WO2022012058A1

WO2022012058A1 - Fused ring compound, and intermediate thereof, preparation method therefor, and application thereof

Info

Publication number: WO2022012058A1
Application number: PCT/CN2021/079576
Authority: WO
Inventors: 朱健; 蒋青
Original assignee: 江苏凯迪恩医药科技有限公司
Priority date: 2020-07-16
Filing date: 2021-03-08
Publication date: 2022-01-20

Abstract

Provided are a compound represented by formula Ia or Ib, and a pharmaceutically acceptable salt, a hydrate, a solvate, a polymorph, a metabolite, a stereoisomer, a tautomer, or a prodrug thereof. The compound is used as a Wnt inhibitor and a mesenchymal stem cell inducer, and has a good induction effect on one or more of sox9, type-I collagen, type-II collagen, an osteogenic transcription gene, and alkaline phosphatase, and thus can be used to prepare a medication for treating and/improving relevant diseases and disorders.

Description

Condensed ring compound and its intermediate, preparation method and application

This application claims the priority of Chinese patent application PCT/CN2020/102369 with the filing date of 2020/7/16. This application cites the full text of the above Chinese patent application.

technical field

The present invention relates to a condensed ring compound and its intermediate, preparation method and application.

Background technique

During vertebrate and invertebrate development, members of the Wnt family of signaling molecules mediate many short- and long-range patterning processes. The Wnt signaling pathway is known to play a role in the inductive interactions regulating growth and differentiation, and also in the homeostatic maintenance of postembryonic tissue integrity. Wnt stabilizes cytoplasmic leukin, which stimulates the expression of genes including cmyc, c-jun, fra-1 and cyclin D1.

Furthermore, dysregulation of Wnt signaling can lead to developmental defects and is implicated in the development of a variety of human cancers. The Wnt pathway has also been found to be involved in the maintenance of stem or progenitor cells in a growing list of adult tissues including skin, blood, gut, prostate, muscle and nervous system.

Mesenchymal stem cells (MSCs) present in the bone marrow and most adult tissues are capable of self-renewal and differentiation into a variety of cell lineages, including chondrocytes, osteoblasts, and adipocytes (Pittenger, MF et al., Science, 1999.284 (5411). ):p.143-7), among which mesenchymal stem cells include synovial mesenchymal stem cells, bone marrow mesenchymal stem cells, etc. The study found that adult articular cartilage contains MSCs capable of multi-lineage differentiation (about 3% of cells). In view of the above characteristics of mesenchymal stem cells, how to realize the osteogenic and chondrogenic directional differentiation of mesenchymal stem cells and provide basis and ideas for the treatment of related diseases has become the focus of everyone's attention.

Therefore, it is of great interest to explore Wnt inhibitors that can be used in stem cell research or treatment of any disease characterized by aberrant activation of Wnt, as well as to induce mesenchymal stem cells for the treatment and/or amelioration of related diseases and disorders.

SUMMARY OF THE INVENTION

The present invention provides a condensed ring compound and its intermediate, preparation method and application which are different from the prior art. The compounds of the present invention are useful as Wnt inhibitors and mesenchymal stem cell inducers for the treatment and/or amelioration of related diseases and disorders.

The present invention provides a compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its interconversion Isomers or their prodrugs:

^{^{Wherein, R 1, R 2, R}} 3, R 1 ', R 2' and R ³ 'is independently H, C ₆ ~ C ₁₈ aryl group or substituted with one or more of R ^1-1 is C ₆ ~ Aryl of C ₁₈ (when there ^{are multiple R 1-1s} , the R ^{1-1s are the} same or different);

Each R ^{1-1 is} independently halogen or C ₁ -C ₄ alkyl;

R ⁴ , R ⁵ , R ⁴ ' and R ⁵ ' are independently H or C ₁ -C ₄ alkyl, but R ⁴ and R ^{5 are} not H at the same time;

R ⁶ and R ⁶ 'are independently C ₆ ~ C ₁₈ aryl group, and one or more of R ^6-1 is a substituted C ₆ ~ C ₁₈ aryl group (when R ^6-1 is a plurality of said R ^{6-1 is the} same or different), a 5- to 10-membered heteroaryl group, or a 5- to 10-membered heteroaryl group substituted by ^{one or more R 6-2} (when there are more than one ^{R 6-2, the R 6-2} ^6-2 same or different); the heteroatoms in the 5-10-membered heteroaryl group are independently selected from one or more of N, O and S, and the number of heteroatoms is 1, 2, 3 or 4 ; heteroatom one or more of the R ^6- ^{2-substituted} 5 to 10-membered heteroaryl group in the 5 to 10-membered heteroaryl group is independently selected from N, O and S, one kind or Various, the number of heteroatoms is 1, 2, 3 or 4;

Each of R ^6-1 and R ^{6-2 is} independently

amino or

and

R ^6-1-1 is a C ₁ -C ₄ alkyl group or a C ₃ -C ₆ cycloalkyl group.

In the present invention, the definitions of some substituents in the compound represented by formula Ia or Ib may be as follows, and the definitions of unmentioned substituents are as described in any of the above schemes.

In a certain preferred embodiment, when R ¹ , R ² , R ³ , R ¹ ', R ² ' and R ³ ' are independently C ₆ -C ₁₈ aryl or one or more R ^1-1 substituted When the C ₆ -C ₁₈ aryl group is, the C ₆ -C ₁₈ aryl group is a C ₆ -C ₁₄ aryl group (such as phenyl, naphthyl, anthracenyl or phenanthrenyl), and further may be benzene base.

In a certain preferred embodiment, when R ¹ , R ² , R ³ , R ¹ ', R ² ' and R ³ ' are independently one or more R ^1-1 substituted C ₆ -C ₁₈ aryl groups , the ^{number of the R 1-1} is one.

In a certain preferred embodiment, when R ^1-1 is halogen, the halogen is F, Cl, Br or I, and it can also be F.

In a certain preferred embodiment, when R ¹ , R ² , R ³ , R ¹ ', R ² ' and R ³ ' are independently one or more R ^1-1 substituted C ₆ -C ₁₈ aryl groups , the one or more aryl groups of C ₆ -C ₁₈ ^{substituted by R 1-1} are fluorophenyl groups, which may further be

In a preferred embodiment, when R ⁴ , R ⁵ , R ⁴ ' and R ⁵ ' are independently C ₁ -C ₄ alkyl groups, the C ₁ -C ₄ alkyl groups are methyl, Ethyl, n-propyl, isopropyl, n-butyl,

or tert-butyl, which can be methyl or isopropyl.

In a preferred embodiment, when R ⁶ and R ⁶ ' are independently 5-10-membered heteroaryl groups or 5-10-membered heteroaryl groups ^{substituted by one or more R 6-2} , the 5- to 10-membered heteroaryl groups The heteroatom in the 10-membered heteroaryl group may be N, and the number of heteroatoms may be one.

In a preferred embodiment, when R ⁶ and R ⁶ ' are independently a 5- to 10-membered heteroaryl or a 5- to 10-membered heteroaryl substituted by ^{one or more R 6-2, the 5- to 10-membered heteroaryl} The 10-membered heteroaryl group is a 5- to 6-membered heteroaryl group.

In a preferred embodiment, when R ⁶ and R ⁶ ' are independently 5-10-membered heteroaryl groups or 5-10-membered heteroaryl groups ^{substituted by one or more R 6-2} , the 5- to 10-membered heteroaryl groups The heteroatom in the 10-membered heteroaryl group may be N, and the number of heteroatoms may be 1; the 5-10-membered heteroaryl group may further be a pyridyl group, such as

In one preferable embodiment, when R ⁶ and R ⁶ 'is independently substituted with one or more R ^6-2 is 5 to 10-membered heteroaryl, said R ^6-2 is a number .

