed groups; and 4) Ring A is substituted with both _Ra and oxoed groups.

In some preferred embodiments of the present application, some fragment structures can be connected to other structures at the left end, and can be connected to other structures at the right end at the same time. When a dotted line or a solid line represents a connecting bond, those skilled in the art can understand by reading this application that the dotted line or solid line directionally indicates the connection state of the fragment structure with the other structures. For example, if ring A is selected from a left-directional dotted line, it indicates that the N atom is connected to the L group of the compound shown in formula (I), and a right-directional dotted line indicates that the C atom is connected to the carbonyl portion of the compound shown in formula (I); for another example, if ring A is selected from a left-directional dotted line, it indicates that the C atom is connected to the L group of the compound shown in formula (I), and a right-directional dotted line indicates that the N atom is connected to the carbonyl portion of the compound shown in formula (I). The left-directional dotted line or solid line refers to starting from the fragment structure itself and extending to the upper left, left or lower left; the right-directional dotted line or solid line refers to starting from the fragment structure itself and extending to the upper right, right or lower right.

In some preferred embodiments of the present application, the chain length refers to the number of atoms used to form the open-chain group, and the atoms include C atoms, N atoms, O atoms or S atoms of different valence states. For example, when L ₂ is selected from -CH ₂ CH ₂ CH ₂ -, the chain length is 3; when L ₂ is selected from -(CH ₂ ) ₂ -O-(CH ₂ ) ₂ -, the chain length is 5; when L ₂ is selected from -(CH ₂ ) ₂ -CH＝CH-(CH ₂ ) ₂ -, the chain length is 6.

The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

The term "hydroxy" refers to an -OH group.

The term "cyano" refers to a -CN group.

The term "mercapto" refers to a -SH group.

The term "amino" refers to a _-NH2 group.

The term "nitro" refers to a _-NO2 group.

The term "alkoxy" refers to an -O-alkyl group.

The term "alkylamino" refers to an -NH-alkyl group.

The term "dialkylamino" refers to -N(alkyl) ₂ .

The term "alkylsulfonyl" refers to a _-SO2 -alkyl group.

The term "alkylthio" refers to an -S-alkyl group.

The term "alkyl" refers to a hydrocarbon group of the general formula _{CnH2n +1} . The alkyl group can be straight-chain or branched. For example, the term " _C1-6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Similarly, the alkyl portion (i.e., alkyl) of alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio groups has the same definition as above.

The term "heteroalkyl" refers to an alkyl structure containing heteroatoms. Unless otherwise indicated, the heteroalkyl group is typically an alkyl group containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen and/or nitrogen.

The term "alkenyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond. Non-limiting examples of alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1,3-butadienyl, and the like. The term "alkenyl" refers to a straight chain alkenyl group.

The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. For example, an aryl group can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and 1,2,3,4-tetralin.

The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system containing at least one ring atom selected from N, O, S, with the remaining ring atoms being C, and having at least one aromatic ring. Preferred heteroaryl groups have single 4 to 8-membered rings, especially 5 to 8-membered rings, or multiple fused rings containing 6 to 14, especially 6 to 10, ring atoms. Non-limiting examples of heteroaryl groups include, but are not limited to, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, etc.

The term "cycloalkyl" refers to a fully saturated carbocyclic ring that can exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is typically a 3- to 10-membered ring. Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, and the like.

The term "non-aromatic heterocyclic radical" refers to a non-aromatic ring that is fully saturated or partially undersaturated (but not fully unsaturated heteroaromatic) and can exist as a monocyclic, bicyclic or spirocyclic ring. Unless otherwise indicated, the heterocycle is typically a 3 to 6-membered ring containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulphur, oxygen and/or nitrogen. The limiting examples of heterocyclic radicals include but are not limited to oxiranyl, tetrahydrofuranyl, dihydrofuranyl, pyrrolidinyl, N-methylpyrrolidinyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyrazolidinyl, 4H-pyranyl, morpholinyl, thiomorpholinyl, tetrahydrothienyl etc.

The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and/or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, prepared by reacting a compound having a specific substituent discovered by the present invention with a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable salts include metal salts, ammonium salts, salts formed with organic bases, salts formed with inorganic acids, salts formed with organic acids, salts formed with basic or acidic amino acids, and the like.

Preferably, the neutral form of the compound is regenerated by contacting the salt with a base or acid in a conventional manner and isolating the parent compound. The parent form of the compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents.

Certain compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are all within the scope of the present invention.

The diagrammatic representations of racemic, ambiscalemic, scalemic, or enantiomerically pure compounds herein are adapted from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise indicated, wedge-shaped and dashed bonds are used to represent the absolute configuration at a stereocenter. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they are intended to include both E and Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are intended to be encompassed within the scope of this invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms comprising the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( ³ H), iodine-125 ( ¹²⁵ I), or C-14 ( ¹⁴ C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic side effects on the host or patient. Representative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, viscosity increasing agents, transdermal enhancers, etc. Their preparation is well known to those skilled in the art of cosmetics or topical medicine. For additional information on carriers, reference can be made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

The term "excipient" generally refers to a carrier, diluent and/or vehicle required to formulate an effective pharmaceutical composition.

With respect to a drug or pharmacologically active agent, the term "effective amount" or "therapeutically effective amount" refers to a non-toxic amount of the drug or agent sufficient to achieve the intended effect. For the oral dosage forms of the present invention, an "effective amount" of an active substance in the composition means the amount required to achieve the intended effect when used in combination with another active substance in the composition. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active substance. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a target disorder, disease, or condition.

When the number of a linking group is 0, such as -(CRR) ₀ -, it means that the linking group is a single bond.

When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A. When the bond of a substituent can cross-link to two atoms on a ring, such substituent can be bonded to any atom on the ring. When the listed substituents do not indicate which atom is connected to the compound included in the chemical structure but not specifically mentioned, such substituent can be bonded through any atom thereof. Combinations of substituents and/or their variants are only allowed when such combinations will produce stable compounds. For example, the structural unit or indicates that it can be substituted at any position on the cyclohexyl group or cyclohexadiene.

Unless otherwise specified, the term "halo" or "halogen," by itself or as part of another substituent, means a fluorine, chlorine, bromine, or iodine atom. Additionally, the term "haloalkyl" is intended to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C ₁ -C ₄ )alkyl" is intended to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, and 3-bromopropyl, among others.

Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl. "Alkoxy" represents an alkyl group as described above with the specified number of carbon atoms attached through an oxygen bridge. _C1-6 alkoxy groups include _C1 , _C2 , _C3 , _C4 , _C5 , and _C6 alkoxy groups. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and S-pentoxy. "Cycloalkyl" includes saturated cyclic groups such as cyclopropyl, cyclobutyl, or cyclopentyl. C3-7 cycloalkyl groups include _C3 , _C4 , _C5 , _C6 , and _C7 cycloalkyl groups. "Alkenyl" includes hydrocarbon chains in a straight or branched configuration with one or more carbon-carbon double bonds present at any stable position along the chain, such as ethenyl and propenyl.

The term "protecting group" includes, but is not limited to, an "amino protecting group," a "hydroxy protecting group," or a "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions at the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl, and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like. An important consideration in planning any synthetic route in this area is the selection of a suitable protecting group for a reactive functional group such as the amino group in this application. For trained practitioners, Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons, 1991) is an authority in this regard. All references cited in this application are incorporated herein in their entirety.

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

The solvent used in the present invention is commercially available. The present invention uses the following abbreviations: aq represents water; HATU represents O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate; EDC represents N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride; m-CPBA represents 3-chloroperoxybenzoic acid; eq represents equivalent; CDI represents carbonyldiimidazole; DCM represents dichloromethane; PE represents petroleum ether; DIAD represents diisopropyl azodicarboxylate; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOAc represents ethyl acetate; EtOH represents ethanol; MeOH represents methanol; CBz represents benzyloxycarbonyl, which is an amine protecting group; BOC represents tert-butylcarbonyl, which is an amine protecting group; HOAc represents acetic acid; NaCNBH ₃ represents sodium cyanoborohydride; rt represents room temperature; O/N represents overnight; THF represents tetrahydrofuran; Boc ₂ represents methyl benzoate; O represents di-tert-butyl dicarbonate; TFA represents trifluoroacetic acid; DIPEA represents diisopropylethylamine; SOCl2 represents thionyl chloride; _CS2 represents carbon disulfide; _TsOH represents p-toluenesulfonic acid; NFSI represents N-fluoro-N-(phenylsulfonyl)benzenesulfonamide; NCS represents 1-chloropyrrolidine-2,5-dione; n- _Bu4NF represents tetrabutylammonium fluoride; iPrOH represents 2-propanol; mp represents melting point; LDA represents lithium diisopropylamide.

DETAILED DESCRIPTION

In order to illustrate the present invention in more detail, the following examples are given, but the scope of the present invention is not limited thereto.

Reference Example 1 Preparation of INT-747

Reference example 1A

To a solution of chenodeoxycholic acid (60.0 g, 152.8 mmol) in methanol/acetic acid/water/ethyl acetate (360/120/30/780 ml) were added tetrabutylammonium bromide (81.0 g, 251.3 mmol) and sodium bromide (9.0 g, 87.5 mmol) portionwise. Sodium hypochlorite (210 ml, 3.4 mol) was then added dropwise at 0°C over 30 minutes. After stirring at 28°C for 16 hours, the mixture was quenched by the addition of saturated sodium bisulfite solution (500 ml). The aqueous layer was extracted with ethyl acetate (1000 ml x 2), and the combined organic layers were washed with water (1000 ml x 5). The organic layer was dried over sodium sulfate, filtered, and spin-dried. The residue was recrystallized (dichloromethane, 200 ml) to give Reference Example 1A (yellow solid, 41 g, 69% yield). ¹ H NMR (400MHz, METHANOL-d ₄ )δ3.44-3.60 (m, 1H), 2.99 (dd, J=5.77, 12.30Hz, 1H), 2.54 (t, J=11.29Hz, 1H), 2.08-2.41 (m, 3H), 2.00-2 .08 (m, 1H), 1.75-1.96 (m, 6H), 1.28-1.69 (m, 9H), 1.09-1.27 (m, 8H), 0.97 (d, J=6.53Hz, 3H), 0.71 (s, 3H).

