CN113045543A

CN113045543A - PRMT5 inhibitor and application thereof

Info

Publication number: CN113045543A
Application number: CN202011543588.7A
Authority: CN
Inventors: 赵传武; 杨金路; 康杰琼; 陈立谦; 刘慧杰; 魏苗苗; 王佳; 耿佳
Original assignee: CSPC Zhongqi Pharmaceutical Technology Shijiazhuang Co Ltd
Current assignee: CSPC Zhongqi Pharmaceutical Technology Shijiazhuang Co Ltd
Priority date: 2019-12-26
Filing date: 2020-12-23
Publication date: 2021-06-29
Anticipated expiration: 2040-12-23
Also published as: CN113045543B

Abstract

The invention provides a compound with a novel structure and PRMT5 inhibitor activity, and a tautomer, an optical isomer, a solvate, an isotope derivative or a pharmaceutically acceptable salt thereof, wherein the compound has a structure shown in a formula (I). The compound has the medical application of treating cancer, can obviously inhibit the activity of PRMT5 and has no toxic or side effect on MV 4; 11 has strong proliferation inhibiting effect.

Description

PRMT5 inhibitor and application thereof

Technical Field

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

Background

Protein arginine methyltransferases (PRMTs) are enzymes that catalyze protein arginine methylation reactions, a class of S-adenosylmethionine-dependent methyltransferases that are responsible for the transfer of the methyl group from AdoMet to the guanidino nitrogen atom at the end of an arginine residue in histones or other proteins. PRMTs play important roles in protein methylation, such as participation in variable cleavage, post-transcriptional regulation, RNA processing, cell proliferation, cell differentiation, apoptosis, and tumor formation. Members of the PRMTs family can be classified into three groups, depending on the way in which arginine methylation is catalyzed: PRMT1-4, PRMT6 and PRMT8 belong to type I, and the catalytic forms are monomethyl and asymmetric dimethyl; PRMT5 and PRMT9 belong to type ii, the catalytic form of which is symmetrical bis-methyl; PRMT7 is of type iii, capable of monomethyl catalysis.

The expression of PRMTs is associated with various diseases of human, especially cancers, for example, PRMT1, PRMT 2, PRMT 3, PRMT 4 and PRMT7 are overexpressed in breast cancer, PRMT1 is overexpressed in lung cancer and leukemia, and PRMT5 plays an important role in the occurrence of various hematological and solid malignancies (gastric cancer, breast cancer, colon cancer, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, bladder cancer, skin cancer, melanoma, glioblastoma, cervical cancer, prostate cancer, etc.), the expression level of which is closely related to the occurrence, development and prognosis of tumors. PRMT6 is highly expressed in bladder and lung cancers, and PRMT9 is implicated in lymphoma, melanoma, testicular and pancreatic cancers. The importance of PRMTs in the development of tumorigenesis is becoming increasingly important, and among these specific inhibitors of PRMT1, PRMT 3, PRMT 4, PRMT5 and PRMT6, small molecule inhibitors of PRMT1 and PRMT5 have entered the clinical stage.

PRMT5 was first isolated by Pollack et al in a yeast two-hybrid study by binding to Jak2(Janus tyrosine kinase 2) in a protein complex, and was therefore also referred to as JBP1(Jak-binding protein 1). The PRMT5 can regulate the gene transcription and protein modification process, has the functions of regulating cell proliferation, differentiation and apoptosis in the growth process of tumor cells, and is a potential tumor treatment target. To date, PRMT5 inhibitors have been developed in an early stage, most rapidly in phase I/II clinical, with GSK3326595 introduced by GSK corporation as indicated for myelodysplastic syndrome (MDS), Acute Myeloid Leukemia (AML), solid tumors including breast cancer, glioblastoma multiforme (GBM), renal cell carcinoma, bladder cancer, and non-hodgkin's lymphoma. JNJ-64619178, first released by Janssen, was in phase I and indicated primarily as relapsed/refractory B-cell non-hodgkin lymphoma (NHL) or advanced solid tumors. PF-06939999 issued by Pfizer is in phase I, and its indications are mainly advanced or metastatic non-small cell lung cancer, squamous cell carcinoma of the head and neck, esophageal cancer, endometrial cancer, cervical cancer and bladder cancer. PRT-543 issued by Prelue Therapeutics is in stage I, and the indications are mainly advanced or metastatic solid tumors. Currently, no structural formulae have been published for PF-06939999 and PRT-543. The structural formulas of GSK3326595 and JNJ-64619178 are as follows:

besides, the published patent applications of selective inhibitors of PRMT5 also include WO2014100730, WO2016034671, WO2016034675, WO2016034673, WO2018167276, US2018298010, WO2018161922, WO2019002074, WO2019178368, WO2019110734, WO2019112719 and the like, but no good PRMT5 inhibitor is found as a drug on the market at present. In order to achieve the aim of better tumor treatment effect and better meet the market demand, a series of selective PRMT5 inhibitors are designed and synthesized.

Disclosure of Invention

The invention provides a PRMT5 inhibitor compound with a novel structure.

The invention provides a compound shown as a formula (I), a tautomer, an optical isomer, a solvate, an isotope derivative or a pharmaceutically acceptable salt thereof, wherein the compound has the following structure:

wherein Y is:

z is selected from

Wherein R is₁₁Is selected from-C_1-6An alkyl group or a group-H,

x is selected from the following connecting fragments:

ring A is selected from substituted or unsubstituted C_3-10Cycloalkyl, substituted or unsubstituted 4-10 membered heterocycloalkyl, substituted or unsubstituted saturated 4-12 membered bridged cyclic group, substituted or unsubstituted saturated 4-12 membered heterobridged cyclic group, substituted or unsubstituted saturated monospirocyclic group, substituted or unsubstituted saturated heteromonospirocyclic group, substituted or unsubstituted saturated condensed cyclic group; wherein, with R₉The linking N atom is attached to the pyrimidine ring;

the saturated monospirocyclic and saturated heteromonospirocyclic groups have ring atom number 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 each ring count includes spiro atoms;

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

the term "substituted" means that the substituents are each independently selected from the group consisting of halogen, hydroxy, cyano, nitro, amino, azido, carbonyl, carboxy, ethynyl, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Haloalkyl, C_3-10Cycloalkyl, 5-10 membered heterocycloalkyl, C_6-14One or more of aryl or 5-10 membered heteroaryl ring groups;

the R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₉And R₁₀Each occurrence is independently selected from hydrogen, halogen, hydroxy, amino, carboxy, nitro, cyano, carbonyl, azido, oxo, ethynyl, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Haloalkyl, C_1-6Alkoxyamino group, C_1-6Alkyl ester group, C_1-6Alkylamino radical, C_1-6Amide group, C_3-10Cycloalkyl radical, C_3-10Cycloalkylamino, C_3-10Cycloalkyl amide group, C_3-10Epoxyalkylamide, 5-to 10-membered heterocycloalkyl, C_6-14Aryl or 5-to 10-membered heteroaromatic ring group, wherein C is_3-10Cycloalkyl, 5-10 membered heterocycloalkyl, C_6-14Aryl or 5-10 membered heteroaryl ring groups may be independently substituted with one or more groups selected from halogen, hydroxy, amino, nitro, C_1-6Alkoxy or C_1-6Alkyl is substituted by a substituent; when R is₄、R₅When one of them is oxo, the other is absent; when R is₆、R₇When one of them is oxo, the other is absent; the oxo group means that two H at the same substitution position are replaced by the same O to form a double bond;

R₃at any substitutable position on the tetrahydroisoquinoline ring, n is 0, 1,2, 3,4, 5 or 6;

the R is₈Independently selected from hydrogen, halogen, hydroxyl, amino, carboxyl, nitro, cyano, carbonyl, azido, ethynyl, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Haloalkyl, C_1-6Alkoxyamino group, C_1-6Alkyl ester group, C_1-6Alkylamino, -NR₁₂R₁₃、C_1-6Amide group, C_3-10Cycloalkyl radical, C_3-10Cycloalkylamino, C_3-10Cycloalkyl amide group, C_3-10Epoxyalkylamide group, 5-to 10-membered heterocycloalkyl group, 5-to 10-membered heterocycloalkylamino group, C_6-14Aryl radical, C_6-14Arylamino, 5-10 membered heteroaromatic ring group amino, wherein said C_3-10Cycloalkyl, 5-10 membered heterocycloalkyl, C_6-14Aryl or 5-10 membered heteroaryl ring groups may be independently substituted with one or more groups selected from halogen, hydroxy, amino, nitro, C_1-6Alkoxy or C_1-6Alkyl is substituted by a substituent; r₁₂、R₁₃Each independently selected from C_1-6An alkyl group;

the definition of "aryl", "heteroaryl ring", "carbocyclyl", "cycloalkyl", "heterocyclyl", "heterocycloalkyl", "heterospirocyclyl", "heterobridged ring" and "heterofused ring" in the context of the present invention is as defined below in the section "definitions".