In a certain preferred embodiment, when R ^6-2 is

When the R ^6-1-1 is a C ₁ -C ₄ alkyl group, the C ₁ -C ₄ alkyl group can be methyl, ethyl, n-propyl, isopropyl, n-butyl ,

or tert-butyl, and can also be

or tert-butyl.

In a certain preferred embodiment, when R ⁶ and R ⁶ ' are independently one or more R ^6-2 substituted 5-10-membered heteroaryl groups, the one or more R ^6-2 substituted heteroaryl groups 5-10-membered heteroaryl group is

In a certain preferred embodiment, the compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer In its isomer, its tautomer or its prodrug:

R ¹ and R ¹ 'are independently C ₆ ~ C ₁₈ aryl group, or substituted with one or more of R ^1-1 is C ₆ ~ C ₁₈ aryl group; and

R ² , R ³ , R ² ′ and R ³ ′ are independently H;

R ^1-1 is halogen;

One of R ⁴ and R ⁵ is a C ₁ -C ₄ alkyl group, and the other is H;

R ⁴ ' and R ⁵ ' are H;

R ⁶ and R ⁶ ' are independently 5-10-membered heteroaryl or 5-10-membered heteroaryl substituted by ^{one or more R 6-2;}

Each R ^{6-2 is} independently

and

R ^6-1-1 is a C ₁ -C ₄ alkyl group.

In a certain preferred embodiment, R ¹ and R ¹ ' are independently one or more _{C 6} -C ₁₈ aryl groups substituted by ^{R 1-1.}

In a certain preferred embodiment, R ⁶ and R ⁶ ′ are independently one or more R ^6-2 substituted 5-10-membered heteroaryl groups.

In a certain preferred embodiment, the compound represented by Formula Ia, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, Among its tautomers or prodrugs:

R ¹ is a C ₆ -C ₁₈ aryl group or a _{C 6} -C ₁₈ aryl group substituted by ^{one or more R 1-1s;}

R ² and R ³ are H;

R ^1-1 is halogen;

R ⁴ is a C ₁ -C ₄ alkyl group;

R ⁵ is H or C ₁ -C ₄ alkyl;

R ⁶ is a 5- to 10-membered heteroaryl group or a 5- to 10-membered heteroaryl group substituted by ^{one or more R 6-2;}

Each R ^{6-2 is} independently

and

R ^6-1-1 is a C ₁ -C ₄ alkyl group.

R ² and R ³ are H;

R ^1-1 is halogen;

R ⁴ is H or C ₁ -C ₄ alkyl;

R ⁵ is a C ₁ -C ₄ alkyl group;

Each R ^{6-2 is} independently

and

R ^6-1-1 is a C ₁ -C ₄ alkyl group.

R ² and R ³ are H;

R ^1-1 is halogen;

R ⁴ is H;

R ⁵ is a C ₁ -C ₄ alkyl group;

Each R ^{6-2 is} independently

and

R ^6-1-1 is a C ₁ -C ₄ alkyl group.

In a certain preferred embodiment, the compound shown in formula Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, Among its tautomers or prodrugs:

R ¹ _{' is a C 6} -C ₁₈ aryl group substituted by one or more R ^1-1;

R ² ' and R ³ ' are H;

R ^1-1 is halogen;

R ⁴ ' and R ⁵ ' are H;

R ⁶ ' is a 5-10-membered heteroaryl group or a 5-10-membered heteroaryl group substituted by ^{one or more R 6-2;}

Each R ^{6-2 is} independently

and

R ^6-1-1 is a C ₁ -C ₄ alkyl group.

In a certain preferred embodiment, the compound represented by the formula Ia or Ib is any of the following compounds:

The present invention also provides a preparation method of the above-mentioned compound shown in formula Ia or Ib,

The described preparation method of the compound shown in formula Ia is method 1 or method 2:

The method 1 includes the following steps: in a solvent, in the presence of a deprotection reagent, the compound represented by the formula IIa is subjected to the deprotection reaction of the following formula to obtain the compound represented by the formula Ia,

Wherein, Q is an amino protecting group; R ¹ , R ² , R ³ , R ⁴ and R ⁶ are as defined above;

Method 2 includes the following steps: in a solvent, in the presence of a cyclization reagent, the compound shown in formula IIIa and the compound shown in formula IVa are subjected to the following cyclization reaction to obtain the compound shown in formula Ia. That's it,

Wherein, ^{the definitions of R 1} , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are the same as above, and R ^{5 is} not hydrogen;

The described preparation method of the compound shown in formula Ib is method 3 or method 4:

Method 3 includes the following steps: in a solvent, in the presence of a deprotection reagent, the compound shown in formula IIb is subjected to the deprotection reaction of the following formula to obtain the compound shown in formula Ib,

Wherein, Q' is an amino protecting group; R ¹ ', R ² ', R ³ ', R ⁴ ' and R ⁶ ' are defined as described above;

Method 4 includes the following steps: in a solvent, in the presence of a cyclization reagent, the compound shown in formula IIIb and the compound shown in formula IVb are subjected to the following cyclization reaction to obtain the compound shown in formula Ib. You can;

Wherein, R ¹ ', R ² ', R ³ ', R ⁴ ', R ⁵ ' and R ⁶ ' are defined as above, and R ⁵ ' is not hydrogen.

The conditions and operations of the deprotection reactions described in methods 1 and 3 can be conventional conditions and operations in the art, and the following conditions and operations are particularly preferred in the present invention.

Wherein, when Q or Q' is an amino protecting group, the amino protecting group can be a conventional amino protecting group in the art, preferably tetrahydropyranyl (THP).

Wherein, the solvent can be a conventional solvent in the art, or a chlorinated hydrocarbon solvent, and further can be dichloromethane.

Wherein, the deprotection reagent may be a conventional deprotection reagent in the art, or may be an organic silane compound (eg, triethylsilane) and an organic acid (eg, trifluoroacetic acid). The molar ratio of the organosilane compound to the organic acid may be 1:1 to 1:5, for example, 1:2.5.

Wherein, the temperature of the deprotection reaction may be 10˜40° C., for example, room temperature.

The progress of the deprotection reaction can be monitored by conventional methods in the art (eg TLC, HPLC, nuclear magnetic resonance), and usually the end point of the reaction is that the compound represented by formula IIa or IIb no longer reacts or disappears. The time of the deprotection reaction can be 10-18 hours, such as overnight.

After the reaction is completed, the following post-processing steps can be adopted: the reaction solution after the reaction is concentrated, and the solid is precipitated and column chromatography is performed.

In the post-processing step, the conditions and operations of the concentration can all adopt the conventional conditions and operations in the art.

In the post-processing step, the conditions and operations for the precipitation of solids can all be conventional conditions and operations in the art. The reagent used for the described solid precipitation can be an inorganic base, more preferably ammonia water (for example, the concentration of ammonia water is 5 mol/L).

In the post-processing step, the conditions and operations of the column chromatography can all adopt the conditions and operations conventional in the art. The eluents used in the column chromatography can be alcohol reagents and chlorinated hydrocarbon reagents, and further can be dichloromethane and methanol (for example, the volume ratio of dichloromethane and methanol is 20:1-10:1).

The conditions and operations of the cyclization reaction described in methods 2 and 4 can be conventional conditions and operations in the art, and the following conditions and operations are particularly preferred in the present invention.

Wherein, the solvent can be a conventional solvent in the art, or an amide solvent, and further can be DMF.

Wherein, the cyclization reagent may be a conventional reagent in the art, or may be an alkali metal metabisulfite (eg, sodium metabisulfite).

Wherein, the molar ratio of the cyclization reagent to the compound represented by formula IIIa or IIIb may be 4:1 to 1:1, for example, 2:1.

