CN108191733B - Benzene-pyrrolidine compound, preparation method thereof and application thereof in resisting hepatic fibrosis - Google Patents

Abstract

The invention provides a benzene-pyrrole compound with a novel structure, a preparation method thereof and anti-hepatic fibrosis application, which is a compound shown in a formula (I), and an enantiomer, a diastereoisomer, a racemate, a pharmaceutically acceptable salt, a crystal hydrate or a solvate thereof. Part of the compounds with the structure show good affinity to vitamin D receptors and I-type collagen synthesis resistance, and can be applied to the field of preparing medicaments for treating hepatic fibrosis diseases.

Description

Benzene-pyrrolidine compound, preparation method thereof and application thereof in resisting hepatic fibrosis

Technical Field

The present invention relates to the fields of medicinal chemistry and pharmacotherapeutics. In particular, the invention relates to a benzene-pyrrolidine compound (I) with a novel structure and derivatives thereof, a preparation method thereof, a pharmaceutical composition and application thereof in preparing medicaments for treating diseases related to vitamin D receptors, especially liver fibrosis diseases.

Background

Hepatic fibrosis (hepatic fibrosis) is a compensatory response in the course of tissue repair following inflammation or injury of the liver due to various causes, pathologically characterized by excessive deposition of extracellular matrix (ECM) in the liver^[1]. Common causes of liver fibrosis are damage to hepatocytes induced by factors such as alcohol, ischemia, parasites, HBV and HCV viral infections, autoimmune attack, non-alcoholic fatty liver disease, drug therapy, and hepatotoxin, leading to activation and release of large amounts of cytokines and soluble factors by other types of hepatocytes such as hepatic Kupffer cells, hepatic sinus endothelial cells, hepatic fibroblasts, and migratory inflammatory cells, while these factors lead to activation, differentiation and proliferation of Hepatic Stellate Cells (HSCs) or hepatic fibroblasts (MFBs)), synthesis of large amounts of ECM, and gradual deposition of ECM, leading to liver fibrosis.

Chronic liver disease and cirrhosis have become health concerns worldwide. Chronic liver disease has become the fifth leading killer following heart disease, cancer, stroke and chest disease in australia and the uk. China is a high incidence region of viral hepatitis, only the carriers of hepatitis B patients reach 1.2 hundred million, wherein more than 3000 ten thousand of chronic hepatitis B patients exist, and about 25 percent of chronic hepatitis B patients finally develop cirrhosis, even liver cancer^[2]. Other diseases such as alcoholic liver disease, nonalcoholic steatohepatitis (NASH), autoimmune hepatitis, and Primary Biliary Cirrhosis (PBC) are not rare in China, and hepatic fibrosis and cirrhosis may also occur. Chronic liver disease caused by various reasons is accompanied with the occurrence of hepatic fibrosis in the course of disease, while progressive hepatic fibrosis can not be effectively controlledThe liver cirrhosis and even liver cancer finally develop, and the health and the life of the human beings are greatly threatened.

In recent years, with the extensive and intensive research on the pathogenesis of liver fibrosis, it has been clarified that liver fibrosis is completely reversible^[3]2013, Salk biological research finds that hepatic stellate cells causing hepatic fibrosis have high-level vitamin D receptors, and after vitamin D is given, the hepatic stellate cells can simultaneously resist the expression of various hepatic fibrosis factors by acting on a genome of a TGF- β 1/Smad signal pathway^[22]Thereby inhibiting the formation of liver fibrosis. Therefore, the development of agonists targeting vitamin D receptors opens a new door to the treatment of liver fibrosis.

1,25(OH)₂D₃As natural Vitamin D Receptor Agonists (VDRAs), have strong activity and a wide range of physiological functions. But because of its ability to bind to the vitamin D Binding Protein (DBP) in blood^[17]Resulting in an extended half-life, chronic administration can cause elevation of blood calcium and damage to the neuromuscular system, cardiovascular system, gastrointestinal system, urinary system and skeletal system, known as hypercalcemia (hypercalcemia). This side effect also becomes limiting 1,25(OH)₂D₃The key problem of wide application in clinic. Therefore, attempts to solve the problem of 1,25(OH) have been made₂D₃The structural modification of the vitamin D aims to find the analogues of the active vitamin D, so that the calcium-phosphorus metabolism regulation function of the vitamin D is effectively separated from other functions, and the clinical adverse reactions such as hypercalcemia and the like are reduced while certain non-calcium-phosphorus regulation physiological activity is maintained.

Up to now, there are about 3000 more 1,25(OH)₂D₃The steroid analogue of (a) is synthesized. The improved physiological activity of the steroid compounds is separated from the ability of increasing blood calcium to a certain extent, but for the treatment of oral administration, the harm of the side effect of the steroid compounds still can not ensure that the steroid compounds can be used for the treatment of osteoporosis, tumors and severe psoriasis, and the steroid compounds have complicated structures,The stability is low and the synthesis difficulty is large, so the research and development of a VDR receptor agonist with high efficiency and low toxicity is still urgently needed in the field.

In 1999, a dual benzene ring non-steroidal VDR agonist-LG 190178 was first reported. The compound not only retains the differentiation promoting capability of steroid compounds on HL-60 cells and the anti-proliferation capability of various tumor cells, but also overcomes the binding capability on DBP. The results of further in vivo experiments show that the compounds can achieve physiological activity and can be well separated from the ability of increasing blood calcium, and a new direction is opened for developing selective VDR agonists with novel structures.

Disclosure of Invention

The invention aims to provide a benzene-pyrrole compound with VDR (VDR) agonistic activity shown as a general formula (I), and pharmaceutically acceptable salts, enantiomers, diastereomers, racemates or mixtures thereof, and pharmaceutically acceptable salts, crystal hydrates and solvates thereof.

Another object of the present invention is to provide a process for the preparation of said compounds of general formula (I).

Another object of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the group consisting of the compounds of the general formula (I), enantiomers, diastereomers, racemates thereof and mixtures thereof, and pharmaceutically acceptable salts, crystalline hydrates and solvates thereof; and one or more pharmaceutically acceptable carriers.

The invention also aims to provide application of the compound shown in the general formula (I), enantiomers, diastereomers, racemes and mixtures thereof, and pharmaceutically acceptable salts, crystal hydrates and solvates thereof in preparing medicaments for treating diseases related to vitamin D receptors, in particular application in preparing medicaments for treating hepatic fibrosis diseases.

The object of the invention can be achieved by the following measures:

the invention relates to a compound shown as a formula (I), and an enantiomer, a diastereoisomer, a racemate, a pharmaceutically acceptable salt, a crystal hydrate or a solvate thereof,

wherein the content of the first and second substances,

r or R' are each independently hydrogen, an isotope of hydrogen, halogen, C_1-6Alkyl or C_1-6Haloalkyl, or R and R' are joined together to form a substituted or unsubstituted, saturated or unsaturated, 3-to 8-membered cycloalkane, wherein the substituents are selected from halogen, amino, hydroxy, nitrile or nitro;

R_Por R_TEach independently selected from hydrogen, hydrogen isotope, halogen, amino, hydroxyl, nitrile group, nitro, hydroxymethyl and C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Alkoxy radical, C_1-6Haloalkoxy, C_1-6Alkylthio radical, C_1-6Haloalkylthio, C_1-6Alkanoyl radical, C_1-6Haloalkanoyl group, C_2-6Alkenyl or C_2-6A haloalkenyl group;

(L_P) Or (L)_T) Each independently is a divalent linking group independently selected from the group consisting of:

m is 0, 1 or 2, X₁Is oxygen or sulfur, R₄₀Independently hydrogen, isotopes of hydrogen, halogen, C_1-6Alkyl radical, C_1-6Haloalkyl or C_3-6A cycloalkyl group;

Z_Por Z_TEach independently selected from: I. br, Cl, F, X₃、-X₃-X₄、

X₃Or X₄Is C_1-5Alkyl radical, C_1-5Haloalkyl, C_3-5Cycloalkyl or C_3-5Halogenated cycloalkyl radicals，X₂Is oxygen or sulfur.

