WO2021088992A1

WO2021088992A1 - Tetrahydroisoquinoline spiro compound as prmt5 inhibitor

Info

Publication number: WO2021088992A1
Application number: PCT/CN2020/127166
Authority: WO
Inventors: 刘磊; 刘扬; 赵春艳; 唐任宏; 任晋生
Original assignee: 江苏先声药业有限公司
Priority date: 2019-11-07
Filing date: 2020-11-06
Publication date: 2021-05-14
Also published as: CN114728938A

Abstract

The present invention discloses a tetrahydroisoquinoline spiro compound represented by formula (I) as a PRMT5 inhibitor, a preparation method therefor, a pharmaceutical composition containing the compound, and use thereof in the treatment of PRMT5-mediated diseases, wherein A is a 5-14 membered spiro group optionally substituted by R⁶.

Description

Tetrahydroisoquinoline spiro compounds as PRMT5 inhibitors

Technical field

This application relates to a tetrahydroisoquinoline spiro compound as a PRMT5 inhibitor, its preparation method, a pharmaceutical composition containing the compound, and its use in the treatment of PRMT5-mediated diseases.

Background technique

Epigenetic changes are key mediators that drive and maintain the malignant phenotype of tumors. DNA methylation, histone acetylation and methylation, non-coding RNA, and post-translational modifications are all epigenetic driving forces for cancer, and have nothing to do with changes in DNA sequence. Arginine methylation is an important type of post-translational modification that affects cell growth and proliferation, apoptosis, angiogenesis, and metastasis by regulating transcription and post-transcriptional RNA processing. There are three types of methylarginine, ω-NG-monomethylarginine (MMA), ω-NG, N'G-asymmetric dimethylarginine (ADMA) and ω-NG, N 'G-Symmetric dimethylarginine (SDMA). This modification is catalyzed by the protein arginine methyltransferase (PRMT) family, which transfers methyl groups from S-adenosylmethionine (AdoMet) to the arginine side chains of histones and non-histone proteins. Nine PRMT genes are annotated in the human genome, which are classified into type I (PRMT1,2,3,4,6 and 8), type II (PRMT5 and PRMT9), and type III enzymes based on the type of methylarginine produced (PRMT7). PRMT5 is mainly a type II enzyme, which can catalyze the symmetric dimethylation of arginine. PRMT5 was discovered for the first time in a two-hybrid experiment to detect proteins interacting with Janus tyrosine kinase (Jak2).

PRMT5 is a universal transcription repressor that forms a complex with other transcription factors, including BRG1 and hBRM, Blimp1 and Snail. PRMT5 participates in a variety of different cell biological processes by methylating a variety of substrates in the cytoplasm and nucleus, including histone H4 residue Arg3 (H4R3) and H3 residue Arg8 (H3R8). H4R3 methylation is related to transcriptional inhibition, whereas H3R8 methylation is considered to be related to both transcriptional activation and transcriptional inhibition. In addition to direct induction of inhibitory histone markers, PRMT5's role in gene silencing is also mediated by the formation of multiple inhibitory protein complexes, including NuRD components, HDACs, MDB proteins and DNA methyltransferases. PRMT5 affects its substrate specificity by interacting with some binding proteins. The core component of this protein complex is MEP50. MEP50 is necessary for the enzymatic activity of PRMT5. Studies have found that PRMT5 can methylate proteins involved in RNA splicing, such as SmD3, which can be used to track the chemical activity of cellular biological PRMT5.

PRMT5 plays an important role in tumorigenesis. Studies have found that the expression of PRMT5 is up-regulated in a variety of tumors, including lymphoma, lung cancer, breast cancer and colorectal cancer. In addition, PRMT5 expression is increased in mantle cell lymphoma (MCL) patient samples, and PRMT5 knockout inhibits MCL cell proliferation, indicating that PRMT5 plays an important role in MCL. PRMT5 overexpression promotes cell proliferation. On the contrary, in melanoma, breast cancer and lung cancer cell lines, PRMT5 knockout inhibits the proliferation of these cells. Therefore, PRMT5 is a potential target for cancer treatment.