Reference Example 1B

To a solution of Reference Example 1A (246.0 g, 629.9 mmol) in methanol (2 L) was added p-toluenesulfonic acid (10.9 g, 63.0 mmol) in one portion, and the mixture was reacted at 80°C for 4 hours. After cooling to room temperature, the mixture was evaporated to dryness and quenched with saturated sodium bicarbonate solution (1500 ml). The aqueous layer was extracted with ethyl acetate (1500 ml x 3). The combined organic layers were washed with brine (1000 ml x 3), dried over sodium sulfate, filtered, and spin-dried. The residue was recrystallized (ethyl acetate, 500 ml) to give Reference Example 1B (white solid, 202 g, 79% yield). ¹ H NMR (400MHz, CHLOROFORM-d) δ3.66 (s, 3H), 3.55-3.62 (m, 1H), 2.85 (dd, J=6.02, 12.55Hz, 1H), 2.28-2.43 (m, 2H) , 2.13-2.26(m, 2H), 1.64-2.04(m, 10H), 1.19-1.51(m, 11H), 1.00-1.17(m, 3H), 0.86-0.97(m, 3H), 0.65(s, 3H).

Reference Example 1C

To a solution of trimethylsilyl chloride (107.5 g, 989.5 mmol) in tetrahydrofuran (500 mL) was added dropwise lithium diisopropylamide (87.4 g, 815.6 mmol) under nitrogen at -78°C. After stirring for 40 minutes, a solution of Reference Example 1B (50 g, 123.6 mmol) in tetrahydrofuran (300 mL) was added dropwise. After the addition was complete, stirring was continued at -78°C for 40 minutes, followed by the addition of triethylamine (182.5 g, 1.8 mol). After 1 hour, the mixture was quenched with saturated sodium bicarbonate (1000 mL), and the aqueous layer was extracted with ethyl acetate (1000 mL x 3). The combined organic layers were washed with water (100 ml × 6) and saturated brine (1000 ml × 2), and the organic layer was dried over sodium sulfate, filtered and spin-dried to obtain Reference Example 1C (brown oil, 68 g, yield 100%), which could be used directly in the next step without further purification. ¹ H NMR (400MHz, CHLOROFORM-d) δ4.75 (dd, J=1.38, 5.90Hz, 1H), 3.69 (s, 3H), 3.48-3.59 (m, 1H), 2.13-2.42 (m, 2H), 1.52-2.04 (m, 10H), 1.29-1.48(m, 7H), 0.99-1.23(m, 5H), 0.95(d, J=6.53Hz, 3H), 0.85(s, 3H), 0.70(s, 3H), 0.17-0.20(m, 9H), 0.13(s, 9H).

Reference example 1D

To a solution of Reference Example 1C (68.0 g, 123.9 mmol) in dichloromethane (500 mL) was added anhydrous acetaldehyde (10.1 g, 229.2 mmol). A solution of boron trifluoride-diethyl ether (64.4 g, 453.4 mmol) in dichloromethane (300 mL) was then added dropwise under nitrogen at -78°C. The addition rate was such that the internal temperature remained -78°C. After stirring for 1 hour, the temperature was raised to 30°C and stirring was continued for 2 hours. The solution was quenched with saturated sodium bicarbonate (1000 mL). The aqueous layer was extracted with dichloromethane (1000 mL x 3). The combined organic layers were washed with saturated brine (1000 mL x 2), dried over sodium sulfate, filtered, and spin-dried. The residue was purified by column chromatography to afford Reference Example 1D (yellow solid, 43 g, 81% yield). ¹ H NMR (400MHz, CHLOROFORM-d) δ6.12 (q, J=7.03Hz, 1H), 3.52-3.66 (m, 4H), 2.54 (dd, J=4.02, 13.05Hz, 1H), 2.13-2.40 (m , 5H), 1.68-1.98 (m, 7H), 1.65 (d, J=7.03Hz, 3H), 1.00-1.52 (m, 11H), 0.97 (s, 3H), 0.89 (d, J=6.53Hz, 3H), 0.61 (s, 3H).

Reference Example 1E

To a solution of Reference Example 1D (212.0 g, 492.3 mmol) in methanol (500 ml) was added a solution of NaOH (39.4 g, 984.6 mmol) in water (50 ml), and the mixture was stirred at 50°C for 2 hours. After the solvent was evaporated, water (500 ml) was added, and the mixture was extracted with ethyl acetate (500 ml x 2). The aqueous phase was adjusted to pH 3 with dilute HCl and then extracted with dichloromethane (600 ml x 2). The combined organic layers were concentrated, and the residue was purified by recrystallization (ethanol, 200 ml) to give Reference Example 1E (yellow solid, 147 g, 72% yield). ¹ H NMR (400MHz, CHLOROFORM-d) δ6.19 (q, J=7.36Hz, 1H), 3.60-3.74 (m, 1H), 2.58 (dd, J=4.02, 13.05Hz, 1H), 2.40 (tt, J=5.02, 10.29Hz, 3H), 2.19-2.32(m, 2H), 1.61-2.06(m, 10H), 1.04-1.54(m, 14H), 1.01(s, 3H), 0.95(d, J=6.53Hz, 3H), 0.65(s, 3H).

Reference Example 1F

To a solution of Reference Example 1E (140.0 g, 336.1 mmol) in NaOH (0.5 mol) in water (600 mL) was added 10% Pd—C (19.9 g, 134.4 mmol) in one portion, and hydrogen was introduced at 15 psi. The reaction was carried out at 100°C for 16 hours. The mixture was filtered, and the pH of the filtrate was adjusted to 3 with dilute hydrochloric acid. The aqueous layer was extracted with dichloromethane (1500 mL x 3). The combined organic layers were washed with brine (1000 mL x 3), dried over sodium sulfate, filtered, and spin-dried to obtain Reference Example 1F (white solid, 101 g, 72% yield), which was used directly in the next step without further purification. ¹ H NMR (400MHz, CHLOROFORM-d) δ3.49-3.60 (m, 1H), 2.70 (q, J=6.02Hz, 1H), 2.12-2.45 (m, 4H), 1.65-2.02 (m , 9H), 1.29-1.52 (m, 6H), 1.05-1.24 (m, 8H), 0.93 (d, J=6.53Hz, 5H), 0.81 (t, J=7.53Hz, 3H), 0.66 (s, 3H).

Reference compound INT-747

To a solution of Reference Example 1F (16.0 g, 38.2 mmol) in sodium hydroxide (2 mol, 100 ml) was added sodium borohydride (8.7 g, 229.3 mmol) portionwise, and the mixture was stirred at 100°C for 2 hours. After cooling to room temperature, saturated aqueous ammonium chloride (150 ml) was added, and the pH was adjusted to 3 with dilute hydrochloric acid. The aqueous layer was extracted with dichloromethane (300 ml x 3), and the combined organic layers were washed with brine (200 ml x 3), dried over sodium sulfate, filtered, and evaporated. The residue was purified by column chromatography to give reference compound INT-747 (white solid, 14.5 g, 90% yield). ¹ H NMR (400MHz, CHLOROFORM-d) δ3.71 (br.s, 1H), 3.36-3.48 (m, 1H), 2.18-2.47 (m , 2H), 1.56-2.01(m, 10H), 1.06-1.54(m, 15H), 0.86-0.97(m, 9H), 0.66(s, 3H).

Route 1

Example 1

Example 1A

Under nitrogen, perchloric acid (6.0 g, 60.0 mmol) was added to a solution of reference compound INT-747 (2.7 g, 6.4 mmol) and formic acid (0.3 g, 6.4 mmol) in tetrahydrofuran (40 mL). The mixture was stirred at 55°C for 6 hours. The solution was concentrated under reduced pressure and diluted with water (100 mL). The aqueous layer was extracted with ethyl acetate (100 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The residue was purified by column chromatography to afford Example 1A as a white solid (2.8 g, 92% yield). ¹ H NMR (400MHz, CHLOROFORM-d) δ8.16 (s, 1H), 8.05 (s, 1H), 5.20 (br.s., 1H), 4.77-4.65 (m, 1H), 2.45-2.34 (m, 1H) , 2.31-2.21(m, 1H), 2.01-1.58(m, 11H), 1.55-1.30(m, 8H), 1.22-1.05(m, 6H), 0.97-0.88(m, 9H), 0.66(s, 3H).

Example 1B

To a solution of Example 1A (100.0 mg, 0.2 mmol) and lead acetate (186.0 mg, 0.4 mmol) in carbon tetrachloride (2 mL) was added elemental iodine (106.0 mg, 0.4 mmol), and the reaction system was reacted under light for 12 hours. The reaction was quenched by adding sodium thiosulfate solution (1 mL). The aqueous layer was extracted with dichloromethane (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and spin-dried. The residue was purified by preparative thin layer chromatography (petroleum ether:ethyl acetate = 5:1) to afford Example 1B (colorless solid, 50.0 mg, 38% yield). 1H-NMR (CDCl3, 400MHz) δ8.14 (s, 1H), 8.03 (s, 1H), 5.19 (br.s., 1H), 4.62-4 .77(m, 1H), 3.23-3.32(m, 1H), 3.00-3.12(m, 1H), 1.96-2.02(m, 2H), 1.85-1 .92(m,2H),1.70-1.82(m,7H),1.66-1.72(m,2H),1.39-1.45(m,2H),1.23-1 .32(m, 5H), 1.09-1.19(m, 6H), 0.96(s, 3H), 0.90-0.93(m, 6H), 0.67(s, 3H).

