EA038580B1 - Steroid derivative fxr agonist - Google Patents

Abstract

The invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, and relates to applications thereof in the preparation of drugs for treating FXR related diseases.

Description

The technical field to which the invention relates

The present invention relates to a compound represented by formula (I), a tautomer or a pharmaceutically acceptable salt thereof, and relates to its use for the preparation of medicaments for the treatment of diseases associated with FXR.

Background of the invention

The farnesoid X receptor (FXR) is an orphan nuclear receptor originally identified from the rat liver cDNA library (V.M. Forman, et al., Cell, 81: 687-693 (1995)) that is most closely related to the ecdysone receptor insect. FXR is a member of the ligand-activated transcription factor nuclear receptor family that includes receptors for steroid, retinoid and thyroid hormones (D. J. Mangelsdorf, et al., Cell, 83: 841850 (1995)). Northern blot analysis and in situ analysis show that FXR is highly expressed in the liver, intestinal tract, kidneys and adrenal gland (V.M. Forman et al., Cell, 81: 687-693 (1995) and W. Seol et al., Mol. Endocrinnol. 9: 72-85 (1995)). FXR binds to DNA after heterodimer formation with the 9-cis retinoic acid receptor (RXR). Heterodimer FXR / RXR preferentially binds to elements that consist of two hemisites of the nuclear receptor with the consensus sequence AG (G / T) TCA, which is formed as an inverted repeat and is separated by a single nucleotide (IR-1 motif) (V.M. Forman, et al., Cell 81: 687-693 (1995)). However, such compounds do not activate mouse and human FXRs, so the nature of the endogenous FXR ligand remains uncertain. Several naturally occurring bile acids bind and activate FXR at physiological concentrations (PCT WO 00/37077, published June 29, 2000). As discussed herein, bile acids that act as ligands for FXR include chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), and tauric and glycine conjugates of such bile acids.

WO-2005082925 discloses the use of INT747 for the preparation of drugs for the treatment of diseases associated with FXR o

INT-747

Brief description of the invention

An object of the present invention is to provide a compound represented by formula (I)

where ring A is selected from 5-12 membered aryl, 5-12 membered heteroaryl containing from 1 to 2 heteroatoms, 5-6 membered non-aromatic heterocyclyl containing from 1 to 2 heteroatoms, or 5-6 membered cycloalkyl, and wherein said ring A is optionally substituted with 1, 2 or 3 R _a or 1, 2 or 3 oxo groups, and wherein said heteroatom is selected from N, O or S;

L is selected from C _1-4 αalkyl, C _1-4 α alkoxy, C _1-4 alkylthio, C _1-4 alkylamino, C _1-4 alkyl-C (= O) NH-, C _2-4 α-alkenyl, or C _{1- 3} alkyl-O-C _1-3 α-alkyl, and wherein said L is optionally substituted with 1, 2 or 3 Rb;

L1 is selected from O, N (R), N (Rd) S (= O) 2, or N (R) S (= O);

R1 is selected from H, C _1-6 αalkyl, C _1-6 heteroalkyl containing one or more heteroatoms selected from sulfur, oxygen and / or nitrogen, 5-6 membered aryl, 4-6 membered heteroaryl containing one or more heteroatoms selected from sulfur, oxygen and / or nitrogen, C _3-6 cycloalkyl or 3-6 membered heterocycloalkyl containing one or more heteroatoms selected from sulfur, oxygen and / or nitrogen, and wherein said C _1-6 alkyne, C _1-6 heteroalkyl, 5-6 membered aryl, 4-6 membered heteroaryl, C ₃₆ cycloalkyl or 3-6 membered heteropycloalkyl are optionally substituted with 1, 2 or 3 Rc or 1, 2 or 3 oxo groups;

each of Ra, Rb, Rc and Rd is independently selected from H, 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, and wherein said C _1-3 alkylamino, N, N-di (C _1-3 alkyl) amino, C _1-3 alkyl, C _1-3 alkyloxy, or C _1-3 α alkylthio optionally

- 1,038580 are 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 ) or N (CH ₃ ) ₂ ;

or a pharmaceutically acceptable salt thereof.

In some embodiments, implementation of the present invention in the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, each of the above R _a , Rb, R _c and Rd is independently selected from H, F, Cl, Br, I, COOH, S (= O) ₂ OH, Me, CF ₃ , CHF ₂ , CH ₂ F, Et, OMe, NH (CH ₃ ) or N (CH3) 2.

In some embodiments, in the compound represented by formula (I) or in a pharmaceutically acceptable salt thereof, ring A is selected from phenyl, pyridyl, pyridin-2 (1H) -onyl, pyrimidyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, isoxazolyl, isothiazolyl, bicyclo [1.1.1] pentyl, benzoxazolyl, benzo [d] isoxazolyl, indazolyl, indolyl, quinolinyl, isoquinolinyl] 2,3 pyridyl, indolizinyl, benzothiazolyl or benzothienyl, and wherein said ring A is optionally substituted with 1, 2 or 3 R _a .

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

or wherein said Ring A is optionally substituted with 1, 2 or 3 Ra.

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

- 2 038580

In some embodiments, in the compound represented by formula (I) or in a pharmaceutically acceptable salt thereof, said 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 alkenyl, or C _1-3 alkylO-C _1-3 alkyl, and wherein said L is optionally substituted with 1, 2 or 3R _b .

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

H .N '' and wherein said L is optionally substituted with 1, 2 or 3 R _b .

In some embodiments, implementation of the present invention in the compound represented by the agent of formula (I), or in its pharmaceutically acceptable salt, L is selected from

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

In some embodiments, implementation of the present invention, the above L1 is selected from O, NH, NHS (= O) 2, or NHS (= O).

In some embodiments, implementation of the present invention in the compound represented by formula (I), or in a pharmaceutically acceptable salt thereof, the above R1 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 wherein said C _1-3 alkyl, C ₃₆ cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, or thienyl 1 , 2 or 3 Rc, and wherein said Rc is as defined above.

In some embodiments, implementation of the present invention in the compound represented by formula (I), or in a pharmaceutically acceptable salt thereof, the above R1 is selected from H,. , ^ COOH Me,

0 ^he or

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

where

L2 is selected from C _1-6 alkyl, C _1-6 heteroalkyl, or C _2-6 alkenyl;

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

said ring B is optionally substituted with 1, 2, or 3 R ₂ or 1 oxo, and wherein said R2 is selected from F, Cl, Br, I, OH, CN, NO2, NH2, COOH, S (= O) ₂ OH, C _1-3 alkylamino, N, N-di (C _1-3 alkyl) amino, C _1-3 alkyl, C _1-3 lyloxy or C _1-3 alkylthio;

- 3 038580

L1 is selected from O, NH, NHS (= O) 2, or NHS (= O);

R1 is selected from H, C1-6alkyl, S1-6geteroalkila, 5-6-membered aryl, 4-6 membered heteroaryl, C _3-6 cycloalkyl or a 3-6-membered heterocycloalkyl, and wherein said 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 are optionally substituted with 1, 2, or 3 Rc or 1, 2 or 3 oxo groups, and wherein said Rc is selected from F, Cl, Br, I, OH, CN, NO2, NH2, COOH, or S (= O) 2OH;

or a pharmaceutically acceptable salt thereof.

In some embodiments, implementation of the present invention in the compound represented by formula (II), or in its pharmaceutically acceptable salt L2, 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 -, and wherein said X is selected from NH, O or S, and wherein said Y is selected from -CH = CH-, provided that the chain length L _{2 is} selected from values from 1 to 6, preferably from 2 to 4.

In some embodiments, the compound represented by formula (II) or a pharmaceutically acceptable L2 salt thereof is selected from -CH2CH2-, -CH2CH2CH2-, -CH = CH-, -CH2CH = CH-, -CH = CHCH2- , -CH2CH2O-, -CH2CH2CH2O, -CH2CH2NH-, -CH2CH2CH2NH-, -CH2CH2S-, or -CH2CH2CH2S-, preferably from -CH2CH2-, -CH2CH2CH2-, -CH2 = CH-, -CH2CH2CH2- CH2 = CH-, -CH2CH2CH2 -.

In some embodiments, in the compound represented by formula (II) or in a pharmaceutically acceptable salt thereof, ring B is selected from a benzene ring, a 5-membered heteroaromatic ring containing 1 to 2 heteroatoms selected from N, O, or S, or a 6-membered heteroaromatic ring containing 1 to 2 heteroatoms selected from N, wherein said Ring B is optionally substituted with 1, 2, or 3 R2 or 1 oxo, and wherein said R2 is as defined above.

In some embodiments of the present invention, in the compound represented by formula (II) or in a pharmaceutically acceptable salt thereof, ring B is selected from benzene ring, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, or pyrazinyl, and wherein said Ring B is optionally substituted with 1,2 or 3 R ₂ or 1 oxo, and wherein said R ₂ is as defined above.

In some embodiments, implementation of the present invention in a compound represented by formula (II), or in a pharmaceutically acceptable salt thereof, said ring B is optionally substituted with 1 R2 or 1 oxo, and wherein said R2 is selected from F, Cl, Br, I, OH, CN, NO2, NH2, COOH or S (= O) ₂ OH, preferably from F, Cl or Br.

In some embodiments, implementation of the present invention in the compound represented by formula (II), or in a pharmaceutically acceptable salt thereof, the R ₂ -substituted or oxo-substituted position is in the ortho position relative to L2 on ring B (α-position).

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

In some embodiments, implementation of the present invention in the compound represented by formula (II), or in a pharmaceutically acceptable salt thereof, said R1 is selected from H, Me,

I I '- COOH, D. ,, t> U or

In some embodiments, implementation of the present invention in the compound represented by formula (II), or in a pharmaceutically acceptable salt thereof, the structural fragment L1-R1 is selected from

-OH, -NHSO ₂ CH ₃ , n

-N-so ₂ ^ q, -NHSO ₂ C (CH ₃ ) ₃ , -nhch ₂ ch ₂ so ₂ oh, -nhch ₂ cooh, n / = \ n n / = \ -N-SO ₂ - ^ b _N - <AND - ^Ν —Y #, -NHSO ₂ CH (CH ₃ ) ₂ ,, -NHCH ₃ , -OCH ₃ or.

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

- 4 038580

and wherein said L2 and R2 are as defined above.

In some embodiments, implementation of the present invention in the compound represented by formula (II), or in

its pharmaceutically acceptable salt, the structural fragment is selected from

- 5 038580

The present invention also provides a compound represented by formula (III)

where Ring A is 5-12 membered aryl, 5-12 membered heteroaryl, or 5-6 membered non-aromatic heterocyclyl, 5-6 membered cycloalkyl optionally substituted with 1, 2 or 3 R;

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

R is F, Cl, Br, I, OH, CN, NO2, NH2, or R is selected from C _1-3 alkylamino, Ν, Ν-di

- 6 038580 (C1-3alkyl) amino, C _1-3 αlkila, C _1-3 alkyloxy, C _1-3 alkylthio optionally substituted by 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 (CH3) 2;

said hetero is a heteroatom or a heteroatom group selected from -C (= O) NH-, N, -NH -, - C (= NH) -, -S (= O) ₂ NH-, -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;

or a pharmaceutically acceptable salt thereof.

In some embodiments, implementation of the present invention in the compound represented by formula (III), or in a pharmaceutically acceptable salt thereof, the above R is selected from F, Cl, Br, I, Me, CF3, CHF2, CH2F, Et, OMe, NH (CH3) , N (CH3) 2.

In some embodiments, implementation of the present invention in the compound represented by formula (III), or in a pharmaceutically acceptable salt thereof, the above ring A is phenyl, pyridyl, pyridin-2 (III) -onyl, pyrimidyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, isoxazolyl, isothiazolyl, bicyclo [1.1.1] pentyl, benzoxazolyl, benzo [b] isoxazolyl, indazolyl, indolyl, quinolinyl, isoquinolinyl, quinazolinyl, quinazolinyl pyridyl, indolizinyl, benzothiazolyl and benzothienyl optionally substituted with 1, 2 or 3 R.

In some embodiments, in the compound represented by Formula (III) or in a pharmaceutically acceptable salt thereof, Ring A is selected from optionally substituted 1, 2, or 3 R.

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

- 7 038580

In some embodiments, implementation of the present invention in the compound represented by formula (III), or in a pharmaceutically acceptable salt _{thereof, the above L is selected from C 1} _ ₄ α alkyl, C ₁ . ₄ alkoxy, C ₁ . ₄ alkylthio, C ₁ . ₄ alkylamino, C ₁ _ ₄ α-alkyl-C (= O) NH-, C ₂ . ₄ alkenyl optionally substituted with 1, 2 or 3 R.

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

'Optionally substituted with 1, 2 or 3 R.