Wherein, the temperature of the cyclization reaction can be 100-140°C, for example, 120°C.

The progress of the cyclization reaction can be monitored by conventional methods in the art (eg, TLC, HPLC, NMR), and usually the end point of the reaction is that the compound represented by formula IIIa or IIIb no longer reacts or disappears. The time of the deprotection reaction may be 3 to 9 hours, for example, 6 hours.

After the reaction is completed, the following post-processing steps can be adopted: the reaction solution after the reaction is cooled to room temperature, and solid is precipitated.

In the post-processing step, the conditions and operations for the precipitation of solids can all be conventional conditions and operations in the art. The reagent used for the described solid precipitation can be water.

The present invention also provides a compound of formula IIa, IIb or IIIb:

Wherein, Q or Q 'is an amino protecting ^{^{group; R 1, R 2, R}} 3, R 4, R 6, R 1', R 2 ', R 3', R 4 ', R 5' and R ⁶ 'is The definitions are the same as above.

In a preferred embodiment, the compound represented by formula IIb is

The present invention also provides a pharmaceutical composition comprising a compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereo Isomers, tautomers or prodrugs thereof, and pharmaceutical excipients.

The present invention also provides a compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual Use of the variant or its prodrug, or the above-mentioned pharmaceutical composition in the preparation of an inducer.

In the above application, the above-mentioned inducer can induce proteoglycan (Aggrecan), type II collagen (Collagen II), sox9, type I collagen (collagen I), type X collagen (collagen X), osteogenic transcription gene ( Elevated expression of one or more genes in RUNX2) or alkaline phosphatase (ALP).

In the above application, the inducer can induce synovial stem cells to differentiate into chondroblasts and/or osteoblasts. The inducer can also be used as a bone building agent, or in the treatment or prevention of periodontal disease, arthritis (such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, polyarticular juvenile idiopathic joints). one or more of osteoarthritis, osteoarthritis), joint damage, cartilage repair, osteogenesis regeneration, osteoporosis, osteopenia, spinal fusion, periodontal disease, and bone tumors.

The bone building agents can enhance fracture recovery and stimulate bone growth at the site of bone, joint or dental implants.

The present invention also provides a compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual Use of the variant or its prodrug, or the above-mentioned pharmaceutical composition, in the preparation of an inhibitor of one or more proteins in the Wnt pathway.

The present invention also provides a compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual Use of the variant or its prodrug, or the above-mentioned pharmaceutical composition, in the preparation of a medicament for treating and/or preventing a disease associated with one or more proteins in the Wnt pathway.

In the above application, the disease associated with one or more proteins in the Wnt pathway may be cancer, a disease associated with abnormal angiogenesis, a disease associated with cell proliferation and cell cycle, or a Wnt signaling component. A disease caused by one or more variants or disorders.

Wherein, the cancer can be one or more of colon cancer, hepatocellular carcinoma, lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, bone cancer and leukemia.

Wherein, the diseases related to abnormal angiogenesis can be hematopoietic system tumors of lymphoid lineage, hematopoietic system tumors of myeloid lineage, tumors of mesenchymal origin, tumors of central and peripheral nervous system and melanoma, seminoma, Kaposi's sarcoma, benign prostatic hyperplasia, familial adenomatous polyposis, neurofibromas, atherosclerosis, arthritis, nephritis, restenosis after angioplasty or vascular surgery, inflammatory bowel disease, transplant rejection, endotoxin One or more of shock and fungal infection.

Wherein, the disease caused by the variation or disorder of one or more of the Wnt signaling components can be colon polyposis, osteoporosis, pseudoglioma syndrome, familial exudative vitreous Retinopathy, retinal angiogenesis, early coronary heart disease, congenital limb amputation syndrome, paramesonephric reversion and virilization, SERKAL syndrome, type 2 diabetes, Fuhrman syndrome, AARRS short limb syndrome, dental nails Cutaneous dysplasia, obesity, split hand/foot deformity, caudal duplication syndrome, congenital missing teeth, Wilms tumor, skeletal dysplasia, focal skin hypoplasia, neural tube defect, alpha thalassemia syndrome, fragility One or more of Syndrome X, ICF Syndrome, Angelman Syndrome, Parr-Williams Syndrome, Belle-Williams Syndrome, Norrie Disease and Rett Syndrome.

The present invention also provides a compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual Use of the variant or its prodrug, or the above-mentioned pharmaceutical composition in the preparation of a medicine.

In the above application, the drug can make synovial stem cells differentiate into chondroblasts and/or osteoblasts. The medicament can also be used as a bone building agent, or for the treatment or prevention of periodontal disease, arthritis (such as rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, polyarticular juvenile idiopathic arthritis). , osteoarthritis), joint damage, cartilage repair, osteogenesis regeneration, osteoporosis, osteopenia, spinal fusion, periodontal disease and one or more of bone tumors.

In the above applications, the medicament can be used to treat and/or prevent diseases associated with one or more proteins in the Wnt pathway.

Wherein, the mentioned diseases are the same as those mentioned above.

In the above-mentioned applications, in the medicines, the medicines can be used to treat and/or prevent cancer, diseases related to abnormal angiogenesis, diseases related to cell proliferation and cell cycle, or one of Wnt signaling components. Disease caused by one or more variants or disorders.

wherein the cancer, the disease associated with abnormal angiogenesis, the disease associated with cell proliferation and cell cycle, or the mutation or dysregulation of one or more of the Wnt signaling components The diseases caused are the same as before.

The present invention also provides a method for inducing synovial mesenchymal stem cells to differentiate into chondroblasts, inducing osteoblast differentiation or inhibiting osteoclast formation, comprising administering to a subject (preferably, a therapeutically effective amount to the subject) The above-mentioned compound represented by formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its tautomerism need to be treated Construct or its prodrug, or the above-mentioned pharmaceutical composition.

The subject can be a mammal, and further can be a human.

The chondroblasts and/or osteoblasts are derived from the differentiation of synovial mesenchymal stem cells. MSCs are pluripotent stem cells that can differentiate into several different types of cells, including but not limited to osteoblasts, chondrocytes and adipocytes. Differentiation is the process of forming specialized cell types from less specialized cell types, such as chondrocytes from MSCs.

Said chondrocytes are produced or maintained by the expression of one or more genes in proteoglycan (Aggrecan), type II collagen (Collagen II) and sox9.

The described promotion of osteogenic differentiation and inhibition of osteoclast formation are composed of one of type I collagen (collagen I), type X collagen (collagen X), osteogenic transcription gene (RUNX2) or alkaline phosphatase (ALP). Or the expression of multiple genes is generated or maintained.

The present invention also provides a method of treating or preventing a disease associated with one or more proteins in the Wnt pathway, comprising administering to a subject (preferably a therapeutically effective amount to the subject) the above-described formula Ia for a subject in need of treatment Or the compound shown in Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its tautomer or its prodrug, or The above-mentioned pharmaceutical composition.

The diseases described are the same as before.

The term "plurality" refers to 2, 3, 4 or 5.

The term "pharmaceutically acceptable" means that salts, solvents, excipients, etc. are generally non-toxic, safe, and suitable for patient use. The "patient" is preferably a mammal, more preferably a human.