Example 1

To a solution of methyl 6-ketopiperidine-3-carboxylate (21.0 mg, 134 μmol) in N,N-dimethylformamide (1 ml) was added sodium hydroxide (7.0 mg, 179 μmol, 60%) at zero degrees Celsius. After half an hour, a solution of Example 1B (50.0 mg, 89.5 μmol) in N,N-dimethylformamide (1 ml) was slowly added dropwise at zero degrees Celsius. After the addition was complete, the reaction system was slowly warmed to room temperature and allowed to react for 12 hours. Water (3 ml) and lithium hydroxide monohydrate (4 mg, 89.5 mmol) were added to the reaction system, and stirring was continued for 3 hours. Water (10 ml) was added, and the reaction system was adjusted to pH 6 with hydrochloric acid (1 M). The mixture was extracted with dichloromethane:methanol = 10:1 (10 ml x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and dried by rotary evaporation. The residue was purified by preparative thin layer plate (dichloromethane:methanol=10:1) to give Example 1 (15 mg, 32.4% yield). ¹ H NMR (400MHz, METHANOL-d ₄ )δ3.67(br.s., 1H), 3.58-3.48(m, 2H), 3.45-3.34(m, 2H), 3.31-3.27(m, 1H), 2.87(d, J=13.1Hz, 1H), 2.49-2.29(m, 2H), 2.11(br.s., 2H), 2.00-1.35(m, 19H), 1.30-1.10(m, 6H), 1.04(d, J=6.5Hz, 3H), 0.95-0.90(m, 6H), 0.72(s, 3H).

The operation was the same as in Example 1, and Example Compound 2-16 was synthesized as follows:

Route 2

Example 17 and Example 18

Example 2A

To a solution of Example 1A (500.0 mg, 1 mmol) in tetrahydrofuran (5 mL) were added triethylamine (153.0 mg, 1.5 mmol) and ethyl chloroformate (167.0 mg, 1.5 mmol), and the reaction was allowed to react at 25°C for 2 hours. The system was cooled to 0°C, and a solution of sodium borohydride (210.0 mg, 5.6 mmol) in methanol (5 mL) was slowly added to the reaction system. The reaction was allowed to proceed at 0°C for 15 minutes and then at 25°C for 15 minutes. The reaction was quenched with 0.2 M dilute hydrochloric acid (1 mL). The aqueous layer was extracted with ethyl acetate (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and dried by rotary evaporation. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 4:1) to afford Example 2A (250.0 mg, 70%). ¹ H NMR (400MHz, CHLOROFORM-d) δ8.15 (s, 1H), 8.04 (s, 1H), 5.19 (br.s., 1H), 4.77-4.66 (m, 1H) , 3.66-3.57(m, 2H), 2.06-1.77(m, 7H), 1.45-1.06(m, 21H), 0.97-0.88(m, 9H), 0.66(s, 3H).

Example 2B

To a mixed solution of Example 2A (280.0 mg, 605 μmol), triphenylphosphine (476.0 mg, 1.8 mmol) and imidazole (124.0 mg, 1.8 mmol) in toluene (4 mL) and acetonitrile (1 mL) was added iodine (461.0 mg, 1.8 mmol) at zero degree Celsius, and the reaction system was reacted at 25 degrees Celsius for 3 hours. Saturated sodium sulfite solution (10 mL) was added to the reaction system, and the aqueous layer was extracted with ethyl acetate (10 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and spin-dried. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to give Example 2B (250.0 mg, 70%). ¹ H-NMR (CDCl ₃ , 400MHz) δ8.16 (s, 1H), 8.07-8.02 (m, 1H), 5.20 (br.s., 1H), 4.76-4.66 (m, 1H), 3.24- 3.08(m, 2H), 2.01-1.72(m, 10H), 1.46-1.06(m, 17H), 0.97-0.89(m, 9H), 0.66(s, 3H).

Example 17 and Example 18

To a solution of methyl 6-hydroxynicotinate (80.0 mg, 523 μmol) in N,N-dimethylformamide (3 mL) was added sodium hydroxide (21.0 mg, 523 mmol, 60%) at zero degrees Celsius. Half an hour later, a solution of Example 2B (150.0 mg, 262 μmol) in N,N-dimethylformamide (5 mL) was added dropwise at zero degrees Celsius. After the addition was complete, the reaction system was slowly warmed to room temperature and allowed to react for 12 hours. Water (3 mL) and lithium hydroxide monohydrate (55 mg, 1.31 mmol) were added to the reaction system, and stirring was continued for 3 hours. Water (10 mL) was added, and the system was adjusted to pH 6 with hydrochloric acid (1 M). The mixture was extracted with dichloromethane:methanol = 10:1 (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and dried by rotary evaporation. The residue was separated and purified by preparative thin layer plate (dichloromethane:methanol=10:1) to give Example 17 (40 mg, 29%). ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 8.44 (d, J=2.5 Hz, 1H), 7.97 (dd, J=2.5, 9.5 Hz, 1H), 6.55 (d, J=9.5 Hz, 1H), 4.10-3.96 (m, 2H), 3.70-3.63 (m, 1H), 3.37-3.34 (m, 1H), 2.02-1.11 (m, 27H), 0.98 (d, J=6.0 Hz, 3H), 0.95-0.88 (m, 6H), 0.71 (s, 3H). and Example 18 (20 mg, 14%), ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 8.77 (s, 1H), 8.21 (dd, J=1.8, 8.8 Hz, 1H), 6.84 (d, J=8.5 Hz, 1H), 4.34 (d, J=2.5 Hz, 2H), 3.78-3.58 (m, 1H), 3.38-3.33 (m, 1H), 2.02-1.22 (m, 27H), 1.01 (d, J=6.5 Hz, 3H), 0.95-0.90 (m, 6H), 0.72 (s, 3H).

Route 3

Example 19

Example 3A

To a solution of Example 2A (500.0 mg, 1.1 mmol), copper acetate (99.0 mg, 0.6 mmol), and lead acetate (2.4 g, 11.0 mmol) in toluene (5 mL) was added pyridine (980.0 mg, 12.4 mmol), and the reaction system was reacted at 110°C for 12 hours. The reaction system was filtered, the filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to give the title compound (120.0 mg, 27.0%). ¹ H NMR (400MHz, CHLOROFORM-d) δ8.15 (s, 1H), 8.05 (s, 1H), 5.71-5.59 (m, 1H), 5.19 (br.s., 1H), 4.93-4.80 (m, 2H), 4.76-4.6 7 (m, 1H), 2.12-1.61 (m, 11H), 1.52-1.09 (m, 13H), 1.04 (d, J=6.8Hz, 3H), 0.97 (s, 3H), 0.91 (t, J=7.4Hz, 3H), 0.68 (s, 3H).

Example 3B

To a solution of Example 3A (100.0 mg, 0.2 mmol), sodium carbonate (74.0 mg, 0.7 mmol), and tetrabutylammonium bromide (66.0 mg, 0.2 mmol) in N,N-dimethylformamide (2 ml) were added palladium acetate (5.0 mg, 23.0 μmol) and methyl 2-iodobenzoate (60.8 mg, 0.2 mmol) under nitrogen. The reaction system was reacted at 85°C for 12 hours. The solvent was evaporated, and water (10 ml) was added to the reaction system. The aqueous layer was extracted with dichloromethane (20 ml x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and evaporated. The residue was purified by thin-layer preparative chromatography (petroleum ether:ethyl acetate = 10: ¹ ) to give the title compound (80.0 mg, 61.0%). NMR (400MHz, CHLOROFORM-d) δ8.16 (s, 1H), 8.07 (s, 1H), 8.01 (s, 1H), 7.90-7.84 (m, 1H), 7 .51 (d, J=7.8Hz, 1H), 7.41-7.33 (m, 1H), 6.40-6.31 (m, 1H), 6.16 (dd, J=8.8, 15.8Hz, 1H), 5 .21 (br.s., 1H), 4.79-4.63 (m, 1H), 3.94 (s, 3H), 2.29 (d, J=6.8Hz, 1H), 2.08-1.62 (m, 11H ), 1.52-1.20 (m, 11H), 1.15 (d, J=6.5Hz, 3H), 1.00 (s, 3H), 0.95-0.90 (m, 3H), 0.74 (s, 3H).

Example 3C

To a solution of Example 3B (100.0 mg, 0.2 mmol) in methanol (3 mL) was added wet palladium on carbon (50 mg, 10%) under nitrogen. The reaction was stirred at 25°C under hydrogen (15 psi) for 12 hours. The reaction was filtered and dried to afford Example 3C (90.0 mg, 90.0%). ¹ H NMR (400MHz, CHLOROFORM-d) δ 8.18-8.11 (m, 1H), 8.06-8.00 (m, 1H), 7.85-7.81 (m, 2H), 7.35-7.31 (m, 2H), 5.18 (br.s., 1H), 4.7 6-4.51 (m, 1H), 2.77-2.64 (m, 1H), 2.54-2.44 (m, 1H), 1.95-1.20 (m, 25H), 1.01 (d, J=6.5Hz, 3H), 0.95-0.87 (m, 6H), 0.64 (s, 3H).

Example 19

To a solution of Example 3C (160.0 mg, 282.0 μmol) in tetrahydrofuran (1 mL), methanol (1 mL), and water (1 mL) was added lithium hydroxide (59.0 mg, 1.4 mmol). The reaction was incubated at 20°C for 12 hours. Water (5 mL) was added to the reaction system, and the pH was adjusted to 1-2 with hydrochloric acid (1 M). The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and spun down to dryness. The residue was purified by preparative separation (HCl) to afford Example 19 (80 mg, 57%). ¹ H NMR (400MHz, METHANOL-d ₄ )δ7.93-7.76(m, 2H), 7.45-7.35(m, 2H), 3.67(br.s., 1H), 3.37-3.35(m, 1H), 2.84-2.73(m, 1H), 2 .60 (d, J=10.5Hz, 1H), 2.06-1.27 (m, 25H), 1.09 (d, J=6.3Hz, 3H), 0.94-0.91 (m, 6H), 0.70 (s, 3H).