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

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

The compound of the present invention is selected from

- 8 038580

- 9 038580

- 10 038580

The present invention also provides a pharmaceutical composition for the treatment of diseases associated with the farnesoid X-receptor, containing a therapeutically effective amount of the above compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The present invention also provides for the use of the above compound or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition for the preparation of a medicament for the treatment of diseases associated with the farnesoid X receptor, fibrotic diseases, chronic liver disease, diseases associated with high cholesterol levels, diseases associated with high triglyceride levels, or cardiovascular disease.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition for the preparation of a medicament for the treatment of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), cholestatic hepatopathy, chronic liver disease, hepatitis C virus 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, diabetes Type 2, atherosclerosis, arteriosclerosis, liver dysfunction caused by hypercholesterolemia or hyperlipidemia.

The compound of the present invention is an FXR agonist. The compound of the present invention can be used for methods of preventing or treating dyslipidemia or diseases associated with dyslipidemia, including administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof.

The compound of the present invention can be used to lower total cholesterol, lower LDL cholesterol, lower VLDL cholesterol, lower HDL cholesterol, and / or lower triglycerides. A decrease in triglyceride levels in accordance with the present invention means that the triglyceride level in subjects in need thereof is reduced below the baseline triglyceride level in subjects to be prevented or treated prior to taking the compounds of this application. For example, the compound of the present invention can reduce liver triglyceride production or liver triglyceride secretion by decreasing fat absorption. The compound of the present invention can also reduce the amount of triglycerides in the blood serum and the amount of triglycerides in the liver.

The compound 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, in particular a human), such as, without limitation, atherosclerosis, arteriosclerosis, hypercholesterolemia, hyperlipidemia, thrombosis, coronary artery disease, stroke or hypertension.

The compound of the present invention can be used to prevent or treat diseases of the liver / biliary system in a subject (e.g., a mammal, in particular a human), such as, without limitation, diseases associated with high cholesterol levels, diseases associated with high triglycerides, or fibrotic diseases such as, but not limited to, non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), gallbladder stones, non-alcoholic cirrhosis, biliary atresia, cholestatic hepatopathy, chronic liver disease, hepatitis caused by a virus (type B or C), alcoholic liver disease, or liver fibrosis.

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

The compound of the present invention can be used to prevent or treat diabetes or diseases associated with insulin resistance, glucose intolerance in a subject (eg, a mammal, in particular a human).

A compound of the present invention can be used to prevent or treat a lower urinary tract (surrounding area) symptom and benign prostatic hyperplasia (BPH) or ureteral stones in a subject (eg, a mammal, particularly a human).

Another aspect of the present invention provides for the use of a compound represented by Formula I, II or III, a pharmaceutically acceptable salt thereof, or the aforementioned pharmaceutical composition for the manufacture of a medicament for the prevention or treatment of a disease in which the FXR agonist action is beneficial, and wherein said Diseases in which FXR agonist is beneficial includes cardiovascular disease, liver / biliary system disease, obesity, diabetes, lower urinary tract (surrounding area) symptom, and benign prostatic hyperplasia (BPH) or ureteral stones ...

Another aspect of the present invention provides a method for preventing or treating a disease, wherein the FXR agonist effect is beneficial, comprises administering a therapeutically effective amount of a compound of Formula I, II or III, a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition to a patient, and wherein specified disease in which the FXR agonist effect is beneficial includes cardiovascular disease, liver / biliary system disease, obesity, diabetes, lower urinary tract symptom (surrounding area) and benign prostatic hyperplasia (BPH) or stones in ureter.

Cardiovascular diseases include cardiovascular diseases associated with hypertriglyceridemia and / or hypercholesterolemia. Cardiovascular diseases further include atherosclerosis, arteriosclerosis, hypercholesterolemia, hyperlipidemia, thrombosis, coronary artery disease, stroke, or hypertension. Diseases of the liver / biliary system include diseases associated with high cholesterol levels, diseases associated with high triglyceride levels, or fibrotic diseases. Liver / biliary system diseases additionally include non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), gallstones, non-alcoholic cirrhosis, biliary atresia, cholestatic hepatopathy, chronic liver disease, hepatitis, (type B or C), alcoholic liver disease, or liver fibrosis.

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Definitions and representations.

Unless otherwise stated, the following terms and expressions used in this document have the following meanings. A specific term or expression should not be considered vague or unclear in the absence of a specific definition, but should be understood in accordance with its generally accepted meaning. Whenever a trade name is mentioned in this document, it is assumed that it refers to the respective product or active ingredient.

The term “substituted” means that any one or more hydrogen atoms on a particular atom are replaced with a substituent, provided that the valence state of the particular atom is normal and the substituted compound is stable.

The term oxo means that two hydrogen atoms on a particular atom are replaced by = O. If the aromatic ring is oxo-substituted, the oxo-substituted ring may remain aromatic or lose aromaticity. It will be understood that the oxo-substituted group is stable. For example, if the benzene ring is oxo-substituted, it can be selected from; if pyridine O nA is oxo-substituted, it can be selected from

The term is optional or optional means that the event or situation described below may or may not occur, and the description includes cases where the event or situation occurs and if not. For example, the phrase that ethyl is optionally substituted halogen means that ethyl can be unsubstituted (CH2CH3), monosubstituted (e.g. CH2CH2F), polysubstituted (e.g. CHFCH2F, CH2CHF2, etc.), or completely substituted (CF2CF3). Also, for example, optionally said Ring B is substituted with 1, 2 or 3 R2 or 1 oxo means that Ring B is substituted with 1, 2 or 3 R2 or 1 oxo or Ring B is not substituted with R2 or oxo. One skilled in the art will understand that for any groups containing one or more substituents, any substitutions or substitution patterns that cannot spatially exist and / or cannot be synthesized will not be introduced.

C _mn as used herein means that the moiety contains an integer number of carbon atoms in the indicated range. For example C1. ₆ means that the group may contain 1, 2, 3, 4, 5 or 6 carbon atoms.

Numeric range in this document means every integer in the specified range. For example, a C1 _-3 alkyl group means that the group may be selected from C1, C ₂ and C ₃ alkyl; C. _3- 10tsikloalkil means that the group may be selected from C _3, C _4, C _5, C _6, C _7, C _8, C ₉ and C ₁₀ cycloalkyl; 5-6 membered cycloalkyl means that the group can be selected from 5 and 6 membered heterocycloalkyl.

If any variable (for example, R) occurs more than once in a composition or compound structure, its definition is independent in each case. Thus, for example, if a group is substituted with 0-2 R, the group may optionally be substituted with no more than two Rs, and R in each case has independent parameters. In addition, a combination of substituents and / or their variants is allowed only if such a combination leads to the formation of stable compounds.

Where a portion of the groups may be substituted or oxo-substituted herein, the substitution or oxo-substitution is independent of each other, including cases where the groups are either only substituted or only oxo-substituted, or substituted and oxo-substituted at the same time. For example, if ring A is optionally substituted with 1, 2, or 3 Ra or 1, 2 or 3 oxo groups, it includes the following: 1) there are no substituents and oxo groups on ring A, 2) ring A is only substituted with 1, 2 or 3 Ra, 3) ring A is substituted with only 1, 2 or 3 oxo groups, 4) ring A is substituted with Ra and oxo groups at the same time.

In some preferred embodiments of the present application, a portion of the fragment structure may be connected to other structures at the left end and may be connected to other structures at the right end at the same time. If the dashed line or bold line represents a connecting bond, one skilled in the art can understand by reading this application that the dashed line or bold line directly indicates the bond position of the fragment structure with other structures. For example, if ring A is selected from '· the left dashed line indicates that the N atom is connected to the L-group of the compound represented by the formula (I), and the right dashed line indicates that the C atom is connected to the carbonyl moiety of the compound represented by the formula ( I); also, for example, if ring A is selected from '', the left dashed line indicates that the C atom is connected to the L-group of the compound represented by the formula (I), and the right dashed line indicates that the N atom is connected to the carbonyl moiety of the compound represented by formula (I). The left dashed or bold line refers to the elements of the structure of the fragment, protruding from the left above, left or left below, per se; and the right dashed or bold line refers to the elements of the structure of the fragment, I stand

- 13 038580 top right, bottom right or right per se.

In some preferred embodiments of the present invention, the chain length refers to the number of atoms used to form an open chain group and the atoms include a C atom, an N atom, an O atom, or an S atom with different valences. For example, if L2 is selected from —CH2CH2CH2—, the chain length is 3; if L2 is selected from- (CH ₂ ) ₂ -O- (CH ₂ ) 2-, the chain length is 5; if L2 is selected from - (CH ₂ ) ₂ -CH = CH- (CH ₂ ) ₂ -, the chain length is 6.

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

The term hydroxy refers to the -OH group.

The term cyano refers to the -CN group.

The term thiol refers to the -SH group.

The term amino refers to a —NH2 group.

The term nitro refers to the -NO2 group.

The term alkoxy refers to —O-alkyl.

The term alkylamino refers to —NH-alkyl.

The term dialkylamino refers to —N (alkyl) ₂ .

The term alkylsulfonyl refers to —SO ₂ -alkyl.

The term alkylthio refers to -S-alkyl.

The term alkyl refers to hydrocarbyl with the general formula C _n H _{2n + 1} . Alkyl can be linear or branched. For example, the term C ₁ _ ₆ alkyl refers to an alkyl containing from 1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, npentil, 1- methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Likewise, the alkyl moieties (i.e., alkyl) alkoxy, alkylamino, dialkylamino, alkylsulfonyl, and alkylthio have the same definition as above.

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

The term alkenyl refers to a linear or branched unsaturated aliphatic hydrocarbyl that has at least one double bond of carbon and hydrogen atoms. Non-limiting examples of alkenyl include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1,3-butadienyl, etc. The term linear alkenyl refers to a straight chain alkenyl.

The term aryl refers to a monocyclic group containing only carbon atoms with a conjugated π-electron system or a fused polycyclic aromatic ring group. For example, aryl can contain 6 to 20, 6 to 14, or 6 to 12 carbon atoms. Non-limiting examples of aryl include, but are not limited to, phenyl, naphthyl, anthryl, and 1,2,3,4tetrahydronaphthalene, etc.

The term heteroaryl refers to a monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, S, the remaining ring atoms being C, and containing at least one aromatic ring. A preferred heteroaryl contains one 4-8, in particular 5-8 membered ring, or contains several fused rings containing from 6 to 14, in particular from 6 to 10 ring atoms. Non-limiting examples of heteroaryl include, but are not limited to, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, etc.

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

The term non-aromatic heterocyclyl refers to a fully saturated or partially unsaturated (but not fully unsaturated heteroaromatic) non-aromatic ring, which may be present as a monocyclic ring, bicyclic ring, or spirocyclic ring. Unless otherwise indicated, a heterocyclic ring is typically a 3-6 membered ring containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen, and / or nitrogen. Non-limiting examples of heterocyclyl include, but are not limited to, oxiranyl, tetrahydrofuranyl, dihydrofuranyl, pyrrolidinyl, N-methylpyrrolidinyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyrazolidinyl, 4H-pyranyl, morpholinyl, thiomorpholinyl, etc.

As used herein, the term pharmaceutically acceptable refers to those compounds, materials, compositions and / or dosage forms that, within the scope of a reasonable medical judgment, are suitable for use in contact with human tissue and

- 14,038580 animals without undue toxicity, irritation, allergic reaction or other problems or difficulties commensurate with an acceptable benefit / risk ratio.

The term “pharmaceutically acceptable salt” refers to a salt of a compound of the present invention which is derived from a compound with specific substituents found in the present invention and a relatively non-toxic acid or base. If the compound of the present invention contains relatively acidic functional groups, base addition salts thereof can be prepared by contacting the neutral form of such a compound with a sufficient amount of base in a neat solution or a suitable inert solvent. If the compound of the present invention contains relatively basic functional groups, its acid addition salts can be obtained by contacting the neutral form of such a compound with a sufficient amount of acid in a neat solution or a suitable inert solvent. The pharmaceutically acceptable salts which may be mentioned are metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, etc.

Preferably, the salt is contacted with a base or acid in a conventional manner and then the starting compound is isolated, thereby reducing the compound's neutral form. The initial form of the compound differs from the various forms of its salt in some physical properties, for example, different values of solubility in polar solvents.

Some of the compounds of the present invention may contain asymmetric carbon atoms (optical centers) or double bonds. All racemates, diastereomers, geometric isomers and single isomers are included within the scope of the present invention.

Graphical representations of racemic, ambiscalemic and scalemic or enantiomerically pure compounds herein are taken from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, the absolute configuration of the stereocenter is represented by solid and dotted wedges. If the compounds described herein contain an olefinic double bond or other geometric asymmetric centers, unless otherwise indicated, they include the geometric isomers E, Z. Likewise, all tautomeric forms are included within the scope of the present invention.

The compound of the present invention can exist in specific geometric or stereoisomeric forms. All such compounds contemplated in the present invention include cis and trans isomers, pairs of (-) - and (+) enantiomers, (R) - and (S) enantiomers, diastereomers, (D) isomers, (b) β-isomers and racemic mixtures and other mixtures thereof such as enantiomeric or diastereomeric enriched mixtures, all of which are within the scope of the present invention. Other asymmetric carbon atoms may be present on substituents such as alkyl. All such isomers and mixtures thereof are included within the scope of the present invention.

