CN112778275B

CN112778275B - Adamantyl PRMT5 inhibitor and application thereof

Info

Publication number: CN112778275B
Application number: CN202011209671.0A
Authority: CN
Inventors: 赵传武; 齐非; 雷凯; 张莉; 韩强; 张瑞兴; 张阳; 李冰
Original assignee: CSPC Zhongqi Pharmaceutical Technology Shijiazhuang Co Ltd
Current assignee: CSPC Zhongqi Pharmaceutical Technology Shijiazhuang Co Ltd
Priority date: 2019-11-11
Filing date: 2020-11-03
Publication date: 2023-05-12
Anticipated expiration: 2040-11-03
Also published as: CN112778275A

Abstract

The invention provides a compound with a novel structure and PRMT5 inhibitor activity, and tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein the compound has the structure shown in a formula (I). The compound has medical application for treating cancers, can obviously inhibit PRMT5 activity and has stronger proliferation inhibition effect on HGC-27.

Description

Adamantyl PRMT5 inhibitor and application thereof

Technical Field

The invention relates to the technical field of medicines, in particular to a compound serving as a PRMT5 inhibitor and application of the compound or a pharmaceutical composition in preparation of medicines.

Background

PRMT5 (alias: hs17, jbp, skb1, capsule or Dart 5) was first discovered as a homolog SKBlHs of the merozoite Shk1 Kinase binding protein 1 (Shk 1 Kinase-binding protein, SKB 1) (Journal of Biological Chemistry,2001.276 (14): 11393-401), the earliest identified class II member. The manner in which PRMT5 acts biologically is generally divided into two layers, one of which is epigenetic regulation of expression of a target gene; and secondly, the PRMT5 can methylate different proteins to regulate different physiological processes, such as assembly of a Golgi apparatus, ribosome synthesis, cell differentiation, germ cell specialization, cell proliferation apoptosis and the like.

PRMT5 is both an important epigenetic enzyme and an important oncogene mediating the development of solid tumors. In mammalian cells, it is localized mainly to the cytoplasm, partly to the nucleus, where it is widely expressed in tissues. PRMT5 has been found in many studies to play an important role in the development of various hematological diseases and solid malignant tumors (gastric cancer, breast cancer, renal cancer, colon cancer, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, bladder cancer, skin cancer, cervical cancer, prostate cancer, etc.), and its expression level is closely related to the occurrence, development and prognosis of tumors. For example, in breast cancer, the tumor-inhibiting effect of programmed death factor 4 (programmed cell death protein, PDCD 4) is reduced by PRMT5, and in an in vitro model of orthotopic engraftment tumor, the interaction of PDCD4 and PRMT5 accelerates the proliferation of tumor cells, and this growth phenotype is dependent on the catalytic activity of PRMT5 and the methylation level of PDCD4 therein, while PRMT5 is also closely correlated with prognosis of breast cancer patients. PRMT5 is highly expressed in prostate cancer cells, promoting proliferation of prostate cancer cells by epigenetic activation of transcribed Androgen Receptor (AR). PRMT5 expression is significantly higher in prostate cancer tissue than in benign prostatic hyperplasia tissue, and PRMT5 expression is positively correlated with AR expression at both protein and mRNA levels. PRMT5 epigenetic activation of AR transcription is through symmetrical dimethyl H4R3. Whereas Sp1, PRMT5 and Brg1 also activate AR transcription by forming a complex at the AR proximal promoter region. Upon knocking down PRMT5, AR expression was significantly inhibited and growth of xenograft tumors in mice was inhibited. In addition, PRMT5 has been demonstrated as a tumor therapeutic target in glioma and mantle cell lymphoma diseases (Nature Chemical Biology,2015, (11): 432-437). Numerous data results confirm that targeted PRMT5 inhibitors have become one of the hot spots in current anti-tumor drug research.

To date, PRMT5 inhibitors have been developed early, with the fastest progression being that GSK3326595, proposed by GSK corporation, is in phase II clinic, with indications of myelodysplastic syndrome, acute myelogenous leukemia. JNJ-64619178 at stage I was first prescribed in 2018, month 6, 29, and was mainly relapsed/refractory B-cell non-hodgkin lymphoma or advanced solid tumors. The structural formulas of GSK3326595 and JNJ-64619178 are as follows:

in addition, published PRMT5 selective inhibitor patent applications include WO2014100730, WO2016034671, WO2016034675, WO2016034673, WO2018167276, US2018298010, WO2018161922, WO2019002074, etc., but no good PRMT5 inhibitors are found to be commercially available. In order to achieve the aim of better tumor treatment effect and better meet the market demand, we design and synthesize a series of selective PRMT5 inhibitors of adamantyl derivatives.