The loss of methylthioadenosine phosphorylase (MTAP) gives cells a selective dependence on PRMT5 and its binding protein WDR77. MTAP is often lost due to its proximity to the normally missing tumor suppressor gene CDKN2A. The concentration of intracellular methylthioadenosine (MTA, a metabolite cleaved by MTAP) in cells carrying the MTAP deletion increases. In addition, MTA specifically inhibits the enzymatic activity of PRMT5. Compared with MTAP-expressing cells, MTA or PRMT5 small molecule inhibitors significantly inhibited the cell viability of cancer cell lines lacking MTAP.

Therefore, there is a need in the art to develop small molecules that can inhibit the activity of PRMT5 and treat various PRMT5-related diseases.

Summary of the invention

This application relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof,

among them,

A is a 5-14 membered spirocyclic group ^{optionally substituted by R 6;}

R ¹ and R ^{2 are} independently selected from H, C _1-4 alkyl, halogen, C _1-4 alkoxy, or

R ¹ , R ² and the C atom to which they are connected together form a C _3-8 cycloalkyl or 3-8 membered heterocycloalkyl, the cycloalkyl or heterocycloalkyl is optionally substituted by halogen;

R ³ , R ⁴ , and R ^{5 are} independently selected from H, halogen and C _1-4 alkyl;

m is 1, 2, 3 or 4;

R ^{6 is} independently selected from halogen, hydroxyl, cyano, amino, C _1-3 alkylamino, di(C _1-3 alkyl)amino, oxo, C _1-4 alkyl, C _1-4 alkoxy Group, C _1-4 alkoxycarbonyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl.

In some embodiments, A is C _6-13 spirocycloalkyl or 6-13 membered spiroheterocycloalkyl, which is optionally substituted with ^{R 6.}

In some embodiments, A is optionally substituted with ^{R 6}

among them

n, n', p, and q are independently selected from 1, 2, 3, and 4, and n+n'+p+q≤10;

W is selected from CH or N;

X and Y are independently selected from CH ₂ , NH or O;

Z is selected from CH ₂ , NH, O or a bond.

In some embodiments, n, n', p, q are independently selected from 1, 2, and 3, and n+n'+p+q≤10.

In some embodiments, n, n'are independently selected from 1, 2, and 3, and p, q are independently selected from 1 and 2.

In some embodiments, W is N.

In some embodiments, A is selected from optionally substituted with ^{R 6}

In some embodiments, A is selected from optionally substituted with ^{R 6}

In some embodiments, A is selected from

In some embodiments, A is selected from

In some embodiments, R ^{6 is} independently selected from halogen, hydroxy, cyano, amino, C _1-3 alkylamino, oxo, C _1-4 alkyl, C _1-4 alkoxy, C _{1- 4} Alkoxycarbonyl and C _3-6 cycloalkyl.

In some embodiments, R ^{6 is} independently selected from halogen, hydroxy, cyano, amino, oxo, C _1-4 alkyl, C _1-4 alkoxy, and C _1-4 alkoxycarbonyl.

In some embodiments, R ^{6 is} independently selected from fluoro, hydroxy, cyano, amino, oxo, methyl, methoxy, and methoxycarbonyl.

In some embodiments, R ¹ and R ^{2 are} independently selected from H, C _1-4 alkyl, or R ¹ , R ² and the C atom to which they are attached together form a C _3-6 cycloalkyl group, and the C _{3- The 6} cycloalkyl group is optionally substituted with halogen.

In some embodiments, R ¹ and R ^{2 are} independently selected from H, methyl, ethyl, or R ¹ , R ² and the C atom to which they are attached together form a cyclopropyl or cyclobutyl, the cyclopropyl or Cyclobutyl is optionally substituted with fluorine.

In some embodiments, R ¹ and R ² are methyl groups.