Optically active (R) - and (S) -isomers, as well as D- and L-isomers, can be obtained using chiral synthesis, or chiral reagents, or other conventional methods. If an enantiomer of a particular compound of the present invention is required, it can be obtained by asymmetric synthesis or by derivatization with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group is cleaved off to give the pure desired enantiomers. Alternatively, if the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl), it forms a diastereomeric salt with a suitable optically active acid or base, and then the separation of the diastereomers is carried out by a conventional method well known in the art. techniques, followed by extraction to obtain enantiomers. In addition, the separation of enantiomers and diastereomers in general is achieved through the use of chromatography, which uses a chiral stationary phase, and optionally in combination with a chemical derivatization method (for example, forming carbamates from amines).

The compound of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that form the compound. For example, the compound can be labeled with a radioactive isotope such as tritium ( ³ H), iodine-125 ( ¹²⁵ I), or 14-C ( ¹⁴ C). Any changes in the isotopic composition of a compound of the present invention, whether they are radioactive or not, are included within the scope of the present invention.

The term pharmaceutically acceptable carrier refers to any formulation or vehicle capable of delivering an effective amount of an active agent of the present invention without adversely affecting the biological activity of the active agent and without toxic side effects to the host or patient. Illustrative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, and the like. Such bases include suspensions, tackifiers, transdermal penetration enhancing agents, etc. Their formulations are well known to those skilled in the cosmetic or topical drug arts. For other media information see Remington: The Science and Practice of Pharmacy, 21 ^st Ed., Lippincott.

- 15 038580

Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

The term excipient generally refers to a carrier, diluent and / or medium that is necessary to formulate an effective pharmaceutical composition.

For drugs or pharmacologically active agents, the term effective amount or therapeutically effective amount refers to a sufficient amount of a drug or agent that is not toxic but can achieve the desired effect. For an oral dosage form according to the present invention, the term effective amount of one active agent in a composition means the amount required to achieve the desired effect when used in combination with another active agent in the composition. The determination of the effective amount differs for each individual, depending on the age and general condition of the subject, as well as the particular active substance. An appropriate effective amount in each case can be determined by a person skilled in the art in accordance with routine experimentation.

The terms active ingredient, therapeutic agent, active agent, or active agent refer to a chemical compound that can effectively treat a targeted disorder, disease, or condition.

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

If the substituent is free, this means that the substituent does not exist. For example, if X is free in A-X, the structure is actually A. If a bond in one substituent can provide cross-linking between two atoms on the same ring, that substituent can be bonded to any atom on the ring. Unless indicated by means of which atom said substituent is linked to the compound in which it is included, but is not specifically indicated in the chemical structural formula, a given substituent may be linked through any of its atoms. A combination of substituents and / or their variants is allowed only if such a combination leads to

OS / persistent connections. For example, the structural unit \ = / \ - 'or \ = / \ - / indicates that substitution can occur at any position on cyclohexyl or cyclohexadiene.

Unless otherwise indicated, the term halogenated element or halogen per se or as part of another substituent denotes a fluorine, chlorine, bromine or iodine atom. In addition, the term haloalkyl is intended to include monohaloalkyl and polyhaloalkyl. For example, the term halo (C _g C4) alkyl is intended to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, etc.

Examples of haloalkyl include, without limitation, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl. Alkoxy is the aforementioned alkyl containing a specific number of carbon atoms linked by an oxygen bridge. C1 _-6 alkoxy include C 1, C _2, C _3, C _4, C ₅ and C ₆ alkoxy. Examples of alkoxy include, without limitation, methoxy, ethoxy, n-propoxy, isopropoxy, nbutoxy, sec-butoxy, tert-butoxy, n-pentoxy, and S-pentoxy. Cycloalkyl includes saturated ring groups such as cyclopropyl, cyclobutyl, or cyclopentyl. 3-7 membered cycloalkyl includes C ₃ , C ₄ , C ₅ , C6 and C ₇ cycloalkyl. Alkenyl includes a straight or branched hydrocarbon chain in which one or more carbon-carbon double bonds are present at any stable position along the chain, for example vinyl 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 nitrogen position of the amino group. Illustrative amino protecting groups include, but are not limited to, formyl; acyl such as alkanoyl (eg acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethyloxycarbonyl (Fmoc); arylmethyl such as benzyl (Bn), trityl (Tr), 1,1-di- (4'-methoxyphenyl) methyl; silyl such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term hydroxy protecting group refers to protecting groups suitable for preventing side reactions of hydroxyl groups. Illustrative protecting groups for hydroxy groups include, but are not limited to, alkyl such as methyl, ethyl, and tert-butyl; acyl such as alkanoyl (eg acetyl); arylmethyl such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (benzhydryl, DPM); silyl such as trimethylsilyl (TMS), tert-butyldimethylsilyl (TBS), etc. An important factor to consider in any synthetic route planning in the art is the selection of suitable protecting groups for reactive functional groups (such as the amino group in this application). For the skilled practitioner, (Protective Groups In Organic Synthesis, Wiley and Sons, 1991) of Greene and Wuts is a reference in this regard. All references cited in this application are incorporated herein in their entirety.

The compound of the present invention can be prepared by a variety of methods

16,038580 syntheses well known to those skilled in the art, including the following exemplary embodiments, embodiments formed by combining them with other chemical synthetic methods and equivalent alternatives known to those of ordinary skill in the art. Preferred embodiments include, without limitation, examples of the present invention.

The solvents used in the present invention are commercially available. In the present invention, the following abbreviations are used: aq is water; HATU means O- (7-α-zabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate; EDC is N (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride; m-CPBA means 3-chloroperoxybenzoic acid; eq denotes equivalent, equal amount; CDI stands for carbonyldiimidazole; DCM stands for dichloromethane; PE stands for petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF stands for N, N-dimethylformamide; DMSO stands for dimethyl sulfoxide; EtOAc means ethyl acetate; EtOH stands for ethanol; MeOH stands for methanol; CBz is carbobenzyloxy, which is an amino protecting group; BOC is tert-butylcarbonyl, which is an amino protecting group; HOAc stands for acetic acid; NaCNBH ₃ is sodium cyanoborohydride; Ph. D. indicates room temperature; O / N means during the night; THF stands for tetrahydrofuran; Boc2O is di-tert-butyl dicarbonate; TEA stands for trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl2 is thionyl chloride; CS2 is carbon disulfide; TsOH stands for p-toluenesulfonic acid; NFSI stands for N-fluoroN- (phenylsulfonyl) benzenesulfonamide; NCS represents 1-chloropyrrolidine-2,5-dione; n-Bu4NF is tetrabutylammonium fluoride; iPrOH is 2-propanol; t. pl. denotes the melting point; LDA stands for lithium diisopropylamide.

Detailed description of the invention

For the purpose of illustrating the present invention in more detail, the scope of the present invention is not limited thereto.

Reference Example 1. Retrieving INT-747.

the following examples are given,

Reference example 1A

acid (60.0

152.8 g,

To a solution of chenodeoxycholic mmol) in methanol / acetic acid / water / ethyl acetate (360/120/30/780 ml), tetrabutylammonium bromide (81.0 g, 251.3 mmol) and sodium bromide (9.0 g, 87 , 5 mmol) and then sodium hypochlorite (210 ml, 3.4 mol) was added dropwise at 0 ° C over 30 min. After stirring at 28 ° C for 16 h, saturated sodium bisulfite solution (500 ml) was added to quench. The aqueous layer was extracted with ethyl acetate (1000 ml x 2) and the combined organic layer was washed with water (1000 ml x 5). The organic layer was dried over sodium sulfate, filtered and evaporated to dryness. The residue was recrystallized (dichloromethane, 200 ml) to give Reference Example 1A (yellow solid, 41 g, 69% yield). 'H NMR (400 MHz, methanol ^) δ 3.44-3.60 (m, 1H), 2.99 (dd, J = 5.77, 12.30 Hz, 1H), 2.54 (t, J = 11.29 Hz, 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 = 6.53 Hz, 3H), 0.71 (s, 3H).

Reference example 1B

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To a solution of reference example 1A (246.0 g, 629.9 mmol) in methanol (2 L) p-toluenesulfonic acid (10.9 g, 63.0 mmol) was added in one portion and allowed to react between them at

80 ° C for 4 h. After cooling to room temperature, this was evaporated to dryness and quenched with saturated sodium bicarbonate solution (1500 ml). Then the aqueous layer was extracted with ethyl acetate (1500 ml x 3). The combined organic layer was washed with brine (1000 ml x 3), dried over sodium sulfate, filtered and evaporated to dryness. The residue was recrystallized (ethyl acetate, 500 ml) to give reference example 1B (white solid, 202 g, 79% yield). 1H NMR (400 MHz, chloroform-d) δ 3.66 (s, 3H), 3.55-3.62 (m, 1H), 2.85 (dd, J = 6.02, 12.55 Hz, 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 chlorotrimethylsilane (107.5 g, 989.5 mmol) in tetrahydrofuran (500 ml) was added lithium diisopropylamide (87.4 g, 815.6 mmol) dropwise at -78 ° C under nitrogen atmosphere and after stirring for 40 min additionally added dropwise a solution of reference example 1B (50 g, 123.6 mmol) in tetrahydrofuran (300 ml). After completion of the dropwise addition, this was stirred at -78 ° C for an additional 40 min and then triethylamine (182.5 g, 1.8 mol) was added. This was quenched with saturated sodium bicarbonate (1000 ml) after 1 h and the aqueous layer was extracted with ethyl acetate (1000 ml × 3). The combined organic layer was further washed with water (100 ml x 6) and brine (1000 ml x 2). The organic layer was dried over sodium sulfate, filtered and evaporated to dryness, thus obtaining Reference Example 1C (brown-yellow oil, 68 g, 100% yield), which can be used directly in the next step without further purification. ¹ H NMR (400 MHz, chloroform-d) δ 4.75 (dd, J = 1.38, 5.90 Hz, 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 = 6.53 Hz, 3H), 0.85 (s, 3H), 0.70 (s, 3H), 0.17-0.20 (m, 9H), 0.13 (s, 9H).

Reference example 1D y o—

I 1 ^H Iz

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 complex (64.4 g, 453.4 mmol) in dichloromethane (300 ml) was added dropwise at -78 ° C under nitrogen atmosphere. For the dropwise addition, it was necessary to maintain the internal temperature at -78 ° C. After stirring for 1 hour, it was heated to 30 ° C. and stirred for an additional 2 hours. The above solution was quenched with saturated sodium hydrogen carbonate (1000 ml). The aqueous layer was extracted with dichloromethane (1000 ml x 3). The combined organic layer was washed with brine (1000 ml x 2), dried over sodium sulfate, filtered and evaporated to dryness. The residue was purified by column chromatography to give Reference Example 1D (yellow solid, 43 g, 81% yield). 1H NMR (400 MHz, chloroform-d) δ 6.12 (q, J = 7.03 Hz, 1H), 3.52-3.66 (m, 4H), 2.54 (dd, J = 4, 02, 13.05 Hz, 1H), 2.13-2.40 (m, 5H), 1.68-1.98 (m, 7H), 1.65 (d, J = 7.03 Hz, 3H ), 1.00-1.52 (m, 11H), 0.97 (s, 3H), 0.89 (d, J = 6.53 Hz, 3H), 0.61 (s, 3H).

Reference example 1E h he. Mr G X

[ΊηΤη

HO '' ^ -

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 this was stirred at 50 ° C in over 2 h. After evaporation of the solvent to dryness, water (500 ml) was added and ethyl acetate (500 ml × 2) was used for extraction. The aqueous phase was brought to pH 3 with diluted HCl and extracted with dichloromethane (600 ml x 2). The combined organic layer was concentrated. The residue was purified by recrystallization (ethanol, 200 ml) to give Reference Example 1E (yellow solid, 147 g, 72% yield). 1H NMR (400 MHz, chloroform-d) δ 6.19 (q, J = 7.36 Hz, 1H), 3.60-3.74 (m, 1H), 2.58 (dd, J = 4, 02, 13.05 Hz, 1H), 2.40 (tt, J = 5.02, 10.29 Hz, 3H), 2.19-2.32 (m, 2H),

- 18 038580

1.61-2.06 (m, 10H), 1.04-1.54 (m, 14H), 1.01 (s, 3H), 0.95 (d, J = 6.53 Hz, 3H) , 0.65 (s, 3H).