Disclosure of Invention

The invention provides a PRMT5 inhibitor with a novel structure.

The invention provides a compound shown as a formula (I), and tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, which have the following structures:

wherein Y is:

l is selected from the following linker fragments:

wherein, the right end of L and R ₁₇ The N atom connected with the pyrimidine ring;

the A ring is selected from substituted or unsubstituted saturated C _3-10 Cycloalkyl, substituted or unsubstituted saturated 5-10 membered heterocyclyl, substituted or unsubstituted saturated 4-12 membered bridged ring, substituted or unsubstituted saturated 4-12 membered heterobridged ring, substituted or unsubstituted saturated Shan Luohuan, substituted or unsubstituted saturated hetero Shan Luohuan, substituted or unsubstituted saturated fused ring;

the number of ring atoms of the saturated Shan Luohuan groups and saturated hetero Shan Luohuan groups is selected from 3-membered/5-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered, and 5-membered/6-membered rings, wherein the count of each ring includes a spiro atom;

the saturated fused ring groups and saturated heterofused ring groups are selected from 5-membered/5-membered, 5-membered/6-membered or 6-membered/6-membered rings, wherein the count of each ring includes a common atom;

the term "substituted" means that the substituents are each independently selected from halogen, hydroxy, cyano, nitro, amino, azido, carbonyl, carboxyl, ethynyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Haloalkyl, C _3-10 Cycloalkyl, 5-10 membered heterocycloalkyl, C _6-14 Aryl or C _5-10 One or more of the heteroaryl ring radicals;

the R is ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ 、R ₉ 、R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each occurrence is independently selected from hydrogen, halogen, hydroxy, amino, carboxy, nitro, cyano, carbonyl, azido, oxo, ethynyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Haloalkyl, C _1-6 Alkoxy groupAmino, C _1-6 Alkyl ester group, C _1-6 Alkylamino, C _1-6 Amide group, C _3-10 Cycloalkyl, C _3-10 Cycloalkylamino, C _3-10 Cycloalkyl amide group, C _3-10 Epoxy alkylamide group, 5-10 membered heterocycloalkyl group, C _6-14 Aryl or C _5-10 Heteroaryl ring radical, wherein, the C _6-14 Aryl or C _5-10 The heteroaryl groups being independently substituted by one or more groups selected from halogen, hydroxy, amino, nitro, C _1-6 Alkoxy or C _1-6 Substituted by alkyl; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond;

when R is ₁₃ 、R ₁₄ Wherein when one of them is oxo, the other is absent;

when R is ₁₅ 、R ₁₆ Wherein when one of them is oxo, the other is absent;

R ₁₂ at any substitutable position of the tetrahydroisoquinoline ring, n is 0,1, 2, 3,4, 5 or 6;

wherein the hetero atoms in the heterocyclic group, the heteroaromatic ring group, the hetero Shan Luohuan group, the hetero bridge ring group and the hetero condensed ring group are independently selected from O, N or S, and the number of the hetero atoms is 1,2 or 3.

Preferably, the compound of the present invention or a tautomer, an optical isomer, a solvate, an isotopic derivative or a pharmaceutically acceptable salt thereof, is characterized by having a structure as shown in formula (II):

therein, Y, R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ And R is ₁₆ Is defined as compound (I).

Further preferably, the compound of the present invention or a tautomer, an optical isomer, a solvate, an isotopic derivative or a pharmaceutically acceptable salt thereof, is characterized by having a structure as shown in formula (III):

Further preferably, the compound of the present invention or a tautomer, an optical isomer, a solvate, an isotopic derivative or a pharmaceutically acceptable salt thereof, is characterized by having a structure represented by formula (IV):

Further, in certain embodiments, the compounds of the present invention or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein ring A is selected from substituted or unsubstituted saturated C _4-7 Cycloalkyl, substituted or unsubstituted saturated 5-7 membered heterocyclyl, substituted or unsubstituted saturated 4-12 membered bridged ring, substituted or unsubstituted saturated 4-12 membered heterobridged ring, substituted or unsubstituted saturated Shan Luohuan, substituted or unsubstituted heteromonospirocyclic.

Further, in certain embodiments, the compounds of the present invention or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein the heteroatom of the saturated 5-10 membered heterocyclyl is an N atom.

Further, in certain embodiments, the compounds of the present invention or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein the heteroatom of saturated hetero Shan Luohuan group is selected from N or O atoms, and the number of heteroatoms is 1.

Further, in certain embodiments, the compounds of the invention or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein the heteroatom of the saturated heterobridge ring group is 1N atom.