The term "pharmaceutically acceptable salts" refers to salts of compounds of the present invention prepared with relatively non-toxic, pharmaceutically acceptable acids or bases. When compounds of the present invention contain relatively acidic functional groups, base additions can be obtained by contacting neutral forms of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent. A salt. Pharmaceutically acceptable base addition salts include, but are not limited to, lithium, sodium, potassium, calcium, aluminum, magnesium, zinc, bismuth, ammonium, diethanolamine. When compounds of the present invention contain relatively basic functional groups, acid addition can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent. A salt. The pharmaceutically acceptable acids include inorganic acids, including but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like. Described pharmaceutically acceptable acid includes organic acid, described organic acid includes but is not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid , fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citric acid, oleic acid , tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, sugar acid, formic acid, ethanesulfonic acid, pamoic acid (i.e. 4,4'-methylene-bis( 3-hydroxy-2-naphthoic acid), amino acids (eg, glutamic acid, arginine), and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. For details, see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66:1-19 (1977), or, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

The terms "compound" and "pharmaceutically acceptable salt" can exist in crystalline or amorphous form. The term "crystal form" means that the ions or molecules in it are arranged in a strictly periodic manner in three-dimensional space in a certain manner, and have the regularity of periodic repetition at a certain distance; crystal form, or polymorphism.

The terms "compound" and "pharmaceutically acceptable salt", if stereoisomers exist, may exist as single stereoisomers or as mixtures thereof (eg, racemates).

The term "stereoisomer" refers to a cis-trans isomer or an optical isomer. These stereoisomers can be separated, purified and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin layer chromatography, spin chromatography, column chromatography, gas chromatography, high pressure liquid chromatography, etc.) It can be obtained by chiral resolution by forming bonds with other chiral compounds (chemical bonding, etc.) or forming salts (physical bonding, etc.).

The term "single stereoisomer" means that the mass content of one stereoisomer of the compound of the present invention relative to all stereoisomers of the compound is not less than 95%.

The terms "compound" and "pharmaceutically acceptable salt", if tautomers exist, may exist as single tautomers or as mixtures thereof, preferably as more stable tautomers The main form exists.

Atoms in the terms "compound" and "pharmaceutically acceptable salt" may exist in their natural or unnatural abundance. Taking a hydrogen atom as an example, its natural abundance means that about 99.985% of it is protium and about 0.015% is deuterium; its unnatural abundance means that about 95% of it is deuterium. That is, one or more atoms in the terms "compound" and "pharmaceutically acceptable salt" may be atoms that exist in unnatural abundance.

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

The term "alkyl" refers to a straight or branched chain alkyl group having the indicated number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and It resembles an alkyl group.

The term "alkoxy" refers to the group -OR ^X , wherein R ^X is an alkyl group as defined above.

The term "cycloalkyl" refers to a monovalent saturated cyclic alkyl group, preferably a monovalent saturated cyclic alkyl group having 3-7 ring carbon atoms, more preferably 3-6 carbon atoms, such as cyclopropyl, cyclobutyl cyclopentyl, cyclopentyl or cyclohexyl.

The term "heterocycloalkyl" refers to a saturated monocyclic group having a heteroatom, preferably a 3-7 membered saturated one containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S. single ring. Examples of heterocycloalkyl groups are: pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl , piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, oxazepanyl and the like. Preferred heterocyclyl groups are morpholin-4-yl, piperidin-1-yl, pyrrolidin-1-yl, thiomorpholin-4-yl and 1,1-dioxo-thiomorpholin-4 -base.

The term "aryl" refers to a group having a 4n+2 aromatic ring system (eg, with 6, 10, or 14 shared p electrons in a cyclic array). Examples of the above-mentioned aryl unit include phenyl, naphthyl, phenanthryl, or anthracenyl.

The term "heteroaryl" refers to an aromatic group containing a heteroatom, preferably containing 1, 2 or 3 aromatic 5-6 membered monocyclic or 9-10 membered bicyclic rings independently selected from nitrogen, oxygen and sulfur , such as furanyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, diazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazole base, thiazolyl, isothiazolyl, thiadiazolyl, benzimidazolyl, indolyl, indazolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinoline olinyl, isoquinolinyl, etc.

The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the formulation of prescriptions, and are all substances contained in pharmaceutical preparations other than active ingredients. See the Pharmacopoeia of the People's Republic of China (2015 edition) four, or, Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009 Sixth Edition).

The term "therapeutically effective amount" refers to a dose that produces a therapeutic effect. The exact dosage will depend on the purpose of the treatment and can be determined by one of skill in the art using known techniques (see, eg, Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Pharmaceutical, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

On the basis of not violating common sense in the art, the above-mentioned preferred conditions can be combined arbitrarily to obtain the preferred examples of the present invention.

The reagents and raw materials used in the present invention are all commercially available.

The positive improvement effect of the present invention is that: as a Wnt inhibitor and a mesenchymal stem cell inducer, the compound of the present invention can inhibit one or more of sox9, type I collagen, type X collagen, osteogenic transcription gene and alkaline phosphatase. This species has better induction effect, and then can be used to treat and/or improve related diseases and disorders.

detailed description

Synthesis of Intermediates

Synthesis of III-24000-C

To a solution of compound 4 (1.8 g, 5.6 mmol) and TEA (1.7 g, 16.7 mmol) in DMF (20 mL) was added dropwise compound 5 (0.8 g, 6.69 mmol) under ice bath. The reaction solution was warmed to room temperature and stirred at room temperature for 2 hours. The reaction solution was poured into 100 mL of water. The solution was extracted twice with ethyl acetate, the organic phases were combined, dried, the solvent was removed, and purified by reverse-phase flash preparative chromatography to give 1.09 g of a white solid with a yield of 48%.

MS(ESI) m/z=407[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ) δ 10.32(s, 1H), 10.22(s, 1H), 8.86(s, 1H), 8.73(d, J=1.4Hz, 1H), 8.44(s, 1H), 8.27 (d, J=8.6Hz, 2H), 7.72 (dd, J=8.3, 1.5Hz, 1H), 6.23 (dd, J=9.4, 2.0Hz, 1H), 3.95–3.78 (m, 3H) ), 2.28(d, J＝7.1Hz, 2H), 2.19-1.99(m, 3H), 1.84-1.75(m, 1H), 1.64(br, 2H), 0.98(s, 3H), 0.96(s, 3H).

Preparation of intermediate compound IV-20000-6

The first step: 3-bromo-5-nitropyridin-4-amine

To a solution of 3-nitropyridin-4-amine (100 g, 0.716 mol) and sodium acetate (177.2 g, 2.21 mol) in acetic acid (800 mL) was added dropwise bromine (44.4 mL, 0.864 mol) at room temperature. The reaction solution was stirred at 110°C overnight. TLC showed that the reaction was completed, the reaction solution was concentrated, adjusted to pH 9-10 with aqueous sodium carbonate solution, stirred for 15 minutes, filtered, and the filter cake was washed with n-hexane/ethyl acetate (200 mL x 2, 10/1), and dried to obtain 3 -Bromo-5-nitropyridin-4-amine (116 g, 74% yield), yellow solid.

MS(ESI) m/z=219.7[M+1].

¹ H NMR (400MHz, DMSO-d ₆ ) ppm 8.97(s,1H), 8.54(s,1H), 7.82(s,2H).

Step 2: 3-(3-Fluorophenyl)-5-nitropyridin-4-amine

In the three-necked reaction flask, successively add 3-bromo-5-nitropyridine-4-amine (50.0g, 230.41mmol), (3-fluorophenyl)boronic acid (38.71g, 276.50mmol), sodium carbonate (61.1g, 576.0 mmol), bistriphenylphosphine palladium dichloride (8.06 g, 11.52 mmol), 1,4-dioxane/water (600 mL, 3/1), under nitrogen protection, stirred at 95°C overnight. TLC showed that the reaction was complete, the filtrate was filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column (eluent: 10%-50% ethyl acetate/n-hexane) to give (3-fluorophenyl)-5-nitropyridine (30.5 g , yield 59%), yellow solid.

MS(ESI)m/z=234.2[M+1].