In one embodiment, R or R' are each independently hydrogen, halogen, C_1-4Alkyl or C_1-4Haloalkyl, or R and R' together are linked to form a saturated or unsaturated 3-to 8-membered cycloalkane.

In a preferred embodiment, R or R' are each independently hydrogen, C_1-4Alkyl or C_1-4Haloalkyl, or R and R' together are linked to form a saturated or unsaturated 3-to 8-membered cycloalkane.

In another preferred embodiment, R or R' are each independently C_1-4Alkyl or C_1-4A haloalkyl group.

In another preferred embodiment, R or R' are each independently C_1-4Alkyl, in particular methyl, ethyl or propyl.

In one embodiment, R_POr R_TEach independently selected from hydrogen, halogen, amino, hydroxy, hydroxymethyl, C_1-4Alkyl radical, C_1-4Haloalkyl, C_1-4Alkoxy or C_1-4A haloalkoxy group.

In a preferred embodiment, R_POr R_TEach independently selected from hydrogen, halogen, C_1-4Alkyl or C_1-4A haloalkyl group.

In another preferred embodiment, R_POr R_TEach independently selected from hydrogen or C_1-4An alkyl group.

In another preferred embodiment, R_POr R_TEach independently selected from C_1-4Alkyl, in particular methyl, ethyl or propyl.

In one embodiment, (L)_P) Or (L)_T) Each independently selected from the following groups:

in a preferred embodiment, (L)_P) Or (L)_T) Are independently selected fromFrom:

in another preferred embodiment, (L)_P) Is selected from

(L_T) Is selected from

M in the present invention is 0, 1 or 2, preferably 1 or 2.

X in the invention₁Is oxygen or sulfur.

In one embodiment, Z_POr Z_TEach independently selected from: x₃、-X₃-X₄、

In a preferred embodiment, Z_POr Z_TEach independently selected from: x₃、-X₃-X₄、

In another preferred embodiment, Z_PIs selected from

In another advantageIn option, Z_PIs selected from

In another preferred embodiment, Z_TIs selected from X₃、-X₃-X₄、

In another preferred embodiment, Z_TIs selected from X₃、-X₃-X₄、

X in the invention₃Is C_1-5Alkyl radical, C_1-5Haloalkyl or C_3-5A cycloalkyl group.

In a preferred embodiment, X is₃Is C_1-5Alkyl or C_1-5A haloalkyl group.

In a preferred embodiment, X is₄Is C_1-4Alkyl or cyclopropyl.

X in the invention₂Is oxygen or sulfur.

The present invention relates to compounds, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts, crystalline hydrates or solvates thereof, wherein the compounds are preferably selected from the group consisting of:

5- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (2-hydroxy-2-methyl) -1H-pyrrole-2-carboxamide (A01),

5- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-pentyl-1H-pyrrole-2-carboxamide (A02),

n- (cyclopropyl) -5- (3- (4- (2, 3-dihydroxypropyl) -3-methylphenyl) -3-yl) -ethyl-1-2-pyrrolidinamide (A03),

5- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (2-hydroxyethyl) -1H-pyrrole-2-carboxamide (A04),

5- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (2,2, 2-trifluoroethyl) -1H-pyrrole-2-carboxamide (A05),

5- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -N- (3- (dimethylamino) propyl) -1-ethyl-1H-pyrrole-2-carboxamide (A06),

n- (2- (dimethylamino) ethyl) -1-ethyl-5- (3- (4-hydroxy-3-pentylphenyl) -3-yl) -1H-pyrrole-2-carboxamide (A07),

5- (3- (4- (2, 3-dihydroxypropyl) -3-pentylphenyl) -3-yl) -1-pentyl ethyl-1H-pyrrole-2-carboxamide (A08),

4- (3- (4- (2, 3-dihydroxypropyl) -3-pentylphenyl) -3-yl) -1-ethyl-N- (2-hydroxy-2-methylpropyl) -1H-pyrrole-2-carboxamide (A09),

4- (3- (4- (2, 3-dihydroxypropyl) -3-pentylphenyl) -3-yl) -1-ethyl-N-neopentyl-1H-pyrrole-2-carboxamide (A10),

n- (cyclopropylmethyl) -4- (3- (4- (2, 3-dihydroxypropyl) -3-pentylphenyl) -3-yl) ethyl-1H-pyrrole-2-carboxamide (A11),

4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (2-hydroxyethyl) -1H-pyrrole-2-carboxamide (A12),

4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (2,2, 2-trifluoroethyl) -1H-pyrrole-2-carboxamide (A13),

4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (1-hydroxypropan-2-yl) -1H-pyrrole-2-carboxamide (A14),

4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl-N- (1-hydroxy-3-methylbutan-2-yl) -1H-pyrrole-2-carboxamide (A15),

4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-pentylethyl-1H-pyrrole-2-carboxamide (A16),

4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentane-3-yl) -1-ethyl- (3-hydroxypropyl) -1H-pyrrole-2-carboxamide (A17),

4- (4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrole) -3-pentyl) -2-methylphenoxy) -butyric acid (A18),

5- (4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrole) -3-pentyl) -2-methylphenoxy) -pentanoic acid (A19),

4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrole) -3-pentyl) -methylphenylalanine (A20),

4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrole) -3-pentyl) -methylphenyl 3-aminopropionic acid (A21),

4- (4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrole) -3-pentyl) -2-methylphenoxy) -oxobutanoic acid (a 22).

The invention also provides a preparation method of the compound of the general formula (I) and an intermediate thereof, and raw materials and reagents used in the invention are all purchased commercially if no special description is provided.

Reagents and reaction conditions: a boron trifluoride diethyl etherate, dichloromethane and 0 ℃; b, alkyl halide, sodium hydride, N-dimethylformamide and 0-25 ℃; c palladium on carbon, ammonium formate, methanol/ethyl acetate (10:1), 25 ℃; d 2mol/L potassium hydroxide, ethanol and room temperature; e organic amine or alcohol, condensing agent, triethylamine, dichloromethane and room temperature; f electrophilic reagent, alkali, N-dimethyl formamide, 80 ℃.

The preparation according to scheme one is described in detail below:

step a: preparation of compound 3: under the catalysis of boron trifluoride diethyl etherate, carrying out Friedel-crafts alkylation reaction on the compound 1 and the compound 2 to generate a compound 3;

step b: preparation of compound 4: under the alkaline condition, pyrrole nitrogen generates a compound 4 through alkylation reaction;

step c: preparation of compound 5: reducing the compound 4 to generate a compound 5;

step d: preparation of compound 6: hydrolyzing the compound 5 to generate a compound 6;

step e: preparation of compound 7: carrying out condensation reaction on the compound 6 and organic amine or alcohol to generate a compound 7;

step f: preparation of compound 8: compound 7 is reacted with an electrophile under basic conditions to form compound 8.

The "pharmaceutically acceptable inorganic or organic salt" in the present invention is a salt of the compound represented by the general formula (I) with an inorganic acid such as hydrochloric acid, hydrobromic acid, hydroiodic acid, hydrofluoric acid, sulfuric acid, nitric acid or phosphoric acid, a salt with an organic acid such as formic acid, oxalic acid, propionic acid, oxalic acid, malonic acid, maleic acid, tartaric acid, malic acid, fumaric acid, methanesulfonic acid, citric acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, aspartic acid or glutamic acid, or a sodium, potassium, calcium or ammonia salt with a base such as sodium hydroxide, potassium hydroxide, calcium hydroxide or ammonia water.

The "isotope of hydrogen" in the present invention is deuterium (D or 2H) or tritium (T or 3H).

The halogen in the invention is F, Cl, Br or I.

As used herein, "alkyl" refers to saturated aliphatic groups containing 1 to 20 carbon atoms, including straight and branched chain groups (the numerical ranges mentioned in this application, e.g., "1 to 20", refer to groups, in this case alkyl, which may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms). Preferably, the alkyl group is a medium size alkyl group having 1 to 10 carbon atoms, such as C_1-10Alkyl, more preferably, the alkyl is a medium size alkyl having 1 to 6 carbon atoms, such as C_1-6An alkyl group; such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, and the like. Preferably, the alkyl group is a lower alkyl group having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, or the like.