Preferably, the heteroatoms in the heterocycloalkyl, heteroaryl, heteromonospiro, heterobridged, or heterofused ring groups are independently selected from O, N or S, and the number of heteroatoms 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, has a structure represented by formula (II):

wherein, Y, R₁、R₂、R₃、R₄、R₅、R₆、R₇And n is as defined for 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, has a structure represented by formula (III):

wherein, X, Z, R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈And n is as defined for compound (I).

Further, in certain embodiments, the compounds of the present invention or tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein ring a is selected from substituted or unsubstituted C_5-7Cycloalkyl, substituted or unsubstituted 4-7 membered heterocycloalkyl, substituted or unsubstituted saturated 4-10 membered bridged ring group, substituted or unsubstituted saturated 4-10 membered heterobridged ring group, substituted or unsubstituted saturated monospirocyclic group and substituted or unsubstituted saturated heteromonospirocyclic group.

Further, in certain embodiments, the compounds of the present invention, or tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein ring a is selected from substituted or unsubstituted 4-6 membered heterocycloalkyl, substituted or unsubstituted saturated 4-or 6-membered monospirocyclyl, substituted or unsubstituted saturated 4-or 6-membered heteromonospirocyclyl.

Further, in certain embodiments, the compound of the present invention, or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein the heteroatom of the heterocyclyl, heterobridged ring, or heteromonospiro ring in the definition of ring a is selected from N or O atoms, preferably N atoms, with a heteroatom number of 1.

Further, in certain embodiments, the compounds of the present invention, or tautomers, optical isomers, solvates, isotopic derivatives thereof, or pharmaceutically acceptable salts thereof, wherein the heteroatom of heterocycloalkyl in the definition of ring a is a N atom.

Further, in certain embodiments, the compounds of the present invention or tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein "substituted" in the definition for ring a means that the substituent is selected from halogen or C_1-6An alkyl group.

Further, in certain embodiments, the compounds of the present invention or tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein X is selected from the group consisting of

The A ring is substituted or unsubstituted 4-10 membered heterocycloalkyl, substituted or unsubstituted saturated 4-12 membered heterobridged ring group, substituted or unsubstituted saturated heteromonospirocyclic group, substituted or unsubstituted saturated hetero condensed ring group, and the A ring is bonded with

Attached is a heteroatom.

Preferably, in the above embodiment, the A ring is substituted with

The heteroatom attached is an N atom.

Ring A is a substituted or unsubstituted saturated 4-membered/4-membered or 4-membered/6-membered spirocyclic group.

Further, in certain embodiments, the compounds of the present invention or tautomers, optical isomers thereofA structure, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, wherein R is₈Independently selected from-H,

Further, in certain embodiments, the compounds of the present invention or tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein X is independently selected from substituted or unsubstituted

The substituted position is any position on the ring of the group, and the substituted substituent is selected from-F, -Cl and-CH₃or-OH, the number of substituents being selected from 1,2 or 3.

Further, in certain embodiments, the compounds of the present invention or tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, wherein X is independently selected from the group consisting of

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 is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₉And R₁₀Each independently selected from hydrogen, halogen, hydroxyl, amino, nitro, carbonyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6A haloalkyl group.

Still 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 is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₉And R₁₀Each independently selected from hydrogen.

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

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

The invention also provides the use of the compound.

The application of the compound or the tautomer, the optical isomer, the solvate, the isotopic derivative or the pharmaceutically acceptable salt thereof or the pharmaceutical composition in the invention in preparing PRMT5 inhibitor medicines.

The invention also provides a pharmaceutical composition for preventing and/or treating cancer, which comprises the compound or the tautomer, the optical isomer, the solvate, the isotopic derivative or the pharmaceutically acceptable salt thereof.

Further, in the above use, 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 carcinoma, T-cell lymphoma, chronic and acute leukemia.

In another aspect, the invention also provides methods for preparing the compounds.

The preparation method of the compound comprises the following condensation reaction:

wherein, Y, X, R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈And n is as defined for compound (I).

Definition of

Unless otherwise specified, the term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, a straight or branched chain group containing 1 to 20 carbon atoms, e.g., "C_1～6Alkyl 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, tert-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.

Unless otherwise specified, "carbocyclyl" or "carbocycle" refers to a non-aromatic cyclic hydrocarbon radical having from 3 to 14 ring carbon atoms ("C_3-14Carbocyclyl ") and have no heteroatoms in the non-aromatic ring system. In some embodiments, carbocyclyl groups have 3-12 ring carbon atoms ("C)_3-12Carbocyclyl "), or 4 to 12 ring carbon atoms (" C)_4-12Carbocyclyl "), or 3 to 10 ring carbon atoms (" C)_3-10Carbocyclyl "). In some embodiments, carbocyclyl groups have 3 to 8 ring carbon atoms ("C)_3-8Carbocyclyl "). In some embodiments, carbocyclyl groups have 3 to 7 ring carbon atoms ("C)_3-7Carbocyclyl "). In some embodiments, carbocyclyl groups have 4 to 6 ring carbon atoms ("C)_4-6Carbocyclyl "). In some embodiments, carbocyclyl groups have 5 to 10 ring carbon atoms ("C5-10 carbocyclyl"), or 5 to 7 ring carbon atoms ("C)_5-7Carbocyclyl "). Exemplary C_3-6Carbocyclyl groups include, but are not limited to, cyclopropyl (C)₃) Cyclopropenyl group (C)₃) Cyclobutyl (C)₄) Cyclobutenyl radical (C)₄) Cyclopentyl (C)₅) Cyclopentenyl group (C)₅) Cyclohexyl (C)₆) Cyclohexenyl (C)₆) Cyclohexadienyl (C)₆) And the like. Exemplary C_3-8Carbocyclyl groups include, but are not limited to, the aforementioned C_3-6Carbocyclyl group and cycloheptyl (C)₇) Cycloheptenyl (C)₇) Cycloheptadienyl (C)₇) Cycloheptatrienyl (C)₇) Cyclooctyl (C)₈) Cyclooctenyl (C)₈) Bicyclo [2.2.1]Heptylalkyl radical (C)₇) Bicyclo [2.2.2]Octyl radical (C)₈) And the like. Exemplary C_3-10Carbocyclyl groups include, but are not limited to, the aforementioned C_3-8Carbocyclyl group and cyclononyl (C)₉) Cyclononenyl (C)₉) Cyclodecyl (C)₁₀) Cyclodecenyl (C)₁₀) octahydro-1H-indenyl (C)₉) Decahydronaphthyl (C)₁₀) Spiro [4.5 ]]Decyl (C)₁₀) And the like. As illustrated by the examples above, in certain embodiments, the carbocyclyl group is monocyclic ("monocyclic carbocyclyl") or is a fused (fused ring), bridged (bridged ring), or spiro-fused (spiro ring) ring system, such as a bicyclic ring system ("bicyclic carbocyclyl") and may be saturated or may be partially unsaturated. "carbocyclyl" also includes ring systems in which the carbocyclyl ring as defined above is fused by one or more aryl or heteroaryl groups, whichIs on the carbocyclic ring, and in such cases, the number of carbons continues to indicate the number of carbons in the carbocyclic ring system. In certain embodiments, each instance of a carbocyclyl group is independently optionally substituted, e.g., unsubstituted (an "unsubstituted carbocyclyl") or substituted with one or more substituents (a "substituted carbocyclyl"). In certain embodiments, the carbocyclyl group is unsubstituted C_3-10A carbocyclic group. In certain embodiments, the carbocyclyl group is a substituted C_3-10A carbocyclic group.