¹ H NMR (400MHz, DMSO-d ₆ )ppm 9.04(s,1H), 8.12(s,1H), 7.61-7.55(m,1H), 7.41(s,2H), 7.34-7.26(m,2H) .

Step 3: 5-(3-Fluorophenyl)pyridine-3,4-diamine

3-(3-fluorophenyl)-5-nitropyridin-4-amine (30.5g, 130.90mmol), Pd/C (5.0g), methanol (500mL), Pd/C (5.0g), methanol (500mL) were added successively to a 500-mL there-necked flask, and the Hydrogen, stirred at room temperature overnight. TLC showed that the reaction was complete, filtered, and the filtrate was concentrated, and the crude product was washed with diethyl ether (50 mL x 2) to give 5-(3-fluorophenyl)pyridine-3,4-diamine (20.1 g, 75% yield) as a yellow solid.

MS(ESI)m/z=203.9[M+1].

¹ H NMR (400MHz, DMSO-d ₆ ) ppm 7.68(s,1H), 7.53-7.47(m,1H), 7.45(s,1H), 7.24-7.17(m,3H), 5.08(s,2H) ,4.72(s,2H).

¹³ C NMR (400 MHz, DMSO-d ₆ ) ppm 164.1 161.6, 140.0, 139.9, 139.8, 138.3, 134.9, 131.4, 131.3, 131.2, 125.6, 120.3, 116.3, 116.1, 114.5, 114.3.

Example 1 Synthesis of I-24003

Add III-24000-C (203mg, 0.5mmol, 1eq), IV-20000-6 (102mg, 0.5mmol, 1eq), sodium metabisulfite (191mg, 1mmol, 2eq), 5ml N-formaldehyde to a 100ml single-necked bottle in turn pyrrolidone, under nitrogen protection, and magnetically stirred at 120 °C overnight. Cool to room temperature, add 20 ml of water, stir for 1 h, and precipitate 310 mg of gray solid.

The above-mentioned solid was transferred to a 100-ml single-neck flask, and dichloromethane (10 ml), triethylsilane (0.15g, 1.25mmol, 2.5eq), trifluoroacetic acid (1.71g, 15mmol, 30eq) were successively added, and the mixture was stirred at room temperature. overnight. Concentrate under reduced pressure, add 10 mL of ammonia water (5 mol/L), stir, and precipitate a solid, which is purified by column chromatography (eluent: dichloromethane/methanol=20:1-10:1) to obtain 100 mg of brown solid (purity 97.07 %).

MS(ESI) m/z=506.1[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ) ppm: 14.30 (s, 1H), 10.42 (s, 1H), 10.30 (s, 1H), 9.02-7.50 (m, 12H), 2.20 (d, 2H), 2.05 (m, 1H), 0.95 (dd, 6H).

Example 2 Synthesis of I-25001

Intermediate synthesis

Intermediate 1-2: Add raw material 1-1 (5.0g, 36mmol) into a 100ml single-necked bottle, add potassium acetate (5.28g, 1.5eq), acetic acid (50ml), dropwise add liquid bromine (2.75ml, 1.5 eq). The reaction was carried out at room temperature for 20h. Saturated aqueous sodium bicarbonate solution was added to quench, extracted with dichloromethane (100 ml×2), the organic layers were combined, dried over anhydrous sodium sulfate and spin-dried to obtain crude intermediate 1-2. The crude product was recrystallized from ethanol:tetrahydrofuran (10:1) to obtain Intermediate 1-2 as a yellow solid with a yield of 67%.

Intermediate 1-4a: Intermediate 1-2 (2.18 g, 10 mmol), Intermediate 1-3 (1.68 g, 1.2 eq), Pd(PPh ₃ ) ₄ (0.57 g, 0.05 eq), sodium bicarbonate ( 2.12g, 2eq), toluene (15ml), water (6ml), and ethanol (3ml) were added to a 50ml single-neck flask, replaced with nitrogen 5 times, heated to reflux for 8h reaction. After the reaction is completed, suction filtration, spin dry, and directly purify by silica gel column chromatography to obtain intermediate 1-4a as a yellow solid, and the yield is greater than 90%.

Intermediate 1-5a: Intermediate 1-4a was dissolved in methanol, a catalytic amount of palladium on carbon was added, the air was replaced with hydrogen, the reaction was carried out at room temperature for 6 hours, suction filtration, and spin-dried to obtain dark red oily substance 1-5a, yield 95% .

Intermediate 1-4b: Intermediate 1-4a (0.47g, 2mmol) was added to a 50ml single-neck flask, DMF (18ml), NaH (0.12g, 1.5eq) were added, and iodomethane (0.57g, 2eq) was added under ice bath ), and reacted in an ice bath for 30 min. After the reaction was completed, water was added, extracted with ethyl acetate, and then subjected to silica gel column chromatography to obtain the monomethylated product Intermediate 1-4b as a yellow solid with a yield of 41%.

Intermediate 1-5b: its preparation method is the same as that of intermediate 1-5a. A pale red oil was obtained with a yield of 96%.

Intermediate 2-2: R ^1a is i-Bu: The raw material 2-1 (3.4 g, 20 mmol) was dissolved in dichloromethane, cooled to 0°C in an ice bath, isovaleric acid chloride (1.5 eq) was dissolved in dichloromethane Triethylamine (2eq) was added after post-dropping. Stir under ice bath for 30 min. Spin dry, add ethyl acetate to dissolve, wash the organic layer with dilute hydrochloric acid and saturated sodium bicarbonate, spin dry, and beat with dichloromethane to obtain 3.8 g of a white solid with a yield of 74%.

Intermediate 2-2: ^{The preparation method of the compound whose R 1a} is t-Bu is the same as the preparation method of the compound whose R ^1a is i-Bu, white solid, yield 70%.

Intermediate 3-2: Add raw material 3-1 (20g, 0.1mol) and acetone (450ml) into a 1L three-necked flask, cool down to 0°C in an ice-salt bath, add dropwise an aqueous solution of sodium nitrite (55.2g, 8eq solution) in 100 ml of water), keeping the temperature below 0 °C. Dilute hydrochloric acid (2N HCl, 240 ml) was added dropwise. Then the temperature was raised to room temperature for 4 h. Spin dry, add ethyl acetate for extraction (100 ml×2), combine the organic layers, spin dry to obtain crude product. The crude product was slurried with dichloromethane to obtain Intermediate 3-2 as a brown-red solid with a yield of 90%.

Intermediate 3-3: R ^2a =CH ₃ : Intermediate 3-2 (3.0 g, 13.3 mmol) was added to a 100 ml single-neck flask, potassium carbonate (3.6 g, 2 eq), DMF (30 ml) were added, and nitrogen replaced air, The reaction was carried out at 100°C for 4h. The reaction was quenched by adding 2N sodium hydroxide solution, extracted with ethyl acetate (50ml×2), the organic layers were combined, spin-dried and purified by silica gel column chromatography to obtain Intermediate 3-3 as a brown solid with a yield of 70%.

Intermediate 3-3: The preparation method of the compound in which ^{R 2a} ^{is i-Pr is the same as the preparation method in which R 2a} is CH ₃ to obtain a brown solid with a yield of 54%. The preparation method of the compound whose R ^2a ^{is THP is the same as the preparation method of R 2a} is CH ₃ above.

Intermediate 3-3: R ² is THP: Intermediate 3-2 (R ² is THP) was added to a 250ml three-necked flask, tetrahydrofuran (180ml), p-toluenesulfonic acid (0.69g, 0.1eq) were added, room temperature 3,4-Dihydropyran (6.72 g, 2 eq) was added dropwise and reacted at 40° C. for 8 h. Then, the solvent was evaporated under reduced pressure, and the crude product of Intermediate 3-3 was obtained by silica gel column chromatography. After beating with PE, a white solid was obtained, and the yield was 32%.