In some embodiments, R ³ , R ⁴ , R ^{5 are} independently selected from H, F, Cl, methyl, and ethyl.

In some embodiments, R ³ , R ⁴ , and R ⁵ are H.

In some embodiments, m is 1 or 2.

In some embodiments, m is 1.

In some embodiments, the compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof is selected from a compound of formula (II) or a pharmaceutically acceptable salt thereof,

Wherein A, R ¹ , R ² , R ³ , R ⁴ , R ⁵ and m are as defined above.

In some embodiments, the compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof is selected from the compound of formula (III) or a pharmaceutically acceptable salt thereof,

Wherein A, R ¹ , R ² , R ³ , R ⁴ , R ⁵ and m are as defined above.

In some embodiments, the compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

In another aspect, the present application relates to a pharmaceutical composition comprising the compound of formula (I) of the present application or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition of the present application further includes pharmaceutically acceptable excipients.

On the other hand, this application relates to a method for treating a disease mediated by PRMT5 in a mammal, including administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a mammal in need of such treatment, preferably a human, Or its pharmaceutical composition.

On the other hand, this application relates to a method for inhibiting PRMT5 activity in vivo or in vitro, the method comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a subject in need thereof, Or its pharmaceutical composition.

On the other hand, this application relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the preparation of a medicine for preventing or treating PRMT5-mediated diseases.

On the other hand, this application relates to the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the prevention or treatment of PRMT5-mediated diseases.

On the other hand, the present application relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preventing or treating PRMT5-mediated diseases.

In some embodiments, the PRMT5-mediated disease is a proliferative disease, a metabolic disease or a blood disease, preferably a proliferative disease or a metabolic disease.

In some embodiments, the proliferative disease is selected from cancer, autoimmune disease or inflammatory disease, preferably cancer.

In some embodiments, the PRMT5-mediated disease is cancer.

definition

Unless otherwise specified, the following terms used in this application have the following meanings. A specific term should not be considered uncertain or unclear without a special definition, but should be understood according to the ordinary meaning in the field. When a trade name appears in this article, it is meant to refer to its corresponding commodity or its active ingredient.

The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, as long as the valence of the specific atom is normal and the substituted compound is stable. When the substituent is oxo (ie =O), it means that two hydrogen atoms are replaced, and the oxo will not occur on the aromatic group. The "stable" means that when the optional range of substituents includes optional options that cannot be used to replace a specific group due to valence requirements, chemical stability, or other reasons, the optional range should be understood as the context according to the context Including those options suitable for substituting specific groups. For example, when considering the degree of optional substitution of a particular part, it should be understood that the number of substituents does not exceed the valence suitable for that part.

The term "optional" or "optionally" means that the event or situation described later can occur or not occur, and the description includes occurrence of said event or situation and non-occurrence of said event or situation. For example, the ethyl group is "optionally" substituted by halogen, meaning that the ethyl group can be unsubstituted (CH ₂ CH ₃ ), monosubstituted (such as CH ₂ CH ₂ F), or polysubstituted (such as CHFCH ₂ F, CH ₂ CHF ₂ etc.) or completely substituted (CF ₂ CF ₃ ). Those skilled in the art can understand that for any group containing one or more substituents, any substitution or substitution pattern that is impossible to exist in space and/or cannot be synthesized will not be introduced.

_{C mn in} this document means that the part has an integer number of carbon atoms in the given range, that is, from an integer m to an integer n (inclusive). For example, "C _1-6 "means that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.

When any variable (such as R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. For example, if a group is replaced by 2 Rs, then each R has independent options.

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

When the bond of a substituent is cross-linked to two atoms on a ring, the substituent can be bonded to any atom on the ring. For example, the structural unit

It means that R ⁵ can be substituted at any position on the benzene ring.

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

The term "hydroxy" refers to the -OH group.

The term "cyano" refers to the -CN group.

The term "amino" refers to the -NH ₂ group.

