WO2017174023A1 - Biphenyl compound serving as ezh2 inhibitor - Google Patents

Abstract

Disclosed is a novel type of compounds serving as an EZH2 inhibitor; specifically disclosed are a compound as represented by formula (III) and a pharmaceutically acceptable salt thereof.

Description

Biphenyl compound as an EZH2 inhibitor Field of invention

The present invention relates to a novel class of compounds as EZH2 inhibitors, in particular to compounds of formula (III) or pharmaceutically acceptable salts thereof.

Background of the invention

In eukaryotic cells, DNA and histone packaging form chromatin. Approximately 150 base pairs of DNA are entangled in two rounds of histone octamers (two of histones 2A, 2B, 3 and 4) to form the basic unit of chromatin, the nucleosome. Changes in the ordered structure of chromatin can cause changes in the transcription of related genes. This process is highly controlled because the depth of change in gene expression patterns affects basal cellular processes such as differentiation, proliferation, and apoptosis. By covalently modifying histones, most notably its N-terminal tail mediates the control of changes in chromatin structure (and thus transcription). These modifications are often referred to as epigenetic modifications because they can cause heritable changes in gene expression without affecting the sequence of the DNA itself. Covalent modifications (eg, methylation, acetylation, phosphorylation, and ubiquitination) of amino acid side chains are enzymatically mediated.

The selective addition of a methyl group to a particular amino acid site on a histone is controlled by the action of a unique family of enzymes called histone methyltransferase (HMT). The level of expression of a particular gene is affected by the presence or absence of one or more methyl groups at the associated histone site. The specific action of the methyl group at a particular histone site continues until the methyl group is removed by histone demethylase, or until the modified histone is replaced by nucleosome turnover.

Biochemical and genetic studies provide evidence that the Drosophila PcG protein plays a role in at least two different protein complexes, the polycomb inhibiting complex 1 (PRC1) and the ESC-E (Z) complex. (Also known as Multi-Comb Inhibition Complex 2 (PRC2)), although the composition of the complex can be dynamic. Otte et al. (2003) CurrOpinGenet Dev13: 448-54. Studies in Drosophila and studies in mammalian cells have demonstrated that ESC-E(Z)/EED-EZH2 (ie, PRC2) complexes have intrinsic histones Methyltransferase activity. Although the composition of the complexes isolated by the different groups is slightly different, it generally includes EED, EZH2, SUZ12 and RbAp48 or their Drosophila homologs. However, only the reconstituted complex including EED, EZH2, and SUZ12 retained histone methyltransferase activity against the 27th lysine of histone H3.

EZH2 (enhancer of zeste homolog 2; human EZH2 gene: Cardoso et al, European J of Human Genetics, Vol. 8, 174-180, 2000) is a catalytic structure of polycomb inhibiting complex 2 (PRC2), its function is through the pair of groups Protein H3 lysine 27 is trimethylated (H3K27me3) to silence target gene function.

Histone H3 is one of the five major histone proteins involved in chromatin of eukaryotic cells. Histone H3 has a major globular domain and a long N-terminal tail, and histones are involved in the structure of nucleosomes, like pearls on necklaces. Histones are highly modified after translation and histone H3 is the most extensive modification of five histones. Histone H3 is an important protein in the emerging field of epigenetics, and its sequence variation and multiple modification states are thought to play a role in dynamic and long-term regulation of genes.

In human cancer, there is increasing evidence that dysregulation of epigenetic enzyme activity leads to cancer-associated cell proliferation uncontrolled and other cancer-related phenotypes, such as enhanced cell migration and invasion. In addition to cancer, there is increasing evidence of the role of epigenetic enzymes in many other human diseases, including metabolic diseases (such as diabetes), inflammatory diseases (such as Crohn's disease), and neurodegenerative diseases ( For example, Alzheimer's disease) and cardiovascular disease. Therefore, the selective regulation of the abnormal effects of epigenetic enzymes has broad prospects for the treatment of a range of diseases.

Recently, somatic mutations in EZH2 and germinal centers of follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL) have been reported. B cell-like (GCB) subtypes are associated. Morin et al. (2010) NatGenet 42: 181-5. In all cases, the presence of the mutant EZH2 gene was found to be heterozygous and expression of wild-type and mutant alleles was detected in the mutant samples by transcriptome sequencing. Currently, the standard of care for the treatment of most DLBCL is the R-CHOP method. However, the results of this method are far from satisfactory. Therefore, there is an urgent need in the medical field to identify new and effective treatments that are optionally dependent on the genetic profile of the subject.

EZH2 inhibitors have a therapeutic effect on cancer, and patents that have been filed by PCT are PCT/US2011/035336, PTC/US2011/035340, PCT/US2011/035344 and U.S. Patent No. 8,410,088. It is an ideal tumor recognition feature or subgroup (eg genotype).

WO2000001666A1 reports compounds

Summary of the invention

a compound of the formula (III), a pharmaceutically acceptable salt thereof and a tautomer thereof,

among them,

R _{31 is} selected from H, halogen, CN, OH, amino, or selected from, optionally substituted by 1, 2 or 3 R: 5- to 6-membered aryl, 5- to 12-membered heteroaryl, C _1-6 alkane a group, a C _1-6 heteroalkyl group, a C _3-9 cycloalkyl group, and a _3-9 membered heterocycloalkyl group;

D _{3 is} selected from -[C(R) ₂ ] _1-3 -, -O, -S-, -N(R)-, -C(=O)-, -C(=O)O-, -S (=O)-, -S(=O) ₂ -, -C(=O)N(R)-, -N(R)C(=O)N(R)-; T are independently selected from N Or CH;

R is selected from H, OH, CN, NH ₂ , COOH, halogen, or selected from, optionally substituted by 1, 2 or 3 R's: 5- to 6-membered aryl, 5- to 12-membered heteroaryl, C _{1 a -6} alkyl group, a C _1-6 heteroalkyl group, a C _3-9 cycloalkyl group, and a _3-9 membered heterocycloalkyl group;

R' is selected from the group consisting of F, Cl, Br, I, OH, CN, NH ₂ , COOH, Me, Et, CH ₂ F, CHF ₂ , CF ₃ , NH(CH ₃ ), N(CH ₃ ) ₂ ; "" denotes a hetero atom or a hetero atom selected from -C(=O)N(R)-, -N(R)-, -C(=NR)-, -S(=O) ₂ N(R)-, -S(=O)N(R)-, -O-, -S-, =O, =S, -ON=, -C(=O)O-, -C(=O)-, -C( =S)-, -S(=O)-, -S(=O) ₂ - and -N(R)C(=O)N(R)-;

In either case, the number of heteroatoms or heteroatoms is independently selected from 1, 2 or 3.

In some embodiments of the invention, the above R is selected from the group consisting of H, OH, CN, NH ₂ , COOH, halogen, or selected from the group consisting of 1, 2 or 3 R's substituted: 5-6 aryl, 5~ 6-membered heteroaryl, C _1-3 alkyl, C _1-3 alkyl-NH-, N,N-di(C _1-3 alkyl)amino, C _1-3 alkyl-O-, C _{1 -3} alkyl-S-, C _3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl.