Intermediate 3-4 (boronic esterification): Intermediate 3-3 (R ^2a is THP, 3.1 g, 10 mmol) was added to a 100 ml single-neck flask, and pinacol biboronate (3.0 g, 1.2 eq) was added , potassium acetate (2.94g, 3eq), Pd ( dppf) Cl 2 (0.36 g, 0.05eq), dioxane (26ml). The temperature was raised to reflux for 6 h, and after cooling, the intermediate 3-4 was filtered with suction, spin-dried, and purified by silica gel column chromatography as a white solid with a yield of 90%.

Intermediate 4-1: intermediate 3-4 (R ^2a is THP, 1.07 g, 3 mmol), intermediate 2-2 (isovaleric acid, 0.93 g, 1.2 eq), sodium carbonate (0.95 g, 3 eq), Pd(dppf)Cl ₂ (0.1g, 0.05eq), dioxane (9ml) and water (1.5ml) were added to a 50ml single-neck flask, nitrogen was replaced with air, and the temperature was raised to reflux and reacted for 8h. After cooling, suction filtration, spin drying, and purification by silica gel column chromatography to obtain Intermediate 4-1 as a yellow oil with a yield of 69%. For the preparation method of the intermediate compound in which R ^2a is methyl or isobutyl, refer to the preparation method of the intermediate compound in which R ^2a is THP.

I-25001:

Intermediate 4-1 (R ^1a =i-Bu, R ^2a =CH ₃ , 0.18g, 0.5mmol), Intermediate 1-5 (R ^3a =H, 0.1g, 1eq) were added to a 50ml single-necked bottle, Sodium metabisulfite (0.14g, 1.5eq) and DMF (5ml) were added, and the reaction was carried out at 120°C for 6h. After cooling, add water to quench, filter with suction and wash with water to obtain the crude product of KDN25001, which is then beaten with methanol to obtain I-25001, which is an off-white solid.

¹ H NMR (DMSO-d ₆ , 300MHz): ppm.δ=10.27(s, 1H), 9.09(s, 1H), 8.77(s 2H), 8.67(s, 1H), 8.51(s, 1H), 8.25–8.19(m, 2H), 8.03(d, J=8.8Hz, 1H), 7.90(d, J=8.8Hz, 1H), 7.65(d, J=6.2Hz, 1H), 7.33(s, 1H) ), 4.31(s, 3H), 2.28(s, 2H), 2.12(s, 1H), 0.97(s, 6H).

MS(ESI)m/z=520.22[M+H] ⁺ .

Example 3 Synthesis of I-25002

I-25002: the same operation as I-25001, using intermediate 4-1 (R ^1a =t-Bu, R ^2a =CH ₃ ), intermediate 1-5 (R ^3a =H). Off-white solid.

¹ H NMR (DMSO-d ₆ , 300MHz): δ=9.66 (s, 1H), 9.11 (s, 1H), 8.98 (d, J=1.9 Hz, 1H), 8.88 (s, 1H), 8.70 (d , J＝5.8Hz, 2H), 8.56(s, 1H), 8.20(m, 2H), 8.01(d, J＝8.8Hz, 1H), 7.89(d, J＝8.8Hz, 1H), 7.66(q , J=7.8Hz, 1H), 7.36(t, J=8.1Hz, 1H), 4.29(s, 3H), 1.30(s, 9H).

MS(ESI)m/z=520.22[M+H] ⁺ .

Example 4 Synthesis of I-25004

I-25004: the same operation as I-25001, using intermediate 4-1 (R ^1a =i-Bu, R ^2a =i-Pr), intermediate 1-5 (R ^3a =H). Off-white solid.

¹ H NMR (DMSO-d ₆ , 300MHz): ppm.δ=10.26(s, 1H), 9.07(s, 1H), 8.84-8.79(m, 3H), 8.67(s, 1H), 8.50(s, 1H), 8.31–8.20 (m, 2H), 8.10 (d, J=8.7Hz, 1H), 7.88 (d, J=8.7Hz, 1H), 7.68–7.60 (m, 1H), 7.35–7.29 (m ,1H),5.32–5.23(m,1H),2.31(d,J=7.0Hz,2H),2.17–2.09(m,1H),1.67(d,J=6.4Hz,6H),0.99(d, J=6.5Hz, 6H).

MS(ESI) m/z=548.26[M+H] ⁺ .

Example 5 Synthesis of I-25005

I-25005: the same operation as I-25001, using intermediate 4-1 (R ^1a =i-Bu, R ^2a =THP), intermediate 1-5b (R ^3a =CH ₃ ). And then deprotected _{(TFA: 10eq, Et 3 SiH} : 2.5eq, DCM) to give I-25005, as a white solid.

¹ H NMR (DMSO-d ₆ , 300MHz): ppm.δ=14.27(s,1H), 10.38(s,1H), 9.60(s,1H), 8.85-8.48(m,5H), 7.89-7.48( m, 6H), 3.96 (s, 3H), 2.28 (s, 2H), 2.13 (s, 1H), 0.98 (d, J=5.2 Hz, 6H).

MS(ESI)m/z=520.22[M+H] ⁺ .

Example 6 Synthesis of I-25006

I-25006: Intermediate 4-1 (R ^1a =i-Bu, R ^2a =CH ₃ , 0.18 g, 0.5 mmol), Intermediate 1-5b (R ^3a =CH ₃ , 0.1 g, 1 eq) were added to In a 50ml single-neck bottle, sodium metabisulfite (0.14g, 1.5eq) and DMF (5ml) were added, and the reaction was carried out at 120°C for 6h. After cooling, water was added to quench, suction filtration, and water washing to obtain crude product, which was then slurried with methanol to obtain 50 mg of off-white solid.

¹ HNMR (DMSO-d ₆ , 300MHz): ppm.δ=14.27(s, 1H), 10.38(s, 1H), 9.30(s, 1H), 8.80-8.41(m, 4H), 7.93-7.45(m ,6H),4.20(s,3H),3.90(s,3H),2.28(s,2H),2.10(s,1H),0.98(d,J＝5.2Hz,6H).

MS(ES ⁺ ): 534.24.

Effect Example

In vitro chondrogenic and osteogenic differentiation experiments of human-derived synovial stem cells

Human-derived synovial mesenchymal stem cells (SMSCs) were seeded into each well of a six-well plate _{, and cultured at 5% CO 2} at 37° C. until the cell density per well reached 95%. Was added to a final concentration of 10μM of the compound to the cells and the cells were cultured at 5% CO _2, 37 ℃ (in DMSO), for 21 days. RNA was extracted by Trizol method and reverse transcribed into cDNA. Detection of cartilage-forming genes proteoglycan (Aggrecan), type II collagen (Collagen II) and sox9, and osteogenic genes type I collagen (collagen I), type X collagen (collagen X), osteogenic transcription gene (RUNX2), basic Relative expression levels of genes such as phosphatase (ALP) in cells. Relative expression levels were detected using the LightCycler 480 II system (Roche). Basal levels of chondrogenic osteoblast differentiation were determined using vehicle (DMSO) as a control. The target genes to be tested in this experiment are Aggrecan, Collagen II, sox9, Collagen I, Collagen X, RUNX2, and ALP, and the internal reference gene is GAPDH. Among them, three genes, Aggrecan, Collagen II, and sox9, are cartilage genes, and collagen type I (collagen I), collagen X (collagen X), osteogenic transcription gene (RUNX2), and alkaline phosphatase (ALP) are osteogenic genes. The gene, sox2, is a transcription factor that is a hallmark of induced stem cells. The specific results are shown in Table 1:

Table 1

Wherein, the structural formula of compound number SM04690 in the table is

Remarks: A to D represent different activity levels, see Table 2 for details:

Table 2

活性级别activity level	AA	BB	CC	DD
mAGGRECANmAGGRECAN	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0
mCOL2A1mCOL2A1	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0
SOX9SOX9	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0
COL1COL1	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0
COL10COL10	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0
RUNX2RUNX2	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0
ALPALP	0-1.00-1.0	1.0-3.01.0-3.0	3.0-5.03.0-5.0	5.0-10.05.0-10.0

Note: The values in Table 2 refer to the relative expression levels of genes induced by the compounds of the present application compared to the control group (DMSO).