The term "5-14 membered spirocyclic group" refers to a cyclic group with at least two rings sharing one ring atom and forming a total of 5-14 ring atoms, in which all ring atoms can be carbon atoms or ring atoms It may contain at least one heteroatom selected from N, O or S. The spirocyclic group may be saturated or partially saturated.

The term "alkyl" refers to a hydrocarbon group of the _{general formula C n} H _2n+1. The alkyl group may be linear or branched. For example, the term "C ₁ - ₆ alkyl" refers to (e.g., methyl, ethyl, n-propyl, isopropyl, alkyl containing 1 to 6 carbon atoms, n-butyl, isobutyl, sec-butyl, Tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Similarly, the alkyl moiety (ie, alkyl) of alkoxy, alkylamino, and dialkylamino has the same definition as described above.

The term "alkoxy" refers to -O-alkyl.

The term "alkylamino" refers to -NH-alkyl.

The term "dialkylamino" refers to -N(alkyl) ₂ .

The term "cycloalkyl" refers to a carbocyclic ring that is fully saturated and may exist as a monocyclic, bridged, or spiro ring. Unless otherwise indicated, the carbocyclic ring is usually a 3 to 10 membered ring. Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, diamond Alkyl, spiro[4.5]decane, etc. Spirocycloalkyl refers to a cycloalkyl that exists as a spiro ring.

The term "cycloalkenyl" refers to a non-aromatic carbocyclic ring that is not fully saturated and may exist as a single ring, a bridged ring, or a spiro ring. Unless otherwise indicated, the carbocyclic ring is usually a 5- to 8-membered ring. Non-limiting examples of cycloalkenyl include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, and the like.

The term "heterocycloalkyl" refers to a cyclic group that is fully saturated and may exist as a monocyclic, bridged, or spiro ring. Unless otherwise indicated, the heterocyclic ring is generally a 3 to 7 membered ring containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen, and/or nitrogen. Examples of 3-membered heterocycloalkyl groups include, but are not limited to, oxirane, sulfidene, and azaethylenyl groups, and non-limiting examples of 4-membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetane Examples of cyclic group, thiabutanyl, 5-membered heterocycloalkyl include but are not limited to tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidine Examples of 6-membered heterocycloalkyl groups include but are not limited to piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1, Examples of 4-thiaxanyl, 1,4-dioxanyl, thiomorpholinyl, 1,3-dithianyl, 1,4-dithianyl, and 7-membered heterocycloalkyl include But it is not limited to azepanyl, oxepanyl, and thieppanyl. Preferably, it is a monocyclic heterocycloalkyl group having 5 or 6 ring atoms. Spiroheterocycloalkyl refers to a heterocycloalkyl that exists as a spiro ring.

Although the present application may refer to any compound or specific group of compounds defined herein by optional, preferred or suitable features or in other ways according to specific embodiments, the present application may also involve expressly excluding said optional, Any compound or specific group of compounds of a preferred or suitable feature or specific embodiment.

When used herein in conjunction with a measurable value (e.g., a quantity or a period of time, etc.), the term "about" is intended to encompass reasonable variations in that value, for example, to allow for experimental error in measuring the value.

The term "treatment" means administering the compound or formulation described in this application to prevent, ameliorate or eliminate a disease or one or more symptoms related to the disease, and includes:

(i) Preventing the occurrence of a disease or disease state in mammals, especially when such mammals are susceptible to the disease state, but have not been diagnosed as having the disease state;

(ii) Suppress the disease or disease state, that is, curb its development;

(iii) Alleviate the disease or disease state, even if the disease or disease state subsides.

It should be understood that the term "prevention" is not absolute, but refers to where the administration of the compound or composition reduces the likelihood or severity of the condition, symptom, or disease state, and/or delays the onset of the condition, symptom, or disease state for a period of time Uses and results.

As used herein alone or in combination with another term or terms, the terms "subject" and "patient" suitably refer to mammals, particularly humans.