In some embodiments of the invention, the above R is selected from the group consisting of H, OH, CN, NH ₂ , COOH, halogen, or selected from the group consisting of 1, 2 or 3 R' substitutions: Me, Et, isopropyl, ring Propyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

Phenyl, pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolidinyl, piperidine Pyridyl, morpholinyl, piperazinyl, azetidinyl.

In some aspects of the invention, the above R is selected from the group consisting of: H, OH, CN, NH ₂ , COOH, F, Cl, Br, I, Me, Et, CH ₂ F, CHF ₂ , CF ₃ , NH(CH ₃ ) , N(CH ₃ ) ₂ ,

Cyclopropyl,

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of 1, 2 or 3 R: 5- to 6-membered aryl , 5-6-membered heteroaryl, C _1-3 alkyl, C _1-3 alkyl-NH-, N,N-di(C _1-3 alkyl)amino, C _1-3 alkyl-O- C _1-3 alkyl-S-, C _3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl.

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of 1, 2 or 3 R: Me, Et, Isopropyl Base, NH(CH ₃ ), N(CH ₃ ) ₂ ,

Phenyl, pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolidinyl, piperidine Pyridyl, morpholinyl, piperazinyl, azetidinyl, cyclopropyl.

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of 1, 2 or 3 R: Me, Et, Isopropyl Base, NH(CH ₃ ), N(CH ₃ ) ₂ ,

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, Me, Et, isopropyl, NH(CH ₃ ),

In some aspects of the invention, the above D _{3 is} selected from the group consisting of: -CH ₂ -, -CH(OH)-, -O, -S-, -NH-, -C(=O)-, -C(=O) O-, -S(=O)-, -S(=O) ₂ -, -C(=O)NH-, -NHC(=O)NH-.

In some embodiments of the invention, the above compounds, pharmaceutically acceptable salts thereof, and tautomers thereof, are selected from the group consisting of:

among them,

R ₃₁ and D _{3 are} as defined above.

In some embodiments of the invention, the above R is selected from the group consisting of H, OH, CN, NH ₂ , COOH, halogen, or selected from the group consisting of 1, 2 or 3 R's substituted: 5-6 aryl, 5~ 6-membered heteroaryl, C _1-3 alkyl, C _1-3 alkyl-NH-, N,N-di(C _1-3 alkyl)amino, C _1-3 alkyl-O-, C _{1 -3} alkyl-S-, C _3-6 cycloalkyl and 3 to 6 membered heterocycloalkyl, the other variables are as defined above.

In some embodiments of the invention, the above R is selected from the group consisting of H, OH, CN, NH ₂ , COOH, halogen, or selected from the group consisting of 1, 2 or 3 R' substitutions: Me, Et, isopropyl, ring Propyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

Phenyl, pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolidinyl, piperidine Pyridyl, morpholinyl, piperazinyl, azetidinyl, other variables are as defined above.

In some aspects of the invention, the above R is selected from the group consisting of: H, OH, CN, NH ₂ , COOH, F, Cl, Br, I, Me, Et, CH ₂ F, CHF ₂ , CF ₃ , NH(CH ₃ ) , N(CH ₃ ) ₂ ,

Cyclopropyl,

Other variables are as defined above.

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of 1, 2 or 3 R: 5- to 6-membered aryl , 5-6-membered heteroaryl, C _1-3 alkyl, C _1-3 alkyl-NH-, N,N-di(C _1-3 alkyl)amino, C _1-3 alkyl-O- C _1-3 alkyl-S-, C _3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl, and other variables are as defined above.

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of 1, 2 or 3 R: Me, Et, Isopropyl Base, NH(CH ₃ ), N(CH ₃ ) ₂ ,

Phenyl, pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolidinyl, piperidine Pyridyl, morpholinyl, piperazinyl, azetidinyl, cyclopropyl, other variables are as defined above.

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of 1, 2 or 3 R: Me, Et, Isopropyl Base, NH(CH ₃ ), N(CH ₃ ) ₂ ,

Other variables are as defined above.

In some embodiments of the invention, the above R _{31 is} selected from the group consisting of: H, F, Cl, Br, I, CN, OH, amino, Me, Et, isopropyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

Other variables are as defined above.

In some aspects of the invention, the above D _{3 is} selected from the group consisting of: -CH ₂ -, -CH(OH)-, -O, -S-, -NH-, -C(=O)-, -C(=O) O-, -S(=O)-, -S(=O) ₂ -, -C(=O)NH-, -NHC(=O)NH-, other variables are as defined above.

Still other aspects of the invention are arbitrarily combined by the above variables.

The compounds of the invention are selected from:

Another object of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of the above compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

Another object of the present invention is to provide the use of the above compound or a pharmaceutically acceptable salt thereof or the above pharmaceutical composition for the preparation of a medicament for treating various conditions associated with EZH2 receptor.

Definition and description

Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered undefined or unclear without a particular definition, but should be understood in the ordinary sense. When a trade name appears in this document, it is intended to refer to its corresponding commodity or its active ingredient.

C _{1-12 is} selected from C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ and C ₁₂ ; C _{3-12 is} selected from C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ and C ₁₂ .

C _1-12 alkyl or heteroalkyl, C _3-12 cyclo or heterocycloalkyl, C _1-12 alkyl or heteroalkyl substituted by C _3-12 cycloalkyl or heterocycloalkyl includes, but is not limited to:

C _1-12 alkyl, C _1-12 alkylamino, N,N-di(C _1-12 alkyl)amino, C _1-12 alkoxy, C _1-12 alkanoyl, C _1-12 alkoxy Carbonyl, C _1-12 alkylsulfonyl, C _1-12 alkylsulfinyl, C _3-12 cycloalkyl, C _3-12 cycloalkylamino, C _3-12 heterocycloalkylamino, C _3-12 Cycloalkoxy, C _3-12 cycloalkyl acyl, C _3-12 cycloalkyloxycarbonyl, C _3-12 cycloalkylsulfonyl, C _3-12 cycloalkylsulfinyl, 5-12 aryl Or a heteroaryl group, a 5- to 12-membered aralkyl group or a heteroarylalkyl group;

Methyl, ethyl, n-propyl, isopropyl, -CH ₂ C(CH ₃ )(CH ₃ )(OH), cyclopropyl, cyclobutyl, propylmethylene, cyclopropionyl, benzyloxy Base, trifluoromethyl, aminomethyl, hydroxymethyl, methoxy, formyl, methoxycarbonyl, methylsulfonyl, methylsulfinyl, ethoxy, acetyl, ethylsulfonyl, ethoxycarbonyl , dimethylamino, diethylamino, dimethylaminocarbonyl, diethylaminocarbonyl;