As can be seen from the above Table 1, under the induction of the compound of the present application, the gene expression of one or more of the sox9 gene, collagen I, type X collagen, osteogenic transcription gene (RUNX2) and alkaline phosphatase increased, This proves that synovial mesenchymal stem cells have a tendency to transform into chondroblasts and/or osteoblasts under the induction of the drug of the present application.

Although the specific embodiments of the present invention have been described above, those skilled in the art should understand that these are only examples, and various changes may be made to these embodiments without departing from the principle and essence of the present invention. Revise. Accordingly, the scope of protection of the present invention is defined by the appended claims.

Claims

A compound as shown in formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its tautomer or Its prodrugs:

Wherein, R 1, R 2, R 3, R 1 ', R 2' and R 3 'is independently H, C 6 ~ C 18 aryl group or substituted with one or more of R 1-1 is C 6 ~ C 18 aryl;

Each R 1-1 is independently halogen or C 1 -C 4 alkyl;

R 4 , R 5 , R 4 ' and R 5 ' are independently H or C 1 -C 4 alkyl, but R 4 and R 5 are not H at the same time;

R 6 and R 6 ′ are independently C 6 -C 18 aryl, one or more R 6-1 substituted C 6 -C 18 aryl, 5-10-membered heteroaryl, or one or more R 6-2 substituted 5-10-membered heteroaryl; the heteroatoms in the 5-10-membered heteroaryl are independently selected from one or more of N, O and S, and the number of heteroatoms is 1 , 2, 3 or 4; the heteroatoms in the 5-10-membered heteroaryl group in the one or more R 6-2 substituted 5-10-membered heteroaryl groups are independently selected from N, O and One or more of S, the number of heteroatoms is 1, 2, 3 or 4;

Each of R 6-1 and R 6-2 is independently
amino or
and

R 6-1-1 is a C 1 -C 4 alkyl group or a C 3 -C 6 cycloalkyl group.
The compound represented by formula Ia or Ib according to claim 1, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual Variants or prodrugs thereof, wherein when R 1 , R 2 , R 3 , R 1 ', R 2 ' and R 3 ' are independently C 6 -C 18 aryl or one or more R 1 When -1 substituted C 6 -C 18 aryl group, the C 6 -C 18 aryl group is a C 6 -C 14 aryl group, and may also be a phenyl group, a naphthyl group, an anthracenyl group or a phenanthryl group, Further can be phenyl;

And/or, when R 1 , R 2 , R 3 , R 1 ', R 2 ' and R 3 ' are independently one or more C 6 -C 18 aryl groups substituted by R 1-1, the The number of R 1-1 is one;

And/or, when R 1-1 is halogen, the halogen is F, Cl, Br or I, or F;

And/or, when R 4 , R 5 , R 4 ' and R 5 ' are independently C 1 -C 4 alkyl groups, the C 1 -C 4 alkyl groups are methyl, ethyl, n- propyl, isopropyl, n-butyl,
Or tert-butyl, or methyl or isopropyl;

And/or, when R 6 and R 6 ' are independently 5-10-membered heteroaryl or 5-10-membered heteroaryl substituted by one or more R 6-2 , the 5-10-membered heteroaryl The heteroatom in the aryl group is N, and the number of heteroatoms can be 1;

And/or, when R 6 and R 6 ' are independently 5-10-membered heteroaryl groups or 5-10-membered heteroaryl groups substituted by one or more R 6-2 , the 5-10-membered heteroaryl groups The base is a 5- to 6-membered heteroaryl group;

And/or, when R 6 and R 6 ' are independently 5-10-membered heteroaryl groups or 5-10-membered heteroaryl groups substituted by one or more R 6-2 , the 5-10-membered heteroaryl groups The heteroatom in the base is N, and the number of heteroatoms can be 1; the 5-10-membered heteroaryl group can further be a pyridyl group or a pyridyl group.

And / or, when R 6 and R 6 'is independently substituted with one or more R 6-2 is 5 to 10-membered heteroaryl, said R is a number of 6-2;

and/or, when R 6-2 is
When the R 6-1-1 is a C 1 -C 4 alkyl group, the C 1 -C 4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl,
or tert-butyl, or
or tert-butyl.
The compound represented by formula Ia or Ib according to claim 1 or 2, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, Its tautomer or its prodrug, wherein,

When R 1 , R 2 , R 3 , R 1 ′, R 2 ′ and R 3 ′ are independently one or more C 6 -C 18 aryl groups substituted by R 1-1 , the one or more The C 6 -C 18 aryl group substituted by each R 1-1 is a fluorophenyl group, which can also be

And/or, when R 6 and R 6 ' are independently one or more R 6-2 substituted 5-10-membered heteroaryl groups, the one or more R 6-2 substituted 5-10-membered heteroaryl groups Heteroaryl is
The compound represented by formula Ia or Ib according to claim 1, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual A variant or a prodrug thereof, wherein,

R 1 and R 1 'are independently C 6 ~ C 18 aryl group, or substituted with one or more of R 1-1 is C 6 ~ C 18 aryl group; and

R 2 , R 3 , R 2 ′ and R 3 ′ are independently H;

R 1-1 is halogen;

One of R 4 and R 5 is a C 1 -C 4 alkyl group, and the other is H;

R 4 ' and R 5 ' are H;

R 6 and R 6 ' are independently 5-10-membered heteroaryl or 5-10-membered heteroaryl substituted by one or more R 6-2;

Each R 6-2 is independently
and

R 6-1-1 is a C 1 -C 4 alkyl group.
The compound represented by formula Ia or Ib according to any one of claims 1-4, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer isomer, its tautomer or its prodrug, wherein,

R 1 and R 1 ' are independently one or more C 6 -C 18 aryl groups substituted by R 1-1;

And/or, R 6 and R 6 ′ are independently 5-10-membered heteroaryl groups substituted by one or more R 6-2 .
The compound represented by formula Ia or Ib according to claim 4, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer, its mutual A variant or a prodrug thereof, wherein,

In the compound shown in the formula Ia: R 1 is an aryl group of C 6 -C 18 or an aryl group of C 6 -C 18 substituted by one or more R 1-1 ; R 2 and R 3 are H R 1-1 is halogen; R 4 is C 1 -C 4 alkyl; R 5 is H or C 1 -C 4 alkyl; R 6 is 5-10-membered heteroaryl or one or more R 6-2 substituted 5-10-membered heteroaryl; each R 6-2 is independently
and R 6-1-1 is a C 1 -C 4 alkyl group;

Or, in the compound shown in the formula Ia: R 1 is an aryl group of C 6 -C 18 or an aryl group of C 6 -C 18 substituted by one or more R 1-1 ; R 2 and R 3 is H; R 1-1 is halogen; R 4 is H or C 1 -C 4 alkyl; R 5 is C 1 -C 4 alkyl; R 6 is 5-10-membered heteroaryl or one or more 5-10-membered heteroaryl substituted by R 6-2 ; each R 6-2 is independently
and R 6-1-1 is a C 1 -C 4 alkyl group;

Or, R 1 is a C 6 -C 18 aryl group or a C 6 -C 18 aryl group substituted by one or more R 1-1;

R 2 and R 3 are H;