Route 4

Example 20

To a solution of Example 3B (40.0 mg, 70.8 μmol) in tetrahydrofuran (0.5 mL), methanol (0.5 mL), and water (0.5 mL) was added lithium hydroxide (15 mg, 0.35 mmol). The reaction was allowed to react at 20°C for 12 hours. Water (5 mL) was added to the reaction system, and the pH was adjusted to 1-2 with hydrochloric acid (1 M). The aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and spin-dried. The residue was purified by preparative thin layer chromatography (dichloromethane:methanol = 10:1) to afford Example 20 (25.0 mg, 71.0%). ¹ H NMR (400MHz, METHANOL-d ₄ )δ8.00 (br.s., 1H), 7.84 (d, J=7.0Hz, 1H), 7.55 (d, J=7.5Hz, 1H), 7.42-7.34 (m, 1H), 6.46-6.32 (m, 1H), 6.19 (dd, J=8.8, 15.6Hz, 1H), 3 .72-3.62 (m, 1H), 3.39-3.37 (m, 1H), 2.32 (d, J=6.0Hz, 1H), 2.07-1.29 (m, 22H), 1.17 (d, J=6.3Hz, 3H), 0.95-0.89 (m, 6H), 0.78 (s, 3H).

Route 5

Example 21

Example 5A

A mixture of Reference Example 1 (100.0 mg, 0.2 mmol), triethylamine (48.0 mg, 0.5 mmol) and O,N-dimethylhydroxylamine hydrochloride (23.0 mg, 0.2 mmol) in acetonitrile (2 ml) was stirred at 25 degrees Celsius for 0.5 hours, followed by the addition of O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (95.0 mg, 0.3 mmol). The resulting mixture was stirred at 25 degrees Celsius for 12 hours. After removing the solvent by rotary evaporation in vacuo, the residue was purified by column chromatography to give Example 5A as a white solid (90 mg, 82% yield). ¹ H NMR (400MHz, CHLOROFORM-d) δ3.73-3.67(m, 4H), 3.45-3.36(m, 1H), 3.18(s, 3H), 2.51-2.40(m, 1H), 2.38-2.27(m, 1H), 2.00-1.89(m, 2H), 1.87- 1.74(m, 5H), 1.71-1.56(m, 5H), 1.53-1.31(m, 11H), 1.23-1.14(m, 3H), 1 .06-0.99 (m, 1H), 0.96 (d, J=6.3Hz, 3H), 0.93-0.88 (m, 6H), 0.67 (s, 3H).

Example 5B

To a solution of Example 5A (100.0 mg, 0.2 mmol) in tetrahydrofuran (5 mL) was added a solution of methylmagnesium bromide (0.4 mL, 1.1 mmol, 3N) in diethyl ether at 0°C, and stirring was continued at 0°C for 30 minutes, then warmed to room temperature and stirred for 12 hours. Ice water was added to quench the reaction, and then extracted with ethyl acetate (60 mL×2). After washing with water, the organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed in vacuo. The residue was purified by preparative TLC to give Example 5B as a white solid (6 mg, 66% yield). ^1H NMR (400MHz, CHLOROFORM-d) δ3.71 (br.s., 1H), 3.48-3.35 (m, 1H), 2.51-2.41 (m, 1H), 2.39-2.29 (m, 1H), 2.14 (s, 3H), 1.99-1.79 (m, 5H), 1.76-1.56 (m, 5H), 1.51-1.30 (m, 10H), 1.22-1.11 (m, 3H), 1.00 (dt, J=3.3, 14.2Hz, 1H), 0.94-0.87 (m, 9H), 0.66 (s, 3H).

Example 5C

Example 5B (1.0 g, 2.4 mmol) was dissolved in 1,4-dioxane (20.0 mL), and 1,4-dihydropyran (2.0 g, 23.9 mmol, 2.2 mL) and p-toluenesulfonic acid (90.9 mg, 478.0 μmol) were added. The reaction mixture was stirred at 30°C for 36 hours. The solvent was removed by concentration, and water (5 mL) was added to the reaction mixture. The mixture was then extracted with ethyl acetate (10 mL x 3). The organic layer was washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was separated by column chromatography to obtain Example 5C ( ^450.0 mg, 32.1% yield, colorless oil). NMR (400MHz, CHLOROFORM-d) δ4.73 (d, J=3.3Hz, 1H), 4.56 (d, J=4.3Hz, 1H), 3.95-3.81 ( m, 2H), 3.76-3.72 (m, 1H), 3.55-3.31 (m, 4H), 2.52-2.41 (m, 1H), 2.34 (ddd, J=5.8, 9.9, 16.0Hz, 1H), 1.97-1.80 (m, 7H), 1.74-1.67 (m, 4H), 1.55 (br.s., 4H), 1.41 (d, J=7.3Hz, 3H), 1.33-1.27(m, 3H), 1.17-1.08(m, 3H), 0.90(s, 3H), 0.88(s, 3H), 0.69-0.59(m, 3H).

Example 5D

Sodium metal (68.2 mg, 3.0 mmol) was dissolved in anhydrous ethanol (5 mL). After cooling to 0°C, Example 5C (170.0 mg, 296.8 μmol, dissolved in 2 mL of anhydrous ethanol) was slowly added and stirring continued at 0°C for half an hour. Diethyl oxalate (65.0 mg, 445.1 μmol, 60.8 μL, dissolved in 1 mL of anhydrous ethanol) was then added dropwise. The reaction solution was stirred at 0°C for half an hour, then warmed to 25°C and stirred for 35 hours. Ethanol (6 mL) was added to the reaction solution, and the solvent was concentrated to remove it. Ethyl acetate (40 mL) was added, and a dimolar aqueous solution of citric acid (to pH 5-6) was added with stirring in an ice bath. The aqueous phase was extracted with ethyl acetate (10 mL x 2). The organic layer was washed with water (10 mL), brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was separated by column chromatography to give Example 5D (90.0 mg, 44.2% yield). ¹ H NMR (400 MHz, CHLOROFORM-d) δ 4.73 (d, J = 3.3 Hz, 1H), 4.57 (d, J = 4.0 Hz, 1H), 4.36 (q, J = 7.0 Hz, 2H), 3.97-3.80 (m, 2H), 3.70 (br. s., 1H), 3.52-3.34 (m, 3H), 2.54 (ddd, J = 5.1, 10.3, 15.4 Hz, 1H), 2.44-2 .29(m, 1H), 1.98-1.90(m, 2H), 1.82(br.s., 3H), 1.74-1.67(m, 4H), 1.53(br.s., 7H), 1.38(s, 3H), 1.26 (t, J=7.0Hz, 2H), 1.16 (d, J=9.8Hz, 2H), 0.95 (d, J=6.5Hz, 2H), 0.89 (s, 3H), 0.71-0.59 (m, 3H).

Example 5E

Example 5D (90.0 mg, 131.0 μmol) was dissolved in ethanol (3.0 mL), and hydroxylamine hydrochloride (27.3 mg, 393.0 μmol) was added. The reaction mixture was stirred at 80°C for five hours. The solvent was concentrated and removed. Water (5 mL) was added to the reaction mixture, and the aqueous phase was extracted with ethyl acetate (10 mL x 3). The organic layer was washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was decompressed and dried by rotary evaporation. The crude product was obtained without ^further purification as Example 5E (86.0 mg, crude product, yellow oil). HNMR (400MHz, CHLOROFORM-d) δ6.39 (s, 1H), 4.43 (q, J=7.3Hz, 2H), 3.72 (br.s., 1H), 3. 43-3.40 (m, 1H), 2.90-2.64 (m, 2H), 2.42 (t, J=6.9Hz, 2H), 2.00-1.95 (m, 2H), 1.75 (d, J =6.8Hz, 3H), 1.69 (br.s., 4H), 1.60 (d, J = 3.3Hz, 3H), 1.48 (d, J = 7.5Hz, 4H), 1.41 (s, 3H ), 1.28(br.s., 4H), 1.20(d, J=8.5Hz, 3H), 0.90(s, 3H), 0.89-0.88(m, 1H), 0.65(s, 3H).

Example 21

Example 5E (115.0 mg, 222.9 μmol) was dissolved in tetrahydrofuran (1.0 mL), water (1.0 mL), and methanol (1.0 mL). Lithium hydroxide monohydrate (93.6 mg, 2.2 mmol) was added, and the reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was acidified with monomolar hydrochloric acid (to pH = 5-6) and extracted with ethyl acetate (10 mL x 3). The organic layer was washed with water (10 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was evaporated to dryness under reduced pressure. The crude product was separated by thin layer chromatography to give Example 21 (28.0 mg, 25.8% yield). ¹ H NMR (400 MHz, METHANOL-d ₄ )δ6.41(s, 1H), 3.67(br.s., 1H), 3.31(br.s., 1H), 2.94-2.83(m, 1H), 2.80-2.70(m , 1H), 2.03 (d, J=12.0Hz, 1H), 1.96-1.83 (m, 5H), 1.80-1.73 (m, 2H), 1.64 (br.s., 1H) ,1.58-1.50(m,5H),1.48-1.42(m,2H),1.42-1.34(m,3H),1.32-1.27(m,3H),1.24( d, J=9.5Hz, 2H), 1.05 (d, J=6.0Hz, 3H), 0.93 (s, 3H), 0.92-0.89 (m, 3H), 0.71 (s, 3H).

Route 6

Rb＝H or X, for example:

Example 22

Example 6A

To a solution of methyl 6-hydroxynicotinate (438.7 mg, 2.9 mmol) and Example 1B (800 mg, 1.4 mmol) in chloroform (20 mL) was added silver carbonate (790 mg, 2.9 mmol). The reaction system was allowed to react at 60°C for 72 hours. The residue was filtered and concentrated, and then separated by column chromatography (petroleum ether:ethyl acetate = 10:1) to afford the title compound 6A (400 mg, 47.9%). ¹ HNMR (400MHz, CHLOROFORM-d) δ 8.82 (d, J = 2.3Hz, 1H), 8.16 (s, 1H), 8.16-8.12 (m, 1H), 8.05 (s, 1H), 6.74 (d, J = 8.5Hz, 1H), 5.21 (br.s., 1H), 4.80-4.6 1(m, 1H), 4.48-4.27(m, 2H), 3.91(s, 3H), 2.02-1.63(m, 11H), 1.46-1.09(m , 14H), 1.02 (d, J=6.5Hz, 3H), 0.97 (s, 3H), 0.94-0.89 (m, 3H), 0.68 (s, 3H).