_{N (CH 3) 2, NH} (CH 3), - CH 2 CF 3, -CH2CH 2 CF 3, -CH 2 CH 2 F, -CH 2 CH 2 S (= O) 2 CH 3, -CH 2 CH ₂ CN, -CH ₂ CH(OH)(CH3) ₂ , -CH ₂ CH(F)(CH ₃ ) ₂ , -CH ₂ CH ₂ F, -CH ₂ CF ₃ , -CH ₂ CH ₂ CF ₃ ,- CH ₂ CH ₂ NH ₂ , -CH ₂ CH ₂ OH, -CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ N(CH ₃ ) ₂ , -S(=O) ₂ CH ₃ , -CH ₂ CH ₂ S(=O) ₂ CH ₃ ,; and

Phenyl, thiazolyl, biphenyl, naphthyl, cyclopentyl, furyl, 3-pyrrolyl, pyrrolidinyl, 1,3-oxapentacyclyl, pyrazolyl, 2-pyrazolyl, Pyrazolidinyl, imidazolyl, oxazolyl, thiazolyl, 1,2,3-oxazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4- Thiadiazolyl, 4H-pyranyl, pyridyl, piperidinyl, 1,4-dioxolyl, morpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, 1,3 , 5-trithiazide, 1,3,5-triazinyl, benzofuranyl, benzothienyl, fluorenyl, benzimidazolyl, benzothiazolyl, fluorenyl, quinolinyl, iso a quinolyl group, a porphyrin group or a quinoxaline group;

The term "pharmaceutically acceptable" as used herein is intended to mean that those compounds, materials, compositions and/or dosage forms are within the scope of sound medical judgment and are suitable for use in contact with human and animal tissues. Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the invention prepared from a compound having a particular substituent found in the present invention and a relatively non-toxic acid or base. When a relatively acidic functional group is contained in the compound of the present invention, a base addition salt can be obtained by contacting a neutral amount of such a compound with a sufficient amount of a base in a neat solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts. When a relatively basic functional group is contained in the compound of the present invention, an acid addition salt can be obtained by contacting a neutral form of such a compound with a sufficient amount of an acid in a neat solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogencarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and an organic acid salt, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine, etc.) And salts of organic acids such as glucuronic acid (see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds of the invention contain both basic and acidic functional groups which can be converted to any base or acid addition salt.

Preferably, the salt is contacted with a base or acid in a conventional manner, and the parent compound is separated, thereby regenerating the neutral form of the compound. The parent form of the compound differs from the form of its various salts by certain physical properties, such as differences in solubility in polar solvents.

As used herein, a "pharmaceutically acceptable salt" is a derivative of a compound of the invention wherein the parent compound is modified by salt formation with an acid or with a base. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of bases such as amines, alkali metal or organic salts of acid groups such as carboxylic acids, and the like. Pharmaceutically acceptable salts include the conventional non-toxic salts or quaternary ammonium salts of the parent compound, for example salts formed from non-toxic inorganic or organic acids. Conventional non-toxic salts include, but are not limited to, those derived from inorganic acids and organic acids selected from the group consisting of 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, Benzenesulfonic acid, benzoic acid, hydrogencarbonate, carbonic acid, citric acid, edetic acid, ethane disulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptose, gluconic acid, glutamic acid, glycolic acid, Hydrobromic acid, hydrochloric acid, hydroiodide, hydroxyl, hydroxynaphthalene, isethionethane, lactic acid, lactose, dodecylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, nitric acid, oxalic acid, Pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturaldehyde, propionic acid, salicylic acid, stearic acid, acrylic acid, succinic acid, sulfamic acid, p-aminobenzenesulfonic acid, sulfuric acid, tannin, Tartaric acid and p-toluenesulfonic acid.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by conventional chemical methods. In general, such salts are prepared by reacting these compounds in water or an organic solvent or a mixture of the two via a free acid or base form with a stoichiometric amount of a suitable base or acid. Generally, a nonaqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is preferred.

In addition to the form of the salt, the compounds provided herein also exist in the form of prodrugs. Prodrugs of the compounds described herein are readily chemically altered under physiological conditions to convert to the compounds of the invention. Furthermore, prodrugs can be converted to the compounds of the invention by chemical or biochemical methods in an in vivo setting.

Certain compounds of the invention may exist in unsolvated or solvated forms, including hydrated forms. In general, the solvated forms are equivalent to the unsolvated forms and are included within the scope of the invention.

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

Graphical representations of racemates, ambiscalemic and scalemic or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62: 114-120. 1985, 62: 114-120. Unless otherwise stated, the wedge key and the dashed key indicate a solid center For the configuration. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include the E and Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the invention.

The compounds of the invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including the cis and trans isomers, the (-)- and (+)-p-enantiomers, the (R)- and (S)-enantiomers, and the diastereomeric a conformation, a (D)-isomer, a (L)-isomer, and a racemic mixture thereof, and other mixtures, such as enantiomerically or diastereomeric enriched mixtures, all of which belong to It is within the scope of the invention. Additional asymmetric carbon atoms may be present in the substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the invention.

The optically active (R)- and (S)-isomers as well as the D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary wherein the resulting mixture of diastereomers is separated and the auxiliary group cleaved to provide pure The desired enantiomer. Alternatively, when a molecule contains a basic functional group (e.g., an amino group) or an acidic functional group (e.g., a carboxyl group), a diastereomeric salt is formed with a suitable optically active acid or base, followed by conventional methods well known in the art. The diastereomers are resolved and the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomers is generally accomplished by the use of chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (eg, formation of an amino group from an amine). Formate).

The compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms that make up the compound. For example, radiolabeled compounds can be used, such as tritium ⁽³ H), iodine -125 ⁽¹²⁵ I) or ^C-14 (14 C). Alterations of all isotopic compositions of the compounds of the invention, whether radioactive or not, are included within the scope of the invention.

The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium that is capable of delivering an effective amount of an active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic side effects to the host or patient, including water, oil, Vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, tackifiers, transdermal enhancers and the like. Their formulations are well known to those skilled in the cosmetic or topical pharmaceutical arts. For additional information on vectors, reference is made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are hereby incorporated by reference.

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

The term "effective amount" or "therapeutically effective amount" with respect to a pharmaceutical or pharmacologically active agent refers to a sufficient amount of a drug or agent that is non-toxic but that achieves the desired effect. For oral dosage forms in the present invention, an "effective amount" of an active substance in a composition refers to the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount will vary from person to person, depending on the age and general condition of the recipient, and also on the particular active substance, and a suitable effective amount in a case can be determined by one skilled in the art based on routine experimentation.

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

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, including variants of heavy hydrogen and hydrogen, as long as the valence of the particular atom is normal and the substituted compound is stable. . When the substituent is a keto group (ie, =0), it means that two hydrogen atoms are substituted. Ketone substitution does not occur on the aryl group. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemically achievable.

When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted by 0-2 R, the group may optionally be substituted at most by two R, And each case has an independent option for R. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

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

When one of the variables is selected from a single bond, it means that the two groups to which it is attached are directly linked. For example, when L represents a single bond in A-L-Z, the structure is actually A-Z.