Reference example 1F g he

I 1 ⁿ 1 /

To a solution of reference example 1E (140.0 g, 336.1 mmol) in a solution of NaOH (0.5 mol) in water (600 ml) was added 10% Pd-C (19.9 g, 134.4 mmol) in one portion and introduced hydrogen at 15 psi. and allowed to react therebetween at 100 ° C. for 16 hours. After suction filtration, the filtrate was adjusted to pH 3 with diluted hydrochloric acid. The aqueous layer was extracted with dichloromethane (1500 ml x 3). The combined organic layer was washed with brine (1000 ml x 3) and dried over sodium sulfate, filtered and evaporated to dryness, thus obtaining Reference Example 1F (white solid, 101 g, 72% yield) which can be directly applied to the following stages without additional purification. 1H NMR (400 MHz, chloroform-d) δ 3.49-3.60 (m, 1H), 2.70 (q, J = 6.02 Hz, 1H), 2.122.45 (m, 4H), 1 , 65-2.02 (m, 9H), 1.29-1.52 (m, 6H), 1.05-1.24 (m, 8H), 0.93 (d, J = 6.53 Hz , 5H), 0.81 (t, J = 7.53 Hz, 3H), 0.66 (s, 3H).

Reference connection INT-747

To a solution of reference example 1F (16.0 g, 38.2 mmol) in sodium hydroxide (2 mol, 100 ml) sodium borohydride (8.7 g, 229.3 mmol) was added in portions and this was stirred at 100 ° C for 2 h. After cooling to room temperature, saturated aqueous ammonium chloride solution (150 ml) was added. Its pH was adjusted to 3 with diluted hydrochloric acid. The aqueous layer was extracted with dichloromethane (300 ml x 3). The combined organic layer was washed with brine (200 ml x 3), dried over sodium sulfate, filtered and evaporated. The residue was purified by column chromatography, thereby obtaining the reference compound INT-747 (white solid, 14.5 g, 90% yield). ¹ H NMR (400 MHz, chloroform-d) δ 3.71 (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), 0.860.97 (m, 9H), 0.66 (s, 3H).

Path 1

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Example 1

Example 1A

Perchloric acid (6.0 g, 60.0 mmol) in a nitrogen atmosphere. After stirring at 55 ° C. for 6 hours and concentrating in vacuo, the above solution was diluted with water (100 ml). The aqueous layer was extracted with ethyl acetate (100 ml x 2). The combined organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was purified by column chromatography, thus obtaining Example 1A as a white solid (2.8 g, 92% yield). 1H NMR (400 MHz, chloroform-d) δ 8.16 (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 iodine (106.0 mg, 0.4 mmol) and ensured the reaction in the reaction system for 12 h under light irradiation. The reaction in the above solution was quenched by the addition of sodium thiosulfate solution (1 ml). The aqueous layer was extracted with dichloromethane (10 ml x 3), dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was purified using a preparative thin layer chromatography plate (petroleum ether: ethyl acetate = 5: 1), thus obtaining Example 1B (colorless solid, 50.0 mg, 38% yield). Ή NMR (CDCl ₃ , 400 MHz) δ 8.14 (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-oxopiperidine-3-carboxylate (21.0 mg, 134 μmol) in N, N-dimethylformamide (1 ml) was added sodium hydride (7.0 mg, 179 μmol, 60%). 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 0 ° C. After completion of the dropwise addition, the reaction system was slowly warmed to room temperature and allowed to react for 12 h. Water (3 mL) and lithium hydroxide monohydrate (4 mg, 89.5 mmol) were added to the reaction system and stirred for an additional 3 h. Water (10 ml) was added and the reaction system was brought to pH 6 with hydrochloric acid (1M), extracted with a mixture of dichloromethane: methanol = 10: 1 (10 ml × 3). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was purified using a preparative thin layer chromatography plate (dichloromethane: methanol = 10: 1), thus obtaining Example 1 (15 mg, 32.4% yield). 1H NMR (400 MHz, methanol-d ₄ ) δ 3.67 (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 = 13.1 Hz, 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 = 6.5 Hz, 3H), 0.95- 0.90 (m, 6H), 0.72 (s, 3H).

The compounds of Examples 2-16 were synthesized by the same procedure as for Example 1 and are shown below.

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Connection No. Output, % Connection structure 'NMR Example 2 6% BUT T I ^N T /] RJR o ^ o I ¹ H NMR (400 MHz, CHLOROFORM-7) δ 3.97-3.88 (m, IH), 3.77-3.66 (m, 2H), 3.47-3.37 (m, IH) , 3.17-3.04 (m, IH), 2.83 (q, 7 = 9.1 Hz, IH), 2.37-2.31 (m, 2H), 2.08 (d, J = 12.8 Hz, 7H), 1.93-1.79 (m, 4H), 1.69-1.45 (m, 7H), 1.23-1.11 (m, 4H), 1, 00 (d, J = 4.5 Hz, 3H), 0.92-0.90 (m, 8H), 0.88 (d, J = 2.0 Hz, 4H), 0.68 (s, 3H ) Example 3 6% O ^ OH ¹ H NMR (400 MHz, CHLOROFORM-7) δ 4.03-3.91 (m, IH), 3.73 (t, J = 7.2 Hz, IH), 3.68 (br. S., IH), 3.41 (d, J = 4.8Hz, IH), 3.25-3.16 (m, IH), 3.05 (br. S., IH), 2.87-2, 80 (m, IH), 2.39-2.25 (m, 2H), 2.06-1.95 (m, 2H), 1.92-1.77 (m, 6H), 1.77- 1.76 (m, IH), 1.72-1.56 (m, 4H), 1.50 (d, J = 5.0 Hz, 3H), 1.44 (d, J = 12.3 Hz , 3H), 1.19-1.11 (m, 4H), 1.06-1.00 (m, IH), 0.96 (d, J = 4.5Hz, 3H), 0.91- 0.89 (m, IH), 0.90 (br. S., 3H), 0.88 (br. S., 2H), 0.86 (d, J = 2.5 Hz, 4H), 0 , 84 (d, J = 3.0 Hz, 2H), 0.68-0.62 (m, 3H) Example 4 52.7% NO'U । 1 ⁿ I / ΌΗ \ Xi ^ COOH ^X H NMR (400 MHz, CHLOROFORM-7) δ 7.96 (s, IH), 7.93 (s, IH), 4.26-4.16 (m, IH), 4.15-4.06 (m, IH), 3.70 (br. s., IH), 3.46-3.33 (m, IH), 2.10-2.02 (m, IH), 1.99-1, 86 (m, 2H), 1.84-1.77 (m, 3H), 1.71-1.56 (m, 4H), 1.50-1.40 (m, 5H), 1.34- 1.30 (m, 2H), 1.26 (s, 3H), 1.22-1.17 (m, 3H), 1.02 (d, J = 6.5Hz, 4H), 0.92 (s, IH), 0.89 (s, 3H), 0.88-0.85 (m, 2H), 0.64 (s, 3H) Example 5 57% 1 ^Mr. [/ IR | R < O I ^X H NMR (400 MHz, METHANOL-7 /) δ 8.40 (s, IH), 7.97 (dd, J = 2.0, 9.5 Hz, IH), 6.54 (d, J = 9.5 Hz, IH), 4.20-3.98 (m, 2H), 3.67 (br. S., IH), 3.32-3.28 (m, IH), 2.05- 1.73 (m, 8H), 1.63-1.21 (m, 16H), 1.12 (d, J = 6.0 Hz, 3H), 1.07-0.99 (m, IH) , 0.95-0.90 (m, 6H), 0.71 (s, 3H).

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Example 6 10.9% he BUT '' ΌΗ ¹ H NMR (400 MHz, METHANOL - ^) δ 7.66 (d, 7 = 5.0 Hz, 1H), 7.15-6.97 (m, 1H), 6.82 (br. S., 1H), 4.17-3.92 (m, 2H), 3.66 (br.s., 1H), 3.32-3.29 (br, 1H), 2.04-1.25 (m , 25H), 1.26-1.25 (m, 1H), 1.11 (d, J = 6.0 Hz, 3H), 0.95-0.89 (m, 6H), 0.70 ( s, ЗН). Example 7 53% syO NO '' ^ - '' ^he ° ^ he ¹ H NMR (400 MHz, METHANOL-7 ₄ ) δ 0.75 (s, 3 Η) 0.87-0.97 (m, 6 Η) 1.07 (d, J = 6.53 Hz, 3 Η ) 1.12-2.11 (m, 25 Η) 3.27-3.32 (m, 1 Η) 3.69 (br. S., 1 H) 4.01-4.17 (m, 2 H) 7.16 (dd, 7 = 8.16, 2.13 Hz, 1 H) 7.38 (t, 7 = 7.91 Hz, 1 H) 7.54 (s, 1 H) 7.61 (d, 7 = 7.53 Hz, 1H). Example 8 16% \ ° V ^OH NO '''''' OH ¹ H NMR (400 MHz, METHANOL-7 ₄ ) δ 0.74 (s, 3) 0.90-0.96 (m, 6 Η) 1.06 (d, 7 = 6.53 Hz, 3 Η ) 1.21-2.13 (m, 25 Η) 3.31 (br. S., 1 Η) 3.68 (br. S., 1 H) 4.07-4.27 (m, 2 H ) 7.03 (t, 7 = 7.53 Hz, 1 H) 7.15 (d, 7 = 8.53 Hz, 1 H) 7.46-7.66 (m, 1 H) 7.82 ( dd, 7 = 7.78, 1.25 Hz, 1 H). Example 9 10.9% y ^^ he | ] A | A BUT '' ^ - 'Gt' ΌΗ ¹ H NMR (400 MHz, CD ₃ OD) δ 8.71 (br. S. 1H), 8.37 (br. S., 1H), 7.89 (br. S., 1H), 4, 16 (d, 7 = 6.8 Hz, 2H), 3.67 (br. S., 1H), 3.37 (s, 1H), 2.01-1.20 (m, 25H), 1, 06 (d, 7 = 6.5 Hz, ZN), 0.94-0.90 (m, 6H), 0.73 (s, 3H). Example 10 16% H0 '''% x7X /''oH ^N ° Ή NMR (400 MHz, METHANOL- ^) δ 8.57 (br. S., 1H), 8.47 (d, 7 = 8.3 Hz, 1H), 8.19 (d, 7 = 8.0 Hz, 1H), 4.37 (d, 7 = 5.3 Hz, 2H), 3.68 (br. S., 1H), 3.37 (s, 1H), 2.03-1.28 ( m, 25H), 1.10 (d, 7 = 6.3 Hz, 3H), 0.95-0.91 (m, 6H), 0.75 (s, 3H). Example 11 72% ... / —V-ο о I 1 ⁿ Г / </ ΌΗ 1 ⁱ 1 ⁱ ΗΟ '''% ^ 4Ν <' ΌΗ ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 8.71 (br. S., 1H), 8.37 (br. S., 1H), 7.89 (br. S., IH), 4 , 16 (d, 7 = 6.8 Hz, 2H), 3.67 (br. S., IH), 3.37 (s, IH), 2.01-1.20 (m, 25H), 1 , 06 (d, 7 = 6.5 Hz, 3H), 0.94-0.90 (m, 6H), 0.73 (s, 3H). Example 12 61% <Ta] a ^Ν ^ γ ° Η ΗΟ '·% Τ ⁴ <-ΌΗ О ¹ H NMR (400 MHz, METHANOL- ^) δ 8.62 (s, 2H), 4.38-4.26 (m, 2H), 3.68 (br. S., IH), 3.31- 3.25 (m, IH), 2.13-1.67 (m, 9H), 1.65-1.14 (m, 15H), 1.12-0.98 (m, 4H), 0, 96-0.89 (m, 6H), 0.75 (s, 3H). Example 13 31% ct ^ / b HO '^ dSr ΌΗ ¹ H NMR (400 MHz, METHANOL - ^) δ 8.40 (s, IH), 8.22 (br. S., IH), 7.52 (br. S., IH), 4.22 (dd , 7 = 7.5, 15.8 Hz, 2H), 3.68 (br. S., IH), 3.35-3.34 (m, IH), 2.01-1.37 (m, 25H), 1.06 (d, 7 = 6.3 Hz, 3H),

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0.94-0.89 (m, 6H), 0.74 (s, 3H). Example 14 16% ΗΟ '··· ^ —T'r' Ό ^ ¹ H NMR (400 MHz, METHANOL - ^) δ 3.72-3.63 (m, 3H), 3.54-3.39 (m, 4H), 2.26 (s, 3H), 1.96 (s, 3H), 1.88-1.04 (m, 25H), 0.99-0.87 (m, 9H), 0.70 (s, 3H) Example 15 75% ΗΟ υ ^ ΗΧ - / οΗ ¹ H NMR (400 MHz, METHANOL- ^) δ 7.67-7.48 (m, 1H), 7.06-6.94 (m, 1H), 4.34-4.12 (m, 2H) , 3.66 (br s, 1H), 3.30-3.26 (m, 1H), 2.06-1.14 (m, 25H), 1.04 (d, 7 = 6.5 Hz, 3H), 0.94-0.86 (m, 6H), 0.72 (s, 3H) Example 16 34% Y ^he go and G TyT a HO '''^UbX'OH ¹ H NMR (400 MHz, METHANOL- ^) δ 8.40 (s, 1H), 8.22 (br s, 1H), 7.52 (br s, 1H), 4.22 (br dd, 7 = 7.5, 15.8 Hz, 2H), 3.68 (br s, 1H), 3.35-3.34 (m, 1H), 2.01-1.37 (m, 25H), 1, 06 (br d, 7 = 6.3 Hz, 3H), 0.94-0.89 (m, 6H), 0.74 (s, 3H)

Path 2

Examples 17 and 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 allowed to react in the reaction system at 25 ° C for 2 h. 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 and the reaction between them was ensured at 0 ° C for 15 min and additionally allowed the reaction to proceed at 25 ° C for 15 min. The reaction was quenched with 0.2 M diluted 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 evaporated to dryness. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 4: 1), thus obtaining Example 2A (250.0 mg, 70%). ¹ H NMR (400 MHz, chloroform-d) δ 8.15 (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).