The term "therapeutically effective amount" means (i) treatment or prevention of a particular disease, condition or disorder, (ii) reduction, amelioration or elimination of one or more symptoms of a particular disease, condition or disorder, or (iii) prevention or delay The amount of the compound of the present application for the onset of one or more symptoms of a specific disease, condition, or disorder described herein. The amount of the compound of the present application that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but it can be routinely determined by those skilled in the art. Determined by its own knowledge and this disclosure.

The term "PRMT5-mediated disease" refers to any disease, disorder, or other pathological condition in which PRMT5 is known to play a role. Therefore, in some embodiments, the application relates to treating or reducing the severity of one or more diseases in which PRMT5 is known to play a role.

The term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and/or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues, but not Many toxicity, irritation, allergic reactions or other problems or complications are commensurate with a reasonable benefit/risk ratio.

As pharmaceutically acceptable salts, for example, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, etc. can be mentioned. . In some embodiments, the compound of formula (I) is isolated as a pharmaceutically acceptable salt.

The term "pharmaceutical composition" refers to a mixture of one or more of the compounds of the application or their salts and pharmaceutically acceptable excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound of the present application to the organism.

The term "pharmaceutically acceptable excipients" refers to those excipients that have no obvious stimulating effect on the organism and will not damage the biological activity and performance of the active compound. Suitable auxiliary materials are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and/or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water and the like.

The word "comprise" or "comprise" and its English variants such as comprises or comprising should be understood as an open, non-exclusive meaning, that is, "including but not limited to".

The compounds and intermediates of the present application may also exist in different tautomeric forms, and all such forms are included in the scope of the present application. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also called proton transfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerization. A specific example of a proton tautomer is the imidazole moiety, in which protons can migrate between two ring nitrogens. Valence tautomers include interconversion through the recombination of some bonding electrons.

The present application also includes compounds of the present application that are the same as those described herein, but have one or more atoms replaced by an isotope-labeled atom having an atomic weight or mass number different from the atomic weight or mass number commonly found in nature. Examples of isotopes that can be bound to the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I and ³⁶ Cl, etc.

Certain isotope-labeled compounds of the application (such as ^{those labeled with 3} H and ¹⁴ C) can be used in compound and/or substrate tissue distribution analysis. Tritiated (i.e. ³ H) and carbon-14 (i.e. ¹⁴ C) isotopes are especially preferred due to their ease of preparation and detectability. Positron emission isotopes such as ¹⁵ O, ¹³ N, ¹¹ C, and ¹⁸ F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Generally, the isotopically-labeled compounds of the present application can be prepared by the following procedures similar to those disclosed in the schemes and/or examples below, by replacing non-isotopically-labeled reagents with isotope-labeled reagents.

In addition, substitution with heavier isotopes (such as deuterium (ie ² H)) can provide certain therapeutic advantages resulting from higher metabolic stability (for example, increased in vivo half-life or reduced dosage requirements), and therefore in certain situations The following may be preferred, where the deuterium substitution can be partial or complete, and partial deuterium substitution refers to the substitution of at least one hydrogen with at least one deuterium.

The compounds of the present application may be asymmetric, for example, have one or more stereoisomers. Unless otherwise specified, all stereoisomers include, for example, enantiomers and diastereomers. Enantiomers are characterized by the absolute configuration of their asymmetric centers, and are described by the R- and S-ordering rules of Cahn and Prelog, or by the way the molecule rotates the plane of polarized light, and is called right-handed or left-handed . The compounds of the present application containing asymmetric carbon atoms can be isolated in an optically pure form or in a racemic form. The optically active pure form can be resolved from the racemic mixture or synthesized by using chiral raw materials or chiral reagents. Non-limiting examples of stereoisomers include, but are not limited to:

Certain compounds of formula (I) may have one or more asymmetric centers and therefore may exist in multiple stereoisomeric configurations. Therefore, these compounds can be synthesized and/or separated as a mixture of enantiomers and/or as a single (pure) enantiomer, and in the case of two or more asymmetric centers, as a single non- Synthesis and/or separation of mixtures of enantiomers and/or diastereomers. It should be understood that this application includes all these enantiomers and diastereomers and all unfavorable mixtures thereof.