When a substituent is vacant, it means that the substituent is absent. For example, when X is vacant in AX, the structure is actually A. When a bond of a substituent can be cross-linked to two atoms on a ring, the substituent can be bonded to any atom on the ring. When the recited substituents do not indicate which atom is attached to a compound included in the chemical structural formula including but not specifically mentioned, such a substituent may be bonded through any atomic phase thereof. Combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds. For example, a structural unit

It is indicated that it can be substituted at any position on the cyclohexyl or cyclohexadiene.

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

Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl. "Alkoxy" represents the above alkyl group having a specified number of carbon atoms attached through an oxygen bridge. The C _1-6 alkoxy group includes a C ₁ , C ₂ , C ₃ , C ₄ , C ₅ and C ₆ alkoxy groups. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy and S- Pentyloxy. "Cycloalkyl" includes saturated cyclic groups such as cyclopropyl, cyclobutyl or cyclopentyl. The 3-7 cycloalkyl group includes C ₃ , C ₄ , C ₅ , C ₆ and C ₇ cycloalkyl groups. "Alkenyl" includes hydrocarbon chains in a straight or branched configuration wherein one or more carbon-carbon double bonds, such as vinyl and propylene groups, are present at any stable site on the chain.

The term "halo" or "halogen" refers to fluoro, chloro, bromo and iodo.

Unless otherwise specified, the term "hetero" denotes a hetero atom or a hetero atomic group (ie, a radical containing a hetero atom), including atoms other than carbon (C) and hydrogen (H), and radicals containing such heteroatoms, including, for example, oxygen (O). ), nitrogen (N), sulfur (S), silicon (Si), germanium (Ge), aluminum (Al), boron (B), -O-, -S-, =O, =S, -C (= O) O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, and optionally substituted -C(=O)N (H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- or -S(=O)N(H)-.

Unless otherwise specified, "ring" means substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl or heteroaryl. So-called rings include single rings, interlocking rings, spiral rings, parallel rings or bridge rings. The number of atoms on the ring is usually defined as the number of elements of the ring. For example, "5 to 7-membered ring" means 5 to 7 atoms arranged in a circle. Unless otherwise specified, the ring optionally contains from 1 to 3 heteroatoms. Thus, "5- to 7-membered ring" includes, for example, phenyl, pyridine, and piperidinyl; on the other hand, the term "5- to 7-membered heterocycloalkyl ring" includes pyridyl and piperidinyl, but does not include phenyl. The term "ring" also includes ring systems containing at least one ring, each of which "ring" independently conforms to the above definition.

Unless otherwise specified, the term "heterocycle" or "heterocyclyl" means a stable monocyclic, bicyclic or tricyclic ring containing a hetero atom or a heteroatom group which may be saturated, partially unsaturated or unsaturated ( Aromatic) which comprise a carbon atom and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S, wherein any of the above heterocycles may be fused to a phenyl ring to form a bicyclic ring. The nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). The nitrogen atom may be substituted or unsubstituted (ie N or NR, where R is H or this article has Other substituents that have been defined). The heterocyclic ring can be attached to the side groups of any hetero atom or carbon atom to form a stable structure. If the resulting compound is stable, the heterocycles described herein can undergo substitutions at the carbon or nitrogen sites. The nitrogen atom in the heterocycle is optionally quaternized. A preferred embodiment is that when the total number of S and O atoms in the heterocycle exceeds 1, these heteroatoms are not adjacent to each other. Another preferred embodiment is that the total number of S and O atoms in the heterocycle does not exceed one. The term "aromatic heterocyclic group" or "heteroaryl" as used herein means a stable 5, 6, or 7 membered monocyclic or bicyclic or aromatic ring of a 7, 8, 9 or 10 membered bicyclic heterocyclic group, It contains carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S. The nitrogen atom can be substituted or unsubstituted (i.e., N or NR, wherein R is H or other substituents as already defined herein). The nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O)p, p is 1 or 2). It is worth noting that the total number of S and O atoms on the aromatic heterocycle does not exceed one. Bridged rings are also included in the definition of heterocycles. A bridged ring is formed when one or more atoms (ie, C, O, N, or S) join two non-adjacent carbon or nitrogen atoms. Preferred bridged rings include, but are not limited to, one carbon atom, two carbon atoms, one nitrogen atom, two nitrogen atoms, and one carbon-nitrogen group. It is worth noting that a bridge always converts a single ring into a three ring. In the bridged ring, a substituent on the ring can also be present on the bridge.

Examples of heterocyclic compounds include, but are not limited to, acridinyl, octanoyl, benzimidazolyl, benzofuranyl, benzofuranylfuranyl, benzindenylphenyl, benzoxazolyl, benzimidin Oxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, oxazolyl, 4aH-carbazolyl, Porphyrin, chroman, chromene, porphyrin-decahydroquinolinyl, 2H, 6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b] Tetrahydrofuranyl, furyl, furfuryl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-carbazolyl, nonenyl, indanyl, mesoindolyl, fluorenyl, 3H-indole Mercapto, isobenzofuranyl, isodecyl, isoindoline, isoquinolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl , octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, hydroxymethyl, pyrimidinyl, phenanthryl, phenanthroline, phenazine, phenothiazine , benzoxanthyl, phenoloxazinyl, pyridazinyl, piperazinyl, piperidinyl, piperidinone, 4-piperidinone, piperonyl, pteridinyl, fluorenyl, pyranyl, Pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridylthiazole, pyridyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl , pyrrolyl, quinazolinyl, quinolyl, 4H-quinazinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolyl, tetrazolyl, 6H -1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4 -thiadiazolyl, thiazolidine, thiazolyl, isothiazolylthiophenyl, thienooxazolyl, thienothiazolyl, thienoimidazolyl, thienyl, triazinyl, 1,2,3-triazole Base, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl and xanthene. Also included are fused ring and spiro compounds.

Unless otherwise specified, the term "hydrocarbyl" or its subordinate concept (such as alkyl, alkenyl, alkynyl, phenyl, etc.) by itself or as part of another substituent means straight-chain, branched or cyclic The hydrocarbon radical or a combination thereof may be fully saturated, mono- or polyunsaturated, may be monosubstituted, disubstituted or polysubstituted, and may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (methine), may include a divalent or polyvalent radical having the specified number of carbon atoms (e.g., C ₁ -C ₁₀ represents 1 to 10 carbons). "Hydrocarbyl" includes, but is not limited to, aliphatic hydrocarbyl groups including chain and cyclic, including but not limited to alkyl, alkenyl, alkynyl groups including, but not limited to, 6-12 members. An aromatic hydrocarbon group such as benzene, naphthalene or the like. In some embodiments, the term "hydrocarbyl" means a straight or branched chain radical or a combination thereof, which may be fully saturated, unitary or polyunsaturated, and may include divalent and multivalent radicals. Examples of saturated hydrocarbon radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, isobutyl, cyclohexyl, (cyclohexyl). A homolog or isomer of a methyl group, a cyclopropylmethyl group, and an atomic group such as n-pentyl, n-hexyl, n-heptyl, n-octyl. The unsaturated alkyl group has one or more double or triple bonds, and examples thereof include, but are not limited to, a vinyl group, a 2-propenyl group, a butenyl group, a crotyl group, a 2-isopentenyl group, and a 2-(butadienyl group). ), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and Structure.