The alkyl group in the present invention includes an alkylene group.

"haloalkyl" as used herein denotes halogen-substituted alkyl, preferably halogen-substituted lower alkyl as defined above, which is substituted by one or more of the same or differentSubstituted by halogen atoms, e.g. -CH₂Cl、-CF₃、-CH₂CF₃、-CH₂CCl₃And the like.

"cycloalkane" as used herein means a monocyclic or fused ring all carbon (by "fused" ring is meant that each ring in the system shares an adjacent pair of carbon atoms with other rings in the system) group in which one or more rings do not have a fully attached pi-electron system, examples of cycloalkyl groups being, without limitation, cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane, and cycloheptatriene.

The "alkoxy group" as used herein means-O- (alkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.

"haloalkoxy" as used herein means an alkoxy group containing one or more substituents of the same or different halogen atoms.

"alkylthio" as used herein means an-S- (alkyl) group. Representative examples include, but are not limited to, methylthio, ethylthio, propylthio, butylthio, cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio, and the like.

"haloalkylthio" as referred to herein is an alkylthio group substituted by one or more same or different halogen atoms.

"alkanoyl" as used herein refers to the group-C (O) - (alkyl).

"haloalkanoyl" as used herein means an alkanoyl group containing one or more substituents which may be the same or different halogen atoms.

The "hydroxyl group" as referred to in the present invention means an-OH group.

The "cyano group" as referred to herein denotes a-CN group.

The "amino group" as referred to in the present invention means-NH₂A group.

The "nitro group" in the present invention means-NO₂A group.

List of "alkenyl" groups in the present inventionUnsaturated aliphatic radicals having from 2 to 20 carbon atoms and containing one or more C ═ C double bonds are illustrated, including straight and branched chain radicals. Preferably, the alkenyl group is a medium size alkenyl group having 2 to 10 carbon atoms, such as C_2-10Alkenyl, more preferably, alkyl is a medium size alkenyl having 2 to 6 carbon atoms, such as C_2-6An alkyl group; such as vinyl, propenyl, allyl, and the like.

"haloalkenyl" as used herein refers to an alkenyl group containing one or more substituents of the same or different halogen atoms.

The invention discloses a pharmaceutical composition, which takes the compound, enantiomer, diastereoisomer, raceme, pharmaceutically acceptable salt, crystal hydrate or solvate thereof as an active ingredient or a main active ingredient.

In addition, the inventor finds that the compound of the general formula (I) has excellent vitamin D receptor binding activity through experiments, and the compound can be used for preparing experimental model tool medicines related to the vitamin D receptor or preparing medicines for treating and preventing diseases related to the vitamin D receptor.

The invention provides a pharmaceutical composition, which takes a compound shown in a formula (I), and enantiomers, diastereoisomers, racemes, pharmaceutically acceptable salts, crystal hydrates or solvates thereof as an active ingredient or a main active ingredient.

The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), enantiomers, diastereomers, racemates thereof and mixtures thereof, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers. The pharmaceutical composition may further comprise conventional additives such as flavoring agents, etc.

The pharmaceutical composition provided by the invention preferably contains 1-99% of the compound of the general formula (I) as an active ingredient by weight, preferably, the compound of the general formula (I) as an active ingredient accounts for 65-99% of the total weight of the pharmaceutical composition, and the balance is a pharmaceutically acceptable carrier and/or a conventional additive.

The compounds and pharmaceutical compositions provided herein may be in a variety of forms such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, and the like, and may be presented in suitable solid or liquid carriers or diluents and in suitable sterile devices for injection or instillation.

Various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional preparation methods in the pharmaceutical field. The unit dosage of the preparation formulation comprises 0.05-200mg of the compound with the general formula (I), and preferably, the unit dosage of the preparation formulation comprises 0.l-100mg of the compound with the general formula (I).

Pharmacodynamic experiments prove that: the compounds of the present invention have strong affinity for VDR receptors and can effectively inhibit the formation of fibrosis in vivo and in vitro. Therefore, the compound of the present invention, its enantiomer, diastereoisomer, racemate, pharmaceutically acceptable salt, crystal hydrate or solvate thereof can be used for treating or preventing diseases related to vitamin D receptor, or various fibrosis and related diseases, especially liver fibrosis diseases.

Drawings

Figure 1 is an HE and Masson staining assay of liver tissue in vivo following compound a01 treatment.

Detailed Description

This is further illustrated in the following examples. The examples are intended to illustrate the invention, but not to limit it in any way. All parameters in the examples and the remaining descriptions are based on mass, unless otherwise specified. Various raw materials of the preparation method which are not described in the specification can be commercially obtained.

Example 1:

1.1.3- (4- (benzyloxy) -3-methylphenyl) pentan-3-ol (Compound 2)

Reference is made to the preparation of Eur.J.Med.chem.2016.107, 48. starting from 4-hydroxy-3-methylbenzoic acid, the carboxyl group of which is first esterified and the phenolic hydroxyl group of which is then protected by a benzyl groupAnd then reacting with a Grignard reagent under the ice bath condition to obtain a target product. The yield in three steps: 80 percent; melting point: 67-70 ℃.¹H NMR(300MHz,CDCl₃)δ:7.49-7.28(5H,m),7.06(1H,s),7.03(1H,d,J＝8.3Hz),6.79(1H,d,J＝8.3Hz),5.04(2H,s),2.27(3H,s),1.77(4H,q,J＝7.5Hz),0.97(6H,t,J＝7.5Hz).

1.2.4- (3- (4- (benzyloxy) -3-methylphenyl) pentan-3-yl) -1H-pyrrole-2-carboxylic acid ethyl ester (Compound 3)

Compound 2(13g) and ethyl 2-pyrrolidinecarboxylate (7.1g) were dissolved in a dichloromethane solution, and BF was slowly added dropwise to the solution at 0 ℃₃·Et₂O (13mL), and the reaction was continued for 1 hour after the completion of the dropwise addition. After the reaction is finished, adding a proper amount of water, fully stirring, layering, separating an organic layer, extracting a water layer by using dichloromethane, combining the organic layers, washing by using a proper amount of saturated saline solution, drying by using anhydrous sodium sulfate, evaporating to remove a solvent, and carrying out column chromatography to obtain the target compound 3.¹H NMR(300MHz,CDCl₃)δ8.49(s,1H),7.53–7.30(m,5H),6.98(d,J＝6.8Hz,2H),6.87(dd,J＝3.7,2.5Hz,1H),6.81(d,J＝9.2Hz,1H),6.20(dd,J＝3.7,2.8Hz,1H),5.09(s,2H),4.28(q,J＝7.1Hz,2H),2.27(s,3H),2.12–1.97(q,J＝7.3Hz,4H),1.35(t,J＝7.1Hz,3H),0.69(t,J＝7.3Hz,6H).MS(TOF)m/z:428.3[M+Na]⁺.

1.3.4- (3- (4- (benzyloxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-1H-pyrrole-2-carboxylic acid ethyl ester (Compound 4)

Compound 3(4.05g) was dissolved in DMF solution and NaH (288mg) was added in portions under ice bath conditions. The reaction mixture was allowed to warm to room temperature, ethyl iodide (1.25g) was added thereto, and the mixture was reacted at room temperature for 2 hours. After the reaction is finished, slowly dropwise adding a proper amount of water under the ice bath condition, then adding a proper amount of ethyl acetate, fully stirring, layering, separating an organic layer, extracting a water layer with ethyl acetate, combining the organic layers, and using a proper amount of waterWashing with saturated saline solution, drying with anhydrous sodium sulfate, evaporating to remove the solvent, and carrying out column chromatography to obtain the target compound 4.¹HNMR(300MHz,CDCl₃)δ(ppm)7.51–7.30(m,5H),7.05(d,J＝4.1Hz,1H),6.90(d,J＝7.0Hz,2H),6.80(d,J＝8.8Hz,1H),6.27(d,J＝4.1Hz,1H),5.08(s,2H),4.25(q,J＝7.1Hz,2H),3.87(q,J＝6.8Hz,2H),2.25(s,3H),2.18–1.98(m,4H),1.36(t,J＝7.1Hz,3H),0.90(t,J＝6.9Hz,3H),0.64(t,J＝7.3Hz,6H).MS(TOF)m/z:456.3[M+Na]⁺.