The pharmaceutical composition of the present application can be prepared by combining the compound of the present application with suitable pharmaceutically acceptable excipients, for example, can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, and powders. , Granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

Typical routes for administering the compound of the present application or a pharmaceutically acceptable salt or pharmaceutical composition thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, Intramuscular, subcutaneous, and intravenous administration.

The pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing method, dissolution method, granulation method, sugar-coated pill method, grinding method, emulsification method, freeze-drying method, etc.

In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These auxiliary materials enable the compound of the present application to be formulated into tablets, pills, lozenges, sugar-coated agents, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to patients.

The solid oral composition can be prepared by conventional mixing, filling or tabletting methods. For example, it can be obtained by the following method: mixing the active compound with solid excipients, optionally grinding the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain tablets Or the core of the dragee. Suitable excipients include, but are not limited to: binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, and the like.

The pharmaceutical composition may also be suitable for parenteral administration, such as a sterile solution, suspension or lyophilized product in a suitable unit dosage form.

In all the administration methods of the compound of general formula I described herein, the daily dose is 0.01 to 100 mg/kg body weight, preferably 0.05 to 50 mg/kg body weight, more preferably 0.1 to 30 mg/kg body weight, in single or divided doses form.

The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent replacement manners, preferred implementation manners include but are not limited to the embodiments of the present application.

The chemical reaction in the specific embodiment of the application is completed in a suitable solvent, and the solvent must be suitable for the chemical change of the application and the required reagents and materials. In order to obtain the compound of the present application, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction schemes on the basis of the existing embodiments.

An important consideration in the planning of synthetic routes in this field is to select suitable protective groups for reactive functional groups (such as amino groups in this application). For example, refer to Greene's Protective Groups in Organic Synthesis (4th Ed). Hoboken, New Jersey: John Wiley & Sons, Inc. All references cited in this application are incorporated into this application as a whole.

In some embodiments, the compound of general formula (I) of the present application can be prepared by those skilled in the art of organic synthesis through Route 1:

Route 1

Compound M1 reacts with M2 to form M3, M3 reacts in the presence of ethanol/CO to form M4, M4 hydrolyzes the ester group in the presence of a base, and forms M5 in the presence of Boc anhydride, M5 reacts with AH, and further removes amino protection The base produces a compound of formula (I), wherein said R ¹ , R ² , R ³ , R ⁴ , R ⁵ , A and m are as defined in formula (I) above.

In some embodiments, the compound M1 and M2 react in the presence of a base to form M3.

In some embodiments, the compound M3 reacts to form M4 in the presence of a palladium catalyst/base/ethanol/CO.

In some embodiments, the compound M5 is reacted with A-H in the presence of a condensing agent, and the amino protecting group is further removed to produce a compound of formula (I).

In some embodiments, the route 1 can be further expressed as:

Route 1

Compound M1 and M2 react in the presence of NaH/DMF to form M3, and M3 is in the presence of [1,1'-bis(diphenylphosphine)ferrocene] palladium dichloride dichloromethane complex/base/ethanol/CO The next reaction generates M4, M4 hydrolyzes the ester group in the presence of a base, and generates M5 in the presence of Boc anhydride. M5 reacts with AH in the presence of a condensing agent, and further removes the amino protecting group to generate the compound of formula (I).

Description of the drawings

Figure 1 shows the ellipsoid of the compound m crystal.

Figure 2 shows the tumor growth curve of the in vivo drug efficacy experiment in mice using the Z-138 subcutaneous tumor model.

Figure 3 shows the mouse body weight change curve of the in vivo drug efficacy experiment in mice using the Z-138 subcutaneous tumor model.

Detailed ways

For the sake of clarity, examples are further used to illustrate the application, but the examples do not limit the scope of the application. All reagents used in this application are commercially available and can be used without further purification.