Example 22

To a solution of Example 6A (800 mg, 1.4 mmol) in methanol (10 ml) was added 10% sodium hydroxide solution (methanol:water ratio of 1:1 by volume) (600 mg, 15 mmol, 6 ml), and the reaction system was reacted at 70°C for 2 hours. The solvent was spin-dried, and the system was adjusted to pH 1-2 with dilute hydrochloric acid (1 M). The system was extracted with dichloromethane:methanol = 10:1 (20 ml x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative chromatography to give title compound 22 (460 mg, 64% yield). ¹ H NMR (400MHz, METHANOL-d ₄ )δ8.78 (d, J=1.8Hz, 1H), 8.21 (dd, J=2.3, 8.5Hz, 1H), 6.84 (d, J=8.8Hz, 1H), 4.56-4.23 (m, 2H), 3.68 (br .s., 1H), 3.35-3.34(m, 1H), 2.04-1.17(m, 25H), 1.07(d, J=6.5Hz, 3H), 0.96-0.89(m, 6H), 0.74(s, 3H).

Procedure for the Preparation of Reference Title Compound 22 in Examples 23-26

Route 7

Example 27

To a solution of Example 22 (100.0 mg, 194.7 μmol), DCC (60.3 mg, 292 μmol), and DMAP (23.8 mg, 194.7 μmol) in dichloromethane (5 mL) was added methylsulfonamide (37 mg, 389.3 μmol). The reaction was allowed to react at 25°C for 12 hours. Water (5 mL) was added, and the pH was adjusted to 2 with dilute hydrochloric acid (1 M). The mixture was extracted with dichloromethane (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative chromatography to afford Example 27 (25 mg, 21.7% yield). ¹ H NMR (400MHz, METHANOL-d ₄ ) δ8.74 (d, J=2.3Hz, 1H), 8.23 (dd, J=2.3, 8.8Hz, 1H), 7.06-6.87 (m, 1H), 4.57-4.40 (m, 2H), 3.67 (br s, 1H), 3.39 (s, 3H), 2.03-1.27 (m, 25H), 1.07 (br d, J=6.5Hz, 3H), 0.96-0.90 (m, 6H), 0.73 (s, 3H).

Procedure for the Preparation of Examples 28-34 Reference Title Compound 27

Route 8

Example 35

The synthesis of Example 35 was carried out using Example 11 as the starting material and the operation was referenced to the synthesis of Example 27, with a yield of 17.4%. ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 8.95-8.70 (m, 2H), 8.48 (s, 1H), 4.41-4.27 (m, 2H), 3.67 (br s, 1H), 3.41 (s, 3H), 3.33-3.32 (m, 1H), 2.02-1.16 (m, 25H), 1.08 (d, J=6.3 Hz, 3H), 0.94-0.88 (m, 6H), 0.74 (s, 3H).

Route 9

Example 36

The synthesis of Example 36 was carried out using Example 23 as the starting material and the operation was referenced to the synthesis of Example 27, with a yield of 17.5%. ¹ H NMR (400 MHz, CHLOROFORM-d) δ = 8.60 (d, J = 2.3 Hz, 1H), 8.18 (d, J = 2.3 Hz, 1H), 4.56-4.33 (m, 2H), 3.69 (br s, 1H), 3.45 (s, 3H), 1.96-1.15 (m, 25H), 1.01 (d, J = 6.5 Hz, 3H), 0.89-0.81 (m, 6H), 0.66 (s, 3H).

Route 10

Example 37

The synthesis of Example 37 was carried out using Example 23 as the starting material and with reference to the synthesis of Example 27. The yield was 60.8%. ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 8.55 (d, J = 1.8 Hz, 1H), 8.17 (d, J = 1.8 Hz, 1H), 4.57-4.38 (m, 2H), 3.87-3.75 (m, 2H), 3.66 (br s, 1H), 3.32-3.20 (m, 1H), 3.11 (t, J = 6.9 Hz, 2H), 2.06-1.23 (m, 25H), 1.06 (br d, J = 6.5 Hz, 3H), 0.96-0.86 (m, 6H), 0.71 (s, 3H).

Route 11

Example 38

The synthesis of Example 38 was carried out using Example 22 as the starting material and the operation was referenced to the synthesis of Example 27, with a yield of 53%. ¹ H NMR (400 MHz, METHANOL-d ₄ ) 8.63-8.62 (m, 1H), 8.09-8.06 (m, 1H), 6.84-6.82 (m, 1H), 4.39-4.28 (m, 2H), 3.82-3.74 (m, 3H), 3.40-3.30 (m, 1H), 3.12-3.08 (m, 2H), 1.99-1.20 (m, 25H), 1.06 (d, J=6.5 Hz, 3H), 0.94-0.90 (m, 6H), 0.73 (s, 3H).

Route 12

Example 39

Example 12A

A solution of glycine hydrochloride (14 mg, 112 μmol) and triethylamine (20.0 μL, 146 μmol) in ethyl acetate (5 mL) was reacted at 20°C for 0.5 h. Example 22 (50 mg, 97 μmol) and 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (36 mg, 146 μmol) were added to the system. The reaction system was reacted at 65°C for 10 h. Water (5 mL) was added to the system, and the aqueous phase was extracted with ethyl acetate (15 mL x 2). The organic phases were combined and washed sequentially with 0.5N aqueous sodium hydroxide solution (15 mL), water (15 mL), 0.5N hydrochloric acid (15 mL), and water (15 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and spun down to dryness. The crude product was purified by preparative separation (TFA) to afford Example 12A (25 mg, 43.9%). ¹ H NMR (400MHz, CDCl ₃ )8.66-8.65(m, 1H), 8.08-8.05(m, 1H), 6.81-6.67(m, 1H), 6.68(s, 1H), 4.42-4.24(m, 2H), 4.35-4.24(m, 2H), 3.81(s, 3H), 3.72-3.71(m, 1H), 3.46-3.44(m, 1H), 1.99-1.20(m, 25H), 1.03(d, J=6.5Hz, 3H), 0.92-0.88(m, 6H), 0.68(s, 3H).

Example 39

To a solution of Example 12A (25.0 mg, 42.7 μmol) in tetrahydrofuran (3 mL), methanol (1 mL), and water (2 mL) was added lithium hydroxide (8.9 mg, 213.7 μmol). The reaction was incubated at 30°C for 4 hours. The pH was adjusted to 5 with hydrochloric acid (1 M). The aqueous phase was extracted with ethyl acetate (20 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and spun down to dryness. The residue was separated by preparative TLC to afford Example 39 (24 mg, 98%). ¹ H NMR (400MHz, METHANOL-d ₄ )8.66-8.65(m, 1H), 8.11-8.02(m, 1H), 6.84-6.82(m, 1H), 4.48-4.29(m, 2H), 4.09-4.08(m, 3H) , 3.66-3.65 (m, 1H), 1.99-1.20 (m, 25H), 1.06 (d, J=6.5Hz, 3H), 0.92-0.88 (m, 6H), 0.72 (s, 3H).

Route 13

Example 40

Example 13A

Using Example 2B as the starting material, the operation of Example 13A was based on the synthesis of Example 6A, with a yield of 16.3%. ¹ H NMR (400 MHz, CHLOROFORM-d) δ 8.69 (d, J = 2.0 Hz, 1H), 8.21 (d, J = 2.0 Hz, 1H), 8.16 (s, 1H), 8.05 (s, 1H), 5.20 (br s, 1H), 4.72 (br s, 1H), 4.46-4.36 (m, 2H), 3.92 (s, 3H), 2.06-1.66 (m, 15H), 1.31-1.04 (m, 10H), 0.97-0.81 (m, 9H), 0.67 (s, 3H).

Example 40

The operation of Example 40 was carried out by referring to the synthesis of Example 22, with a yield of 96.4%. ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ=8.74 (d, J=2.0 Hz, 1H), 8.29 (d, J=1.8 Hz, 1H), 4.51 (t, J=6.4 Hz, 2H), 3.72 (br s, 1H), 3.43-3.39 (m, 1H), 2.11-1.23 (m, 27H), 1.07 (d, J=6.5 Hz, 3H), 1.00-0.93 (m, 6H), 0.77 (s, 3H).

Route 14

Example 14A

To a solution of Reference Example 1 (40.0 g, 95.1 mmol) in methanol (500 mL) was added p-toluenesulfonic acid (4.0 g, 21.0 mmol). The reaction was incubated at 70°C for 3 hours. The solvent was evaporated and the system was diluted with dichloromethane (500 mL). The organic phase was washed sequentially with saturated aqueous sodium carbonate solution (3 x 100 mL), aqueous solution (100 mL), and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Example 14A (41 g, 100% yield) was obtained without purification and used directly in the next step.

Example 14B

Example 14A (41 g, 95.1 mmol) was dissolved in anhydrous tetrahydrofuran (300 mL). The temperature was raised to 50°C under nitrogen, and a solution of methylmagnesium bromide (800 mL, 1 M) in tetrahydrofuran was slowly added dropwise. After the addition was complete, the reaction solution was cooled to room temperature and stirred overnight. After the reaction was completed, cyclohexane (25 mL) was added, filtered, and the residue was dissolved in a mixture of 3N aqueous hydrochloric acid (800 mL) and dichloromethane (200 mL). Stirring was continued for 30 minutes. After the reaction was completed, the organic phase was separated from the aqueous phase, and the aqueous phase was further extracted with dichloromethane (2 x 200 mL). The combined organic phases were washed sequentially with aqueous solution (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Example 14B (51 g, 100% yield) was obtained without purification and used directly in the next step.