Further, in certain embodiments, the compounds of the present invention or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein, the saturated C _3-10 Cycloalkyl, saturated 5-10 membered heterocyclyl, saturated Shan Luohuan, saturated hetero Shan Luohuan, saturated 4-12 membered bridged ring, saturated 4-12 membered heterobridged ring, saturated fused ring and saturated heterofused ring groups, the substituents being halogen, hydroxy or C _1-6 An alkyl group.

Further, in certain embodiments, a compound of the invention or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein R ₁ 、R ₃ 、R ₅ 、R ₇ 、R ₈ And R is ₉ Each independently selected from hydrogen, halogen, hydroxy, amino, and oxo; r is R ₂ 、R ₄ And R is ₆ Each independently selected from hydrogen, halogen, hydroxy and C _1-6 An alkyl group.

Further, the compound of the present invention or a tautomer, an optical isomer, a solvate, an isotopic derivative or a pharmaceutically acceptable salt thereof, wherein R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

The present invention provides the following compounds, or tautomers, optical isomers, solvates, isotopic derivatives, or pharmaceutically acceptable salts thereof, having the following structure:

the invention also provides a pharmaceutical composition comprising the compound or a tautomer, an optical isomer, a solvate, an isotope derivative or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials.

The invention also provides application of the compound.

The compound or tautomer, optical isomer, solvate, isotope derivative or pharmaceutically acceptable salt thereof or the pharmaceutical composition is used for preparing PRMT5 inhibitor drugs.

The compound or tautomer, optical isomer, solvate, isotope derivative or pharmaceutically acceptable salt thereof or the pharmaceutical composition is used for preparing medicines for preventing and/or treating cancers.

Further, in the above uses, the cancer is selected from lung cancer, bone cancer, stomach cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, pancreatic cancer, brain cancer, pituitary adenoma, melanoma, epidermoid cancer, T-cell lymphoma, chronic and acute leukemia.

In another aspect, the invention also provides a method for preparing the compound.

The preparation method of the compound comprises the steps of reacting a parent nucleus compound shown in a formula (I ') with a compound shown in a formula (Y') to obtain the compound shown in the formula (I), wherein the reaction formula is as follows:

wherein the compound of formula (Y') has a structure selected from:

preferably, the preparation method of the present invention, wherein the parent nucleus compound represented by the formula (I') is prepared by the following method: in an organic solvent, carrying out condensation reaction on a compound shown in a formula (M) and a compound shown in a formula (N) to obtain a parent nucleus compound shown in a formula (I'), wherein the reaction formula is as follows:

as a preferable embodiment of the present invention, the production method of the present invention is characterized in that the organic solvent is selected from tetrahydrofuran, acetonitrile, N-dimethylformamide, C _1-4 Halogenated hydrocarbons, ethylene glycol dimethyl ether, dichloroethane, methanol, ethanol, petroleum ether, n-hexane, diethyl ether and ethyl acetate.

Preferably, in the above scheme, the halogenated hydrocarbon is dichloromethane, dichloroethane, chloroform or tetrachloromethane.

As one embodiment of the present invention, the process for producing a compound of the present invention, wherein,

when the structure of the compound of formula (Y') is selected from the following structures:

the compound of formula (Y') is prepared by the following method:

the method comprises the following specific steps:

(1) The compound of the formula (II) and the two forms of the compound of the formula (III) undergo condensation reaction under alkaline conditions to obtain two forms of the compound of the formula (IV);

(2) Acidic hydrolysis of the compound of formula (IV) to give the compound of formula (Y');

wherein the two forms of the compound of formula (III) are:

R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ 、R ₉ 、R ₁₇ and ring A is as defined in formula (I) of the present invention;

and T is an amino protecting group selected from N-benzyloxycarbonyl (Cbz), T-butyloxycarbonyl (Boc), methoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), trimethylsilylethoxycarbonyl (Teoc), phthaloyl (Pht), trifluoroacetyl (Tfa), p-toluenesulfonyl (Tos), trityl (Trt), 2, 4-dimethoxybenzyl (DMb) and p-methoxybenzyl (PMB).

Preferably, the method for preparing the compound of the present invention, wherein T is T-butoxycarbonyl (Boc).

As a preferred embodiment of the present invention, the production method of the present invention, wherein the acid in the step (2) is selected from trifluoroacetic acid, formic acid, methanesulfonic acid, acetic acid, sulfuric acid or hydrochloric acid.

Preferably, the acid is trifluoroacetic acid.

As a preferred embodiment of the present invention, the preparation method of the present invention is characterized in that the base in the step (1) is one or more selected from the group consisting of triethylamine, diisopropylethylamine, piperidine, pyrrole, pyridine, lutidine and dimethylaminopyridine.

Preferably, the base is triethylamine.