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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) iodine (461.0 mg, 1.8 mmol) was added and the reaction system was allowed to react at 25 ° C 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 mlx3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 20: 1), thus obtaining Example 2B (250.0 mg, 70%). 1H NMR (CDCl ₃ , 400 MHz) δ 8.16 (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).

Examples 17 and 18

To a solution of methyl 6-hydroxynicotinate (80.0 mg, 523 μmol) in N, N-dimethylformamide (3 ml) was added sodium hydride (21.0 mg, 523 mmol, 60%) at 0 ° C. After half an hour, a solution of Example 2B (150.0 mg, 262 μmol) in N, N-dimethylformamide (5 ml) was added dropwise at 0 ° C. After completion of the dropwise addition, 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 stirred for an additional 3 hours. Water (10 ml) was added and the system was adjusted to pH 6 with hydrochloric acid (1 M) and extracted with 10: 1 dichloromethane: methanol (10 ml × 3). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was isolated and purified using a preparative thin layer chromatography plate (dichloromethane: methanol = 10: 1), thus obtaining Example 17 (40 mg, 29%), 1H 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%), 1H 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).

Path 3

P b (ОА ^С ) у С υ (ΟΑθ) 2

ONSO '

Example 19 onso

he

- 24 038580

Example FOR

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 allowed to react in the reaction system at 110 ° C. for 12 hours. The reaction system was filtered and the filtrate was concentrated. The residue was purified through column chromatography (petroleum ether: ethyl acetate = 20: 1), thus obtaining the title compound (120.0 mg, 27.0%). ¹ H NMR (400 MHz, chloroform-d) δ 8.15 (s, 1H), 8.05 (s, 1H), 5.71-5.59 (m, 1H), 5.19 (br. S ., 1H), 4.93-4.80 (m, 2H), 4.76-4.67 (m, 1H), 2.12-1.61 (m, 11H), 1.52-1, 09 (m, 13H), 1.04 (d, J = 6.8 Hz, 3H), 0.97 (s, 3H), 0.91 (t, J = 7.4 Hz, 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), palladium acetate (5.0 mg, 23.0 μmol) and methyl 2-iodobenzoate (60.8 mg, 0.2 mmol) were added under a nitrogen atmosphere and the reaction was allowed to proceed in the reaction system at 85 ° C for 12 hours. The solvent was evaporated to dryness 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. The organic phase was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was purified through preparative thin layer chromatography (petroleum ether: ethyl acetate = 10: 1), thus obtaining the title compound (80.0 mg, 61.0%). 1H NMR (400 MHz, chloroform-d) δ 8.16 (s, 1H), 8.07 (s, 1H), 8.01 (s, 1H), 7.90-7.84 (m, 1H) , 7.51 (d, J = 7.8 Hz, 1H), 7.41-7.33 (m, 1H), 6.40-6.31 (m, 1H), 6.16 (dd, J = 8.8, 15.8 Hz, 1H), 5.21 (br. S., 1H), 4.79-4.63 (m, 1H), 3.94 (s, 3H), 2.29 (d, J = 6.8 Hz, 1H), 2.08-1.62 (m, 11H), 1.521.20 (m, 11H), 1.15 (d, J = 6.5 Hz, 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 a wet carbon-supported palladium catalyst (50 mg, 10%) under a nitrogen atmosphere and allowed to react in the reaction system in the presence of hydrogen (15 psi) at 25 ° C for 12 hours. The reaction system was filtered and evaporated to dryness, thus obtaining Example 3C (90.0 mg, 90.0%). 1H NMR (400 MHz, 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), 4.76-4.51 (m, 1H), 2.77-2.64 (m, 1H) , 2.54-2.44 (m, 1H), 1.95-1.20 (m, 25H), 1.01 (d, J = 6.5 Hz, 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 system was allowed to react at 20 ° C. for 12 hours. Water (5 ml) was added to the reaction system and the system was adjusted to pH 1-2 with hydrochloric acid (1 M). The aqueous phase was extracted with ethyl acetate (10 ml x 3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was isolated and purified by preparative isolation (HCl), thus obtaining Example 19 (80 mg, 57%). ¹ H NMR (400 MHz, methanol ^) δ 7.93-7.76 (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 = 10.5 Hz, 1H), 2.06-1.27 (m, 25H), 1.09 (d, J = 6.3 Hz, 3H), 0.94-0 , 91 (m, 6H), 0.70 (s, 3H).

Path 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 system was allowed to react at 20 ° C. for 12 hours. Water (5 ml) was added to the reaction system and the system was adjusted to pH 1-2 with hydrochloric acid (1 M). The aqueous phase was extracted with ethyl acetate (10 ml x 3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was purified by preparative thin layer chromatography (dichloromethane: methanol = 10: 1), thus obtaining Example 20 (25.0 mg, 71.0%). Ή NMR (400 MHz, methanol-d ₄ ) δ 8.00 (br. S., 1H), 7.84 (d, J = 7.0 Hz, 1H), 7.55 (d, J = 7, 5 Hz, 1H), 7.42-7.34 (m, 1H), 6.46-6.32 (m, 1H), 6.19 (dd, J = 8.8, 15.6 Hz, 1H ), 3.72-3.62 (m, 1H), 3.39-3.37 (m, 1H), 2.32 (d, J = 6.0 Hz, 1H), 2.07-1, 29 (m, 22H), 1.17 (d, J = 6.3Hz, 3H), 0.95-0.89 (m, 6H), 0.78 (s, 3H).

Path 5

Example 21

Example 5A

After stirring 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) at 25 ° C for 0.5 h, O-benzotriazole-N, N, N ', N'-tetramethyluronium tetrafluoroborate (95.0 mg, 0.3 mmol) was added thereto. The resulting mixture was stirred at 25 ° C. for 12 hours. After removal of the solvent by evaporation under vacuum, the residue was purified by column chromatography, thereby obtaining Example 5A as a white solid (90 mg, 82% yield).

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1H NMR (400 MHz, chloroform-d) δ 3.73-3.67 (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 = 6.3 Hz, 3H), 0.93-0.88 (m, 6H), 0.67 (s , 3H).

Example 5B. Gn but ^{with OH}

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 this was stirred at 0 ° C for an additional 30 min and then warmed to room temperature and stirred for 12 h. The reaction mixture was quenched by the addition of ice water and then it was extracted with ethyl acetate (60 mlx2). After washing with water, the organic phase was dried over anhydrous sodium sulfate and filtered. After removing the solvent in vacuo, the residue was purified by preparative TLC, thus obtaining Example 5B as a white solid (6 mg, 66% yield). 1H NMR (400 MHz, chloroform-d) δ 3.71 (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, 14.2 Hz, 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) was added thereto and p-toluenesulfonic acid (90.9 mg, 478.0 μmol). The reaction solution was stirred at 30 ° C for 36 hours. The solvent was removed by concentration and water was added to the reaction solution (5 ml), which was extracted with ethyl acetate (10 ml × 3). The organic layer was washed with water (10 ml), dried over anhydrous sodium sulfate, filtered and concentrated to thereby obtain the crude product. The crude product was isolated by column chromatography, thus obtaining Example 5C (450.0 mg, 32.1% yield, colorless oil). 1H NMR (400 MHz, chloroform-d) δ 4.73 (d, J = 3.3 Hz, 1H), 4.56 (d, J = 4.3 Hz, 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, 16.0 Hz, 1H), 1.97-1.80 (m, 7H), 1.74-1.67 (m, 4H), 1.55 (br s., 4H), 1.41 (d, J = 7.3 Hz, 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% ⁰

Metallic sodium (68.2 mg, 3.0 mmol) was dissolved in anhydrous ethanol (5 ml) cooled to 0 ° C, to which Example 5C (170.0 mg, 296.8 μmol, dissolved in 2 ml of anhydrous ethanol) and this was stirred for another half hour at 0 ° C. Then, diethyl oxalate (65.0 mg, 445.1 µmol, 60.8 µl, dissolved in 1 ml of anhydrous ethanol) was added dropwise. After stirring at 0 ° C for half an hour, the reaction solution was warmed to 25 ° C and stirred for another 35 hours. Ethanol (6 ml) was added to the reaction solution, and the solvent was removed by concentration. Ethyl acetate (40 ml) was added and 2 mol of an aqueous citric acid solution was added with stirring in an ice bath (until pH 5-6). 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 thereby obtain the crude product. The crude product was isolated by column chromatography, thus obtaining Example 5D (90.0 mg, 44.2% yield). 1H 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 , 0 Hz, 2H), 1.16 (d, J = 9.8 Hz, 2H), 0.95 (d, J = 6.5 Hz, 2H), 0.89 (s, 3H), 0, 71-0.59 (m, 3H).

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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 thereto and the reaction solution was stirred at 80 ° C for 5 h. The solvent was removed by concentration, and water (5 ml) was added to the reaction solution. 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 evaporated to dryness in vacuo. Example 5E was prepared without purifying the crude product (86.0 mg, crude product, yellow oil). ¹ H NMR (400 MHz, chloroform-d) δ 6.39 (s, 1H), 4.43 (q, J = 7.3 Hz, 2H), 3.72 (br. S., 1H), 3 , 43-3.40 (m, 1H), 2.90-2.64 (m, 2H), 2.42 (t, J = 6.9 Hz, 2H), 2.00-1.95 (m , 2H), 1.75 (d, J = 6.8 Hz, 3H), 1.69 (br. S., 4H), 1.60 (d, J = 3.3 Hz, 3H), 1, 48 (d, J = 7.5 Hz, 4H), 1.41 (s, 3H), 1.28 (br. S., 4H), 1.20 (d, J = 8.5 Hz, 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), and lithium hydroxide monohydrate (93.6 mg , 2.2 mmol). The reaction solution was stirred at 25 ° C for 12 hours. The reaction solution was acidified with 1 mol of hydrochloric acid (to pH 5-6) and extracted with ethyl acetate (10 ml × 3). The organic layer was washed with water (10 ml), dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated to dryness in vacuo. The crude product was isolated by thin layer chromatography, thus obtaining Example 21 (28.0 mg, 25.8% yield). ¹ H NMR (400 MHz, methanol-d ₄ ) δ 6.41 (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 = 12.0 Hz, 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 = 9.5 Hz, 2H), 1.05 (d, J = 6.0 Hz, 3H), 0.93 (s, 3H), 0.92-0.89 (m, 3H), 0.71 (s, 3H).

Path 6

Example 22

Example 6A

To a solution of methyl 6-hydroxynicotinate (438.7 mg, 2.9 mmol) of 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, filtered and concentrated. The residue was isolated from

- 28,038580 using a column (petroleum ether: ethyl acetate = 10: 1), thus obtaining the title compound 6A (400 mg, 47.9%). 1H NMR (400 MHz, chloroform-d) δ 8.82 (d, J = 2.3 Hz, 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), 4.80-4.61 (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 (volume ratio of methanol to water was 1: 1) (600 mg, 15 mmol, 6 ml) and provided the course of the reaction in the reaction system at 70 ° C for 2 hours. The solvent was evaporated to dryness. The system was brought to a pH value of 1-2 with diluted hydrochloric acid (1M) and extracted with a mixture of dichloromethane: methanol = 10: 1 (20 ml × 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative chromatography, thus obtaining the title compound 22 (460 mg, 64% yield). 1H NMR (400 MHz, methanol-d ₄ ) δ 8.78 (d, J = 1.8 Hz, 1H), 8.21 (dd, J = 2.3, 8.5 Hz, 1H), 6, 84 (d, J = 8.8 Hz, 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 = 6.5 Hz, 3H), 0.96-0.89 (m, 6H), 0.74 (s, 3H).

For the preparation of Examples 23-26, the procedure for the title compound 22 was followed.