Example 14C

Acetic anhydride (9.9 mL, 105.1 mmol), pyridine (1.6 mL, 19.8 mmol), and 4-dimethylaminopyridine (0.8 g, 6.6 mmol) were added to a solution of Example 14B (51 g, 95.1 mmol) in anhydrous tetrahydrofuran (300 mL). The reaction was stirred at room temperature overnight. After completion, the mixture was diluted with aqueous solution (100 mL). The aqueous phase was extracted with dichloromethane (3 x 150 mL). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Example 14C (55 g, 100% yield) was obtained without purification and used directly in the next step. 1H-NMR (CDCl ₃ ) δ0.66 (3H, s, CH ₃ -18); 0.77 (3H, s, CH ₃ -26); 1.00 (3H, d, CH ₃ -21); 1.20 (3H, s, CH ₃ -19); 1.96 (3H, s, AcO), 2.18-2.31 (1H, m, CH-22); 3.70 (1H, m, CH-7); 4.55 (1H, m, CH-3); 6.11 (1H, dd, = 6.2Hz, J ₂ =8.3Hz; CH-23); 7.14-7.36 (10H, m, Ph).

Example 14D

Sodium periodate (13.2 g, 61.9 mmol) was dissolved in water (13 ml) and 2N aqueous sulfuric acid (1.7 ml). After stirring for 15 minutes, the reaction solution was cooled to 0 degrees and ruthenium trichloride (71.3 mg, 0.4 mmol) was added. The reaction solution was stirred until the color changed to bright yellow. Ethyl acetate (27 ml) and acetonitrile (20 ml) were added to the reaction solution and stirred for 5 minutes. At 0 degrees, Example 14C (4 g, 6.9 mmol) was added to the above reaction solution. After the reaction was completed, the filtrate was filtered and poured into water and extracted with ethyl acetate (3x100 ml). The organic phases were combined and washed with saturated sodium thiosulfate (200 ml). The mixture was dried over anhydrous sodium sulfate and filtered. The filtrate was spin-dried and separated by silica gel column chromatography to give Example 14D (2.7 g, 89% yield). 1H-NMR (CDCl ₃ ) δ 0.71 (3H, s, CH ₃ -I8); 0.86-1.07 (9H, m, CH ₃ -19, CH ₃ -21, C-24); 2.03 (3H, s, AcO); 4.48-4.61 (1H, m, CH-3).

Example 14E

Triethylamine (6.7 mL, 3.4 mmol) was added to a solution of Example 14D (1 g, 2.2 mmol) and isobutyl chloroformate (3.5 mL, 2.7 mmol) in tetrahydrofuran (20 mL). The reaction was complete after 1 hour and the reaction solution was filtered. Sodium borohydride (847 mg, 22.4 mmol) was added portionwise to the filtrate at 0°C. After the reaction was complete, water (3 mL) was added to quench the reaction. Stirring was continued at room temperature for 2 hours and then acidified with 3N dilute hydrochloric acid aqueous solution and extracted with ethyl acetate (3 x 15 mL). The organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried and separated by silica gel column chromatography to give Example 14E (800 mg, 85% yield). 1H-NMR (CDCI ₃ ) δ0.67 (3H, s, CH ₃ -18); 0.86-0.97 (9H, m, CH ₃ -19, CH ₃ -21, CH ₃ -24); 2.03 (3H, s, AcO); 3.72 (3H, m, (2H, m, CH-7, CH-23); 4.48-4.61 (1H, m, CH-3).

Example 41

To a solution of Example 14E (100 mg, 230.1 μmol) in N,N-dimethylformamide (1 mL) was added sodium hydroxide (18.4 mg, 460.1 μmol, 60%) at zero degrees Celsius. Half an hour later, a solution of methyl-4-chloropyridine-2-carboxylate (78.9 mg, 460.1 μmol) in N,N-dimethylformamide (2 mL) was slowly added dropwise at zero degrees Celsius. After the addition was complete, the reaction system was slowly warmed to room temperature and allowed to react for 12 hours. Water (10 mL) was added, and the reaction system was adjusted to pH 6 with hydrochloric acid (1 M). The mixture was extracted with dichloromethane (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and spun down to dryness. A 10% sodium hydroxide solution (methanol:water volume ratio 1:1) (50 mg, 1.3 mmol, 0.5 mL) was added to a methanol solution of the residue (1 mL). The reaction system was allowed to react at 70°C for 2 hours. The solvent was evaporated, the system was adjusted to pH 1-2 with dilute hydrochloric acid (1 M), and extracted with dichloromethane:methanol = 10:1 (10 ml x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative chromatography to give Example 41 (5 mg, 4% yield). ¹ H NMR (400MHz, METHANOL-d ₄ ) δ 8.67 (br d, J=6.5Hz, 1H), 7.99 (d, J=2.5Hz, 1H), 7.69 (br d, J=6.5Hz, 1H), 4.50 (br d, J=6.5Hz, 2H), 3.78-3.59 (m, 1H), 3.32-3.30 (m, 1H), 2.18-1.17 (m, 25H), 1.10 (d, J=6.5Hz, 3H), 0.95-0.89 (m, 6H), 0.76 (s, 3H).

Route 15

Example 42

To a solution of Example 16D (100.0 mg, 245.9 μmol) and 5-aminopyridine-3-carboxylic acid (50.0 mg, 295 μmol) in DMF (5 mL) were added 1,3-dicyclohexylcarbodiimide (101.5 mg, 492 μmol) and 4-dimethylaminopyridine (1.5 mg, 12.3 μmol). The reaction was incubated at 30°C for 16 hours. A solution of lithium hydroxide (10.3 mg, 246 μmol) in water (2.0 mL) was added. The reaction was incubated at 30°C for 2 hours. The pH was adjusted to 4 with hydrochloric acid (1 M). The aqueous phase was extracted with dichloromethane/methanol (10:1) (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and spun down to dryness. The residue was purified by preparative separation (HCl) to afford Example 42 (20 mg, 15.4%). ¹ H NMR (400MHz, METHANOL-d ₄ )9.14-9.13(m,1H),8.90-8.89(m,1H),8.71-8.70(m,1H),3.68(s,1H),3.40-3.30(m,1H),2.64-2.61 (m, 1H), 2.15-2.06 (m, 2H), 1.99-1.20 (m, 25H), 1.07 (d, J=6.5Hz, 3H), 0.95-0.91 (m, 6H), 0.78 (s, 3H).

Route 16

Example 43

Example 16A

Reference Example 1F (10.0 g, 23.9 mmol) was dissolved in tetrahydrofuran (60.0 ml), and perchloric acid (240.0 mg, 2.4 mmol, 144.6 μL) (approximately 10 drops) was added. Formic acid (40.3 g, 874.7 mmol, 33.0 ml) was added dropwise at 30°C over half an hour. The reaction solution was stirred at 50°C for 11.5 hours. The solvent was removed by concentration, and water (35.0 ml) was added to the reaction solution, followed by extraction with ethyl acetate (30.0 ml x 3). The organic layer was washed with water (10.0 ml x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was separated by column chromatography to obtain Example 16A ( ^7.0 g, 15.7 mmol, 65.6% yield, white solid). NMR (400MHz, CHLOROFORM-d) δ = 8.01 (s, 1H), 4.86-4.73 (m, 1H), 2.82-2.67 (m, 1H), 2.47 -2.35(m, 2H), 2.33-2.16(m, 2H), 2.05-1.93(m, 2H), 1.89(d, J＝13.1Hz, 2H), 1.82(dd, J＝ 5.5, 16.8Hz, 2H), 1.75 (dd, J=6.5, 14.1Hz, 3H), 1.71 (br.s., 1H), 1.58-1.30 (m, 7H), 1. 26 (s, 3H), 1.23-1.02 (m, 4H), 0.95 (d, J = 6.5Hz, 3H), 0.83 (t, J = 7.4Hz, 3H), 0.68 (s, 3H).

Example 16B

Example 16A (5.8 g, 13.0 mmol) was dissolved in trifluoroacetic acid (40.0 mL) and trifluoroacetic anhydride (20.5 g, 97.4 mmol) at 0°C. After the solid dissolved, sodium nitrite (2.7 g, 39.0 mmol) was added portionwise. Stirring was continued at 0°C for one hour. The temperature was then raised to 40°C and stirring continued for another hour. The reaction mixture was cooled to 30°C and neutralized at 0°C with 0.5 M aqueous sodium hydroxide solution (pH = 7-8). The reaction mixture was extracted with ethyl acetate (40 mL x 3). The organic layer was washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by column chromatography (silica gel) to afford Example 16B (3.5 g, 8.5 mmol, 93.0% yield, as a light yellow oil). ¹ H NMR (400MHz, CHLOROFORM-d) δ = 3.59-3.47 (m, 1H), 2.69 (q, J = 6.2Hz, 1H), 2.42-2.31 (m, 2H), 2.29-2.15 (m, 2H), 2.01-1.88 (m, 2H), 1.86 -1.68 (m, 7H), 1.61-1.45 (m, 6H), 1.26 (t, J=7.2Hz, 5H), 1.19-1.12 (m, 5H), 1.02-0.91 (m, 1H), 0.80 (t, J=7.4Hz, 3H), 0.71-0.64 (m, 3H).

Example 16C

Example 16B (3.5 g, 8.5 mmol) was dissolved in methanol (100.0 mL) and aqueous potassium hydroxide (70.0 g, 1.3 mol, dissolved in 100.0 mL of water) was added. The reaction mixture was stirred at 100°C for 12 hours. The solvent was removed by concentration and the mixture was extracted with dichloromethane (30 mL x 3). The aqueous phase was acidified with monomolar hydrochloric acid (pH = 3-4) and extracted with ethyl acetate (30 mL x 3). The organic layer was washed with water (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude product. No purification was required to afford Example 16C (3.2 g, 7.9 mmol, 93.5% yield, yellow oil). ¹ H NMR (400MHz, CHLOROFORM-d) δ = 3.65-3.50 (m, 1H), 2.71 (d, J = 5.8Hz, 1H), 2.48 (dd, J = 2.6, 14.9Hz, 1H), 2.43-2.31 (m, 2H), 2.22-2.15 (m, 1H), 2.07-1 .98(m,2H),1.95-1.86(m,3H),1.82-1.70(m,6H),1.53-1.46(m,3H),1.19 -1.10 (m, 6H), 1.02 (d, J=6.3Hz, 3H), 0.86 (d, J=10.3Hz, 5H), 0.69 (s, 3H).