Unless otherwise specified, the term "cycloalkyl" refers to a monocyclic, saturated "carbocyclyl" or "carbocycle" as defined above, preferably C_3～12Cycloalkyl, more preferably C_3-10Cycloalkyl, further preferably C_3-7Cycloalkyl radical, C_3-6Cycloalkyl radical, C_5-7Cycloalkyl radicals, or C_4-6A cycloalkyl group. Such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

Unless otherwise specified, the term "alkoxy" refers to an-O-alkyl group, said alkyl group being as defined above. Preferably, it has 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms. Representative examples include methoxy, ethoxy, propoxy, t-butoxy, pentyloxy, 1-methylbutyloxy, 2-methylbutyloxy, 3-methylbutyloxy, 1-dimethylpropoxy, 1, 2-dimethylpropoxy, 2-dimethylpropoxy, 1-ethylpropoxy, and the like.

The term "halogen" or "halo" means, unless otherwise specified, F, Cl, Br, I. The term "haloalkyl" refers to an alkyl group as defined herein wherein one, two or more hydrogen atoms are replaced by halogen. Representative examples of haloalkyl groups include CCl₃、CF₃、CHCl₂、CH₂CF₃、CF₂CF₃And the like.

Unless otherwise specified, the term "heterocyclyl" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic non-aromatic substituent having ring carbon atoms and 1 to 4 ring heteroatoms, comprising 3 to 20 ring atoms ("3-20 membered heterocyclyl"), wherein the heteroatoms are each independently selected from N, O or S. Preferably 3 to 12 ring atoms ("3-12 membered heterocyclic group"), further preferably 3 to 10 ring atoms ("3-10 membered heterocyclic group"), further preferably 3 to 8 ring atoms ("3-8 membered heterocyclic group"), further preferably 4 to 7 ring atoms ("4-7 membered heterocyclic group"), further preferably 5 to 10 ring atoms ("5-10 membered heterocyclic group"), further preferably 5 to 6 ring atoms ("5-6 membered heterocyclic group"); the number of heteroatoms is preferably 1,2 or 3. "heterocyclyl" may be monocyclic ("monocyclic heterocyclyl") or a fused ("fused heterocyclyl" or "heterofused ring"), bridged ("heterobridged ring" or "bridged heterocyclyl") or spiro-fused ("hetero-spiro ring" or "spiro heterocyclyl") ring system, such as a bicyclic ring system ("bicyclic heterocyclyl"), and may be saturated or may be partially unsaturated. Heterocyclyl bicyclic ring systems may include one or more heteroatoms in one or both rings. "heterocyclyl" also includes ring systems in which the heterocyclyl ring as defined above is fused by one or more carbocyclyl groups, wherein the point of attachment is on the carbocyclyl or heterocyclyl ring or a ring system in which the heterocyclyl ring as defined above is fused by one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such cases the number of ring members continues to indicate the number of ring members in the heterocyclyl ring system. In certain embodiments, each instance of a heterocyclyl is independently optionally substituted, e.g., unsubstituted (an "unsubstituted heterocyclyl") or substituted (a "substituted heterocyclyl") with one or more substituents.

Exemplary 3-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, aziridinyl, oxacyclopropane (oxalanyl), and thietane (thiorenyl). Exemplary 4-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, azetidinyl, oxetanyl and thietanyl. Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2, 5-dione. Containing 2 hetero atomsExemplary 5-membered heterocyclyl groups include, but are not limited to, dioxolanyl, oxathiolanyl, dithiolanyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thiacyclohexyl (thianyl). Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl. Exemplary 6-membered heterocyclyl groups containing 3 heteroatoms include, but are not limited to, triazacyclohexane, oxadiazinyl, thiadiazinyl, oxathiazinyl, and dioxazazinyl. Exemplary 7-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, azepanyl, oxepinyl, and thiacycloheptyl. Exemplary 8-membered heterocyclyl groups containing 1 heteroatom include, but are not limited to, azacyclooctyl, oxocyclooctyl, and thietanyl. Condensed to a C₆Exemplary 5-membered heterocyclyl groups on the aryl ring (also referred to herein as a 5, 6-bicyclic heterocycle) include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolonyl, and the like. Exemplary 6-membered heterocyclyl groups fused to an aryl ring (also referred to herein as a 6, 6-bicyclic heterocycle) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and the like.

Unless otherwise specified, "heterocycloalkyl" means a monocyclic, saturated "heterocyclyl" or "heterocycle" as defined above, the ring atoms being as defined above, i.e. comprising 3 to 20 ring atoms ("3-20 membered heterocycloalkyl"), the number of heteroatoms being 1,2, 3 or 4 (1-4), preferably 1,2 or 3 (1-3), wherein the heteroatoms are each independently selected from N, O or S. Preferably 3 to 12 ring atoms ("3-12 membered heterocycloalkyl"), more preferably 3 to 10 ring atoms ("3-10 membered heterocycloalkyl"), even more preferably 3 to 8 ring atoms ("3-8 membered heterocycloalkyl"), even more preferably 4 to 7 ring atoms ("4-7 membered heterocycloalkyl"), even more preferably 5 to 10 ring atoms ("5-10 membered heterocycloalkyl"), even more preferably 5 to 6 ring atoms ("5-6 membered heterocycloalkyl"). In certain embodiments, each instance of heterocycloalkyl is independently optionally substituted, e.g., unsubstituted (an "unsubstituted heterocycloalkyl") or substituted (a "substituted heterocycloalkyl") with one or more substituents. Some exemplary "heterocycloalkyl" groups have been given above for "heterocyclyl" or "heterocyclic" moieties, and also include, but are not limited to, oxacyclohexane, thiomorpholinyl, oxathiacyclohexyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, imidazolinidine, and the like.

Unless otherwise specified, the term "spiroheterocyclyl" refers to a polycyclic heterocyclic group in which one atom (referred to as a spiro atom) is shared between monocyclic rings, wherein one or more ring atoms are selected from nitrogen, oxygen, or a heteroatom of S (O) r (where r is an integer 0, 1, 2), the remaining ring atoms being carbon. These may contain one or more double bonds, but none of the rings has a completely conjugated pi-electron system. Spiro heterocyclic groups are classified into a mono-spiro heterocyclic group, a di-spiro heterocyclic group, or a multi-spiro heterocyclic group according to the number of spiro atoms shared between rings.

Unless otherwise specified, the term "fused heterocyclyl" 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 the rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system, in which one or more of the ring atoms is selected from nitrogen, oxygen, or a heteroatom of s (o) r (where r is an integer 0, 1, 2), and the remaining ring atoms are carbon. They can be classified as bicyclic, tricyclic, tetracyclic or polycyclic fused heterocycloalkyl depending on the number of constituent rings.

Unless otherwise specified, the term "bridged heterocyclyl" refers to polycyclic heterocyclic groups in which any two rings share two atoms not directly attached, which may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system, in which one or more ring atoms are selected from nitrogen, oxygen, or heteroatoms of s (o) r (where r is an integer 0, 1, 2), the remaining ring atoms being carbon. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups depending on the number of constituent rings. Unless otherwise specified, the term "aryl" or "aromatic ring group" means monocyclic, bicyclic and tricyclic aromatic carbocyclic ring systems 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 the aryl group may include phenyl, naphthyl, anthryl, phenanthryl, pyrenyl, or the like.

Unless otherwise specified, the term "heteroaryl" or "heteroaryl ring group" 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 heteroatoms being independently selected from O, N or S, the number of heteroatoms being preferably 1,2 or 3. Examples of heteroaryl groups include furyl, thienyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiadiazolyl, triazinyl, phthalazinyl, quinolyl, isoquinolyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, benzothienyl, benzopyridyl, benzopyrimidinyl, benzimidazolyl, phthalizinyl, pyrrolo [2,3-b ] pyridyl, imidazo [1,2-a ] pyridyl, 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.