1.4.1-Ethyl-4- (3- (4-hydroxy-3-methylphenyl) pentan-3-yl) -1H-pyrrole-2-carboxylic acid ethyl ester (Compound 5)

Compound 4(2.0g) was dissolved in a methanol solution, and then Pd/C (0.2g) and ammonium formate (2.9g) were added in this order to conduct a reaction at room temperature overnight. Filtering insoluble substances after the reaction is finished, adding a proper amount of water into the filtrate for dilution, extracting with ethyl acetate, separating an organic layer, extracting a water layer with ethyl acetate, combining the organic layers, washing with a proper amount of saturated saline solution, drying with anhydrous sodium sulfate, evaporating the solvent, and carrying out column chromatography to obtain the target compound 5.¹H NMR(300MHz,DMSO-d₆)δ(ppm)9.17(s,1H),6.89(d,J＝4.0Hz,1H),6.76(s,1H),6.70(s,2H),6.21(d,J＝4.1Hz,1H),4.14(q,J＝7.1Hz,2H),3.86–3.69(m,2H),2.05(s,3H),2.04–1.86(m,4H),1.23(t,J＝7.1Hz,3H),0.75(t,J＝6.7Hz,3H),0.53(t,J＝7.1Hz,6H).MS(TOF)m/z:342.2[M-H]^-.

1.5.1-Ethyl-4- (3- (4-hydroxy-3-methylphenyl) pentan-3-yl) -1H-pyrrole-2-carboxylic acid (Compound 6)

Compound 5(351mg) was dissolved in an ethanol solution, and an aqueous solution of potassium hydroxide was added thereto to conduct a reaction at 80 ℃ for 5 hours. After the reaction is finished, adding a proper amount of diluted hydrochloric acid to adjust the solution to acidity, extracting the solution by using ethyl acetate, separating an organic layer, extracting a water layer by using ethyl acetate, combining the organic layers, washing the organic layer by using a proper amount of saturated saline solution, and drying the organic layer by using anhydrous sodium sulfateDrying, evaporating the solvent, and carrying out column chromatography to obtain the target compound 6.¹HNMR(300MHz,DMSO-d₆)δ(ppm)11.85(s,1H),9.16(s,1H),6.84(d,J＝4.0Hz,1H),6.75(s,1H),6.69(s,2H),6.17(d,J＝4.0Hz,1H),3.76(d,J＝6.8Hz,2H),2.04(s,3H),1.93(dd,J＝19.6,12.7Hz,4H),0.74(t,J＝6.5Hz,3H),0.51(t,J＝7.1Hz,6H).MS(TOF)m/z 314.2[M-H]^-.

1.6.1-Ethyl-N- (2-hydroxy-2-methylpropyl) -4- (3- (4-hydroxy-3-methylphenyl) pent-3-yl) -1H-pyrrole-2-carboxamide (Compound 7a)

Compound 6(250mg) was dissolved in a dichloromethane solution, EDCI (175mg), HOBT (124mg), triethylamine (255. mu.L) and 1-amino-2-methyl-2-propanol (153mg) were added in this order under ice-bath conditions, and after completion of the addition, the mixture was allowed to stand at room temperature and stirred overnight. After the reaction is finished, adding a proper amount of water into the reaction solution, extracting with dichloromethane, separating an organic layer, extracting a water layer with dichloromethane, combining the organic layers, washing with a proper amount of saturated saline solution, drying with anhydrous sodium sulfate, evaporating the solvent, and carrying out column chromatography to obtain the target compound.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.89(s,1H),7.55(t,J＝6.1Hz,1H),6.82(s,1H),6.75(dd,J＝8.4,2.0Hz,1H),6.62(d,J＝1.8Hz,1H),6.56(d,J＝8.4Hz,1H),6.42(d,J＝1.8Hz,1H),4.45(s,1H),4.16(q,J＝7.0Hz,2H),3.03(d,J＝6.1Hz,2H),1.98(s,3H),1.79(dd,J＝14.7,6.7Hz,4H),1.12(t,J＝7.0Hz,3H),0.96(s,6H),0.50(t,J＝7.2Hz,6H).ESI-HRMS calcd for C₂₃H₃₅N₂O₃[M+H]⁺387.2642,found 387.2648.

1.7.4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-N- (2-hydroxy-2-methylpropyl) -1H-pyrrole-2-carboxamide (A01)

Compound 7a (386mg) was dissolved in DMF solution, and sodium hydrogen (80mg) was added to the solution in portions under ice bath conditionsStirring for 1 hour while keeping the temperature, adding glycidol (0.1mL), transferring the reaction solution into an oil bath at 80 ℃ and stirring for 5 hours, after the reaction is finished, adding a large amount of water into the reaction solution, extracting with ethyl acetate, separating an organic layer, extracting a water layer with ethyl acetate, combining the organic layers, washing with an appropriate amount of saturated saline solution, drying with anhydrous sodium sulfate, evaporating the solvent, and purifying by column chromatography to obtain the target compound A01.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.56(t,J＝6.0Hz,1H),6.96(s,2H),6.76(d,J＝9.0Hz,1H),6.69(s,1H),6.47(s,1H),4.80(d,J＝4.8Hz,1H),4.55(t,J＝5.6Hz,1H),4.46(s,1H),4.23(q,J＝7.0Hz,2H),3.96–3.86(m,1H),3.85–3.70(m,2H),3.45(dd,J＝10.4,5.2Hz,2H),3.10(d,J＝6.0Hz,2H),2.09(s,3H),1.89(dd,J＝13.9,6.6Hz,4H),1.20(t,J＝6.9Hz,3H),1.03(s,6H),0.59(t,J＝7.0Hz,6H).ESI-HRMS calcdfor C₂₆H₄₁N₂O₅[M+H]⁺461.301,found 461.3016.

Example 2:

n- (3- (dimethylamino) propyl) -1-ethyl-4- (3- (4-hydroxy-3-methylphenyl) pent-3-yl) -1H-pyrrole-2-carboxamide (Compound 7b)

Prepared by the method of example 1 by replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material N, N-dimethylpropylamine.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.88(s,1H),7.66(t,J＝6.3Hz,1H),6.89(s,1H),6.82(d,J＝8.4Hz,1H),6.62(d,J＝8.7Hz,2H),6.50(s,1H),4.22(q,J＝7.0Hz,2H),2.94(d,J＝6.4Hz,2H),2.05(s,3H),1.86(q,J＝6.9Hz,4H),1.18(t,J＝7.0Hz,3H),0.82(s,9H),0.58(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₄H₃₇N₂O₂[M+H]⁺385.285,found 385.2844.

2.2.4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -N- (3- (dimethylamino) propyl) -1-ethyl-1H-pyrrole-2-carboxamide (A02)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.67(t,J＝6.3Hz,1H),6.96(s,2H),6.76(d,J＝9.1Hz,1H),6.66(s,1H),6.51(s,1H),4.81(s,1H),4.57(s,1H),4.23(q,J＝6.9Hz,2H),3.90(dd,J＝9.1,4.3Hz,1H),3.86–3.71(m,2H),3.45(d,J＝5.3Hz,2H),2.94(d,J＝6.3Hz,2H),2.09(s,3H),1.89(d,J＝7.4Hz,4H),1.19(t,J＝7.0Hz,3H),0.82(s,9H),0.59(t,J＝7.0Hz,6H).ESI-HRMS calcd for C₂₇H₄₃N₂O₄[M+H]⁺459.3217,found 459.3221.