Unless otherwise specified, the term "heterohydrocarbyl" or its subordinate concept (such as heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, etc.), by itself or in combination with another term, means a stable straight chain, branched chain. Or a cyclic hydrocarbon radical or a combination thereof having a number of carbon atoms and at least one heteroatom. In some embodiments, the term "heteroalkyl" by itself or in conjunction with another term refers to a stable straight chain, branched hydrocarbon radical or combination thereof, having a number of carbon atoms and at least one heteroatom. In a typical embodiment, the heteroatoms are selected from the group consisting of B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized. The heteroatom or heteroatom group can be located at any internal position of the heterohydrocarbyl group (including where the hydrocarbyl group is attached to the rest of the molecule). Examples include, but are not limited to, -CH ₂ -CH ₂ -O-CH ₃ , -CH ₂ -CH ₂ -NH-CH ₃ , -CH ₂ -CH ₂ -N(CH ₃ )-CH ₃ , -CH ₂ -S -CH ₂ -CH ₃ , -CH ₂ -CH ₂ , -S(O)-CH ₃ , -CH ₂ -CH ₂ -S(O) ₂ -CH ₃ , -CH=CH-O-CH ₃ ,- CH ₂ -CH=N-OCH ₃ and -CH=CH-N(CH ₃ )-CH ₃ . Up to two heteroatoms may be consecutive, for example, -CH ₂ -NH-OCH _3.

The terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are customary expressions and refer to those alkane which are attached to the remainder of the molecule through an oxygen atom, an amino group or a sulfur atom, respectively. Base group.

Unless otherwise specified, the term "cycloalkyl", "heterocycloalkyl" or its subordinate concept (such as aryl, heteroaryl, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl) A heterocyclic alkynyl group, etc., by itself or in combination with other terms, denotes a cyclized "hydrocarbyl group" or "heterohydrocarbyl group", respectively. Further, in the case of a heterohydrocarbyl group or a heterocycloalkyl group (such as a heteroalkyl group or a heterocycloalkyl group), a hetero atom may occupy a position at which the hetero ring is attached to the rest of the molecule. Examples of cycloalkyl groups include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Non-limiting examples of heterocyclic groups include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl and 2-piperazinyl.

Unless otherwise specified, the term "aryl" denotes a polyunsaturated, aromatic hydrocarbon substituent which may be monosubstituted, disubstituted or polysubstituted, may be monovalent, divalent or polyvalent, it may be monocyclic or Polycyclic (such as 1 to 3 rings; at least one of which is aromatic), which are fused together or covalently linked. The term "heteroaryl" refers to an aryl (or ring) containing one to four heteroatoms. In an illustrative example, the heteroatoms are selected from the group consisting of B, N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a heteroatom. Non-limiting examples of aryl or heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyridyl Azyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxan Azyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thiophene , 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, indolyl, 2-benzimidazolyl, 5-indenyl, 1-isoquinolyl, 5-isoquinolinyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl and 6-quinolinyl. The substituents of any of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.

Unless otherwise specified, aryl groups, when used in conjunction with other terms (e.g., aryloxy, arylthio, aralkyl), include aryl and heteroaryl rings as defined above. Thus, the term "aralkyl" is intended to include those radicals to which an aryl group is attached to an alkyl group (eg, benzyl, phenethyl, pyridylmethyl, and the like), including wherein the carbon atom (eg, methylene) has been, for example, oxygen. Those alkyl groups substituted by an atom, such as phenoxymethyl, 2-pyridyloxymethyl 3-(1-naphthyloxy)propyl and the like.

The term "leaving group" refers to a functional group or atom which may be substituted by another functional group or atom by a substitution reaction (for example, an affinity substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, toluene An acid ester, a p-bromobenzenesulfonate, a p-toluenesulfonate or the like; an acyloxy group such as an acetoxy group, a trifluoroacetoxy group or the like.

The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to, formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, e.g., tert-butoxycarbonyl (Boc) Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1, 1-di -(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) and the like. The term "hydroxy protecting group" refers to a protecting group suitable for use in preventing hydroxy side reactions. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and t-butyl groups; acyl groups such as alkanoyl groups (e.g., acetyl); arylmethyl groups such as benzyl (Bn), Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, combinations thereof with other chemical synthetic methods, and those well known to those skilled in the art. Equivalent alternatives, preferred embodiments include, but are not limited to, embodiments of the invention.

The solvent used in the present invention is commercially available. The present invention employs the following abbreviations: aq for water; HATU for O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate ; EDC stands for N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride; m-CPBA stands for 3-chloroperoxybenzoic acid; eq stands for equivalent, equivalent; CDI stands for Carbonyldiimidazole; DCM stands for dichloromethane; PE stands for petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF stands for N,N-dimethylformamide; DMSO stands for dimethyl sulfoxide; EtOAc stands for acetic acid Esters; EtOH for ethanol; MeOH for methanol; CBz for benzyloxycarbonyl, an amine protecting group; BOC for t-butylcarbonyl is an amine protecting group; HOAc for acetic acid; NaCNBH ₃ for sodium cyanoborohydride ; rt stands for room temperature; O/N stands for overnight; THF stands for tetrahydrofuran; Boc ₂ O stands for di-tert-butyl dicarbonate; TFA stands for trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl ₂ stands for chloride Sulfone; CS ₂ represents carbon disulfide; TsOH represents p-toluenesulfonic acid; NFSI stands for N-fluoro-N-(phenylsulfonyl)benzenesulfonamide; NCS stands for 1-chloropyrrolidine-2,5-dione; n-Bu ₄ NF stands for fluorine Tetrabutylammonium; iPrOH represents 2-propanol; mp mp Representative; Representative LDA lithium diisopropylamide.

Compound by hand or

Software naming, commercially available compounds using the supplier catalog name.

Detailed ways

Example 1

4'-(1,2-Dihydroxyethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-(B (tetrahydro-2H-pyran-4-yl)amino)-4-methyl-[1,1'-diphenyl]-3-carboxamide

Step 1: The compound bromomethyl(triphenyl)phosphorate (21.24 g, 59.46 mmol) was added to THF (100.00 mL). NaH (60%, 1.30 g, 54.05 mmol) was added at 0 ° C and stirred for 30 min. Then p-bromobenzaldehyde 1-1 (10.00 g, 54.05 mmol) was added and stirring was continued at 0 °C for 12 hours. After the reaction mixture was concentrated, EtOAc (EtOAc EtOAc (EtOAc) MS ESI calculated C ₈ H ₇ Br[M+H] ⁺ 183, found 183.