Unless otherwise specified, the term "spiro ring group" refers to all-carbon polycyclic groups sharing a single carbon atom (referred to as a spiro atom) between monocyclic rings, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. Spirocycloalkyl groups are classified as mono-spirocycloalkyl, bis-spirocycloalkyl or multi-spirocycloalkyl depending on the number of spiro atoms shared between rings.

Unless otherwise specified, 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 other rings in the system, wherein one or more rings may contain one or more double bonds, but none of the rings has a fully conjugated pi-electron system. They may be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused ring alkyls depending on the number of constituent rings.

Unless otherwise specified, the term "bridged ring group" refers to all-carbon polycyclic groups in which any two rings share two carbon atoms not directly attached, which may contain one or more double bonds, but none of the rings have a completely conjugated pi-electron system. They can be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl groups depending on the number of constituent rings.

Unless otherwise specified, the term "pharmaceutically acceptable salt" refers to salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of mammals, particularly humans, without excessive toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, such as the medically acceptable salts of amines, carboxylic acids, and other types of compounds, are well known in the art.

Unless otherwise specified, 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 invention are acidic, pharmaceutically acceptable non-toxic bases include salts prepared with inorganic and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, zinc, and the like, and salts derived from organic non-toxic bases include primary, secondary, tertiary, and the like.

The abbreviations used in the preparation examples, examples and elsewhere herein are:

DCM dichloromethane

TEA Triethylamine

DIPEA N, N-diisopropylethylamine

DMF N, N-dimethylformamide

EtOAc ethyl acetate

h hours

ml of

HATU 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate

MeOH methanol

TFA trifluoroacetic acid

DMSO-d₆Deuterated dimethyl sulfoxide

SAM S-adenosylmethionine

DTT dithiothreitol

The invention has the beneficial effects that:

the invention provides a PRMT5 inhibitor with a novel structure, and discloses a preparation method and application thereof in medicine. In particular, the compounds of the invention have utility in the treatment of cancer, with enzymatic and cellular screening results showing: compared with a positive control drug GSK-3326595, the compound provided by the invention can inhibit PRMT5 activity more obviously and can inhibit MV 4; 11 has strong proliferation inhibiting effect.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or under conditions recommended by the manufacturers. 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 to which this invention belongs. 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 embodiments and materials are shown herein for illustrative purposes only.

Preparation example

The preparation of the parent compound and the intermediate of the specific compound of the present invention is described in the following preparation examples.

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

Step 1: (S) -6-chloro-N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide

In a 100mL reaction flask, 6-chloro-pyrimidine 4-carbonyl chloride (0.63g, 3.56mmol) was dissolved in DCM (10mL) and TEA (0.72g, 7.12mmol) was added at 0 ℃. Then (S) -1-amino-3- (3, 4-dihydroisoquinolin-2 (1H) -yl) propan-2-ol (0.66g, 3.20mmol) was added. The reaction solution was stirred at 25 ℃ for 2 h. TLC monitors the reaction is finished, and the reaction solution is H₂O (5mL) was diluted and extracted with DCM (15 mL. times.2). The combined organic layers were washed with saturated brine (10mL), Na₂SO₄Drying, filtering and concentrating under reduced pressure to obtain residue. The residue was purified by column chromatography (DCM: MeOH ═ 10: 1). The compound (S) -6-chloro-N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (I') was obtained as a yellow oil (0.68g, 55.08%).

Step 2: (S) -tert-butyl 4- ((6- ((3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) carbamoyl) pyrimidin-4-yl) amino) piperidine-1-carboxylate

(S) -6-chloro-N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (0.28g, 0.81mmol, 1eq) was dissolved in 20ml of isopropanol, TEA (0.25g, 24.7mmol, 3eq) and tert-butyl 4-aminopiperidine-1-carboxylate (0.24g, 1.2mmol, 1.5eq) were added in this order, and the reaction mixture was heated to 85 ℃ for reaction for 8H. After TLC showed completion of the reaction, the reaction mixture was concentrated to dryness under reduced pressure, and column chromatography (DCM: MeOH ═ 30:1) was carried out to give (S) -tert-butyl 4- ((6- ((3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) carbamoyl) pyrimidin-4-yl) amino) piperidine-1-carboxylate (0.36g, 87.32%).

And step 3: preparation of (S) -N- (3, 4-dihydroisoquinolin-2 (1H) -yl-2-hydroxypropyl) -6- (piperidin-4-ylamino) pyrimidine-4-carboxamide (T-1)

(S) -tert-butyl 4- ((6- ((3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) carbamoyl) pyrimidin-4-yl) amino) piperidine-1-carboxylate (0.36g, 0.71mmol, 1eq) was dissolved in 20ml dichloromethane, and 1ml trifluoroacetic acid was added to react at room temperature for 1H. Concentrating under reduced pressure to dryness to obtain (S) -N- (3, 4-dihydroisoquinoline)-2(1H) -yl-2-hydroxypropyl) -6- (piperidin-4-ylamino) pyrimidine-4-carboxamide (T-1) (0.25g, 86.38%), MS (m/z): 411.35[ M + H]⁺。

Preparation example 2: preparation of (S) -6- ((6-aminospiro [3.3] hept-2-yl) amino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (T-2)

The synthesis was as in preparation 1, except that (6-aminospiro [3.3]]Replacing tert-butyl 4-aminopiperidine-1-carboxylate with tert-butyl hept-2-yl) carbamate to obtain (S) -6- ((6-aminospiro [ 3.3)]Hept-2-yl) amino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (T-2) (0.30g, 85.12%), MS (m/z): 437.62[ M + H]⁺. Preparation example 3: (S) -6- (7-azaspiro [ 3.5)]Preparation of nonan-2-ylamino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (T-3)

The synthesis was the same as in preparation example 1, except that 2-amino-7-Boc-7-azaspiro [3.5]]Replacing 4-aminopiperidine-1-carboxylic acid tert-butyl ester with nonane to obtain (S) -6- (7-azaspiro [ 3.5)]Nonan-2-ylamino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (T-3) (0.35g, 96.33%), MS (m/z): 451.37[ M + H]⁺。

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

Synthesis procedure is as in preparation example 1, except that tert-butyl 4-aminopiperidine-1-carboxylate is replaced with tert-butyl 4-amino-3-fluoropiperidine-1-carboxylate to give N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl)) -2-hydroxypropyl) -6- ((3-fluoropiperidin-4-yl) amino) pyrimidine-4-carboxamide (T-4) (0.16g, 70.49%), MS (m/z): 429.22[ M + H]⁺。

Preparation example 5: preparation of 6- ((3, 3-difluoropiperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (T-5)

The synthesis was identical to preparation 1, except that tert-butyl 4-aminopiperidine-1-carboxylate was replaced with tert-butyl 4-amino-3, 3-difluoro-1-piperidinecarboxylate to give 6- ((3, 3-difluoropiperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (T-5) (0.15g, 69.19%), MS (m/z): 447.54[ M + H]⁺。

The following are specific embodiments for preparing the compounds of the present invention using starting materials or intermediates.

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

(S) -N- (3, 4-dihydroisoquinoline-2 (1H) -yl-2-hydroxypropyl) -6- (piperidin-4-ylamino) pyrimidine-4-carboxamide (T-1) (0.25g, 0.61mmol, 1.0eq) is dissolved in 20mL of DMF, 3mL of TEA (pH & gt, 10) is added sequentially, 2-oxopropanoic acid (0.64g, 0.730mmol, 1.2eq), HATU (0.30g, 0.789mmol, 1.3eq) is added sequentially, and the mixture is reacted at room temperature for 2H. After TLC showed the reaction was complete, water, ethyl acetate and liquid separated. The organic phase was washed with water and saturated brine, and then concentrated to dryness, followed by column chromatography (dichloromethane: methanol ═ 30:1) to give the compound (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (2-oxopropanoyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-1) (0.1g, 34.10%). MS (m/z): 481.25[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.384-1.400(m,2H),1.900-1.966(m,2H),2.349(s,3H),2.645-2.740(m,2H),2.803-2.812(m,2H),2.956-2.994(m,1H),3.180-3.219(m,1H),3.276-3.298(m,2H),3.354-3.420(m,2H),3.542-3.596(m,3H),3.868-3.876(m,1H),4.100-4.121(m,2H),4.966-4.973(d,1H),6.979-6.991(m,1H),7.048-7.099(m,4H),7.783-7.794(d,1H),8.275(s,1H),8.745-8.763(t,1H).