Example 16D

Example 16C (3.2 g, 7.9 mmol) was added to an aqueous sodium hydroxide solution (949.2 mg, 23.7 mmol, dissolved in 10.00 mL of water). The reaction solution was heated to 80°C, and sodium borohydride (1.8 g, 47.5 mmol) was added portionwise. The reaction solution was stirred at 100°C for 12 hours. Methanol (6 mL) was added dropwise, and the solvent was partially removed by concentration. The reaction solution was acidified with monomolar hydrochloric acid (pH = 5-6) and extracted with ethyl acetate (40 mL x 3). The organic layer was washed with water (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude product, Example 16D (3.1 g, 7.6 mmol, 96.4% yield), as a white solid without isolation. ¹ H NMR (400MHz, CHLOROFORM-d) δ = 3.70 (br.s., 1H), 3.46-3.36 (m, 1H), 2.52-2.39 (m, 1H), 2.02-1.88 (m, 3H), 1.85-1.77 (m, 4H), 1.71-1.59 (m, 3 H), 1.53-1.44(m, 4H), 1.41-1.37(m, 1H), 1.36-1.27(m, 4H), 1.24-1.1 3(m, 4H), 1.04(d, J=6.5Hz, 3H), 0.92-0.88(m, 6H), 0.73-0.69(m, 3H).

Example 16E

To a solution of Example 16D (100 mg, 245.8 μmol) in acetonitrile (2 mL) was added triethylamine (49.8 mg, 491.9 mmol, 68.2 μL) and N,O-dimethylhydroxylamine hydrochloride (24.0 mg, 245.9 μmol) under nitrogen. The mixture was stirred at 25°C for 30 minutes, followed by the addition of O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (98.7 mg, 307.4 μmol), and stirring was continued for 11.5 hours. The reaction mixture was poured into cold water (30 mL) and extracted with ethyl acetate (40 mL x 3). The organic layer was washed with water (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford the crude product, Example 16E (90 mg, 91.4% yield), as a white solid. ¹ H NMR (400MHz, CHLOROFORM-d) δ = 3.76 (s, 3H), 3.70 (br.s., 1H), 3.46-3.36 (m, 1H), 2.79 (s, 3H) , 2.43-2.39 (m, 1H), 2.24-2.18 (m, 1H), 2.00-1.97 (m, 41H), 1.03-0.88 (m, 10H), 0.73 (s, 3H).

Example 16F

To a solution of lithium aluminum tetrahydride (6.3 mg, 168.9 μmol) in tetrahydrofuran (2 mL) was added Example 16E (50 mg, 111.2 μmol) at -78 °C, and the mixture was stirred at -78 °C for 2 hours. After the reaction, water (0.006 mL) was added, the reaction solution was filtered, and the filter cake was dried to give Example 16F (50 mg, 100%, white solid). ¹ HNMR (400MHz, CHLOROFORM-d) δ = 9.76 (s., 1H), 3.70 (br.s., 1H), 3.46-3.36 (m, 1H), 2.4 3-2.39 (m, 1H), 2.24-2.18 (m, 1H), 2.00-1.97 (m, 41H), 1.03-0.88 (m, 10H), 0.73 (s, 3H).

Example 16G

To a solution of Example 16F (150.0 mg, 0.3 mmol) and methyl 5-aminopyridine-3-carboxylate (64.0 mg, 0.4 mmol) in ethyl acetate (5 mL) were added trifluoroacetic acid (57 μL, 0.8 mmol) and sodium triacetylborohydride (146.0 mg, 0.7 mmol). The reaction system was reacted at 20°C for 16 hours. Ethyl acetate (30 mL) was added to the reaction system, and the system was washed with saturated sodium bicarbonate (20 mL x 2) and saturated brine (20 mL x 1). The organic layer was dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product. The crude product was subjected to preparative separation (TFA) to afford Example 16G (60.0 mg, 29.7%). ¹ H NMR (400MHz, CHLOROFORM-d) 8.44-8.43 (m, 1H), 8.43-8.42 (m, 1H), 7.92-7.91 (m, 1H), 4.01 (s, 3H), 3.72-3.71 (m, 1H) ), 3.49-3.48(m, 1H), 3.28-3.13(m, 2H), 1.99-1.20(m, 25H), 1.01(d, J=6.5Hz, 3H), 0.95-0.87(m, 6H), 0.67(s, 3H).

Example 43

To a solution of Example 16G (60.0 mg, 113.9 μmol) in tetrahydrofuran (2 mL), methanol (1 mL), and water (2 mL) was added lithium hydroxide (60.0 mg, 1.43 mmol). The reaction was incubated at 40°C for 1 hour. The pH was adjusted to 4 with hydrochloric acid (1 M). The aqueous phase was extracted with dichloromethane/methanol (10:1) (20 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and spin-dried. The residue was purified by preparative separation (HCl) to afford Example 43 (40 mg, 68%). ¹ H NMR (400MHz, METHANOL-d ₄ )8.30-8.29(m, 1H), 8.03-8.02(m, 1H), 7.59-7.58(m, 1H), 3.70-3.65(m, 1H), 3.40-3.30(m, 1H) , 3.12-3.07(m, 2H), 1.99-1.20(m, 25H), 1.05(d, J=6.5Hz, 3H), 0.95-0.87(m, 6H), 0.72(s, 3H).

Route 17

Example 44

To a solution of Example 22 (50.0 mg, 97.3 μmol) in methanol (5 mL) was added concentrated sulfuric acid (368.0 mg, 3.8 mmol). The reaction was allowed to react at 70°C for 12 hours. Water (5 mL) was added, and the pH was adjusted to 7 with sodium hydroxide (1 M). The mixture was extracted with dichloromethane (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by preparative chromatography to afford title compound 44 (15 mg, 29.2% yield). ¹ H NMR (400MHz, METHANOL-d ₄ ) δ8.76 (d, J=2.0Hz, 1H), 8.18 (dd, J=2.5, 8.8Hz, 1H), 6.82 (d, J=8.8Hz, 1H), 4.49-4.28 (m, 2H), 3.89 (s, 3H), 3.65 (br s, 1H), 3.28 (br s, 1H), 2.03-1.15 (m, 25H), 1.04 (d, J=6.5Hz, 3H), 0.93-0.86 (m, 6H), 0.71 (s, 3H)

Route 18

Example 45

Example 18A

The synthesis of Example 18A was carried out using Example 23 as the raw material and the operation was reference to the synthesis of Example 12A, with a yield of 79.7%, 1HNMR (400 MHz, CHLOROFORM-d) δ8.48 (d, J=2.3 Hz, 1H), 8.08 (d, J=2.3 Hz, 1H), 7.26 (s, 1H), 6.55 (t, J=4.8 Hz, 1H), 4.57-4.38 (m, 2H), 4.24 (d, J=5.0 Hz, 2H), 3.81 (s, 3H), 3.71 (s, 1H), 3.46-3.36 (m, 1H), 2.05-1.12 (m, 33H), 1.06-0.84 (m, 14H), 0.68 (s, 3H).

Example 45

The operation of Example 45 was carried out with reference to the synthesis of Example 39, with a yield of 56.8%. 1H NMR (400 MHz, METHANOL-d4) δ 8.80-8.72 (m, 1H), 8.48 (d, J = 2.0 Hz, 1H), 8.10 (d, J = 2.0 Hz, 1H), 4.50-4.33 (m, 2H), 3.99 (d, J = 4.5 Hz, 2H), 3.56 (s, 1H), 1.98-1.03 (m, 31H), 1.01-0.87 (m, 5H), 0.84-0.74 (m, 10H), 0.62 (s, 3H).

Example 1: In vitro evaluation

FXR biochemical assay

Purpose of the experiment:

The FXR binding activation of the compounds was detected by homogeneous luminescence proximity amplification assay (alphascreen).

Experimental Materials:

1. Protein: Glutathione-S-transferase-labeled FXR human protein (Invitrogen)

2. Coactivator: Biotinylated steroid receptor coactivator (Anaspec)

3. Detection reagent: Homogeneous proximity luminescence amplification assay (alphascreen) detection kit (PerkinElmer)

Experimental methods:

1. Compound Dilution: Prepare the test compound as a 40 μM solution in DMSO and then dilute it 3-fold to a total of 10 concentration points. Prepare the reference compound as a 400 μM solution in DMSO and then dilute it 1.5-fold to a total of 10 concentration points. Add 150 nL of the diluted DMSO solution to each well of a 384-well plate.

2. Prepare a mixture of glutathione-S-transferase-labeled FXR human protein and biotin-labeled steroid receptor coactivator at concentrations of 0.4 nM and 30 nM, respectively. Add 15 μL per well to each well of a 384-well plate. Incubate at room temperature for 1 hour.

4. Dilute the receptor bead mixture from the AlphaScreen assay kit 125-fold and add 7.5 μL per well to the microwells of a 384-well plate. Protect from light during the experiment. Incubate at room temperature for 1 hour.

5. Dilute the donor bead mixture from the AlphaScreen assay kit 125-fold and add 7.5 μL per well to each well of a 384-well plate. Protect from light during the experiment. Incubate at room temperature for 1 hour.

6. EC50 test: Use Envision to excite at 680nm and read the absorbance signal at 520-620nm.

7. Data Analysis: Prism 5.0 was used to analyze the data and calculate the EC50 value of the compound's activation effect. The maximum signal value of the compound was then compared with the maximum signal value of the reference compound to obtain the compound's activation efficacy percentage.

FXR cell experiments

Purpose of the experiment:

The effects of compounds on cell function activity were detected by β-lactamase reporter gene technology.