As another embodiment of the present invention, the production method of the present invention, wherein,

the compound of formula (Y') is prepared by the following method:

the method comprises the following specific steps:

(1) The compound of formula (II ') and the compound of formula (III ') undergo condensation reaction under alkaline conditions to obtain the compound of formula (IV ');

(2) Acidic hydrolysis of the compound of formula (IV ') to give a compound of formula Y';

wherein R is ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ 、R ₉ 、R ₁₇ And ring A is as defined in formula (I) of the present invention;

t is an amino protecting group selected from N-carbobenzoxy (Cbz), T-butoxycarbonyl (Boc), fluorenylmethoxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), trimethylsilylethoxycarbonyl (Teoc), phthaloyl (Pht), trifluoroacetyl (Tfa), p-toluenesulfonyl (Tos), trityl (Trt), 2, 4-dimethoxybenzyl (DMb) and p-methoxybenzyl (PMB).

Preferably, the acid is trifluoroacetic acid.

Preferably, the base is triethylamine.

Definition of the definition

The term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, straight or branched chain group containing 1 to 20 carbon atoms, e.g. "C _1～6 Alkyl "means that the group is alkyl and the number of carbon atoms in the carbon chain is between 1 and 6. Including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl, n-pentyl, neopentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, and the like. In the present invention, the term includes substituted or unsubstituted alkyl groups.

The term "cycloalkyl" refers to a monocyclic saturated aliphatic radical having a specified number of carbon atoms, preferably to a substituted or unsubstituted C _3～12 Cycloalkyl, more preferably, substituted or unsubstituted C _3-10 Cycloalkyl groups. Such as cyclopropyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, and the like.

The term "alkoxy" refers to an-O-alkyl group, which may be saturated or unsaturated, branched, straight-chain or cyclic, including substituted or unsubstituted. Preferably having 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms. Representative examples include methoxy, ethoxy, propoxy, t-butoxy and the like.

The term "halogen" or "halo" refers to F, cl, br, I. The term "haloalkyl" means that one, two or more hydrogen atoms in an alkyl group as defined herein are replaced by halogen. Representative examples of haloalkyl groups include CCl ₃ 、CF ₃ 、CHCl ₂ 、CH ₂ CF ₃ 、CF ₂ CF ₃ Etc.

The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, wherein one or more ring atoms are selected from N, O or S and the remaining ring atoms are C. Preferably containing 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably from 3 to 8 ring atoms, of which 1 to 3 are heteroatoms. More preferably from 5 to 10 ring atoms. Examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyranyl and the like. Polycyclic heterocyclyl groups include spiro, fused and bridged heterocyclic groups.

The term "spiroheterocyclyl" or "heterospirocyclyl" refers to a polycyclic heterocyclic group having one atom (referred to as the spiro atom) in common between monocyclic rings, wherein one or more of the ring atoms is selected from nitrogen, oxygen, or S (O) r (where r is an integer of 0,1, 2) heteroatoms, and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. The spiroheterocyclyl groups are classified into a single spiroheterocyclyl group (or hetero-single spirocyclyl group), a double spiroheterocyclyl group (or hetero-double spirocyclyl group) or a multiple spiroheterocyclyl group (or hetero-multiple spirocyclyl group) according to the number of common spiro atoms between rings.

The term "fused heterocyclic group" or "heterofused ring group" refers to a polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system, one or more of which may contain one or more double bonds, but none of which has a fully conjugated pi electron system, wherein one or more ring atoms are selected from nitrogen, oxygen, or a heteroatom of S (O) r (where r is an integer of 0,1, 2) and the remaining ring atoms are carbon. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl group.

The term "bridged heterocyclyl" or "heterobridged heterocyclyl" refers to a polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a fully conjugated pi electron system, wherein one or more of the ring atoms is selected from nitrogen, oxygen, or a heteroatom of S (O) r (where r is an integer of 0,1, 2), and the remaining ring atoms are carbon. The number of constituent rings may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups.

The term "aryl" means a monocyclic, bicyclic and tricyclic aromatic carbocyclic ring system containing 6 to 16 ring atoms, or 6 to 14 ring atoms, or 6 to 12 ring atoms, or 6 to 10 ring atoms, and the term "aryl" may be used interchangeably with the term "aromatic ring". Examples of aryl groups may include phenyl, naphthyl, anthracyl, phenanthryl, pyrenyl, and the like.