Example 2: preparation of (S) -6- ((1- (2-cyclopropyl-2-oxoacetyl) piperidin-4-yl) amino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-2)

The synthesis was performed as in example 1, except that 2-oxopropanoic acid was replaced with 2-cyclopropyl-2-carbonylacetic acid to give (S) -6- ((1- (2-cyclopropyl-2-oxoacetyl) piperidin-4-yl) amino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-2) (0.27g, 87.51%). MS (m/z): 507.27[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.088-1.233(m,5H),1.360-1.378(m,2H),1.935-1.982(m,2H),2.245-2.251(m,1H),2.688-2.834(m,4H),2.994-3.033(m,1H),3.244-3.298(m,2H),3.405-3.426(m,2H),3.511-3.533(m,1H),3.612(m,2H),3.895(m,1H),4.164(m,2H),4.990(s,1H),7.024-7.109(m,5H),7.813-7.822(d,1H),8.301(s,1H),8.778(s,1H).

Example 3: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (3, 3-dimethyl-2-oxobutanoyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-3)

The synthesis was performed as in example 1, except that trimethylpyruvic acid was used instead of 2-oxopropanoic acid, to give (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (3, 3-dimethyl-2-oxobutanoyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-3) (0.27g, 84.85%). MS (m/z): 523.30[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.191(s,9H),1.308-1.369(m,2H),1.918-1.981(m,2H),2.615-2.755(m,2H),2.818-2.827(m,2H),2.970-3.008(m,1H),3.212-3.331(m,5H),3.406-3.427(m,1H),3.572-3.644(m,2H),3.881-3.890(m,1H),4.173(m,2H),4.980-4.987(d,1H),7.007-7.111(m,5H),7.810-7.820(d,1H),8.290(s,1H),8.774-8.782(t,1H).

Example 4: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (3-methyl-2-oxobutanoyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-4)

The synthesis was performed as in example 1, except that 3-methyl-2-oxobutanoic acid was used in place of 2-oxopropanoic acid, to give (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (3-methyl-2-oxobutanoyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-4) (0.24g, 77.42%). MS (m/z): 509.29[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.095(s,6H),1.345-1.360(m,2H),1.920-1.982(m,2H),2.667-2.755(m,2H),2.816-2.825(m,2H),2.990-3.036(m,2H),3.212-3.307(m,4H),3.408-3.486(m,2H),3.572-3.644(m,2H),3.872-3.889(m,1H),4.157-4.175(m,2H),4.971-4.978(d,1H),7.005-7.109(m,5H),7.799-7.810(d,1H),8.291(s,1H),8.756-8.774(t,1H).

Example 5: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (4-methyl-2-oxovaleryl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-5)

The synthesis was performed as in example 1, except that 4-methyl-2-oxopentanoic acid was used in place of 2-oxopropanoic acid to give (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (4-methyl-2-oxopentanoyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-5) (0.24g, 77.42%). MS (m/z): 523.30[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:0.921(s,6H),1.326-1.343(m,2H),1.925-1.948(m,2H),2.081-2.103(m,1H),2.610-2.755(m,4H),2.815-2.824(m,2H),2.990-3.036(m,1H),3.223-3.307(m,4H),3.407-3.429(m,1H),3.525-3.642(m,3H),3.870-3.898(m,1H),4.121-4.142(m,2H),4.970-4.977(d,1H),7.003-7.108(m,5H),7.793-7.804(d,1H),8.289(s,1H),8.755-8.773(t,1H).

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

The synthesis was performed as in example 1, except that glycolic acid was used instead of 2-oxopropanoic acid, to give (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((1- (2-hydroxyacetyl) piperidin-4-yl) amino) pyrimidine-4-carboxamide (PT-6) (0.20g, 70.10%). MS (m/z): 469.25[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.297-1.390(m,2H),1.899-1.912(m,2H),2.682-2.757(m,2H),2.828-2.880(m,3H),3.086-3.122(m,1H),3.305(brs,2H),3.420-3.439(m,2H),3.579-3.677(m,3H),3.894(m,1H),4.083-4.113(m,3H),4.210-4.224(m,1H),4.523(s,1H),4.976(s,1H),7.008-7.099(m,5H),7.771-7.782(m,1H),8.297(s,1H),8.758(s,1H).

Example 7: preparation of 6- ((1- (2-cyclopropyl-2-hydroxypropionyl) piperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-7)

The synthesis was performed as in example 1, except that 2-cyclopropyl-2-hydroxypropionic acid was used in place of 2-oxopropanoic acid, to give 6- ((1- (2-cyclopropyl-2-hydroxypropionyl) piperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-7) (0.18g, 5.49%). MS (m/z): 523.27[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:0.315-0.345(m,3H),0.398-0.407(m,1H),1.154-1.186(m,2H),1.283(s,3H),1.349(m,2H),1.914(m,2H),1.988(m,1H),2.688-2.755(m,2H),2.828(m,2H),3.032(s,1H),3.244-3.298(m,1H),3.409-3.431(m,1H),3.576-3.644(m,2H),3.883-3.889(m,1H),4.010-4.046(m,1H),4.117(s,1H),4.452(m,2H),4.960-4.967(s,1H),5.044(s,1H),7.007-7.111(m,5H),7.755-7.767(d,1H),8.290(s,1H),8.738-8.756(s,1H).

Example 8: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (3, 3-dimethyl-2-oxobutanoyl) -7-azaspiro [3.5] nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-8)

The synthesis method is the same as example 1, except that T-3 is used to replace T-1, and trimethylpyruvic acid is used to replace 2-oxopropanoic acid, thus obtaining (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (3, 3-dimethyl-2-oxobutanoyl) -7-azaspiro [ 3.5)]Nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-8) (0.28g, 89.68%). MS (m/z): 563.33[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.170-1.181(s,9H),1.514(m,2H),1.597(m,2H),1.704(m,2H),2.313(m,2H),2.665-2.754(m,2H),2.816-2.825(m,2H),3.071(m,1H),3.154(m,1H),3.277-3.309(m,2H),3.309(s,1H),3.392-3.474(m,3H),3.572-3.643(m,2H),3.881-3.889(m,1H),4.404-4.430(m,1H),4.980-4.987(d,1H),7.018-7.100(m,5H),8.049-8.059(m,1H),8.271(s,1H),8.767(s,1H).

Example 9: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (4-methyl-2-oxovaleryl) -7-azaspiro [3.5] nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-9)

The synthesis was performed as in example 1 except that T-3 was used instead of T-1 and 4-methyl-2-oxopentanoic acid was used instead of 2-oxopropanoic acid to give (S) -N- (3- (3, 4-bis)Hydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (4-methyl-2-oxovaleryl) -7-azaspiro [ 3.5)]Nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-9) (0.21g, 67.26%). MS (m/z): 563.33[ M + H]+；¹H NMR(600MHz,DMSO-d₆)δ:0.899-0.919(d,6H),1.522-1.533(m,2H),1.609(brs,2H),1.700(brs,2H),2.054-2.111(m,1H),2.309(brs,2H),2.582-2.604(m,3H),2.716-2.825(m,2H),2.839(s,2H),3.175(m,1H),3.249-3.294(m,3H),3.373-3.455(m,3H),3.643(brs,2H),3.902(brs,1H),4.418(brs,1H),4.994(brs,1H),7.026-7.108(m,5H),8.041-8.062(m,1H),8.284(s,1H),8.759(s,1H).