R 1-1 is halogen;

R 4 is H;

R 5 is a C 1 -C 4 alkyl group;

R 6 is a 5-10-membered heteroaryl group or a 5-10-membered heteroaryl group substituted by one or more R 6-2;

Each R 6-2 is independently
and

R 6-1-1 is a C 1 -C 4 alkyl group;

Or, in the compound shown in formula Ib: R 1 ' is a C 6 -C 18 aryl group substituted by one or more R 1-1 ; R 2 ' and R 3 ' are H; R 1- 1 is halogen; R 4 ' and R 5 ' are H; R 6 ' is a 5-10-membered heteroaryl or a 5-10-membered heteroaryl substituted by one or more R 6-2 ; each R 6-2 independently for
and R 6-1-1 is a C 1 -C 4 alkyl group.
The compound represented by formula Ia or Ib according to any one of claims 1-6, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its stereoisomer Construct, its tautomer or its prodrug, wherein, the compound shown in formula Ia or Ib is any of the following compounds:
A preparation method of the compound shown in any one of claims 1-7 as formula Ia or Ib, wherein,

The described preparation method of the compound shown in formula Ia is method 1 or method 2:

The method 1 includes the following steps: in a solvent, in the presence of a deprotection reagent, the compound represented by the formula IIa is subjected to the deprotection reaction of the following formula to obtain the compound represented by the formula Ia,

Wherein, Q is an amino protecting group; R 1 , R 2 , R 3 , R 4 and R 6 are as defined in any one of claims 1-7;

Method 2 includes the following steps: in a solvent, in the presence of a cyclization reagent, the compound shown in formula IIIa and the compound shown in formula IVa are subjected to the following cyclization reaction to obtain the compound shown in formula Ia. That's it,

wherein, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 are as defined in any one of claims 1-7, and R 5 is not hydrogen;

The described preparation method of the compound shown in formula Ib is method 3 or method 4:

Method 3 includes the following steps: in a solvent, in the presence of a deprotection reagent, the compound shown in formula IIb is subjected to the deprotection reaction of the following formula to obtain the compound shown in formula Ib,

Wherein, Q' is an amino protecting group; the definitions of R 1 ', R 2 ', R 3 ', R 4 ' and R 6 ' are as described in any one of claims 1-7;

Method 4 includes the following steps: in a solvent, in the presence of a cyclization reagent, the compound shown in formula IIIb and the compound shown in formula IVb are subjected to the following cyclization reaction to obtain the compound shown in formula Ib. You can;

Wherein, R 1 ', R 2 ', R 3 ', R 4 ', R 5 ' and R 6 ' are as defined in any one of claims 1-7, and R 5 ' is not hydrogen.
A preparation method of the compound shown in formula Ia or Ib as claimed in claim 8, wherein,

When Q or Q' is an amino protecting group, the amino protecting group is tetrahydropyranyl;

And/or, in methods 1 and 3, the solvent is a chlorinated hydrocarbon solvent, or dichloromethane;

And/or, the deprotection reagents are organosilane compounds and organic acids; the organosilane compounds may be triethylsilane; the organic acid may be trifluoroacetic acid; the organosilanes The molar ratio of the compound to the organic acid can be 1:1 to 1:5, or 1:2.5;

And/or, the temperature of the deprotection reaction is 10-40°C, or room temperature;

And/or, the time of the deprotection reaction is 10 to 18 hours, or overnight;

And/or, in methods 2 and 4, the solvent is an amide solvent, or DMF;

And/or, described cyclization reagent is alkali metal metabisulfite, also can be sodium metabisulfite;

And/or, the molar ratio of the cyclization reagent to the compound represented by formula IIIa or IIIb is 4:1 to 1:1, or 2:1;

And/or, the temperature of the cyclization reaction is 100-140°C, or 120°C;

And/or, the time for the deprotection reaction is 3 to 9 hours, or 6 hours.
A compound of formula IIa, IIb or IIIb:

Wherein, Q or Q 'is an amino protecting group; R 1, R 2, R 3, R 4, R 6, R 1', R 2 ', R 3', R 4 ', R 5' and R 6 'is The definition is as in any one of claims 1-7.
A compound shown below:
A pharmaceutical composition, it comprises the compound shown in any one of claim 1-7 such as formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorphism , its metabolites, its stereoisomers, its tautomers or its prodrugs, and pharmaceutical excipients.
A compound as described in any one of claims 1-7 as shown in formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its Use of a stereoisomer, its tautomer or its prodrug, or the pharmaceutical composition according to claim 12, in the preparation of an inducer.
The application of claim 13, wherein,

The inducer induces proteoglycan (Aggrecan), type II collagen (Collagen II), sox9, type I collagen (collagen I), type X collagen (collagen X), osteogenic transcription gene (RUNX2) or alkaline phosphatase Elevated expression of one or more genes in (ALP);

Or, the inducer induces synovial stem cells to differentiate into chondroblasts and/or osteoblasts;

Or, the inducing agent is used as a bone building agent, or, for the treatment or prevention of periodontal disease, arthritis, joint damage, cartilage repair, osteogenesis regeneration, osteoporosis, osteopenia, spinal fusion, periodontal disease and one or more of bone tumors; wherein the bone building agent enhances fracture recovery and stimulates bone growth at the site of bone graft, joint graft or dental graft.
A compound as described in any one of claims 1-7 as shown in formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its Use of a stereoisomer, a tautomer or a prodrug thereof, or a pharmaceutical composition as claimed in claim 12 for the preparation of an inhibitor of one or more proteins in the Wnt pathway.
A compound as described in any one of claims 1-7 as shown in formula Ia or Ib, its pharmaceutically acceptable salt, its hydrate, its solvate, its polymorph, its metabolite, its Stereoisomer, tautomer or prodrug thereof, or pharmaceutical composition as claimed in claim 12 in the manufacture of a medicament for the treatment and/or prevention of a disease associated with one or more proteins in the Wnt pathway Applications.
The application of claim 16, wherein,

The disease associated with one or more proteins in the Wnt pathway is cancer, a disease associated with abnormal angiogenesis, a disease associated with cell proliferation and cell cycle, or one or more of Wnt signaling components. Disease caused by variation or disorder.
The use of claim 17, wherein,

The cancer is one or more of colon cancer, hepatocellular carcinoma, lung cancer, ovarian cancer, prostate cancer, pancreatic cancer, bone cancer and leukemia;

And/or, the disease with abnormal angiogenesis is hematopoietic system tumor of lymphatic lineage, hematopoietic system tumor of myeloid lineage, tumor of mesenchymal origin, tumor of central and peripheral nervous system and melanoma, seminoma , Kaposi's sarcoma, benign prostatic hyperplasia, familial adenomatous polyposis, neurofibromas, atherosclerosis, arthritis, nephritis, restenosis after angioplasty or vascular surgery, inflammatory bowel disease, transplant rejection, endovascular one or more of toxin shock and fungal infection;

And/or, the disease caused by the variation or disorder of one or more of the Wnt signaling components is colon polyposis, osteoporosis, pseudoglioma syndrome, familial exudative Vitreoretinopathy, retinal angiogenesis, early coronary heart disease, congenital limb amputation syndrome, paramesonephric reversion and virilization, SERKAL syndrome, type 2 diabetes, Fuhrman syndrome, AARRS short limb syndrome, dental Onychocutaneous dysplasia, obesity, split hand/foot deformity, caudal duplication syndrome, congenital missing teeth, Wilms tumor, skeletal dysplasia, focal skin hypoplasia, neural tube defect, alpha thalassemia syndrome, One or more of Fragile X Syndrome, ICF Syndrome, Angelman Syndrome, Parr-Williams Syndrome, Belle-Williams Syndrome, Norrie Disease and Rett Syndrome.