Step 2: Compound 1-2 (100.00 mg, 546.30 umol), methyl 3-[ethyl(tetrahydropyran-4-yl)amino]-2-methyl-5-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (146.89 mg, 364.20 umol), Cs ₂ CO ₃ (355.99 mg, 1.09 mmol) and Pd (dppf) Cl ₂ (26.65 mg, 36.42 umol) was added to a mixed solvent of THF (10.00 mL) / H2O (2.00 mL), and the mixture was stirred at 80 ° C for 12 hours under nitrogen atmosphere. 3) Extraction, concentration, and purification with a silica gel column to give Compound 1-3 (110 mg, yield: 79%) as a colorless oily liquid. MS ESI calc. C ₂₄ H ₂₉ NO 3 [M+H] ⁺ 380, 380.

Step 3: Compound 1-3 (110.00 mg, 289.86 umol) was added to a mixed solvent of H ₂ O (5.00 mL) / MeOH (5.00 mL) / THF (5.00 mL), and OsO ₄ (14.74 mg, 57.97 umol) and NMO (67.91 mg, 579.72 umol) were stirred at room temperature for 12 hours. The reaction mixture was concentrated, and the residue was purified mjjjjjjj MS ESI calcd for _{C 24 H 31 NO5 [M +} H] + 414, found 414.

Step 4: Compound 1-4 (100.00 mg, 241.83 umol) was added to a mixed solvent of EtOH (10.00 mL) / H ₂ O (3.00 mL), then NaOH (29.02 mg, 725.50 umol) was added, the reaction was Stir at 60 ° C for 12 hours. After completion of the reaction, the reaction mixture was adjusted to pH 6-7 with EtOAc (2M), and filtered to afford compound 1-5 (60.00 mg, crude) as a brown oil. MS ESI calcd for _{C 23 H 29 NO 5 [M} + H] + 400, found 400.

Step 5: Compound 1-5 (50.00 mg, 125.16 umol) was added to DMSO (3.00 mL), then HOBt (25.37 mg, 187.74 umol), EDCI (35.99 mg, 187.74 umol) and Et ₃ N (38.00 mg) were added. , 375.49 umol), stirred at room temperature for 2 hours, then compound 1-6 was added, and stirred at 50 ° C for 22 hours. The mixture was poured into water, filtered, and the filter cake was washed with water to obtain a target compound 1 which was purified. ^{1 H NMR (400MHz, METHANOL-} d 4) δ8.33 (br s, 1H) 7.58 (d, J = 8.28Hz, 2H) 7.43-7.50 (m, 3H) 7.33 (d, J = 1.25Hz, 1H) 6.13(s,1H)4.69-4.78(m,1H)4.47-4.55(m,2H)3.94(br d,J=11.04Hz, 2H)3.60-3.70(m,2H)3.35-3.45(m,2H) 3.10-3.25(m,3H)2.41(s,3H)2.34(s,3H)2.26(s,3H)1.74-1.86(m,2H)1.57-1.71(m,2H)0.92(t,J=7.03Hz , 3H) .MS ESI calcd for _{C 31 H 39 N 3 O 5} [M + H] + 534, found 534.

Example 2

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-3-(ethyl(tetrahydro-2H-pyran-4-yl) Ammonia)-5-(1-hydroxy-2-morpholinoethyl)-2-methylbenzamide

Step 1: Compound 2-1 (400.00 mg, 1.05 mmol) was added to DCM (15.00 mL). The mixture was then added saturated NaHCO ₃ (40mL) solution, and extracted with DCM (50mL * 3), NaHSO 3 (2M, 70mL) aqueous solution, and purified by silica gel column to give compound 2-2 (200mg, yield 48%) as a yellow Oily liquid. MS ESI calcd for _{C 24 H 29 NO4 [M +} H] + 396, found 396.

Step 2: Compound 2-2 (150.00 mg, 379.28 umol) was added to EtOH (15.00 mL), then EtOAc (66. After the completion of the reaction, the reaction mixture was concentrated to give a purified crystals of 2-3 (100 mg, yield 54%) as a colorless oil. MS ESI calculated for C ₂₈ H ₃₈ N2O5 [M+H] ⁺ 483, found 483.

Step 3: Compound 2-3 (100.00 mg, 207.21 umol) was added to a mixed solvent of dioxane (10.00 mL) / water (2.00 mL), and NaOH (24.87 mg, 621.63 umol) was added. Stir at 60 ° C for 12 hours. After completion of the reaction, the reaction mixture was adjusted to pH 6-7 with EtOAc (2M). MS ESI calcd for C ₂₇ H ₃₆ N ₂ O ₅ [M+H] ⁺ 469, found 469.

Step 4: Compound 2-4 (60.00 mg, 128.04 umol) was added to DMF (3.00 mL), then TBTU (82.22 mg, 256.08 umol), DIEA (33.10 mg, 256.08 umol) and 3-(aminomethyl) 4,6-Dimethyl-1H-pyridin-2-one (38.97 mg, 256.08 umol), stirred at room temperature for 12 hours. The title compound 2 (20.00 mg, yield 25%) was obtained by preparative chromatography. ^{1 H NMR (400MHz, METHANOL-} d 4) δ7.91 (br s, 1H) 7.82 (d, J = 8.28Hz, 1H) 7.71-7.77 (m, 2H) 7.68 (d, J = 8.28Hz, 1H) 7.56(d, J=8.03Hz, 1H) 6.12(s,1H)4.48(s,1H)4.44-4.58(m,1H)4.40-4.44(m,1H)4.07(m,1H)4.03-4.22(m ,1H)3.90-4.04(m,4H)3.83-3.90(m,1H)3.70-3.89(m,4H)3.50-3.91(m,2H)3.30-3.39(m,3H)3.00-3.25(m,2H 2.42 (s, 3H) 2.36 (s, 3H) 2.22 (s, 3H) 1.74-2.01 (m, 3H) 0.96-1.06 (m, 3H). MS ESI calcd for C ₃₅ H ₄₆ N ₄ O ₅ [M+H] ⁺ 603.

Example 3

N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-methyl-4'-(morpholino)-5-( 1-morpholinopropyl)-[1,1'-biphenyl]-3-carboxamide

Step 1: To a solution of compound 3-1 (20.00 g, 92.16 mmol) in MeOH (15 <RTI ID=0.0></RTI><RTIgt;</RTI><RTIgt; The mixture was concentrated under reduced pressure, then extracted with water and dichloromethane. The crude product was purified by column to afford compound 3-2 (19.4 g, yield: 91.11%) as white solid. ^{1 H NMR (400MHz, CHLOROFORM-} d) δ10.70 (s, 1H), 7.96 (d, J = 2.51Hz, 1H), 7.54 (dd, J = 8.78,2.51Hz, 1H), 6.89 (d, J =8.78 Hz, 1H), 3.97 (s, 3H).