Example 10: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (3-methyl-2-oxobutanoyl) -7-azaspiro [3.5] nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-10)

The synthesis method is the same as example 1, except that T-3 is used instead of T-1, and 3-methyl-2-oxobutanoic acid is used instead of 2-oxopropanoic acid, to obtain (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (3-methyl-2-oxobutanoyl) -7-azaspiro [ 3.5)]Nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-10) (0.24g, 78.84%). MS (m/z): 549.31[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.077-1.100(d,6H),1.244-1.267(m,2H),1.525-1.533(m,2H),1.612(brs,2H),1.711(brs,2H),2.321(brs,2H),2.623-2.693(m,1H),2.702-2.793(m,1H),2.825-2.834(m,2H),2.952-3.014(m,1H),3.151(m,1H),3.223-3.242(m,1H),3.286-3.336(m,1H),3.411(brs,1H),3.493(brs,1H),3.582-3.652(brs,3H),3.889-3.898(m,1H),4.428-4.440(m,1H),4.978-4.986(brs,1H),7.016-7.119(m,5H),8.047-8.070(m,1H),8.279(s,1H),8.764(s,1H).

Example 11: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (2- (dimethylamino) -2-oxoacetyl) -7-azaspiro [3.5] nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-11)

The synthesis method is the same as example 1, except that T-3 is used to replace T-1, and N, N-dimethyl oxamic acid is used to replace 2-oxo-propionic acid, to obtain (S) -N- (3- (3, 4-dihydroisoquinoline-2 (1H) -yl) -2-hydroxypropyl) -6- ((7- (2- (dimethylamino) -2-oxo-acetyl) -7-azaspiro [ 3.5)]Nonan-2-yl) amino) pyrimidine-4-carboxamide (PT-11) (0.15g, 49.20%). MS (m/z): 550.31[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.244(m,1H),1.530(m,2H),1.613(m,2H),1.711(m,2H),2.315-2.330(m,2H),2.620-2.623(m,1H),2.693(m,3H),2.740(m,1H),2.900(m,8H),3.148-3.157(m,1H),3.221-3.231(m,1H),3.401-3.410(m,1H),3.475-3.628(m,2H),3.896(m,2H),4.437(m,1H),4.987(brs,1H),7.031-7.237(m,4H),7.962(s，1H),8.064(m,1H),8.284(s,1H),8.770(s,1H)

Example 12: preparation of (S) -6- ((6- (2-cyclopropyl-2-oxoacetamido) spiro [3.3] hept-2-yl) amino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-12)

The synthesis was performed as in example 1 except that T-1 was replaced with T-2 and 2-oxopropanoic acid was replaced with 2-cyclopropyl-2-oxoacetic acid to give (S) -6- ((6- (2-cyclopropyl-2-oxoacetamido) spiro [3.3]Hept-2-yl) amino) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-12) (0.21g, 68.85%). MS (m/z): 533.28[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:0.848-0.860(m,2H),0.871-0.965(m,1H),1.095-1.108(m,1H),1.245(brs,2H),1.892-1.963(m,2H),2.122-2.179(m,3H),2.285-2.2359(m,2H),2.676-2.745(m,2H),2.813-2.847(m,3H),3.283-3.294(m,2H),3.419(brs,1H),3.618-3.643(m,2H),3.878-3.887(m,1H),4.142-4.155(m,1H),4.303-4.317(m,1H),4.963(s,1H),6.997-7.109(m,5H),7.971-7.982(d,1H),8.267(s,1H),8.735-8.788(m,2H)。

Example 13: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (3, 3-dimethyl-2-oxobutanamido) spiro [3.3] hept-2-yl) amino) pyrimidine-4-carboxamide (PT-13)

The synthesis method is the same as example 1, except that T-2 is used to replace T-1, and trimethylpyruvic acid is used to replace 2-oxopropanoic acid, thus obtaining (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (3, 3-dimethyl-2-oxobutanamido) spiro [3.3]Hept-2-yl) amino) pyrimidine-4-carboxamide (PT-13) (0.24g, 76.38%). MS (m/z): 549.31[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.185(s,9H),1.839-2.388(m,8H),2.674-2.745(m,2H),2.817-2.825(m,4H),3.283-3.294(m,1H),3.410(brs,1H),3.617-3.643(m,2H),3.886(brs,1H),4.102-4.142(m,1H),4.313-4.325(m,1H),4.989(brs,1H),7.005-7.110(m,5H),7.992-8.002(d,1H),8.269(s,1H),8.745-8.757(m,2H)。

Example 14: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (4-methyl-2-oxopentanamido) spiro [3.3] hept-2-yl) amino) pyrimidine-4-carboxamide (PT-14)

The synthesis procedure is as in example 1, except that T-2 is used instead of T-1 and 4-methyl-2-oxopentanoic acid is used instead of 2-oxopropanoic acid, to give (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (4-methyl-2-oxopentanamide) spiro [3.3]Hept-2-yl) amino) pyrimidine-4-carboxamide (PT-14) (0.24g, 76.38%). MS (m/z): 549.31[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:0.868-0.879(d,6H),1.235(m,2H),1.872-1.954(m,2H),1.996-2.041(m,1H),2.089-2.167(m,3H),2.282(m,1H),2.346(m,1H),2.662-2.674(m,3H),2.732-2.743(m,1H),2.824(m,2H),3.283-3.327(m,1H),3.301-3.407(m,2H),3.572-3.642(m,2H),3.879-3.887(m,1H),4.095-4.136(m,1H),4.307-4.318(m,1H),4.969-4.974(m,1H),6.994-7.110(m,5H),7.978-7.989(d,1H),8.265(s,1H),8.754-8.790(m,2H)。

Example 15: preparation of (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (2-hydroxyacetamido) spiro [3.3] hept-2-yl) amino) pyrimidine-4-carboxamide (PT-15)

The synthesis procedure is as in example 1, except that T-2 is substituted for T-1 and glycolic acid is substituted for 2-oxopropanoic acid, to give (S) -N- (3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (2-hydroxyacetamido) spiro [3.3]Hept-2-yl) amino) pyrimidine-4-carboxamide (PT-15) (0.17g, 59.94%). MS (m/z): 495.26[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.168-1.235(m,1H),1.890-1.940(m,2H),2.042-2.086(m,2H),2.158(m,1H),2.288-2.342(m,2H),2.543(m,2H),2.737-2.841(m,4H),3.283-3.327(m,1H),3.415(m,2H),3.663-3.751(m,3H),3.909(m,1H).4.166-4.178(m,1H),4.319(m,1H),5.022(brs,1H),5.375(s,1H),7.017-7.108(m,5H),7.849-7.983(d,2H),8.284(s,1H),8.744(s,1H)。

Example 16: (S) -N¹(superscript of 1) - (6- ((6- ((3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) carbamoyl) pyrimidin-4-yl) amino) spiro [3.3]Hept-2-yl) -N²，N²Preparation of (E) -Dimethyloxamide (PT-16)

The synthesis was performed as in example 1 except that T-1 was replaced with T-2 and 2-oxopropanoic acid was replaced with N, N-dimethylacetoamine to give (S) -N¹- (6- ((6- ((3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) carbamoyl) pyrimidin-4-yl) amino) spiro [3.3]Hept-2-yl) -N²，N²Dimethyl oxamide (PT-16) (0.26g, 84.77%). MS (m/z): 536.29[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.971-2.050(m,4H),2.199-2.238(m,1H),2.298(m,1H),2.396(m,1H),2.475-2.504(m,1H),2.668-2.696(m,1H),2.727-2.755(m,1H),2.815-2.873(m,5H),2.937(s,3H),3.290-3.340(m,3H),3.414-3.435(m,1H).3.575-3.643(m,2H),3.881-3.890(m,1H),4.105-4.144(m,1H),4.310(brs,1H),4.965-4.973(d,1H),7.003-7.109(m,5H),7.974-7.985(d,1H),8.269(s,1H),8.742(s,1H),8.816-8.828(d,1H)。

Example 17: preparation of N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (2-hydroxypropionylamino) spiro [3.3] hept-2-yl) amino) pyrimidine-4-carboxamide (PT-17)