The term "heteroaryl" means an aromatic monocyclic or polycyclic ring system containing a 5-12 membered structure, or a 5-10 membered structure, preferably a 5-8 membered structure, more preferably a 5-6 membered structure, wherein at least one ring atom is a heteroatom and the remaining atoms are carbon, the heteroatom being independently selected from O, N or S, the number of heteroatoms preferably being 1,2 or 3. Examples of heteroaryl groups include furan, thiophene, oxazole, thiazole, isoxazole, oxadiazole, thiadiazole, pyrrole, pyrazole, imidazole, triazole, tetrazole, pyridine, pyrimidine, pyrazine, pyridazine, thiodiazole, triazine, phthalazine, quinoline, isoquinoline, pteridine, purine, indole, isoindole, benzofuranyl, benzothienyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, benzimidazolyl, benzophthalazinyl, imidazo [1,2-a ] pyridinyl, pyrazolo [1,5-a ] pyrimidinyl, imidazo [1,2-b ] pyridazinyl, [1,2,4] triazolo [4,3-b ] pyridazinyl, [1,2,4] triazolo [1,5-a ] pyrimidinyl, [1,2,4] triazolo [1,5-a ] pyridinyl, and the like.

The term "spirocyclic group" refers to all-carbon polycyclic groups having a single carbon atom (referred to as a spiro atom) shared between the monocyclic rings, which may contain one or more double bonds, but no ring has a fully conjugated pi-electron system. Spirocycloalkyl groups are classified as mono-, di-or multicycloalkyl groups according to the number of common spiro atoms between rings.

The term "fused ring group" refers to an all-carbon polycyclic group in which each ring in the system shares an adjacent pair of carbon atoms with the other rings in the system, wherein one or more of the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. The number of constituent rings may be classified as a bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyl group.

The term "bridged ring radical" refers to all-carbon polycyclic groups wherein any two rings share two carbon atoms that are not directly attached, which may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. Cycloalkyl groups can be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged, depending on the number of constituent rings.

The term "pharmaceutically acceptable salts" refers to salts suitable for use in contact with mammalian, especially human, tissues without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and are well known in the art.

The term "salt" encompasses salts derived from inorganic acids such as hydrochloric acid, sulfuric acid, sulfurous acid, nitric acid, phosphoric acid, hydrobromic acid, and the like, as well as salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, maleic acid, fumaric acid, salicylic acid, and the like. If the compounds of the present invention are acidic, pharmaceutically acceptable non-toxic bases include salts prepared with inorganic and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, ferric salts, ferrous salts, lithium salts, magnesium salts, zinc salts, and the like, and salts derived from organic non-toxic bases include primary amine salts, secondary amine salts, tertiary amine salts, and the like.

Abbreviations used elsewhere herein for the preparation examples, and are:

DCM dichloromethane

TEA triethylamine

DIPEA N, N-diisopropylethylamine

DMF N, N-dimethylformamide

EtOAc ethyl acetate

h hours

ml milliliter

HATU 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate

MeOH methanol

TFA trifluoroacetic acid

The beneficial effects of the invention are as follows:

the invention provides a PRMT5 inhibitor with a novel structure, and discloses a preparation method and medical application thereof. In particular, the compounds of the present invention relate to use in medicaments for the treatment of cancer. Enzymatic and cell screening results showed that: compared with a positive control medicine GSK-3326595, the compound provided by the invention has the advantages that PRMT5 activity is obviously inhibited, and the proliferation inhibition effect on HGC-27 is stronger.

Detailed Description

The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred methods and materials are presented herein for illustrative purposes only.

Preparation example

The following preparation examples describe the preparation methods of the parent compound and intermediate of the specific compound of the present invention.

Preparation example 1: preparation of (S) -6-chloro-N- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide

In a 100ml reaction flask, 6-chloro-pyrimidine 4-carbonyl chloride (630 mg,3.56 mmol) was dissolved in DCM (10 ml) and TEA (720.38 mg,7.12 mmol) was added at 0deg.C. (S) -1-amino-3- (3, 4-dihydroisoquinolin-2 (1H) -yl) propan-2-ol (660.85 mg,3.20 mmol) was then added. The reaction solution was stirred at 25℃for 2h. TLC monitoring of the reaction was completed, the reaction mixture was purified by H ₂ O (5 mL) was diluted and extracted with DCM (15 mL. Times.2). The combined organic layers were washed with saturated brine (10 ml) and dried over Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain residue. The residue was purified by column chromatography (DCM: meoh=10:1). Obtaining (S) -6-chloro-N- (3, 4-dihydro-isoquinolin-2 (1H) -yl) as a yellow oily compound) -2-hydroxypropyl) pyrimidine-4-carboxamide (680 mg, yield 55.08%).