Example 3:

n- (cyclopropylmethyl) -1-ethyl-4- (3- (4-hydroxy-3-methylphenyl) pent-3-yl) -1H-pyrrole-2-carboxamide (Compound 7c)

Prepared by the method of example 1, replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material cyclopropylmethylamine.¹H NMR(300MHz,DMSO)δ8.89(s,1H),7.84(t,J＝5.6Hz,1H),6.88(s,1H),6.81(d,J＝8.4Hz,1H),6.67(s,1H),6.63(d,J＝8.3Hz,1H),6.42(s,1H),4.24(q,J＝6.8Hz,2H),2.97(t,J＝6.1Hz,2H),2.05(s,3H),1.86(d,J＝7.3Hz,4H),1.20(t,J＝6.9Hz,3H),0.92(dt,J＝11.7,5.8Hz,1H),0.57(t,J＝7.0Hz,6H),0.36(d,J＝8.0Hz,2H),0.14(d,J＝4.5Hz,2H).ESI-HRMS calcd for C₂₃H₃₃N₂O₂[M+H]⁺369.2537,found 369.2538.

N- (cyclopropylmethyl) -4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-1H-pyrrole-2-carboxamide (A03)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.81(t,J＝5.8Hz,1H),6.89(d,J＝7.1Hz,2H),6.69(d,J＝9.1Hz,1H),6.64(d,J＝1.8Hz,1H),6.34(d,J＝1.8Hz,1H),4.82(d,J＝4.7Hz,1H),4.57(t,J＝5.6Hz,1H),4.18(q,J＝7.0Hz,2H),3.83(dd,J＝9.1,4.3Hz,1H),3.71(dt,J＝13.6,5.2Hz,2H),3.38(dd,J＝10.6,5.3Hz,2H),2.89(t,J＝6.3Hz,2H),2.02(s,3H),1.81(dt,J＝10.2,6.4Hz,4H),1.13(t,J＝7.0Hz,3H),0.95–0.76(m,1H),0.50(t,J＝7.2Hz,6H),0.33–0.23(m,2H),0.10–0.01(m,2H).ESI-HRMS calcdforC₂₆H₃₉N₂O₄[M+H]⁺443.2904,found 443.291.

Example 4:

methyl (1-ethyl-4- (3- (4-hydroxy-3-methylphenyl) pentan-3-yl) -1H-pyrrole-2-carbonyl) glycinate (compound 7d)

Prepared by the method of example 1 by replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material glycine methyl ester.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.97(s,1H),8.25(t,J＝5.9Hz,1H),6.89(s,1H),6.83(d,J＝8.4Hz,1H),6.75(s,1H),6.64(d,J＝8.3Hz,1H),6.47(d,J＝1.5Hz,1H),4.23(q,J＝6.9Hz,2H),3.82(d,J＝5.8Hz,2H),3.61(s,3H),2.06(s,3H),1.94–1.78(m,4H),1.20(t,J＝7.0Hz,3H),0.59(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₂H₃₄N₂O₄[M+H]⁺387.2216,found 387.2219.

4.2.4- (3- (4- (2, 3-dihydroxypropyl) -3-methylpentan-3-yl) -1-ethyl-N- (2-hydroxyethyl) -1H-pyrrole-2-carboxamide (A04)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.75(d,J＝5.0Hz,1H),6.98(d,J＝6.8Hz,2H),6.78(d,J＝9.1Hz,1H),6.73(s,1H),6.44(s,1H),4.88(d,J＝4.2Hz,1H),4.63(s,2H),4.26(q,J＝7.0Hz,2H),3.92(dd,J＝9.1,4.3Hz,1H),3.80(dt,J＝14.2,5.3Hz,2H),3.44(dt,J＝11.8,5.7Hz,4H),3.18(dd,J＝11.9,6.0Hz,2H),2.11(s,3H),1.89(dd,J＝7.0,3.7Hz,4H),1.23(t,J＝7.0Hz,3H),0.59(t,J＝7.1Hz,6H).ESI-HRMScalcd for C₂₄H₃₇F₃N₂O₅[M+H]⁺433.2697,found 433.2692.

Example 5:

5.1.1-Ethyl-4- (3- (4-hydroxy-3-methylphenyl) pentan-3-yl) -N- (2,2, 2-trifluoroethyl) -1H-pyrrole-2-carboxamide (Compound 7e)

Prepared by the method of example 1 by replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material 2,2, 2-trifluoroethylamine.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.97(s,1H),8.36(t,J＝6.4Hz,1H),6.89(s,1H),6.86–6.76(m,2H),6.64(d,J＝8.4Hz,1H),6.58(d,J＝1.8Hz,1H),4.25(q,J＝7.0Hz,2H),3.98–3.83(m,2H),2.06(s,3H),1.87(dt,J＝8.6,6.4Hz,4H),1.21(t,J＝7.0Hz,3H),0.58(t,J＝7.2Hz,6H).ESI-HRMS calcd for C₂₁H₂₇F₃N₂NaO₂[M+Na]⁺419.1917,found419.1915.

5.2.4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-N- (2,2, 2-trifluoroethyl) -1H-pyrrole-2-carboxamide (A05)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.38(t,J＝6.2Hz,1H),6.98(d,J＝6.4Hz,2H),6.81(dd,J＝9.4,5.4Hz,2H),6.58(d,J＝1.5Hz,1H),4.90(d,J＝4.7Hz,1H),4.65(t,J＝5.6Hz,1H),4.27(q,J＝6.9Hz,2H),3.92(dt,J＝9.0,7.8Hz,3H),3.86–3.73(m,2H),3.47(dd,J＝10.7,5.4Hz,2H),2.11(s,3H),1.90(dd,J＝7.1,2.9Hz,4H),1.23(t,J＝7.0Hz,3H),0.60(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₄H₃₄F₃N₂O₄[M+H]⁺471.2465,found 471.2468.

Example 6:

(1-Ethyl-4- (3- (4-hydroxy-3-methylphenyl) pentan-3-yl) -1H-pyrrole-2-carbonyl) -D-alanine methyl ester (Compound 7f)

Prepared by the method of example 1 by replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material alanine methyl ester.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.96(s,1H),8.12(d,J＝7.0Hz,1H),6.89(s,1H),6.83(d,J＝8.4Hz,1H),6.74(d,J＝1.7Hz,1H),6.64(d,J＝8.3Hz,1H),6.57(d,J＝1.7Hz,1H),4.36–4.28(m,1H),4.22(q,J＝6.9Hz,2H),3.60(s,3H),2.06(s,3H),1.87(dt,J＝9.4,6.2Hz,4H),1.30(d,J＝7.3Hz,3H),1.19(t,J＝7.1Hz,3H),0.59(t,J＝6.9Hz,6H).ESI-HRMS calcd for C₂₃H₃₃N₂O₄[M+H]⁺401.2435,found 401.2434.

Methyl (4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-1H-pyrrole-2-carbonyl) -D-alanine (A06)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.44(d,J＝8.1Hz,1H),6.97(d,J＝6.7Hz,2H),6.77(d,J＝9.1Hz,1H),6.71(d,J＝1.5Hz,1H),6.45(d,J＝1.5Hz,1H),4.90(d,J＝4.7Hz,1H),4.65(dd,J＝5.6,4.1Hz,2H),4.25(q,J＝6.8Hz,2H),3.96–3.86(m,2H),3.85–3.72(m,2H),3.46(dd,J＝10.7,5.4Hz,2H),3.29–3.17(m,1H),2.10(s,3H),1.97–1.80(m,4H),1.21(t,J＝7.0Hz,3H),1.03(d,J＝6.7Hz,3H),0.58(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₅H₃₉N₂O₅[M+H]⁺447.2853,found 447.2847.