The synthesis procedure is as in example 1, except that T-1 is replaced with T-2 and 2-oxopropanoic acid is replaced with DL-lactic acid to give N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) -6- ((6- (2-hydroxypropionylamino) spiro [3.3]Hept-2-yl) amino) pyrimidine-4-carboxamide (PT-17) (0.20g, 69.01%). MS (m/z): 509.28[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.177(d,3H),1.234-1.302(m,1H),1.872-1.950(m,2H),2.000-2.080(m,2H),2.141-2.179(m,1H),2.284-2.341(m,3H),2.465(m,1H),2.564(m,2H),2.711-2.975(m,5H),3.411(brs,1H).3.879-3.921(m,2H),4.101-4.142(m,1H),4.312-4.324(m,1H),5.050(brs,1H),5.378-5.386(d,1H),7.003-7.115(m,5H),7.791-7.805(d,1H),7.975-7.985(d,1H),8.292(s,1H),8.746(brs,1H)。

Example 18: preparation of 6- ((1- (2-cyclopropyl-2-oxoacetyl) -3, 3-difluoropiperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-18)

The synthesis was performed as in example 1, except that T-5 was used instead of T-1 and 2-oxopropanoic acid was used instead of 2-oxopropanoic acid, to give 6- ((1- (2-cyclopropyl-2-oxoacetyl) -3, 3-difluoropiperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-18) (0.15g, 43.76%). MS (m/z): 543.42[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.075-1.205(m,5H),1.550-1.654(m,1H),1.966-2.002(m,1H),2.243-2.322(m,1H),2.664-2.766(m,3H),2.794-2.834(m,2H),2.882(s,1H),3.125-3.321(m,1H),3.410-3.440(m,2H),3.605-3.626(m,2H),3.826-3.907(m,2H),4.25-4.515(m,1H),4.997(d,2H),7.001-7.214(m,5H),8.083-8.096(d,1H),8.306(s,1H),8.826(t,1H).

Example 19: preparation of 6- ((6- (2-cyclopropyl-2-hydroxypropionylamino) spiro [3.3] hept-2-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-19)

The synthesis was performed as in example 1 except that T-1 was replaced with T-2 and 2-oxopropanoic acid was replaced with 2-cyclopropyl-2-hydroxypropionic acid to give 6- ((6- (2-cyclopropyl-2-hydroxypropionamido) spiro [3.3]Hept-2-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-19) (0.14g, 39.83%). MS (m/z): 549.62[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:0.142(s,1H),0.244-0.311(m,3H),1.062-1.097(m,1H),1.244-1.248(m,3H),1.867-2.083(m,4H),2.158(m,1H),2.281-2.341(m,2H),2.465(m,1H),2.564(m,2H),2.669-2.737(m,2H),2.822(m,2H),3.285(m,1H),3.411(m,1H),3.575-3.644(m,2H),3.881-3.888(m,1H),4.077-4.117(m,1H),4.308-4.320(m,1H),4.915-4.920(d,1H),4.979(s,1H),7.001-7.095(m,5H),7.634-7.647(d,1H),7.974-7.987(d,1H),8.267(s,1H),8.751(s,1H)。

Example 20: preparation of 6- ((1- (2-cyclopropyl-2-oxoacetyl) -3-fluoropiperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinolin-2 (1H) -yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-20)

The synthesis procedure is as in example 1, except that T-4 is used instead of T-1 and 2-oxopropanoic acid is replaced with 2-cyclopropyl-2-carbonylacetic acid, to give 6- ((1- (2-cyclopropyl-2-oxoacetyl) -3-fluoropiperidin-4-yl) amino) -N- ((S) -3- (3, 4-dihydroisoquinoline-2 (1H) -Yl) -2-hydroxypropyl) pyrimidine-4-carboxamide (PT-20) (0.25g, 72.93%). MS (m/z): 507.11[ M + H]⁺；¹H NMR(600MHz,DMSO-d₆)δ:1.107-1.231(m,5H),1.543-1.806(m,2H),2.011(m,1H),2.221(m,1H),2.688-2.834(m,5H),3.325-3.412(m,4H),3.601-3.619(m,3H),3.889(m,1H),4.444-4.624(m,2H),4.978(s,1H),7.014-7.169(m,5H),8.144(d,1H),8.295-8.326(d,1H),8.794(t,1H).

Examples 21 to 67:

according to the methods of preparation example 1 and examples 1 to 20, compounds 21 to 67 were synthesized by selecting the corresponding starting materials, the structures and mass spectra signals of which are as follows:

effect test example 1

1. Cell experiment method

Human acute monocytic leukemia cell MV4 was used for the experiment; 11 (Shanghai cell Bank) the complete medium required for the culture was IMDM (Cat NO.12440-053, gibco) supplemented with 10% serum FBS (Cat NO. SA311.02, cellmax). Cells were incubated at 37 ℃ with 5% CO₂Culturing in an incubator. The test reagents included dimethyl sulfoxide (available from Kemiou Chemicals, Inc., Tianjin). MTT, (Thiazolyl BLUE TETRAZOLIUM Bromide, CAS. NO.2)98-93-1, VWR). The test control GSK-3326595 was obtained by either home-made or commercial purchase. The test article was stored hermetically at 4 ℃.

The test substance and the positive control substance are fully dissolved by using dimethyl sulfoxide as a solvent to prepare the solution with the concentration of 5 multiplied by 10^-2And storing the solution in mol/L. Storing the stock solutions of the test substances at-20 ℃. The complete culture medium is used as a diluent, and the test substance is diluted in a gradient manner to different concentrations for standby. In a 96-well plate, 100. mu.L/well of each test substance was added at respective different concentrations, 8 concentrations were set for each test substance, and 3 multiple wells were set for each concentration. Wells containing only cell suspension without test substance and vehicle were used as control wells. Wells with complete medium only and no cell suspension were used as blank wells. Add 100. mu.L/well (2X 10)³Cell number/well) human acute monocytic leukemia cells MV 4; 11 complete medium suspension, 5% CO at 37 ℃₂Culturing in an incubator. On the sixth day, 20. mu.L/well of MTT was added at 37 ℃ with 5% CO₂Culturing for 4h in an incubator, removing supernatant, adding 150 mu L/hole of dimethyl sulfoxide, shaking and mixing uniformly, and detecting OD value by an enzyme-labeling instrument at 550 nm. The inhibition was calculated according to the formula (test well OD-blank well OD)/(control well OD-blank well OD) × 100%, and the results are shown in table 1.

2. Enzymology experimental method

IC detection of test substances on PRMT5 using the radioisotope FlashPlate technique₅₀。

After the test compounds were dissolved in dimethyl sulfoxide, the test compounds were added to an Echo 384-well plate and diluted to the desired concentration, and the test substances were transferred from the diluted Echo 384-well plate to a 384-well reaction plate using an Echo550 apparatus, and the dimethyl sulfoxide was transferred for both negative and positive controls. PRMT5 was added to a 1-fold reaction buffer (1-fold reaction buffer comprising 10mM Tris-HCl, pH 8.0; 0.01% Tween-20; 1mM DTT) to form a 1.67-fold enzyme solution (enzyme concentration 5 nM). Combining a polypeptide substrate with [3H ]]SAM was added to 1 fold reaction buffer to form a 2.5 fold substrate solution (final substrate concentrations 100nM and 250nM, respectively). To 384 well reaction plate hole add 15 uL/hole of 1.67 times enzyme solution. For the enzyme-inactive control wells, the enzyme solution was replaced with 15uL of 1 fold reaction buffer. Centrifuging at 1000rpm for 1min, and incubating at room temperature for 15min is the same as the formula (I). To each well of a 384-well reaction plate was added 10uL of a 2.5-fold substrate solution. Centrifuge at 1000rpm for 1 min. The reaction was carried out at 25 ℃ for 60 min. The reaction was stopped by adding 5uL of a reaction stop solution to each well of the 384-well reaction plate (125 uM of cold SAM solution). 25uL of each well from the assay plate was transferred to a Flashplate and allowed to stand at room temperature for 1 h. The flashplate plates were then washed 3 times with 0.1% Tween-20 solution. Read with MicroBeta 2. The data were converted to inhibition rate data. Wherein max refers to the conversion rate of a DMSO control, and min refers to the conversion rate of an enzyme-free control. Percent inhibition is (max-conversion)/(max-min) × 100. Fitting of IC with XLFit excel add-in version 5.4.0.8₅₀The value is obtained. Fitting formula Y ═ Bottom + (Top-Bottom)/(1+ (IC)₅₀/X)^HillSlope)。

TABLE 1 cytological and enzymatic Effect test data for Compounds of the invention and Positive controls

Note: -represents no detection.