Preparation example 2: (4-Aminopiperidin-1-yl) (3-hydroxyadamantan-1-yl) methanone (intermediate C1)

Step 1: preparation of tert-butyl (1- (3-hydroxyadamantane-1-carbonyl) piperidin-4-yl) carbamate

In a 100ml reaction flask, 3-hydroxyadamantane-1-carboxylic acid (690 mg,3.52 mmol) was dissolved in DMF (5 ml), followed by DIPEA (1.82 g,14.06 mmol), HATU (1.47 g,3.87 mmol) and 4-t-butoxycarbonylaminopiperidine (1.03 g,3.28 mmol) were added and reacted at 25℃for 10 hours, TLC was monitored for progress of the reaction, and after completion of the reaction, the reaction mixture was reacted with H ₂ O (5 mL) was diluted and extracted with DCM (15 mL. Times.2). The combined organic layers were washed with saturated brine (5 mL) and with Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain residue. The residue was purified by column chromatography (DCM: meoh=30:1). The compound tert-butyl (1- (3-hydroxyadamantane-1-carbonyl) piperidin-4-yl) carbamate (1.00 g,2.64mmol, 75.14% yield) was obtained as a yellow oil. MS (m/z): 379.4[ M+H ]] ⁺ 。

Step 2: preparation of (4-aminopiperidin-1-yl) (3-hydroxyadamantan-1-yl) methanone (intermediate C1)

In a 100ml reaction flask, tert-butyl (1- (3-hydroxyadamantane-1-carbonyl) piperidin-4-yl) carbamate (1 g,2.64 mmol) and TFA (770.00 mg,6.75 mmol) were dissolved in DCM (20 ml) and stirred at 25℃for 2h. TLC was used to monitor the progress of the reaction, after the completion of the reaction, the solvent was removed by rotary evaporation to give crude (4-aminopiperidin-1-yl) (3-hydroxyadamantan-1-yl) methanone (intermediate C1) (1.00 g), which was directly subjected to the next reaction without purification. MS (m/z): 279.3[ M+H] ⁺ 。

Preparation examples 3 to 22: the preparation methods of the compounds of preparation examples 3 to 22 are shown with reference to preparation example 2, and are specifically as follows:

TABLE 1 preparation of intermediates C2 to C21

Example 1: preparation of N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (3-hydroxyadamantane-1-carbonyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (ZY-1)

In a 100ml reaction flask, (S) -6-chloro-N- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (300 mg,865.03 umol), (4-aminopiperidin-1-yl) (3-hydroxyadamantan-1-yl) methanone (intermediate C1) (678.89 mg,1.73 mmol), TEA (350 mg,3.46 mmol) were dissolved in isopropanol (15.0 ml) and nitrogen protected, the reaction solution was stirred at 80℃for 16H, TLC monitored the progress of the reaction, after completion of the reaction the solvent was distilled off under reduced pressure, the crude product was purified by preparative HPLC to give N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (3-hydroxyadamantan-1-carbonyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (ZY-1) (52.11 mg, yield 10.16%).

Examples 2 to 21: preparation of Compound ZY-2-Compound ZY-21

Referring to the preparation method of example 1, only intermediate C1 was replaced with intermediate C2-intermediate C21, respectively, as follows:

TABLE 2 preparation of Compounds ZY-2 to ZY-21

The compounds prepared in examples 1 to 21 were subjected to structural confirmation, and the physical characterization results are shown in the following table:

TABLE 3 Structure confirmation data for compounds ZY-1 to ZY-21

Effect test example 1: cell assay

The complete culture medium for culturing the gastric cancer cells HGC-27 (China center for type culture collection, CCTCC) for experiments is 1640 (Cat NO.SH30809.01, HYCLONE) supplemented with 10% serum FBS (Cat NO. SA311.02, lanzhou BAILING). Cells at 37℃with 5% CO ₂ Culturing in an incubator. The experimental reagents included dimethyl sulfoxide (purchased from the company Miou chemical Co., tianjin). MTT, (THIAZOLYL BLUE TETRAZOLIUM BROMIDE, CAS.NO.298-93-1, amerco). The test control GSK-3326595 was obtained by homemade or commercial purchase. The test object is stored at 4 ℃ in a sealed manner.

Dissolving the test substance and positive control substance with dimethyl sulfoxide as solvent to give a concentration of 5×10 ^-2 mol/L stock solution. The test stock solutions were stored at-20 ℃. Taking complete culture medium as diluent, and gradient diluting test objects to differentThe concentration is kept for later use. In a 96-well culture plate, 10. Mu.L/well of the corresponding test substances with different concentrations are added, 8 concentrations are set for each test substance, and 3 compound wells are set for each concentration. Wells with only cell suspension without test substance and vehicle were used as control wells. The wells with complete medium alone and without cell suspension were used as blank wells. Add 90. Mu.L/well (5.5X103 cells/well) of complete media suspension of gastric cancer cells HGC-27 at 37℃with 5% CO ₂ Culturing in an incubator. On day ten, MTT 20. Mu.L/well was added at 37℃with 5% CO ₂ Culturing in an incubator for 4 hours, removing supernatant, adding 150 mu L/hole of dimethyl sulfoxide, shaking and mixing uniformly, and detecting an OD value at 550nm by an enzyme-labeled instrument. The inhibition ratio was calculated according to the formula = (subject well OD-blank well OD)/(control well OD-blank well OD) ×100% and the results are shown in table 4.