Example 7:

(1-Ethyl-4- (3- (4-hydroxy-3-methylphenyl) pentan-3-yl) -1H-pyrrole-2-carbonyl) -L-valine methyl ester (Compound 7g)

Prepared by the method of example 1 by replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material L-valine methyl ester.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.97(s,1H),7.96(d,J＝7.8Hz,1H),6.90(s,1H),6.86–6.79(m,1H),6.71(s,1H),6.69(s,1H),6.64(d,J＝8.4Hz,1H),4.26–4.17(m,2H),4.13(d,J＝7.7Hz,1H),3.61(s,3H),2.10(dd,J＝13.4,6.4Hz,1H),2.06(s,3H),1.88(d,J＝7.6Hz,4H),1.18(t,J＝7.0Hz,3H),0.89(dd,J＝12.1,6.7Hz,6H),0.59(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₅H₃₇N₂O₄[M+H]⁺429.2748,found 429.2744.

Methyl (4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-1H-pyrrole-2-carbonyl) -L-valine (A07)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.25(d,J＝8.9Hz,1H),6.89(s,2H),6.69(d,J＝9.1Hz,1H),6.61(s,1H),6.43(s,1H),4.80(d,J＝4.8Hz,1H),4.56(t,J＝5.6Hz,1H),4.42(t,J＝5.4Hz,1H),4.15(q,J＝6.7Hz,2H),4.05(d,J＝4.1Hz,1H),3.83(dd,J＝9.1,4.2Hz,1H),3.78–3.66(m,2H),3.60(s,1H),3.40(d,J＝5.4Hz,1H),3.36(d,J＝5.5Hz,2H),2.02(s,3H),1.82(d,J＝7.6Hz,4H),1.72(d,J＝6.5Hz,1H),1.12(t,J＝6.9Hz,3H),0.75(dd,J＝12.4,6.9Hz,16H),0.51(t,J＝6.9Hz,6H).ESI-HRMS calcdforC₂₇H₄₃N₂O₅[M+H]⁺475.3166,found 475.317.

Example 8:

n- (5-Aminopentyl) -1-ethyl-4- (3- (4-hydroxy-3-methylphenyl) pent-3-yl) -1H-pyrrole-2-carboxamide (Compound 7H)

Prepared by the method of example 1 by replacing the starting material 1-amino-2-methyl-2-propanol of example 1.6 with the starting material n-pentylamine.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.89(s,1H),7.72(t,J＝5.5Hz,1H),6.88(s,1H),6.81(d,J＝8.3Hz,1H),6.64(dd,J＝9.8,4.8Hz,2H),6.40(s,1H),4.23(q,J＝6.9Hz,2H),3.08(dd,J＝13.1,6.6Hz,2H),2.04(s,3H),1.85(dd,J＝13.9,6.5Hz,4H),1.50–1.35(m,2H),1.25(d,J＝17.1Hz,4H),1.18(d,J＝6.9Hz,3H),0.83(t,J＝6.8Hz,3H),0.57(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₂H₃₃N₂O₃[M+H]⁺373.2486,found 373.2486.

N- (5-Aminopentyl) -4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-1H-pyrrole-2-carboxamide (A08)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.78(t,J＝5.7Hz,1H),6.98(d,J＝7.7Hz,2H),6.78(d,J＝8.5Hz,1H),6.69(d,J＝1.6Hz,1H),6.43(d,J＝1.6Hz,1H),4.90(d,J＝4.8Hz,1H),4.65(t,J＝5.7Hz,1H),4.26(q,J＝6.9Hz,2H),3.94(dd,J＝8.9,4.1Hz,1H),3.82(dq,J＝16.0,5.2Hz,2H),3.50(dd,J＝10.7,5.4Hz,2H),3.10(dd,J＝13.2,6.6Hz,2H),2.11(s,3H),1.89(dt,J＝8.9,6.5Hz,4H),1.52–1.36(m,2H),1.32–1.23(m,4H),1.22–1.16(m,3H),0.84(t,J＝6.9Hz,3H),0.59(t,J＝7.1Hz,6H).ESI-HRMScalcdfor C₂₇H₄₃N₂O₄[M+H]⁺459.3217,found 459.3215.

Example 9:

9.1.1-Ethyl-4- (3- (4-hydroxy-3-methylphenyl) pent-3-yl) -N- (3-hydroxypropyl) -1H-pyrrole-2-carboxamide (Compound 7i)

The preparation method in example 1 is as followsThe material 1-amino-2-methyl-2-propanol is replaced by the material 3-hydroxy-propylamine.¹H NMR(300MHz,DMSO-d₆)δ(ppm)8.96(s,1H),7.79(t,J＝5.5Hz,1H),6.89(s,1H),6.82(d,J＝8.3Hz,1H),6.68(d,J＝1.7Hz,1H),6.64(d,J＝8.3Hz,1H),6.41(s,1H),4.44(t,J＝5.0Hz,1H),4.24(q,J＝6.9Hz,2H),3.42(d,J＝6.2Hz,2H),3.16(dd,J＝12.7,6.5Hz,2H),2.05(s,3H),1.95–1.76(m,4H),1.58(p,J＝6.6Hz,2H),1.21(t,J＝7.0Hz,3H),0.58(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₂H₃₃N₂O₃[M+H]⁺373.2486,found373.2486.

9.2.4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-N- (3-hydroxypropyl) -1H-pyrrole-2-carboxamide (A09)

The preparation is as described in example 1.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.70(t,J＝5.5Hz,1H),6.89(d,J＝6.8Hz,2H),6.69(d,J＝9.1Hz,1H),6.63(d,J＝1.4Hz,1H),6.32(d,J＝1.5Hz,1H),4.81(d,J＝4.7Hz,1H),4.56(t,J＝5.6Hz,1H),4.36(t,J＝5.3Hz,1H),4.17(q,J＝7.0Hz,2H),3.83(dd,J＝9.2,4.2Hz,1H),3.71(dt,J＝13.7,5.3Hz,2H),3.39(dt,J＝10.8,5.4Hz,2H),3.35–3.30(m,2H),3.07(dd,J＝12.8,6.5Hz,2H),2.02(s,3H),1.89–1.71(m,4H),1.49(p,J＝6.5Hz,2H),1.13(t,J＝7.0Hz,3H),0.50(t,J＝7.1Hz,6H).ESI-HRMScalcdfor C₂₅H₃₉N₂O₅[M+H]⁺447.2853,found 447.2845.

Example 10:

ethyl (4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenoxy) butanoate (compound 7j)

Compound 7a (386mg) was dissolved in DMF solution, sodium hydrogen (80mg) was added to the solution in portions under ice bath conditions, and the mixture was stirred for 1 hour while maintaining the temperatureThen, 4-bromobutyric acid ethyl ester (0.1mL) was added, the reaction solution was stirred in an oil bath at 80 ℃ for 5 hours, after the reaction was completed, a large amount of water was added to the reaction solution, extraction was performed with ethyl acetate, the organic layer was separated, the aqueous layer was extracted with ethyl acetate, the organic layers were combined, washed with an appropriate amount of saturated saline solution, dried over anhydrous sodium sulfate, the solvent was evaporated, and the target compound was obtained by column chromatography purification.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.53(t,J＝6.2Hz,1H),6.90(d,J＝1.9Hz,2H),6.73–6.65(m,1H),6.63(d,J＝1.7Hz,1H),6.41(d,J＝1.7Hz,1H),4.46(s,1H),4.16(dd,J＝13.9,6.8Hz,2H),3.96(q,J＝7.1Hz,2H),3.84(t,J＝6.1Hz,2H),3.03(d,J＝6.1Hz,2H),2.38(t,J＝7.3Hz,2H),2.00(s,3H),1.90(dd,J＝7.8,5.3Hz,2H),1.81(dd,J＝16.5,8.5Hz,4H),1.15-1.09(m,3H),1.07(t,J＝7.1Hz,3H),0.96(s,6H),0.51(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₉H₄₅N₂O₅[M+H]⁺501.3323,found 501.3324.