Experimental Materials:

1. Cell line: FXR HEK 293T DA

2. Cell culture medium: DMEM medium supplemented with 10% serum and Penicillin/Streptomycin (1×)

3. Detection reagent: Reporter gene detection kit (Invitrogen)

Experimental methods:

1. Compound Dilution: Prepare the test compound as a 100 μM solution in DMSO and then dilute it 3-fold to a total of 10 concentration points. Prepare the reference compound as a 100 μM solution in DMSO and then dilute it 1.3-fold to a total of 10 concentration points. Add 200 nL of the diluted DMSO solution to each well of a 384-well plate.

2. Cell seeding: Resuscitate FXR HEK 293T DA cells, resuspend in culture medium, dilute to a density of 5 × 10 ⁵ cells/mL, and add 40 μl per well to the microwells of a 384-well plate.

3. Incubate the 384-well microplate at 37°C, 5% CO2 for 16 hours.

4. Mix 6 μL of 1 mM LiveBLAzer ^™ -FRET B/G (CCF4-AM) substrate, 60 μL of Solution B, and 934 μL of Solution C, and add the resulting mixture to the wells of a 384-well plate at a volume of 8 μL per well.

5. Incubate the 384-well microplate in the dark at room temperature for 2 hours.

6. EC50 test: Use Envision to excite at 409 nm and read the absorbance signals at 460 and 530 nm.

7. Data Analysis: Prism 5.0 was used to analyze the data and calculate the EC50 value of the compound's activation effect. The maximum signal value of the test compound was then compared with the maximum signal value of the reference compound (chenodeoxycholic acid, CDCA) to obtain the compound's activation efficacy percentage.

Table 1 Biochemical test results and cell test results EC ₅₀ :

Conclusion: The compounds of the present invention have significant agonist effects on FXR receptors, and also have relatively significant agonist effects on FXR receptors at the cellular level.

Experimental Example 2: In vivo study

Pharmacokinetics of mice given a single dose:

Twelve C57BL/6J male mice were randomly divided into two groups, six animals each. The first group received intravenous injection of 2 mg/kg (2 mL/kg) of the drug via tail vein (using a 10% HPbCD aqueous solution as the vehicle; a cosolvent was added if drug solubility was unsatisfactory). The second group received oral gavage of 10 mg/kg (10 mL/kg) of the drug (using a 0.5% HPMC aqueous solution as the vehicle). Plasma samples (anticoagulant _K₂ -EDTA) were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration from the intravenous group; and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration from the oral group. Each group consisted of six animals, with three blood samples collected at each time point, with samples from the first batch of three animals interleaved with samples from the second batch of three animals. Plasma samples were analyzed using LC-MS/MS. Plasma concentrations were obtained and plotted against time, and PK parameters were calculated using Phoenix WinNonlin 6.3.

Table 2

Conclusion: As shown in Table 2, after oral administration of the same dose, compound 11 had a higher peak concentration and drug exposure than the control compound INT-747. After oral administration of the same dose, compound 22 had a peak concentration close to that of the control compound INT-747, and drug exposure was higher than that of the control compound INT-747. After oral administration of the same dose, compound 23 had a higher peak concentration and drug exposure than the control compound INT-747.

Cassette-type drug administration mouse liver-blood ratio experiment:

Six C57BL/6J male mice were orally administered with a formulation containing each of the five investigational drugs (2 mg/kg/compound) via gavage in a 0.5% HPMC aqueous solution. Each compound was first dissolved in the vehicle and sonicated or vortexed to a 1 mg/mL solution (either as a clear solution or suspension). The five compound solutions were then mixed in equal volumes (1:1:1:1:1, v:v:v:v:v) in a single glass vial. Following oral administration, plasma and liver tissue samples were collected from three animals 0.5 hours after dosing; from three other animals 3 hours after dosing. Liver tissue was then homogenized in ice-cold homogenization buffer (methanol: 15 mM PBS (pH 7.4) = 1:2, v:v) at a ratio of liver weight to homogenization buffer volume of 1:3. Plasma and liver tissue samples were analyzed using a previously developed five-in-one LC-MS/MS method. Plasma concentrations and liver tissue homogenate concentrations were obtained, and the liver tissue to plasma concentration ratio was calculated using Excel.

Table 3

Note: ND means not detected.

Conclusion: As shown in Table 3, after oral administration of the same dose of the compounds of the present invention, Example 22 had higher liver concentrations at 0.5 and 3 hours than the control compound, and its liver/blood concentration ratios at 0.5 and 3 hours were also higher than the control compound. Example 26 had higher liver concentrations at 0.5 hours than the control compound, and its liver/blood concentration ratios at 0.5 and 3 hours were also higher than the control compound.

Claims (28)

1. The compound represented by formula (Ⅰ) or a pharmaceutically acceptable salt thereof, in, Ring A is selected from 5-12 aryl groups, 5-12 heteroaryl groups containing 1-2 heteroatoms, 5-6 non-aromatic heterocyclic groups containing 1-2 heteroatoms, or 5-6 cycloalkyl groups, wherein ring A is optionally substituted with 1, 2, or 3 _Ra atoms, and the heteroatoms are selected from N, O, or S. L is selected from _C1-4 alkyl, _C1-4 alkoxy, _C1-4 alkylthio, _C1-4 alkylamino, or _C2-4 alkylenyl, wherein L is optionally substituted with 1, 2, or 3 _Rb ; _L1 is selected from O, N(R _d ), N(R _d )S(＝O) ₂ or N(R _d )S(＝O); _R1 is selected from H, _C1-6 alkyl, 5-6 aryl, or _C3-6 cycloalkyl, wherein the _C1-6 alkyl, 5-6 aryl, or _C3-6 cycloalkyl may optionally be substituted by 1, 2, or 3 _Rc . _Ra is selected from F, Cl, Br, I, OH, CN, NO ₂ , or NH ₂ ; _Rb is selected from F, Cl, Br, I, OH, CN, _NO2 , _NH2 , or _C1-3 alkyl groups; R _{_c} is selected from F, Cl, Br, I, OH, CN, NO _₂ , NH _₂ , COOH, or S(=O)_₂OH; R _d is selected from H or _C1-3 alkyl groups. 2. The compound or its pharmaceutically acceptable salt according to claim 1, wherein _Ra is selected from F, Cl, Br, or I. 3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein _Rb is selected from _C1-3 alkyl groups. 4. The compound or its pharmaceutically acceptable salt according to claim 1, wherein _Rc is selected from COOH or S(=O) _₂OH . 5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein _Rd is selected from H. 6. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from phenyl, pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thiophene, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, isoxazolyl, isothiazolyl, bicyclo[1.1.1]pentyl, benzoxazolyl, benzo[d]isoxazolyl, indazole, indolyl, quinolinyl, isoquinolinyl, quinazolinyl, 1H-pyrrolo[2,3-B]pyridinyl, indene, benzothiazolyl, or benzothiaphene, wherein ring A is optionally substituted with 1, 2, or 3 _Ra . 7. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from a benzene ring, or a 5- or 6-membered heteroaromatic ring containing 1 to 2 heteroatoms selected from N, O or S, and ring A is optionally substituted with 1, 2 or 3 Ra _atoms . 8. The compound of claim 7 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from a benzene ring, a 5-membered heteroaromatic ring containing 1 to 2 atoms selected from N, O or S, or a 6-membered heteroaromatic ring containing 1 to 2 atoms selected from N, wherein ring A is optionally substituted with 1, 2 or 3 Ra _atoms . 9. The compound of claim 8 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from benzene ring, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, pyridine, pyridazine, pyrimidine or pyrazine, and ring A is optionally substituted by 1, 2 or 3 _Ra . 10. The compound of claim 9 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from benzene rings, pyrazoles, isoxazoles, pyridines, or pyrimidines, and ring A is optionally substituted with 1, 2, or 3 _Ra . 11. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from ring A optionally substituted with 1, 2 or 3 _Ra . 12. The compound of claim 11 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from... 13. The compound of claim 12 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from... 14. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein L is selected from the compound whose L is optionally substituted with 1, 2 or 3 R _b . 15. The compound of claim 14 or a pharmaceutically acceptable salt thereof, wherein L is selected from... 16. The compound of claim 15 or a pharmaceutically acceptable salt thereof, wherein L is selected from... 17. The compound of claim 16 or a pharmaceutically acceptable salt thereof, wherein L is selected from... 18. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein _L1 is selected from O, NH, NHS (=O) ₂ or NHS (=O). 19. The compound of claim 18 or a pharmaceutically acceptable salt thereof, wherein _L1 is selected from O, NH or NHS (＝O) ₂ . 20. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein _R1 is selected from H, _C1-3 alkyl, _C3-6 cycloalkyl, or phenyl, wherein the _C1-3 alkyl, _C3-6 cycloalkyl, or phenyl is optionally substituted with 1, 2, or 3 _Rc . 21. The compound of claim 20 or a pharmaceutically acceptable salt thereof, wherein _R1 is selected from H, Me, 22. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural fragment _{L1 - R1} is selected _{from -OH, -NHSO2CH3, -NHSO2C(CH3)3 , -NHCH2CH2SO2OH , -NHCH2COOH ,} -NHSO2CH( _CH3 ) _{2 , -NHCH3} , _{-OCH3 , or} 23. The compound according to claim 1, wherein the compound is selected from: 24. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 25. The use of the compound of any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating farnesoid X receptor-related disorders. 26. The use of the compound of any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating chronic liver disease, fibrotic disease, hypercholesterol disease, hypertriglyceridemia and cardiovascular disease. 27. The use of the compound of any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 24, in the preparation of a medicament for treating liver dysfunction caused by non-alcoholic fatty liver disease, cholestatic liver disease, hepatitis C infection, alcoholic liver disease, liver fibrosis, primary sclerosing cholangitis, gallstones, bile duct atresia, lower urinary tract symptoms and benign prostatic hyperplasia, ureteral stones, obesity, type 2 diabetes, arteriosclerosis, hypercholesterolemia, and hyperlipidemia. 28. The use of the compound of any one of claims 1 to 23 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 24 in the preparation of a medicament for treating non-alcoholic steatohepatitis, primary biliary cirrhosis and atherosclerosis.