Effect test example 2: enzymatic experimental method

IC for detecting a test substance on histone methyltransferase PRMT5 using radioisotope Flashplate technology ₅₀ 。

After the test compounds were dissolved in dimethyl sulfoxide, respectively, they were added to an Echo 384-well plate and diluted to a desired concentration, and the test compounds were transferred from the diluted Echo 384-well plate to a 384-well reaction plate using an Echo550 instrument, and dimethyl sulfoxide was transferred from both the negative control and the positive control. PRMT5 was added to 1-fold reaction buffer (1-fold reaction buffer included 10mM Tris-HCl, pH 8.0,0.01% Tween-20,1mM DTT) to form a 1.67-fold enzyme solution (enzyme concentration 5 nM). Polypeptide substrate and [3H]SAM was added to the 1-fold reaction buffer to form a 2.5-fold substrate solution (substrate final concentrations 100nM and 250nM, respectively). To 384-well wells, 15 μl of 1.67-fold enzyme solution was added. For the no enzyme activity control wells, 15. Mu.l of 1-fold reaction buffer was used instead of enzyme solution. Centrifuge at 1000rpm for 1min and incubate at room temperature for 15 min. To 384 well reaction plates 10 μl of 2.5 fold substrate solution was added per well. Centrifuge at 1000rpm for 1min. The reaction was carried out at 25℃for 60 minutes. To each well of 384-well reaction plate, 5. Mu.l of a reaction termination solution (125. Mu.M cold SAM solution) was added to terminate the reaction. 25uL per well from the test plate was transferred to a Flashplate and left at room temperature for 1h. The flashplate was then washed 3 times with 0.1% Tween-20 solution. Read with MicroBeta 2. Converting data into suppressionAnd (5) data of the production rate. Where max refers to the conversion of the DMSO control and min refers to the conversion of the no enzyme activity control. Percent inhibition = (max-version)/(max-min) ×100. Fitting IC with XLFIT exceladd-in version 5.4.0.8 ₅₀ Values. Fitting formula y=bottom+ (Top-Bottom)/(1+ (IC) ₅₀ X)/(HillSlope), the results are shown in Table 4.

TABLE 4 cytological and enzymatic Effect test data for the Compounds of the invention and the Positive control

Compounds of formula (I)	Enzymology (IC) ₅₀ nM)	HGC-27MTT(IC ₅₀ nM)
			GSK3326595	C	B
ZY-1	B	B
			ZY-2	B	B
ZY-3	B	B
			ZY-4	A	A
ZY-5	B	A
			ZY-6	A	A
ZY-7	A	A
			ZY-8	A	A
ZY-9	A	A
			ZY-10	A	A
ZY-11	A	A
			ZY-12	A	A
ZY-13	B	A
			ZY-14	A	A
ZY-15	B	A
			ZY-16	B	B
ZY-17	B	A
			ZY-18	B	A
ZY-19	B	A
			ZY-20	A	A
ZY-21	B	A

Note that: IC (integrated circuit) ₅₀ More than or equal to 50nM is C,10nM is less than or equal to IC ₅₀ < 50nM B, IC ₅₀ < 10nM is A.

The results of enzymology and cell screening show that compared with a positive control medicine GSK-3326595 (208 th compound of patent application WO 2014100719), the compound provided by the invention can remarkably inhibit PRMT5 activity and has a stronger proliferation inhibition effect on HGC-27.

Claims

1. A compound of formula (I), and tautomers, optical isomers or pharmaceutically acceptable salts thereof, characterized by the following structure:

wherein Y is:

l is selected from the following linker fragments:

the term "substituted" means that the substituents are each independently selected from halogen, hydroxy, cyano, nitro, amino, azido, carbonyl, carboxyl, ethynyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Haloalkyl, C _3-10 Cycloalkyl, 5-10 membered heterocycloalkyl, and,C _6-14 Aryl or C _5-10 One or more of the heteroaryl ring radicals;

the R is ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ 、R ₉ 、R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each occurrence is independently selected from hydrogen, halogen, hydroxy, amino, carboxy, nitro, cyano, carbonyl, azido, oxo, ethynyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Haloalkyl, C _1-6 Alkoxyamino, C _1-6 Alkyl ester group, C _1-6 Alkylamino, C _1-6 Amide group, C _3-10 Cycloalkyl, C _3-10 Cycloalkylamino, C _3-10 Cycloalkyl amide group, C _3-10 Epoxy alkylamide group, 5-10 membered heterocycloalkyl group, C _6-14 Aryl or C _5-10 Heteroaryl ring radical, wherein, the C _6-14 Aryl or C _5-10 The heteroaryl groups being independently substituted by one or more groups selected from halogen, hydroxy, amino, nitro, C _1-6 Alkoxy or C _1-6 Substituted by alkyl; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond;

when R is ₁₃ 、R ₁₄ When one of them is oxo, the other is absent;

when R is ₁₅ 、R ₁₆ When one of them is oxo, the other is absent;