(4- (3- (1-Ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenoxy) butanoic acid (A10)

Dissolving a compound 7j (200mg) in an ethanol solution, adding a KOH aqueous solution into the solution at room temperature, transferring the solution into an oil bath at 80 ℃ after the KOH aqueous solution is added, stirring for 5 hours, adjusting the pH to about 3 by using 1M HCl after the reaction is finished, extracting by using ethyl acetate, separating an organic layer, extracting a water layer by using ethyl acetate, combining the organic layers, washing by using a proper amount of saturated saline solution, drying by using anhydrous sodium sulfate, evaporating a solvent, and purifying by using column chromatography to obtain the target compound.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.52(t,J＝6.0Hz,1H),6.90(s,2H),6.69(d,J＝9.2Hz,1H),6.64(d,J＝1.7Hz,1H),6.42(d,J＝1.6Hz,1H),4.16(q,J＝6.8Hz,2H),3.84(t,J＝6.1Hz,2H),3.03(d,J＝6.1Hz,2H),2.31(t,J＝7.2Hz,2H),2.01(s,3H),1.91–1.85(m,2H),1.85–1.74(m,4H),1.13(t,J＝7.0Hz,3H),0.96(s,6H),0.51(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₇H₄₁N₂O₅[M+H]⁺473.301,found473.3016.

Example 11:

ethyl (4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenoxy) pentanoate (Compound 7k)

Prepared by the method of example 1, replacing the glycidol starting material in example 1.7 with ethyl 5-bromovalerate.¹H NMR(300MHz,DMSO-d₆)δ(ppm)7.52(s,1H),6.90(s,2H),6.69(d,J＝9.0Hz,1H),6.63(d,J＝1.4Hz,1H),6.42(d,J＝0.8Hz,1H),4.42(s,1H),4.16(q,J＝6.7Hz,2H),3.95(q,J＝7.1Hz,2H),3.82(d,J＝5.4Hz,2H),3.03(d,J＝5.9Hz,2H),2.28(t,J＝6.5Hz,2H),2.01(s,3H),1.81(d,J＝7.6Hz,4H),1.63(d,J＝3.5Hz,4H),1.14(d,J＝6.6Hz,3H),1.10–1.02(m,4H),0.96(s,6H),0.51(t,J＝7.0Hz,6H).ESI-HRMS calcd for C₃₀H₄₇N₂O₅[M+H]⁺515.3479,found 515.3486.

(4- (3- (1-Ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenoxy) pentanoic acid (A11)

The preparation is as described in example 10.¹H NMR(300MHz,DMSO-d₆)δ(ppm)11.98(s,1H),7.58(t,J＝6.0Hz,1H),7.00(s,2H),6.79(d,J＝9.2Hz,1H),6.72(d,J＝1.5Hz,1H),6.51(s,1H),4.26(q,J＝7.0Hz,2H),3.92(t,J＝5.6Hz,2H),3.12(d,J＝6.1Hz,2H),2.29(t,J＝6.9Hz,2H),2.11(s,3H),1.91(d,J＝7.5Hz,4H),1.82–1.59(m,4H),1.22(t,J＝7.0Hz,4H),1.06(s,6H),0.61(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₈H₄₂N₂O₅[M+H]⁺487.3112,found487.3108.

Example 12:

12.1.4- (3- (1-Ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenyl (tert-butoxycarbonyl) -alanine (Compound 7l)

Compound 7a (386mg) was dissolved in a dichloromethane solution, EDCI (288mg), DMAP (12mg), triethylamine (418. mu.L) and Boc-alanine (153mg) were sequentially added to the solution under ice bath conditions, and the mixture was stirred at room temperature overnight. After the reaction is finished, adding a large amount of water into the reaction liquid, extracting with dichloromethane, separating an organic layer, extracting a water layer with dichloromethane, combining the organic layers, washing with a proper amount of saturated saline solution, drying with anhydrous sodium sulfate, evaporating the solvent, and purifying by column chromatography to obtain the target compound.¹HNMR(300MHz,CDCl₃)δ7.08(d,J＝7.5Hz,2H),6.90(d,J＝8.6Hz,1H),6.60(d,J＝1.5Hz,1H),6.31(s,1H),6.22(d,J＝1.6Hz,1H),5.14(d,J＝6.3Hz,1H),4.56(s,1H),4.32(q,J＝7.1Hz,2H),3.33(d,J＝5.8Hz,2H),2.14(s,3H),1.94(d,J＝7.3Hz,4H),1.56(d,J＝7.3Hz,3H),1.46(s,9H),1.36(t,J＝7.1Hz,3H),1.23(s,6H),0.65(t,J＝7.2Hz,6H).ESI-HRMScalcd for C₃₁H₄₈N₃O₆[M+H]⁺558.3538,found 558.3546.

(3- (1-Ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenyl L-alanine ester (A12)

After the compound (7 l, 116mg) was dissolved in a dichloromethane solution, trifluoroacetic acid was slowly added dropwise under ice-bath conditions, and after completion of the addition, the solution was stirred at room temperature for 2 hours, and the reaction was completed. Adding appropriate amount of water into the reaction solution, extracting with dichloromethane, separating organic layer, extracting water layer with dichloromethane, mixing organic layers, washing with appropriate amount of saturated saline solution, drying with anhydrous sodium sulfate, evaporating solvent, and purifying by column chromatography to obtain the target compound.¹H NMR(300MHz,CDCl₃)δ7.08(d,J＝7.5Hz,2H),6.90(d,J＝8.6Hz,1H),6.60(d,J＝1.5Hz,1H),6.31(s,1H),6.22(d,J＝1.6Hz,1H),5.14(d,J＝6.3Hz,1H),4.56(s,1H),4.32(q,J＝7.1Hz,2H),3.33(d,J＝5.8Hz,2H),2.14(s,3H),1.94(d,J＝7.3Hz,4H),1.56(d,J＝7.3Hz,3H),1.36(t,J＝7.1Hz,3H),1.23(s,6H),0.65(t,J＝7.2Hz,6H).ESI-HRMS calcd for C₃₁H₄₈N₃O₆[M+H]⁺458.3537,found458.3549.

Example 13:

methyl ((tert-butoxycarbonyl) -4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pentan-3-yl) -2-methylphenylcarbonyl) amino) propanoate (compound 7m)

Prepared according to the procedure for preparation of example 12 substituting the starting material Boc-alanine of example 20.1 with the starting material Boc- β -alanine.¹H NMR(300MHz,CDCl₃)δ7.08(d,J＝7.5Hz,2H),6.90(d,J＝8.6Hz,1H),6.60(d,J＝1.5Hz,1H),6.31(s,1H),6.22(d,J＝1.6Hz,1H),5.14(d,J＝6.3Hz,1H),4.56(s,1H),4.32(q,J＝7.1Hz,2H),3.43(m,2H),3.33(d,J＝5.8Hz,2H),2.14(s,3H),1.94(d,J＝7.3Hz,4H),1.46(s,9H),1.36(t,J＝7.1Hz,3H),1.23(s,6H),0.65(t,J＝7.2Hz,6H).ESI-HRMS calcd for C₃₁H₄₈N₃O₆[M+H]⁺558.3527,found 558.3539.

(3- (1-Ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pent-3-yl) -2-methylphenyl 3-aminopropionate (A13)

The preparation is as described in example 12.¹H NMR(300MHz,CDCl₃)δ7.08(d,J＝7.5Hz,2H),6.90(d,J＝8.6Hz,1H),6.60(d,J＝1.5Hz,1H),6.31(s,1H),6.22(d,J＝1.6Hz,1H),5.14(d,J＝6.3Hz,1H),4.56(s,1H),4.32(q,J＝7.1Hz,2H),3.43(m,2H),3.33(d,J＝5.8Hz,2H),2.14(s,3H),1.94(d,J＝7.3Hz,4H),1.36(t,J＝7.1Hz,3H),1.23(s,6H),0.65(t,J＝7.2Hz,6H).ESI-HRMS calcd for C₃₁H₄₈N₃O₆[M+H]⁺558.3527,found 558.3539.