Connection No. Output, % Connection structure ^X NMR Example 23 47.2% BUT '· ’^ I Г И 1 / oh, ^{C1 1} H NMR (400 MHz, METHANOL - ^) δ 8.68 (br.s., 1H), 8.24 (s, 1H), 4.61-4.37 (m, 2H), 3.67 ( br. s., 1H), 3.37 (s, 1H), 2.04-1.18 (m, 25H), 1.08 (d, J = 6.3Hz, 3H), 0.95- 0.89 (m, 6H), 0.73 (s, 3H) Example 24 56.6% BUT '· ’^ ... (nGA X. ¹ H NMR (400 MHz, METHANOL - ^) δ 8.58 (d, J = 1.8 Hz, 1H), 7.94 (dd, 7 = 1.8, 10.5 Hz, 1H), 4, 66-4.37 (m, 2H), 3.74-3.58 (m, 1H), 3.32-3.28 (m, 1H), 2.08-1.20 (m, 25H), 1.08 (d, J = 6.5 Hz, 3H), 0.95-0.87 (m, 6H), 0.73 (s, 3H) Example 25 56.7% BUT '·' ^ I 1 ⁿ I / JRJFI ¹ H NMR (400 MHz, METHANOL- ^) δ = 7.85-7.78 (m, 1H), 7.73 (d, J = 7.3 Hz, 1H), 7.00 (d, J = 8.3 Hz, 1H), 4.53-4.34 (m, 2H), 3.68 (br s, 1H), 2.02-1.10 (m, 25H), 1.08 (br d , J = 6.5 Hz, 3H), 0.97-0.88 (m, 6H), 0.74 (s, 3H) Example 26 75.6% BUT '' ^ I 1 ^Н Т / JRJFI '' ^he % ¹ H NMR (400 MHz, METHANOL- ^) δ = 8.30 (d, J = 5.5 Hz, 1H), 7.54 (dd, J = 1.3, 5.5 Hz, 1H), 7 , 43 (s, 1H), 4.57-4.29 (m, 2H), 3.66 (br s, 1H), 2.08-1.10 (m, 25H), 1.06 (d, J = 6.5 Hz, 3H), 0.95-0.87 (m, 6H), 0.73 (s, 3H)

- 29 038580

Path 7

28 29 30 31 32 33 34

Example 27

Methanesulfonamide (37 mg , 389.3 μmol). The reaction system was allowed to react at 25 ° C. for 12 hours. Water (5 ml) was added to the system. The system was brought to pH 2 with diluted hydrochloric acid (1 M) and extracted with dichloromethane (10 ml × 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative chromatography, thus obtaining example 27 (25 mg, yield 21.7%). ¹ H NMR (400 MHz, methanol-d ₄ ) δ 8.74 (d, J = 2.3 Hz, 1H), 8.23 (dd, J = 2.3, 8.8 Hz, 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 = 6.5 Hz, 3H), 0.96-0.90 (m, 6H), 0.73 (s, 3H).

For the preparation of Examples 28-34, the procedure for the title compound 27 was followed.

Connection No. Output, % Connection structure Ή NMR Example 28 50.0 hhh but '''' ^he ° 0 'H NMR (400 MHz, METHANOL - ^) δ 8.72 (d, 4 = 2.3 Hz, 1H), 8.23 (dd, 4 = 2.4, 8.9 Hz, 1H), 6, 97 (d, 4 = 8.8 Hz, 1H), 4.54-4.32 (br, 2H), 3.65 (br s, 1H), 3.56-3.38 (br, 1H), 3.18-3.07 (br, 1H), 2 , 02-1.15 (br, 29H), 1.05 (d, 4 = 6.3 Hz, ZN), 0.93-0.87 (br, 6H), 0.71 (s, ZN) Example 29 26.0 syChH = - ΗΟ ’^^^ Κ'ΌΗ Oh 'H NMR (400 MHz, METHANOL - ^) δ 8.69 (d, 4 = 2.3 Hz, 1H), 8.16 (dd, 4 = 2.4, 8.9 Hz, 1H), 6, 91 (d, 4 = 8.8 Hz, 1H), 4.56-4.27 (br, 2H), 3.66 (br s, 1H), 3.46 (s, 1H), 2.05-1.57 (br, 15H), 1.50 (s , 9H), 1.33-1.09 (br, YH),

- 30 038580

1.05 (d, 7 = 6.5 Hz, ZN), 0.93-0.86 (br, 6H), 0.71 (s, ZN) Example 30 39.8 Chu Ή NMR (400 MHz, METHANOL- ^) δ = 8.70 (d, 7 = 2.0 Hz, 1H), 8.13 (dd, 7 = 2.5, 8.8 Hz, 1H), 6.86 (d, J = 8.8 Hz, 1H), 4.50-4.33 (m, 2H), 3.91 (quin., 7 = 6.9 Hz, 1H), 3.66 (br s, 1H), 3.50-3.34 (t, 1H), 2.05-1.49 (br, 16H), 1.42 ( d, 7 = 7.0 Hz, 6H), 1.39-1.08 (m, 9H), 1.05 (d, 7 = 6.5 Hz, 3H), 0.93-0.86 (t, 6H), 0.72 (s, 3H) Example 31 68.5 gua Ή NMR (400 MHz, CHLOROFORM-7) δ 8.62 (br s, 1H), 8.09 (br d, 7 = 7.8 Hz, 2H), 7.96 (br d, 7 = 6.0 Hz, 1H), 7.62-7.55 (m, 1H), 7.53-7.46 (m, 2H), 6.78-6.63 (m, 1H), 6.70 (br d , 7 = 5.8 Hz, 1H), 4.41-4.21 (m, 2H), 3.64 (br s, 1H), 3.37 (br s, 1H), 1.98-1.66 (m, 12H), 1.63- 1.48 (m, 5H), 1.45-1.02 (m, 15H), 0.97-0.89 (m, 4H), 0.85-0.78 (m, 6H), 0, 59 (s, 3H). Example 32 76.8 Chu Ή NMR (400 MHz, CHLOROFORM-7) δ 8.74 (br s, 1H), 8.19-8.01 (m, 1H), 8.12 (br s, 1H), 7.60 (br d , J = 7.8 Hz, 2H), 7.35-7.26 (m, 1H), 7.30 (br t, J = 7.5 Hz, 1H), 7.13-7.04 (m , 1H), 6.80 (br s, 1H), 4.46-4.23 (m, 2H), 3.64 (br s, 1H), 3.34 (br s, 1H), 1.97 -1.68 (m, 15H), 1.63-1.49 (m, 5H), 1.46-1.03 (m, 15H), 1.00-0.88 (m, 4H), 0 , 86-0.79 (m, 7H), 0.62 (s, 3H). Example 33 69.6 4l 'H NMR (400 MHz, METHANOL-7D δ 8.62 (s, 1H), 8.21-8.01 (m, 1H), 6.89 (br s, 1H), 4.52-4.30 (m, 2H), 3.68 (br s, 1H), 3.57- 3.44 (m, 1H), 2.93 (s, 3H), 2.04-1.25 (m, 25H), 1.07 (d, 7 = 6.5 Hz, 3H), 0.95-0.94 (m, 1H), 0.95-0.90 ( m, 6H), 0.74 (s, 3H) Example 34 xD Chu NMR (400 MHz, CHLOROFORM-7) δ 9.12 (br s, 1H), 8.63-8.46 (m, 1H), 8.56 (br s, 1H), 8.03 (br s , 1H), 7.17-6.99 (m, 1H), 4.69-4.38 (m, 2H), 3.65-3.28 (m, 5H), 3.03-2.87 (m, 1H), 2.09-1.57 (m, 13H), 1.52-1.13

- 31 038580

(m, 13H), (m, 4H), (m, 11H),

3H).

1.08-0.96

0.93-0.76

0.69 (s,

Path 8

Example 35

Example 35 was synthesized using Example 11 as a starting material and the procedure referred to the synthesis of Example 27. The reaction yield was 17.4%. 1H 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

Example 36 was synthesized using Example 23 as a starting material and the procedure referred to the synthesis of Example 27. The reaction yield was 17.5%. 1H 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).

Path 10

Example 37

Example 37 was synthesized using Example 23 as a starting material and the procedure referred to the synthesis of Example 27. The reaction yield was 60.8%. 1H 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

- 32 038580

Example 38 was synthesized using Example 22 as a starting material and the procedure referred to the synthesis of Example 27. The reaction yield was 53%. 1H NMR (400 MHz, methanol-d ₄ ) δ 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).

Track 12

Example 38

Example 39

Example 12A

Allow the reaction of glycine hydrochloride (14 mg, 112 μmol) with a solution of triethylamine (20.0 μl, 146 μmol) in ethyl acetate (5 ml) at 20 ° C for 0.5 h. To the system was added example 22 (50 mg, 97 μmol) and 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (36 mg, 146 μmol). The reaction system was allowed to react at 65 ° C for 10 hours. 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 hydrochloric acid. an aqueous solution of sodium hydroxide (15 ml), water (15 ml), 0.5 N. hydrochloric acid (15 ml) and water (15 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The crude product was isolated and purified by preparative isolation (TFA), thus obtaining Example 12A (25 mg, 43.9%). 1H NMR (400 MHz, CDCh) δ 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), 1.991.20 (m, 25H), 1.03 (d, J = 6.5 Hz, 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 system was allowed to react at 30 ° C for 4 hours. The system was brought to pH 5 with hydrochloric acid (1 M) and the aqueous phase was extracted with ethyl acetate (20 mlx2). The organic layers were combined and dried over sodium sulfate, filtered and evaporated to dryness. The residue was isolated by preparative TLC, thus obtaining Example 39 (24 mg, 98%). ¹ H NMR (400 MHz, methanol-d ₄ ) δ 8.66-8.65 (m, 1H), 8.11-8.02 (m, 1H), 6.84-6.82 (m, 1H ), 4.48-4.29 (m, 2H), 4.09-33 038580

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).

Track 13

Example 40

Example 13A

Example 2B was used as a starting material, while the procedure for Example 13A referred to the synthesis of Example 6A. The reaction yield was 16.3%. 1H 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

Cl o

For the procedure of Example 40, reference was made to the synthesis of Example 22. The reaction yield was 96.4%. 1H NMR (400 MHz, methanol ^) 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).

Track 14

- 34 038580

Example 14A h °। I ⁿ / [| A | A

BUT '' 'ΌΗ

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 system was allowed to react at 70 ° C. for 3 hours. The solvent was evaporated to dryness and the system was diluted with dichloromethane (500 ml). The organic phase was washed sequentially with saturated aqueous sodium carbonate solution (3x100 ml), aqueous solution (100 ml) and brine (100 ml) and dried over anhydrous sodium sulfate, filtered and concentrated. Example 14A (41 g, 100% yield) was obtained without purification and can be directly used in the next step.

Example 14B

O

G I ή I I '' OH

Example 14A (41 g, 95.1 mmol) was dissolved in anhydrous tetrahydrofuran solution (300 ml) and heated to 50 ° C. under nitrogen and a solution of methyl magnesium bromide (800 ml, 1M) in tetrahydrofuran was slowly added dropwise. After completion of the dropwise addition, the reaction solution was cooled to room temperature and stirred overnight. After completion of the reaction, cyclohexane (25 ml) was added and filtered. The filter residue was dissolved in a mixed solution of 3N hydrochloric acid. aqueous hydrochloric acid (800 ml) and dichloromethane (200 ml). This was stirred for an additional 30 minutes. After completion of the reaction, the organic phase was separated from the aqueous phase. The aqueous phase was additionally extracted with dichloromethane (2x200 ml). The organic phases were combined and washed sequentially with water (100 ml) and brine (100 ml), dried over anhydrous sodium sulfate, filtered and concentrated. Example 14B (51 g, 100% yield) was obtained without purification and can be directly used 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 the solution of Example 14B (51 g, 95.1 mmol) in anhydrous tetrahydrofuran (300 ml). The reaction solution was stirred overnight at room temperature. After completion of the reaction, this was diluted with an aqueous solution (100 ml). The aqueous phase was extracted with dichloromethane (3x150 ml). The organic phases were combined and washed with brine (100 ml), dried over anhydrous sodium sulfate, filtered and concentrated. Example 14C (55 g, 100% yield) was obtained without purification and can be used directly in the next step. 1H NMR (CDCl ₃ ) δ 0.66 (3H, s, CH ₃ -18); 0.77 (3H, s, CH ₃ -26); 1.00 (3H, d, CHs-21); 1.20 (3H, s, CH3-19); 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, J1 = 6.2Hz, J2 = 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 hydrochloric acid. aqueous solution of sulfuric acid (1.7 ml). After stirring for 15 minutes, the reaction solution was cooled to 0 ° C., and ruthenium trichloride (71.3 mg, 0.4 mmol) was added thereto. The reaction solution was continuously stirred until its color turned bright yellow. Ethyl acetate (27 ml) was added to the reaction solution, and

- 35 038580 acetonitrile (20 ml) and the above was stirred for another 5 minutes. Example 14C (4 g, 6.9 mmol) was added to the above reaction solution at 0 ° C, and after completion of the reaction, it was filtered. The filtrate was poured into water and extracted with ethyl acetate (3x100 ml). The organic phases were combined and washed with saturated sodium thiosulfate solution (200 ml), dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated to dryness and then isolated by silica gel column chromatography, thereby obtaining Example 14D (2.7 g, 89% yield). 1H NMR (CDCi ₃ ) δ 0.71 (3H, s, CH ₃ -18); 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