Step 2: To a mixture of compound 3-2 (11.00 g, 47.61 mmol) and compound 3-3 (11.01 g, 119.03 mmol, 11.01 ml), anhydrous AlCl ₃ (19.05 g, 142.83 mmol, 7.81 ml) was added. The suspension was stirred at 80 ° C for 7 hours, extracted with water and dichloromethane at 0 ° C, dried over saturated brine and anhydrous sodium sulfate and evaporated. The crude product was purified by column to afford compound 3-4 (6.90 g, ^{1 H NMR (400MHz, CHLOROFORM-} d) δ12.17 (s, 1H), 8.14 (d, J = 2.51Hz, 1H), 8.07 (d, J = 2.76Hz, 1H), 3.98 (s, 3H), 3.08 (d, J = 7.03 Hz, 2H), 1.20 (t, J = 7.15 Hz, 3H).

Step 3: To a mixture of compound 3-4 (2.00 g, 6.97 mmol) and compound 3-5 (2.32 g, 7.67 mmol) in dioxane (15 ml) / water (3 ml) was added K ₂ CO ₃ ( 1.93 g, 13.9 mmol) and Pd(dppf)Cl ₂ (509.71 mg, 697.00 μmol). The mixture was stirred at 105 ° C for 0.5 hour under a nitrogen atmosphere. The mixture was filtered through celite, and the filtrate was diluted with water and evaporated. The residue was purified by silica gel eluting eluting elute MS ESI calcd for _{C 22 H 25 NO 5 [M} + H] + 384, found 384.

Step 4: Et added to DCM (20ml) Compound 3-6 (1.6g, 3.97mmol) was _{3 N (1.77g, 17.54mmol, 2.43ml} ), and the mixture was cooled to 0 ℃ slowly added dropwise compound 3-7 (2.12 g, 7.51 mmol, 1.24 ml). The suspension was stirred at 0 °C for 2 hours. It was diluted with water and extracted with dichloromethane. The residue was purified by silica gel chromatography eluting elut elut elut elut elut MS ESI calcd for _{C 23 H 24 F 3 NO 7} S [M + H] + 516, found 516.

Step 5: Compound 3-8 (2.20g, 4.27mmol) and Compound 3-9 (510.93mg, 8.54mmol) in tetrahydrofuran (20ml) was added K ₃ PO ₄ (1.81g Under nitrogen, 8.54 mmol And Pd(dppf)Cl ₂ (312.27 mg, 427.00 μmol). Stir at 66 ° C for 4 hours under nitrogen. The mixture was filtered through celite, and the filtrate was evaporated and evaporated. The residue was purified by silica gel column chromatography toield MS ESI calcd for _{C 23 H 24 NO 7 [M} + H] + 382, found 382.

Step 6: To a solution of compound 3-10 (800.00 <RTI ID=0.0></RTI></RTI><RTIgt; The suspension was stirred at 25 ° C for 1 hour. It was diluted with water and extracted with dichloromethane. The residue was purified by silica gel chromatography eluting elut elut elut elut ^{1 H NMR (400MHz, CHLOROFORM-} d) δ8.07 (d, J = 1.76Hz, 1H), 7.67 (d, J = 1.76Hz, 1H), 7.54 (d, J = 8.28Hz, 2H), 7.42 ( d, J=8.03 Hz, 2H), 3.93 (s, 3H), 3.69-3.74 (m, 4H), 3.55 (s, 2H), 2.83-2.95 (m, 2H), 2.55 (s, 3H), 2.48 (br.s., 4H), 1.22 (t, J = 7.28 Hz, 3H). MS ESI calcd for _{C 23 H 29 NO 4 [M} + H] + 384, found 384.

Step 7: Thionyl chloride (46.54 mg, 391.16 μmol, 28.38 μL) was slowly added dropwise to a solution of compound 3-11 (75.00 ml, 195.58 μmol) in DCM (5.00 ml). The suspension was stirred at 20 ° C for 12 hours. Diluted with water and extracted with methylene chloride. EtOAc (EtOAc m. MS ESI calcd for _{C 23 H 28 ClNO 3 [M} + H] + 402, found 402.

Step 8: A solution of compound 3-12 (70.00 mg, 174.16 μmol) in MeCN (5.00 ml) was added to compound 3-13 (22.76 mg, 261.24 μmol, 22.99 μl), and the mixture was stirred at 20 ° C for 2 hours. Then K ₂ CO ₃ (48.14 mg, 348.32 μmol) was added, and the mixture was further stirred at 90 ° C for 2 hours. It was diluted with water and extracted with dichloromethane. The residue was purified by silica gel chromatography eluting elut372 MS ESI calcd for _{C 28 H 37 NO 4 [M} + H] + 452, found 452.

Step 9: To a solution of compound 3-14 (20.00 mg, 44.19 μmol) in MeOH/THF/H ₂ O (1.00 mL, 1.00 mL, 0.5 mL) NaOH (4.42 mg, 110.48 μmol), the suspension at 80 ° C After stirring for 1 hour, the organic phase was concentrated to dryness crystals crystals crystals crystalsssssssssssssssssss MS ESI calcd for _{C 26 H 34 N 2 O 4} [M + H] + 439, found 439.

Step 10: To a solution of compound 3-15 (20.00 mg, 45.60 μmol) and compound 3-16 (10.41 mg, 68.40 μmol) in DMF (5 ml), DIEA (17.68 mg, 136.80 μmol, 23.89 μL) and TBTU (21.96) Mg, 68.40 μmol). The mixture was stirred at 25 ° C for 12 hours, and the crude product was purified by preparative HPLC to afford title compound 3 (15 mg, yield 56.82%). ^{1 H NMR (400MHz, METHANOL-} d 4) δ7.75 (s, 1H), 7.60 (d, J = 8.03Hz, 2H), 7.44-7.50 (m, 2H), 7.43 (s, 1H), 6.14 ( s, 1H), 4.51 (s, 2H), 3.70-3.76 (m, 4H), 3.67 (br.s., 4H), 3.58 (s, 2H), 2.60 (br.s., 2H), 2.50 ( Br.s.,4H),2.41(s,4H), 2.38(s,3H), 2.26(s,3H),1.96-2.10(m,1H),1.69-1.88(m,1H),0.74(t , J = 7.40 Hz, 3H). MS ESI calculated for C ₃₄ H ₄₄ N ₄ O ₄ [M+H] ⁺ 573, found 573.