Example 14:

14.1.4- (4- (3- (1-ethyl-5- ((2-hydroxy-2-methylpropyl) carbamoyl) -1H-pyrrol-3-yl) pent-3-yl) -2-methylphenoxy) -4-oxobutanoic acid (A14)

Dissolving a compound 7a (386mg) in a DMF solution, adding sodium hydrogen (80mg) in batches under the ice bath condition, keeping the temperature and stirring for 1 hour, adding succinic anhydride (150mg) into the reaction solution, moving the reaction solution into an oil bath at 80 ℃ and stirring for 5 hours, after the reaction is finished, adding a large amount of water into the reaction solution, extracting the reaction solution by using ethyl acetate, separating an organic layer, extracting a water layer by using ethyl acetate, combining the organic layers, washing the organic layer by using a proper amount of saturated saline solution, drying anhydrous sodium sulfate, evaporating the solvent, and purifying the mixture by column chromatography to obtain the target compound.¹HNMR(300MHz,DMSO)δ12.29(s,1H),7.63(t,J＝6.1Hz,1H),7.14(s,1H),7.07(d,J＝8.6Hz,1H),6.90(d,J＝8.4Hz,1H),6.78(s,1H),6.55(s,1H),5.75(s,1H),4.27(q,J＝6.8Hz,2H),3.12(d,J＝6.0Hz,2H),2.79(t,J＝6.3Hz,2H),2.59(t,J＝6.3Hz,2H),2.07(s,3H),1.96(dd,J＝15.5,10.3Hz,4H),1.22(t,J＝5.1Hz,4H),1.05(s,6H),0.62(t,J＝7.1Hz,6H).ESI-HRMS calcd for C₂₇H₃₉NO₆[M+H]⁺487.2803,found 487.2808.

The pharmacodynamic test and the result of some compounds of the present invention are shown below, and the structural formulas corresponding to the compound codes are shown in the examples. The structural formula of the positive drug is:

example 15: vitamin D receptor affinity assay

15.1. The experimental method comprises the following steps: the compound concentration was diluted to 1. mu.M with the indicated solvent as required by the Polarscreen Vitamin D Receptor completor Assay kit. After incubation with fluorescent VDR ligand and VDR for 3 hours at room temperature in the dark, the polarized light intensity is detected by a multifunctional microplate reader under the condition of 535nm/590 nm. The affinity of the compound was determined by calcipotriol as a positive control. The affinity ratio of the sample was determined by the following formula, where (%) the affinity ratio (blank-addition group)/(blank-positive control group) × 100%.

15.2. Laboratory apparatus and reagent

An experimental instrument: multifunctional enzyme mark instrument (Diken)

Experimental reagent: polarscreen Vitamin D Receptor completor Assay kit (CatalogNO.: A15907), 384 well plate (Black, opaque)

15.3. The experimental results are shown in table 1, and pharmacodynamic tests prove that part of the compounds have better VDR affinity. Wherein the relative affinity of compound A01 to A05 is higher than that of sw-22(CN201310141958.8) and 19f (Bioorganic & Medicinal Chemistry Letters 27(2017) 1428-1436).

Example 16: anti-type I collagen synthesis activity assay of Compounds

16.1. The experimental method comprises the following steps: taking LX-2 cells in logarithmic growth phase, adjusting cell density to 1 × 10⁵one/mL cell suspension, inoculated into 24-well cell culture plate, each well inoculated with 500. mu.L cell suspension, placed at 37 ℃, 5% CO₂Culturing in a constant temperature incubator. After the cells are inoculated for 24 hours, the supernatant is discarded, and 500uL of the prepared 0.5 mu M compound to be detected is added into each hole by utilizing the incomplete culture solution of Gibco1640, and each compound is provided with 3 parallel holes; at the same time, each plate was equipped with 3 zero wells and 3 control wells (no drug was applied to the cells alone), and the plates were further incubated at 37 ℃ with 5% CO₂Culturing in a constant temperature incubator. After the cells and the drugs are incubated for 24h, 200uL of supernatant is taken, and the content of the type I collagen is determined according to the operation instruction of a human type I collagen (COL1) enzyme-linked immunosorbent assay kit. The OD value corresponding to the compound was measured with a microplate reader (450 nm). The anti-type I collagen synthesis activity of the compound was determined by calcipotriol as a positive control. The ratio of the samples was determined by the following formula, i.e., anti-collagen type I synthesis ratio (%) - (OD)₄₅₀(blank group-additional group)/OD₄₅₀(blank-positive control) x 100%.

16.2. Laboratory instrument and consumable

An experimental instrument: enzyme-linked immunosorbent assay (MK3, Shanghai Zhi Ke apparatus Co., Ltd.), 37 deg.C incubator

Experiment consumables: human type I collagen (COL1) enzyme-linked immunosorbent assay kit (elabsciences), high precision pipette, EP tube and disposable tip: 0.5-10 μ L,2-20 μ L,20-200 μ L,200-

16.3. The experimental results are shown in table 2, and the pharmacodynamic test proves that part of the compounds have better in-vitro anti-hepatic fibrosis capability. Compounds A01, A02 and A05 were more resistant to fibrosis than the positive controls calcipotriol, sw-22(CN201310141958.8) and 19f (Bioorganic & Medicinal Chemistry Letters 27(2017) 1428-. Among them, compound a01 showed the best anti-hepatic fibrosis activity.

Example 17: establishment of hepatic fibrosis animal model

17.1. The experimental method comprises the following steps: male C57BL/6 mice were randomly divided into 4 groups of 5 mice each. Mice were able to freely take food and water, housed in a ventilated room at a temperature of around 23 ℃ and kept in a day-night alternating of 12h light and 12h dark. Carbon tetrachloride (CCl) was used in the experiment₄) The specific operation method of the induced mouse hepatic fibrosis model comprises the following steps of₄Mixing with corn oil at a ratio of 1:50, and performing intraperitoneal injection to each mouse at a ratio of 500 μ L/kg body weight for 4 weeks, 3 times per week. The control group was intraperitoneally injected with corn oil at the same dose every week.

In the administration of CCl₄After 20 days, mice were given calcipotriol or compound by gavage 5 times 1 week. At the last CCl administration₄After 72h, blood was drawn through the orbit and the mice were sacrificed by cervical dislocation.

17.2. Specimen processing

Liver tissues were cut into squares of about 4mm × 4mm, fixed in 4% paraformaldehyde for 24h, and paraffin sections were sequentially sectioned for HE staining and Masson staining.

17.3. The experimental result is shown in figure 1, and the result shows that the compound A01 has stronger in-vivo anti-hepatic fibrosis capability, after the compound A01 is used for treatment, the collagen amount of the hepatic tissue is inhibited, and the tissue vascular structure is repaired to a certain degree.

Table 1: results of a test for the affinity of a Compound for the vitamin D receptor

Table 2: anti-type I collagen synthesis activity results of partial compounds

Claims (4)

1. A compound, enantiomer, diastereomer, racemate or pharmaceutically acceptable salt thereof, wherein the compound is:

4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-N- (2-hydroxy-2-methylpropyl) -1H-pyrrole-2-carboxamide (A01),

n- (cyclopropylmethyl) -4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-1H-pyrrole-2-carboxamide (A03),

4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-N- (2,2, 2-trifluoroethyl) -1H-pyrrole-2-carboxamide (A05), or

4- (3- (4- (2, 3-dihydroxypropoxy) -3-methylphenyl) pentan-3-yl) -1-ethyl-N- (3-hydroxypropyl) -1H-pyrrole-2-carboxamide (A09).

2. A pharmaceutical composition comprising as an active ingredient or as a major active ingredient a compound of claim 1, an enantiomer, a diastereomer, a racemate, or a pharmaceutically acceptable salt thereof.

3. Use of a compound according to claim 1, enantiomers, diastereomers, racemates or pharmaceutically acceptable salts thereof, for the manufacture of a medicament for the treatment or prevention of diseases associated with vitamin D receptors.

4. Use of a compound of claim 1, enantiomers, diastereomers, racemates or pharmaceutically acceptable salts thereof in the manufacture of a medicament for treating or preventing liver fibrosis.

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