To a solution of Example 14D (1 g, 2.2 mmol) and isobutyl chloroformate (3.5 ml, 2.7 mmol) in tetrahydrofuran (20 ml) was added triethylamine (6.7 ml, 3.4 mmol) and the reaction was completed after 1 h. The reaction solution was filtered, and sodium borohydride (847 mg, 22.4 mmol) was added portionwise to the filtrate at 0 ° C. After completion of the reaction, water (3 ml) was added to quench the reaction, and the mixture was stirred at room temperature for another 2 hours, followed by acidification with 3N HCl. diluted aqueous hydrochloric acid and extracted with ethyl acetate (3x15 ml). The organic phases were combined and washed with brine (15 ml), dried over anhydrous sodium sulfate and filtered. After evaporating the filtrate to dryness, Example 14E (800 mg, 85% yield) was obtained by separation by column chromatography on silica gel. ¹ H NMR (CDCi ₃ ) δ 0.67 (3H, s, CH3-I8); 0.86-0.97 (9H, m, CH3-19, CH3-21, CH3-24); 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

Sodium hydride (18.4 mg, 460.1 μmol, 60%) was added to a solution of Example 14E (100 mg, 230.1 μmol) in, Ν-dimethylformamide (1 ml) at 0 ° C. After half an hour, a solution of methyl 4-chloropicolinate (78.9 mg, 460.1 μmol) in, Ν-dimethylformamide (2 ml) was slowly added dropwise at 0 ° C. After completion of the dropwise addition, the reaction system was slowly warmed to room temperature and allowed to react for 12 h. Water (10 ml) was added and the reaction system was adjusted to pH 6 with hydrochloric acid (1 M) and extracted with dichloromethane (10 mlx3). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness. A 10% sodium hydroxide solution (the volumetric ratio of methanol to water was 1: 1) (50 mg, 1.3 mmol, 0.5 ml) was added to a solution of the residue in methanol (1 ml) and the reaction was ensured in the reaction system at 70 ° C for 2 hours. The solvent was evaporated to dryness. The system was brought to a pH value of 1-2 with diluted hydrochloric acid (1 M) and extracted with a mixture of dichloromethane: methanol = 10: 1 (10 ml × 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative chromatography, thus obtaining Example 41 (5 mg, 4% yield). 1H NMR (400 MHz, methanol-d ₄ ) δ 8.67 (br d, J = 6.5 Hz, 1H), 7.99 (d, J = 2.5 Hz, 1H), 7.69 (br d, J = 6.5 Hz, 1H), 4.50 (br d, J = 6.5 Hz, 2H), 3.78-3.59 (m, 1H), 3.32-3.30 ( m, 1H), 2.18-1.17 (m, 25H), 1.10 (d, J = 6.5 Hz, 3H), 0.95-0.89 (m, 6H), 0.76 (s, 3H).

Track 15

Example 42

- 36 038580

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) was added 1,3-dicyclohexylcarbodiimide (101.5 mg , 492 μmol) and 4dimethylaminopyridine (1.5 mg, 12.3 μmol). Allow the reaction to proceed in the reaction system at 30 ° C for 16 hours. To the system was added a solution of lithium hydroxide (10.3 mg, 246 μmol) in water (2.0 ml). The reaction system was allowed to react at 30 ° C for 2 hours. The system was brought to pH 4 with hydrochloric acid (1M) and the aqueous phase was extracted with dichloromethane / methanol (10: 1) (20 ml × 3). The organic layers were combined and dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was isolated and purified by preparative isolation (HCl), thus obtaining Example 42 (20 mg, 15.4%). 1H NMR (400 MHz, methanol-d ₄ ) δ 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 = 6.5 Hz, 3H), 0.950.91 (m, 6H), 0.78 (s, 3H).

Track 16

Example 43

Example 16A

Reference Example 1F (10.0 g, 23.9 mmol) was dissolved in tetrahydrofuran (60.0 ml). To this was added perchloric acid (240.0 mg, 2.4 mmol, 144.6 μL) (approximately 10 drops). Formic acid (40.3 g, 874.7 mmol, 33.0 ml) was added dropwise over half an hour at 30 ° C. and the reaction solution was stirred at 50 ° C. for 11.5 hours. The solvent was removed by concentration. Water (35.0 ml) was added to the reaction solution, which was extracted 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 thereby obtain the crude product. The crude product was isolated by column chromatography, thus obtaining Example 16A (7.0 g, 15.7 mmol, 65.6% yield, white solid). 1H NMR (400 MHz, 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 = 13.1 Hz, 2H) , 1.82 (dd, J = 5.5, 16.8 Hz, 2H), 1.75 (dd, J = 6.5, 14.1 Hz, 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 = 6.5 Hz , 3H), 0.83 (t, J = 7.4 Hz, 3H), 0.68 (s, 3H).

- 37 038580

Example 16B

I. { ⁿ ho-Mv ^ o

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 dissolution of the solid, sodium nitrite (2.7 g, 39.0 mmol) was added in portions, which was stirred at 0 ° C for 1 h, heated to 40 ° C and stirred for another 1 h. The reaction solution was cooled to 30 ° C and neutralized with 0.5 mol of an aqueous sodium hydroxide solution (pH 7-8) at 0 ° C. The reaction solution was extracted with ethyl acetate (40 ml × 3) and the organic layer was washed with water (10 ml), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was isolated by column chromatography (silica gel), thus obtaining Example 16B (3.5 g, 8.5 mmol, 93.0% yield, pale yellow oil). ¹ H NMR (400 MHz, chloroform-d) δ 3.59-3.47 (m, 1H), 2.69 (q, J = 6.2 Hz, 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 = 7.2 Hz, 5H), 1.19-1.12 (m, 5H), 1.02-0.91 (m, 1H), 0, 80 (t, J = 7.4 Hz, 3H), 0.71-0.64 (m, 3H).

Example 16C

I T ^ COOH

[JhTr

Example 16B (3.5 g, 8.5 mmol) was dissolved in methanol (100.0 ml). To this was added an aqueous solution of potassium hydroxide (70.0 g, 1.3 mol, dissolved in 100.0 ml of water) and the reaction solution was stirred at 100 ° C for 12 hours. Part of the solvent was removed by concentration and it was extracted with dichloromethane (30 mlx3). The aqueous phase was acidified with 1 mol hydrochloric acid (pH 3-4) and extracted with ethyl acetate (30 ml × 3). The organic layer was washed with water (20 ml), dried over anhydrous sodium sulfate, filtered and concentrated to thereby obtain the crude product. Example 16C (3.2 g, 7.9 mmol, 93.5% yield, yellow oil) was obtained without purification. 1H NMR (400 MHz, chloroform-d) δ 3.65-3.50 (m, 1H), 2.71 (d, J = 5.8 Hz, 1H), 2.48 (dd, J = 2, 6, 14.9 Hz, 1H), 2.43-2.31 (m, 2H), 2.22-2.15 (m, 1H), 2.07-1.98 (m, 2H), 1.951 , 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 = 6.3 Hz, 3H), 0.86 (d, J = 10.3 Hz, 5H), 0.69 (s, 3H).

Example 16D

I? ⁴ COOH I f ⁿ I /

To an aqueous solution of sodium hydroxide (949.2 mg, 23.7 mmol, dissolved in 10.00 ml of water) was added Example 16C (3.2 g, 7.9 mmol). The reaction solution was heated to 80 ° C and sodium borohydride (1.8 g, 47.5 mmol) was added in portions. The reaction solution was stirred at 100 ° C. for 12 hours. Methanol (6 ml) was added dropwise and concentrated to remove part of the solvent. The reaction solution was acidified with 1 mol of hydrochloric acid (pH 5-6), extracted with ethyl acetate (40 ml × 3). The organic layer was washed with water (20 ml), dried over anhydrous sodium sulfate, filtered and concentrated. Example 16D (3.1 g, 7.6 mmol, 96.4% yield, white solid) was obtained without isolation of the crude product. 1H NMR (400 MHz, 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, 3H), 1.53-1.44 (m, 4H) , 1.41-1.37 (m, 1H), 1.36-1.27 (m, 4H), 1.24-1.13 (m, 4H), 1.04 (d, J = 6, 5 Hz, 3H), 0.92-0.88 (m, 6H), 0.73-0.69 (m, 3H).

Example 16E

I a '° i Г ^Н Г / I' OH

To a solution of Example 16D (100 mg, 245.8 μmol) in acetonitrile (2 mL) were added triethylamine (49.8 mg, 491.9 mmol, 68.2 μL) and N, O-dimethylhydroxylamine hydrochloride (24.0 mg, 245.9 μmol) in a nitrogen atmosphere. Then, after stirring at 25 ° C for 30 min, Obenzotriazole-N, N, N ', N'-tetramethyluronium tetrafluoroborate (98.7 mg, 307.4 μmol) was added and stirred for another 11.5 h.

- 38 038580

The reaction solution was poured into cold water (30 ml) and extracted with ethyl acetate (40 ml × 3). The organic layer was washed with water (20 ml), dried over anhydrous sodium sulfate, filtered and concentrated. Example 16E (90 mg, 91.4% yield, white solid) was obtained without isolation of the crude product. 1H NMR (400 MHz, 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 hydride (6.3 mg, 168.9 μmol) in tetrahydrofuran (2 ml) was added Example 16E (50 mg, 111.2 μmol) at -78 ° C and stirred at -78 ° C for 2 hours. water (0.006 ml) was added to complete the reaction, and the reaction solution was filtered. The filter cake was dried in an oven to give Example 16F (50 mg, 100%, white solid). 1H NMR (400 MHz, chloroform-d) δ 9.76 (s., 1H), 3.70 (br. S., 1H), 3.46-3.36 (m, 1H), 2.43- 2.39 (m, 1H), 2.24-2.18 (m, 1H), 2.001.97 (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) was added trifluoroacetic acid (57 μl, 0 , 8 mmol) and sodium triacetoxyborohydride (146.0 mg, 0.7 mmol). The reaction system was allowed to react at 20 ° C. for 16 hours. Ethyl acetate (30 ml) was added to the reaction system. The system was washed with saturated sodium bicarbonate solution (20 ml x 2) and brine (20 ml x 1). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated to dryness, thereby obtaining a crude product. The crude product was isolated by preparative isolation (TFA) to give Example 16G (60.0 mg, 29.7%). ¹ H NMR (400 MHz, chloroform-d) δ 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 = 6.5 Hz, 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 system was allowed to react at 40 ° C for 1 h. The system was brought to pH 4 with hydrochloric acid (1M) and the aqueous phase was extracted with dichloromethane / methanol (10: 1) (20 ml × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue was isolated and purified by preparative isolation (HCl) to give Example 43 (40 mg, 68%). 1H NMR (400 MHz, methanol-d ₄ ) δ 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 = 6.5 Hz, 3H), 0.95-0.87 (m, 6H), 0.72 (s, 3H).

Track 17

Example 44

- 39 038580

Concentrated sulfuric acid (368.0 mg, 3.8 mmol) was added to a solution of Example 22 (50.0 mg, 97.3 μmol) in methanol (5 ml). The reaction system was allowed to react at 70 ° C. for 12 hours. Water (5 ml) was added to the system. The system was brought to pH 7 with sodium hydroxide (1 M) and extracted with dichloromethane (10 ml × 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative chromatography, thus obtaining the title compound 44 (15 mg, 29.2% yield). ¹ H NMR (400 MHz, methanol ^) δ 8.76 (d, J = 2.0 Hz, 1H), 8.18 (dd, J = 2.5, 8.8 Hz, 1H), 6.82 (d, J = 8.8 Hz, 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 = 6.5 Hz, 3H), 0.93-0.86 (m, 6H), 0.71 (s, 3H).

Track 18

Example 45

Example 18A

Example 18A was synthesized using Example 23 as a starting material and the procedure referred to the synthesis of Example 12A. The reaction yield was 79.7%. ¹ H NMR (400 MHz, chloroform-d) δ 8.48 (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

For the procedure of Example 45, the synthesis of Example 39 was referred to. The reaction yield was 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 assessment.

Biochemical experiment for FXR.

The purpose of the experiment.

The activation effect of the compound on the binding reaction with FXR was determined using AlphaScreen.

Experimental materials.

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

2. Coactivator: Biotin-labeled steroid receptor coactivator (Anaspec).

3. Detection reagent: AlphaScreen detection kit (PerkinElmer).

- 40 038580

Experimental method.

1. Dilution of compounds. The compound to be tested was prepared as a 40 μM solution in DMSO and then diluted 3-fold to 10 concentration points. The reference compound was prepared as a 400 μM solution in DMSO and then diluted 1.5-fold to 10 concentration points. The diluted DMSO solution was added to the wells of a 384-well plate at a volume of 150 nL per well.

2. The human FXR protein labeled with glutathione-S-transferase and the steroid receptor coactivator labeled with biotin were formulated as a mixed solution with concentrations of 0.4 and 30 nM, respectively, added to the wells of a 384-well plate in a volume of 15 μl per well and incubated for 1 h at room temperature.

4. The mixed solution of acceptor beads in the AlphaScreen Detection Kit was diluted 125-fold and added to the wells of a 384-well plate in a volume of 7.5 μl per well. The operation was protected from light during the experimental process. Incubation was carried out for 1 h at room temperature.