Test for inhibition activity of compound EZH2 enzyme (based on micro-radioisotope filter binding assay)

material:

EZH2 reaction buffer; 50 mM Tris-HCl (pH 8.0), 0.01% Brij35, 1 mM EDTA, 1 mM DTT, 1 mM PMSF, 1% DMSO

SET8 and PRMT5 (non-complex) buffer; 50 mM Tris-HCl (pH 8.5), 0.01% Brij35, 1 mM DTT, 1% DMSO NSD3 reaction buffer; 50 mM Bicine (pH 8.5), 0.01% Brij35, 1 mM DTT, 1% DMSO

Other HMTs reaction buffer; 50 mM Tris-HCl (pH 8.5), 5 mM MgCl2, 50 mM NaCl, 0.01% Brij35, 1 mM DTT, 1% DMSO

DNMT1 reaction buffer; 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.01% Brij35, 5 mM DTT, 0.1 mM PMSF, 5% glycerol, 1% DMSO

DNMTs Reaction Buffer; 50 mM Tris-HCl (pH 7.5), 50 mM NaCl, 5 mM EDTA, 5 mM DTT, 0.1 mM PMSF, 5% glycerol, 1% DMSO

Methyl donor: S-adenosine-1-[methyl-3H]methionine (SAM)

Standard reaction conditions;

Substrate: 5M histone, or nucleosome of 0.05 mg/mL HeLa cells or core histone from chicken, S-adenosyl-L-[methyl-3H-]methionine: 1 uM

reaction process

1. Freshly prepared reaction buffer solution of the indicated substrate

2. Add the specified MT to the substrate solution and mix gently.

3. Transfer the nanoliter compound DMSO solution to the MT reaction mixture (by using Acoustic Technology (Echo550, LabCyte Inc. Sunnyvale, CA)) and incubate for 10-15 minutes at room temperature.

4. Add 3H-SAM to the reaction mixture to initiate the reaction

5. Incubate for 30 minutes to 1 hour at 30 ° C.

6. Transfer the reaction mixture to filter paper for testing

7. Analyze data and curve fitting using Excel and GraphPad Prism software IC50 to obtain IC50.

The results of the compound inhibitory activity are shown in Table 1.

Table 1 compound inhibition activity results

A<100nM; 100nM≤B<1000nM

Claims (13)

1. a compound of the formula (III), a pharmaceutically acceptable salt thereof and a tautomer thereof,

   

   among them,

   R _{31 is} selected from H, halogen, CN, OH, amino, or selected from, optionally substituted by 1, 2 or 3 R: 5- to 6-membered aryl, 5- to 12-membered heteroaryl, C _1-6 alkane a group, a C _1-6 heteroalkyl group, a C _3-9 cycloalkyl group, and a _3-9 membered heterocycloalkyl group;

   D _{3 is} selected from -[C(R) ₂ ] _1-3 -, -O, -S-, -N(R)-, -C(=O)-, -C(=O)O-, -S (=O)-, -S(=O) ₂ -, -C(=O)N(R)-, -N(R)C(=O)N(R)-; T are independently selected from N Or CH;

   R is selected from H, OH, CN, NH ₂ , COOH, halogen, or selected from, optionally substituted by 1, 2 or 3 R's: 5- to 6-membered aryl, 5- to 12-membered heteroaryl, C _{1 a -6} alkyl group, a C _1-6 heteroalkyl group, a C _3-9 cycloalkyl group, and a _3-9 membered heterocycloalkyl group;

   R' is selected from the group consisting of F, Cl, Br, I, OH, CN, NH ₂ , COOH, Me, Et, CH ₂ F, CHF ₂ , CF ₃ , NH(CH ₃ ), N(CH ₃ ) ₂ ; "" denotes a hetero atom or a hetero atom selected from -C(=O)N(R)-, -N(R)-, -C(=NR)-, -S(=O) ₂ N(R)-, -S(=O)N(R)-, -O-, -S-, =O, =S, -ON=, -C(=O)O-, -C(=O)-, -C( =S)-, -S(=O)-, -S(=O) ₂ - and -N(R)C(=O)N(R)-;

   In either case, the number of heteroatoms or heteroatoms is independently selected from 1, 2 or 3.
2. The compound according to claim 1, a pharmaceutically acceptable salt thereof, and tautomers thereof, wherein R is selected from the group consisting of H, OH, CN, NH ₂ , COOH, halogen, or selected from the group consisting of 1, 2 Or 3 R' substituted: 5- to 6-membered aryl, 5- to 6-membered heteroaryl, C _1-3 alkyl, C _1-3 alkyl-NH-, N, N-di (C _1-3 Alkyl)amino, C _1-3 alkyl-O-, C _1-3 alkyl-S-, C _3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl.
3. The compound according to claim 2, a pharmaceutically acceptable salt thereof, and tautomer thereof, wherein R is selected from the group consisting of H, OH, CN, NH ₂ , COOH, halogen, or selected from the group consisting of 1, 2 Or 3 R' substituted: Me, Et, isopropyl, cyclopropyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

   

   Phenyl, pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolidinyl, piperidine Pyridyl, morpholinyl, piperazinyl, azetidinyl.
4. The compound according to claim 3, a pharmaceutically acceptable salt thereof, and tautomer thereof, wherein R is selected from the group consisting of: H, OH, CN, NH ₂ , COOH, F, Cl, Br, I, Me, Et, CH ₂ F, CHF ₂ , CF ₃ , NH(CH ₃ ), N(CH ₃ ) ₂ ,

   

   

   Cyclopropyl,

   
5. The compound according to any one of claims 1 to 4, a pharmaceutically acceptable salt thereof, and tautomer thereof, wherein R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, Or selected from the group consisting of 1, 2 or 3 R substituted: 5- to 6-membered aryl, 5- to 6-membered heteroaryl, C _1-3 alkyl, C _1-3 alkyl-NH-, N, N-di(C _1-3 alkyl)amino, C _1-3 alkyl-O-, C _1-3 alkyl-S-, C _3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl.
6. The compound according to claim 5, a pharmaceutically acceptable salt thereof, and tautomer thereof, wherein R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of Substituted by 1, 2 or 3 R: Me, Et, isopropyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

   

   

   Phenyl, pyridyl, pyrimidinyl, pyrazinyl, imidazolyl, pyrazolyl, furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolidinyl, piperidine Pyridyl, morpholinyl, piperazinyl, azetidinyl, cyclopropyl.
7. The compound according to claim 6, a pharmaceutically acceptable salt thereof, and tautomers thereof, wherein R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, or selected from the group consisting of Substituted by 1, 2 or 3 R: Me, Et, isopropyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

   

   
8. The compound according to claim 7, a pharmaceutically acceptable salt thereof, and tautomer thereof, wherein R _{31 is} selected from the group consisting of H, F, Cl, Br, I, CN, OH, amino, Me, Et, Isopropyl, NH(CH ₃ ), N(CH ₃ ) ₂ ,

   

   
9. The compound according to any one of claims 1 to 4, a pharmaceutically acceptable salt thereof, and tautomer thereof, wherein D _{3 is} selected from the group consisting of: -CH ₂ -, -CH(OH)-, -O, -S-, -NH-, -C(=O)-, -C(=O)O-, -S(=O)-, -S(=O) ₂ -, -C(=O)NH- , -NHC(=O)NH-.
10. A compound according to any one of claims 1 to 9, a pharmaceutically acceptable salt thereof, and a tautomer thereof, which are selected from the group consisting of:

    

    among them,

    R ₃₁ and D _{3 are} as defined in claims 1-9.
11. A compound according to any one of claims 1 to 4 which is selected from the group consisting of:

    
12. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
13. Use of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 12, for the manufacture of a medicament for the treatment of various conditions associated with EZH2 receptors.