The results of enzymology and cell screening show that the compound of the invention has more significant inhibition on PRMT5 activity and No. MV4 compared with the positive control drug GSK-3326595 (compound at 208 of patent application WO 2014100719); 11 has strong proliferation inhibiting effect.

Effect test example 2: in vivo efficacy test

Female NOD-SCID mice, SPF grade 4-5 weeks old, were purchased from Peking Wintonlifa laboratory animals technologies, Inc. The mice were injected intraperitoneally one day before cell inoculation with cyclophosphamide at a dose of 100 mg/kg. Inoculating human acute monocytic leukemia cells MV4 subcutaneously in the axilla of the forelimb of the mouse; 11(1X 10)⁷0.1 ml/mouse), a subcutaneous transplantation tumor model is established. The tumor volume is about 110mm³(day 12 after inoculation), mice were divided into 5 groups of 5 mice each with a balanced tumor volume, vehicle control and recipient, respectivelyTest drug group. The dose of the test drug group was set to 100mg/kg, the volume of the drug was 10mL/kg, the frequency of the drug administration was BID, and the tumor size was measured 2 times per week and the data was recorded. The administration was continued for 10 days, and the test was terminated on day 22, and the tumors were weighed by tumor removal.

ˉˉ

Calculating the weight gain rate, tumor volume and tumor weight inhibition rate according to the formula, wherein the weight gain rate is X_(Wi-W0)/X_W0100, wherein W_iThe weight, W, of a particular mouse in each experimental group on the nth day is shown₀Body weight at the time of starting administration for a certain mouse in each experimental group; tumor volume (V) ═ 1/2 × a × b²Wherein a and b represent tumor long and short meridians, respectively; tumor weight inhibition rate (vehicle control group-test drug group)/vehicle control group 100%.

Table 2 MV for compounds of the invention and positive controls; 411 in vivo test data

Note: p <0.05, P <0.01, P <0.001, compared to vehicle control group

MV 4; 11 in vivo efficacy test results show that the compound has obvious tumor inhibition activity and is obviously superior to a positive control drug GSK 3326595.

Claims

1. A compound of formula (I), and tautomers, optical isomers, solvates, isotopic derivatives or pharmaceutically acceptable salts thereof, having the structure:

wherein Y is:

z is selected from

Wherein R is₁₁Is selected from-C_1-6An alkyl group or a group-H,

x is selected from the following connecting fragments:

the R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₉And R₁₀Each occurrence is independently selected from hydrogen, halogen, hydroxy, amino, carboxy, nitro, cyano, carbonyl, azido, oxo, ethynyl, C_1-6Alkyl radical, C_1-6Alkoxy radical,C_1-6Haloalkyl, C_1-6Alkoxyamino group, C_1-6Alkyl ester group, C_1-6Alkylamino radical, C_1-6Amide group, C_3-10Cycloalkyl radical, C_3-10Cycloalkylamino, C_3-10Cycloalkyl amide group, C_3-10Epoxyalkylamide, 5-to 10-membered heterocycloalkyl, C_6-14Aryl or 5-to 10-membered heteroaromatic ring group, wherein C is_3-10Cycloalkyl, 5-10 membered heterocycloalkyl, C_6-14Aryl or 5-10 membered heteroaryl ring groups may be independently substituted with one or more groups selected from halogen, hydroxy, amino, nitro, C_1-6Alkoxy or C_1-6Alkyl is substituted by a substituent; when R is₄、R₅When one of them is oxo, then the other is absent; when R is₆、R₇When one of them is oxo, then the other is absent; the oxo group means that two H at the same substitution position are replaced by the same O to form a double bond;

the R is₈Independently selected from hydrogen, halogen, hydroxyl, amino, carboxyl, nitro, cyano, carbonyl, azido, ethynyl, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Haloalkyl, C_1-6Alkoxyamino group, C_1-6Alkyl ester group, C_1-6Alkylamino, -NR₁₂R₁₃、C_1-6Amide group, C_3-10Cycloalkyl radical, C_3-10Cycloalkylamino, C_3-10Cycloalkyl amide group, C_3-10Epoxyalkylamide group, 5-to 10-membered heterocycloalkyl group, 5-to 10-membered heterocycloalkylamino group, C_6-14Aryl radical, C_6-14Arylamino, 5-10 membered heteroaromatic ring group amino, wherein said C_3-10Cycloalkyl, 5-10 membered heterocycloalkyl, C_6-14Aryl, 5-10 membered heteroaryl ring groups may be independently substituted with one or more groups selected from halogen, hydroxy, amino, nitro, C_1-6Alkoxy or C_1-6Alkyl is substituted by a substituent; r₁₂、R₁₃Each independently selected from C_1-6An alkyl group;

wherein, the heteroatoms in the heterocycloalkyl, the heteroaromatic ring group, the heteromonospiro ring group, the heterobridged ring group and the heterocondensed ring group are independently selected from O, N or S, and the number of the heteroatoms is 1,2 or 3.

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

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

4. a compound according to any one of claims 1 to 3, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: ring A is selected from substituted or unsubstituted C_5-7Cycloalkyl, substituted or unsubstituted 4-7 membered heterocycloalkyl, substituted or unsubstituted saturated 4-10 membered bridged ring group, substituted or unsubstituted saturated 4-10 membered heterobridged ring group, substituted or unsubstituted saturated monospirocyclic group and substituted or unsubstituted saturated heteromonospirocyclic group.

5. The compound of any one of claims 1-4, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: the A ring is selected from substituted or unsubstituted 4-6 membered heterocycloalkyl, substituted or unsubstituted saturated 4-membered/4-membered or 4-membered/6-membered monospirocyclic group, substituted or unsubstituted saturated 4-membered/4-membered or 4-membered/6-membered heteromonospirocyclic group.

6. The compound of any one of claims 1-5, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: the hetero atom of the heterocycloalkyl, heterobridged ring group and heteromonospiro ring group in the definition of the A ring is N or O atom, preferably N atom, and the number of the hetero atom is 1.

7. The compound of any one of claims 1 to 6, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: by "substituted" in the definition of ring A is meant that the substituent is selected from halogen or C_1-6An alkyl group.

8. The compound of any one of claims 1 to 7, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: x is selected from

Attached is a heteroatom.

9. The compound of any one of claims 1 to 8, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: on the A ring with

Connected to each otherThe heteroatom is an N atom.

10. The compound of any one of claims 1 to 7, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: x is selected from

11. The compound of any one of claims 1-10, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: r₈Independently selected from-H,

12. The compound of any one of claims 1-11, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: x is independently selected from substituted or unsubstituted

13. A compound according to any one of claims 1 to 12 or a tautomer, optical isomer, solvate, isotope thereofA derivative or a pharmaceutically acceptable salt thereof, characterized in that: x is independently selected from

14. The compound of any one of claims 1-13, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, wherein: r₁、R₂、R₃、R₄、R₅、R₆、R₇、R₉And R₁₀Each independently selected from hydrogen, halogen, hydroxyl, amino, nitro, carbonyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6A haloalkyl group.

15. The compound of claim 1, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, having the structure:

16. a pharmaceutical composition comprising a compound of any one of claims 1 to 15, or a tautomer, optical isomer, solvate, isotopic derivative, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

17. Use of a compound according to any one of claims 1 to 15, or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16, for the preparation of a medicament for the treatment of a PRMT5 inhibitor.

18. Use of a compound according to any one of claims 1 to 15, or a tautomer, optical isomer, solvate, isotopic derivative or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16, for the preparation of a medicament for the prophylaxis and/or treatment of cancer.

19. The use according to claim 18, 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, pancreatic cancer, brain cancer, pituitary adenoma, melanoma, epidermoid carcinoma, T-cell lymphoma, chronic and acute leukemia.