5. The mixed solution of donor beads in the AlphaScreen Detection Kit was diluted 125-fold and added to the wells of a 384-well plate in a volume of 7.5 μl per well. The operation was protected from light during the experimental process. Incubation was carried out for 1 h at room temperature.

6. Test for EU ₅₀ . Envision was applied at an excitation wavelength of 680 nm with an absorbance readout at 520-620 nm.

7. Analytical data. Data were analyzed using Prism 5.0 and EC ₅₀ values for the activation effects of the compounds were calculated. Then, the ratio of the highest compound signal value to the reference compound value was applied to obtain a percentage of compound activation efficiency.

Cellular experiment for FXR.

The purpose of the experiment.

The effect of the compound on cellular functional activity was determined by the method using the β-lactamase reporter gene.

Experimental materials.

1. Cell line: FXR HEK 293T DA.

2. Cell culture medium: DMEM medium supplemented with 10% serum and penicillin / streptomycin mixture (1x).

3. Detection reagent: GeneBLAzer® reporter gene detection kit (Invitrogen).

Experimental method.

1. Dilution of compounds. The compound to be tested was prepared as a 100 μM solution in DMSO and then the compound was diluted 3-fold to 10 concentration points. The reference compound was prepared as a 100 μM solution in DMSO and then diluted 1.3-fold to 10 concentration points. The diluted DMSO solution was added to the wells of a 384-well plate in a volume of 200 nL per well.

2. Cell inoculation. FXR HEK 293T DA cells were reconstituted, resuspended in culture medium, diluted to 5 x 10 cells / ml and added to the wells of a 384-well plate in a volume of 40 μl per well.

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

4. 6 μl of 1 mM LiveBLAzer ™ -FRET B / G substrate (CCF4-AM) was mixed with 60 μl of solution B and 934 μl of solution C and added to the wells of a 384-well plate in a volume of 8 μl per well.

5. The 384-well microplate was incubated in the dark for 2 hours at room temperature.

6. Test for EU ₅₀ . Envision was applied at an excitation wavelength of 409 nm with absorption readings at 460 and 530 nm.

7. Analytical data. Data were analyzed using Prism 5.0 and EC ₅₀ values were calculated to demonstrate the activation effects of the compounds. Then, the ratio of the highest compound signal to the reference compound (chenodeoxycholic acid, CDCA) was used to obtain the percentage of compound activation efficiency.

- 41 038580

Table 1

_{EC 50} Test Results for Biochemistry Experiment and Cellular Experiment

Test samples (Specified in the title Enzymatic activity of FXR Cellular FXR Activity EC ₅₀ (μM) Efficiency EC50 (μM) Efficiency compound) Chenodeoxycholic acid, CDCA 12.14 100% 10.22 100% Example 1 0.9 320% 17.9 134% Example 2 1.46 114% Example 3 2.62 106% Example 4 0.72 334% 1.87 131% Example 5 0.09 134% 0.57 190% Example 6 0.36 375% 5.16 132% Example 7 0.03 360% 0.06 140% Example 8 oh and 264% Example 9 0.16 407% 0.14 130% Example 10 0.06 188% 0.2 138% Example 11 0.03 241% 0.1 132% Example 12 0.07 240% Example 13 0.39 270% Example 14 0.73 195% Example 16 0.31 247% Example 17 0.60 313% 2.50 131% Example 18 0.07 187% Example 19 0.15 251% 0.37 143% Example 20 0.06 287% Example 22 0.006 249% Example 23 0.0025 248% 0.003 150% Example 24 0.0025 138% Example 25 0.011 233% Example 26 0.13 280% Example 27 0.006 212% Example 28 0.007 219% Example 29 0.012 190% Example 30 0.056 150% Example 31 0.027 204% Example 32 0.650 140% Example 33 0.141 194% Example 34 0.093 191% Example 38 0.02 210% Example 39 0.02 205% Example 40 0.045 197% Example 41 0.08 127% Example 42 1.43 121% Example 43 0.37 193% Example 44 0.330 140%

Output.

The agonist effect of the compound of the present invention on the FXR receptor is significant and the agonist effect on the FXR receptor at the cellular level is also significant.

Experimental example 2. In vivo study.

Pharmacokinetics in mice treated with one compound.

male mice (C57BL / 6J) were randomly divided into two groups, i.e. 6 mice in each group. The first group was the intravenous group, which administered a dose of 2 mg / kg, 2 ml / kg by tail vein injection (the carrier medium was a 10% aqueous solution of HPbCD, and if the solubility of the drug was not satisfactory, a co-solvent was added ); the second group was an oral administration group providing an intragastric dose of 10 mg / kg, 10 ml / kg (the carrier medium was a 0.5% aqueous solution of HPMC). Plasma samples (using K2-EDTA as anticoagulant) were collected at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration in the intravenous group; and plasma samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration in the oral administration group. For 6 animals in each group, blood samples were collected from 3 animals at the same time point. The first batch of 3 animals was taken in turn with the second batch of 3 animals. Plasma samples were analyzed using LC-MS / MS. The resulting plasma concentrations were plotted against time and PK parameters were calculated using Phoenix WinNonlin 6.3.

- 42 038580

table 2

Compound INT-747 Example 11 Example 22 Example 23 Dose (mg / kg) ten ten ten ten Options With max. 1013 1433 936 1777 PK in blood plasma (nM) Tmax. (h.) 0.3 2 0.5 0.5 AUC (n.h.) 993 5173 2337 1109 F% 13% - 34% twenty%

Output.

As shown in table. 2, after oral administration at the same dose, the peak concentration and drug effect of Compound 11 were higher than those of the control compound INT-747. After oral administration at the same dose, the peak concentration of Compound 22 was close to that of the control compound INT-747, and the drug effect of Compound 22 was higher than that of the control compound INT-747. After oral administration at the same dose, the peak concentration of Compound 23 was higher than that of the INT-747 control, and the drug effect of Compound 23 was also higher than that of the INT-747 control.

Liver to Blood Ratio Experiment in Mice Using Cassette Dosing 6 male mice (C57BL / 6J) were grouped into an oral administration group. The composition contained 5 types of developed drugs and intragastric administration was carried out at a dose of 2 mg / kg / compound (the carrier medium was a 0.5% aqueous solution of HPMC). The five compounds were first and respectively dissolved in a carrier medium and sonicated or rotated to form a 1 mg / ml solution (pure solution or suspension), respectively; and then solutions of the five compounds were mixed in equal volumes (1: 1: 1: 1: 1, v: v: v: v: v) in a glass vessel. After intragastric oral administration, blood plasma and liver tissue samples were collected from 3 animals 0.5 h after administration; corresponding samples were collected from the other 3 animals 3 hours after administration. After harvest, liver tissue was homogenized using ice-cold homogenization buffer (methanol: 15 mM PBS buffer (pH 7.4) = 1: 2, v: v) based on the ratio of liver weight: volume of homogenization buffer = 1: 3. Plasma and liver tissue samples were analyzed using a pre-engineered five-in-one LC-MS / MS assay. Plasma and liver tissue homogenate concentrations were obtained, and the ratio of liver tissue concentrations to plasma concentrations was calculated using Excel.

Table 3

Compound INT-747 Example 22 Example 26 Dose (mg / kg) 2 2 2 RK parameters Concentration in the liver (nM) 0.5 h / W h. 711/625 1959/701 3904/358 Plasma concentration (nM) 0.5 h / W h. 151/63 83/45 387/18 The ratio of the concentration in the liver to the concentration in the blood plasma is 0.5 h / W h. 5/10 24/16 10/20

Note: ND indicates undefined.

Output.

As shown in table. 3, after oral administration of the compound of the present invention at the same dose, the liver concentrations of the drug based on Example 22 at 0.5 and 3 hours were higher than those of the control compound, and the liver / blood concentration ratios were also higher. than those for the control compound at 0.5 and 3 hours. The liver drug concentration for Example 26 at 0.5 hour was higher than that for the control compound and the liver / blood concentration ratios for Example 26 were higher than that for the control compound after 0.5 and 3 hours.

Claims (18)

CLAIM 1. Compound of formula (I) - 43 038580 where ring A is selected from 5-12 membered aryl, 5-12 membered heteroaryl containing from 1 to 2 heteroatoms, 5-6 membered non-aromatic heteropyclyl containing from 1 to 2 heteroatoms, or 5-6 membered cycloalkyl, and wherein said ring A is optionally substituted with 1, 2 or 3 Ra or 1, 2 or 3 oxo groups, and wherein said heteroatom is selected from N, O or S; L is selected from C _1-4 αalkyl, C _1-4 α alkoxy, C _1-4 alkylthio, C _1-4 alkylamino, C _1-4 alkyl-C (= O) NH-, C _2-4 alkenyl, or C _{1- 3} α-alkyl-O-C _1-3 α-alkyl, and wherein said L is optionally substituted with 1, 2 or 3 Rb; L1 is selected from O, N (Rd), N (Rd) S (= O) 2, or N (Rd) S (= O); R _{1 is} selected from H, C _1-6 alkyl, C _1-6 heteroalkyl containing one or more heteroatoms selected from sulfur, oxygen and / or nitrogen, 5-6 membered aryl, 4-6 membered heteroaryl containing one or more heteroatoms selected from sulfur, oxygen and / or nitrogen, C _3-6 cycloalkyl or 3-6 membered heterocycloalkyl containing one or more heteroatoms selected from sulfur, oxygen and / or nitrogen, and wherein said C _{1- 6} αalkyl, C _1-6 heteroalkyl, 5-6 membered aryl, 4-6 membered heteroaryl, C _3-6 cycloalkyl or 3-6 membered heteropicloalkyl optionally substituted with 1, 2 or 3 Rc or 1, 2 or 3 oxo groups ; each of Ra, Rb, R _c and Rd is independently selected from H, 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, and wherein said C _1-3 alkylamino, N, N-di (C _{1- 3} alkyl) amino, C _1-3 alkyl, C _1-3 alkyloxy, or C _1-3 αalkylthio are 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 ) or N (CH3) 2; or a pharmaceutically acceptable salt thereof. 2. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein Ra, Rb, R _c and R _{d 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. 3. A compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein ring A is selected from phenyl, pyridyl, pyridin-2 (1H) -onyl, pyrimidyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl, pyrrolidinyl, piperidinyl, morpholinyl , piperazinyl, isoxazolyl, isothiazolyl, bicyclo [1.1.1] pentyl, benzoxazolyl, benzo [d] isoxazolyl, indazolyl, indolyl, quinolinyl, isoquinolinyl, quinazolinyl, 1H-pyrrolo [2,3-B] pyridyl, benzothiazolyl, benzothiazolyl or benzothiazolyl and wherein said Ring A is optionally substituted with 1, 2 or 3 R _a . 4. A compound or pharmaceutically acceptable salt thereof according to claim 3, wherein ring A is selected from and wherein said Ring A is optionally substituted with 1, 2 or 3 R _a . 5. A compound or pharmaceutically acceptable salt thereof according to claim 4, wherein ring A is selected from - 44 038580 6. A compound or pharmaceutically acceptable salt thereof according to claim 5, wherein ring A is selected from 7. A compound or pharmaceutically acceptable salt thereof according to claim 1, wherein L is selected from or 'X- -', wherein said L is optionally substituted with 1, 2 or 3 R _b . 8. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein L is selected from 9. A compound or pharmaceutically acceptable salt thereof according to claim 8, wherein L is selected from 10. A compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein L _{1 is} selected from O, NH, NHS (= O) 2 or NHS (= O). 11. A compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R1 is selected from H, C1 ₃ alkyl, C _3-6 cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl, and wherein said C _1-3 α-alkyl, C ₃ 6 -cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl are optionally substituted with 1, 2 or 3 Rc, and wherein said R _c is as defined above. 12. A compound or a pharmaceutically acceptable salt thereof according to claim 11, wherein R1 is selected from H, Me, - 45 038580 13. The connection according to claim 1, where the connection is selected from but - 46 038580 - 47 038580 - 48 038580 14. A pharmaceutical composition for the treatment of diseases associated with a farnesoid Chreceptor, containing a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13 and a pharmaceutically acceptable carrier. 15. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 13 for the manufacture of a medicament for the treatment of diseases associated with the farnesoid X-receptor. 16. Use according to claim 15, wherein the diseases are selected from fibrotic diseases, chronic liver disease, diseases associated with high cholesterol levels, diseases associated with high triglycerides, or cardiovascular diseases. 17. Use according to claim 15 or 16, wherein the diseases are selected from non-alcoholic fatty liver disease (NAFLD), cholestatic hepatopathy, hepatitis C virus infection, alcoholic liver disease, liver fibrosis, primary sclerosing cholangitis (PSC), gallbladder stone , biliary atresia, lower urinary tract symptom and benign prostatic hyperplasia (BPH), ureteral stones, obesity, type 2 diabetes, arteriosclerosis, liver dysfunction due to hypercholesterolemia or hyperlipidemia. 18. Use according to claim 15 or 16, wherein the diseases are selected from non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), or atherosclerosis.