2. The compound of claim 1, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, having a structure according to formula (II):

3. the compound of claim 1, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, having a structure according to formula (III):

4. a compound according to claim 3, or a tautomer, optical isomer or pharmaceutically acceptable salt thereof, having the structure shown in formula (IV):

5. the compound of any one of claims 1-4, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: the heteroatom of the saturated 5-10 membered heterocyclic group is an N atom.

6. The compound of any one of claims 1-4, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: the heteroatom of the saturated hetero Shan Luohuan group is selected from N or O atoms, and the number of the heteroatom is 1.

7. The compound of any one of claims 1-4, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: the heteroatom of the saturated heterobridge ring group is 1N atom.

8. The compound of any one of claims 1-4, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: the saturated C _3-10 Cycloalkyl, saturated 5-10 membered heterocyclyl, saturated Shan Luohuan, saturated hetero Shan Luohuan, saturated 4-12 membered bridged ring, saturated 4-12 membered heterobridged ring, saturated fused ring and saturated heterofused ring groups, the substituents being halogen, hydroxy or C _1-6 An alkyl group.

9. The compound of any one of claims 1-4, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁ 、R ₃ 、R ₅ 、R ₇ 、R ₈ And R is ₉ Each independently selected from hydrogen, halogen, hydroxy, amino, and oxo; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond; r is R ₂ 、R ₄ And R is ₆ Each independently selected from hydrogen, halogen, hydroxy and C _1-6 An alkyl group.

10. The compound of claim 5, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁ 、R ₃ 、R ₅ 、R ₇ 、R ₈ And R is ₉ Each independently selected from hydrogen, halogen, hydroxy, amino, and oxo; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond; r is R ₂ 、R ₄ And R is ₆ Each independently selected from hydrogen, halogen, hydroxy and C _1-6 An alkyl group.

11. The compound of claim 6, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁ 、R ₃ 、R ₅ 、R ₇ 、R ₈ And R is ₉ Each independently selected from hydrogen, halogen, hydroxy, amino, and oxo; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond; r is R ₂ 、R ₄ And R is ₆ Each independently selected from hydrogen, halogen, hydroxy and C _1-6 An alkyl group.

12. The compound of claim 7, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁ 、R ₃ 、R ₅ 、R ₇ 、R ₈ And R is ₉ Each independently selected from hydrogen, halogen, hydroxy, amino, and oxo; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond; r is R ₂ 、R ₄ And R is ₆ Each independently selected from hydrogen, halogen, hydroxy and C _1-6 An alkyl group.

13. The compound of claim 8, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁ 、R ₃ 、R ₅ 、R ₇ 、R ₈ And R is ₉ Each independently selected from hydrogen, halogen, hydroxy, amino, and oxo; the oxo group refers to that two H at the same substitution position are replaced by the same O to form a double bond; r is R ₂ 、R ₄ And R is ₆ Each independently selected from hydrogen, halogen, hydroxy and C _1-6 An alkyl group.

14. The compound of any one of claims 1-4, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

15. The compound of claim 5, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

16. The compound of claim 6, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

17. The compound of claim 7, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

18. The compound of claim 8, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

19. The compound of claim 9, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogenPlain, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

20. The compound of any one of claims 10-13, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, wherein: r is R ₁₀ 、R ₁₁ 、R ₁₂ 、R ₁₃ 、R ₁₄ 、R ₁₅ 、R ₁₆ And R is ₁₇ Each independently selected from hydrogen, halogen, hydroxy, amino, nitro, carbonyl, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 A haloalkyl group.

21. A compound according to claim 1, or a tautomer, optical isomer or pharmaceutically acceptable salt thereof, having the structure:

22. a pharmaceutical composition characterized by: a compound according to any one of claims 1 to 21, or a tautomer, optical isomer or pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.

23. Use of a compound according to any one of claims 1 to 21, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 22, in the manufacture of a PRMT5 inhibitor medicament.

24. Use of a compound according to any one of claims 1 to 21, or a tautomer, optical isomer, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 22, in the manufacture of a medicament for the prevention and/or treatment of cancer.

25. The use according to claim 24, wherein: the cancer is selected from lung cancer, bone cancer, stomach cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, brain cancer, pituitary adenoma, melanoma, epidermoid carcinoma, T-cell lymphoma, chronic and acute leukemia.