Example 1. Preparation of tert-butyl 3-(oxiran-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (Intermediate 1)

Compound intermediate b:

Add (S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (a) (55g, 200mmol), dimethylhydroxylamine hydrochloride ( 29.4g, 300mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) (91g, 240mmol) was added to 1L single port Into the bottle, add anhydrous N,N-dimethylformamide (500mL), cool in an ice bath after nitrogen protection, and then add N,N-diisopropylethylamine (DIEA) (104mL, 600mmol) dropwise. The reaction solution was reacted at room temperature for 4 hours. After the reaction is complete, the excess N,N-diisopropylethylamine and N,N-dimethylformamide are removed by rotary evaporation, then cooled in an ice bath, diluted with saturated brine (1L), and extracted with ethyl acetate (2 X 200 mL), the organic phases were combined and washed with 5% sodium carbonate aqueous solution (2 X 500 mL), and then washed with saturated brine (500 mL). After drying and filtering with anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure and purified by column chromatography (eluent gradient: petroleum ether/ethyl acetate = 2/1) to obtain the target intermediate tert-butyl (S)-3-(methoxy (Methyl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (b) (62g, yield: 97%), properties: colorless oil.

LCMS: Rt: 1.76min; MS m/z(ESI): 321.3[M+H].

Chiral-HPLC:Rt:3.159

Compound intermediate c:

Add tert-butyl (S)-3-(methoxy(methyl)carbamoyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (b) (20g, 62.5 mmol) was weighed into a 500mL three-necked flask, added to anhydrous tetrahydrofuran (200mL), cooled to -70℃, DIBAL-H toluene solution (1.5M, 83mL, 125mmol) was slowly added dropwise, the reaction solution was stirred at -70℃ for 1 hour. After the reaction was completed, saturated ammonium chloride solution (100 mL) was slowly added to quench at -70°C, and then 0.5N aqueous hydrochloric acid solution was added to dilute (200 mL). After separation, the organic phase was washed with saturated sodium chloride aqueous solution (2 X 200 mL), then dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and column chromatography (eluent gradient: petroleum ether/ethyl acetate = 8/ 1) Purification to obtain the intermediate tert-butyl (S)-3-formyl-3,4-dihydroisoquinoline-2(1H)-carboxylate (c) (15g, yield: 92%), Properties: yellow oil.

LCMS: Rt: 1.93min; MS m/z(ESI): 206.1[M-56+H].

Chiral-HPLC:Rt:2.018

Compound Intermediate 1:

Weigh out sodium hydride (2.1g, 54mmol) in a three-neck flask at room temperature, protect it with nitrogen, add anhydrous DMSO (30mL), and stir at 80°C for 45 minutes. Cool to room temperature, add anhydrous tetrahydrofuran (20 mL) and dilute. Continue to cool to 0°C and add a solution of trimethylsulfur iodide (11.3 g, 55.5 mmol) in anhydrous DMSO (30 mL). After stirring for 10 minutes, add tert-butyl (S)-3-formyl-3,4-dihydroisoquinoline-2(1H)-carboxylate (c) (5g, 19.2mmol) in tetrahydrofuran (20mL) , Slowly rise to room temperature overnight. After the reaction was completed, it was cooled in an ice bath, quenched with saturated ammonium chloride solution (10 mL), diluted with saturated brine (300 mL), extracted with ethyl acetate (2 X 60 mL), the organic phases were combined and dried over anhydrous sodium sulfate and filtered, and the filtrate Concentrate under reduced pressure and purify by reverse phase column (H ₂ O-MeCN-50-100). The separated components are concentrated and extracted with ethyl acetate, dried over anhydrous sodium sulfate and filtered. The filtrate is concentrated under reduced pressure to obtain tert-butyl 3-( Oxide-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (Intermediate 1) (2.3g, yield: 42%) Properties: yellow oil.

LCMS: Rt: 1.10min; MS m/z(ESI): 220.0[M-56+H].

Example 2. Preparation of 6-bromo-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one (Intermediate 2)

Compound intermediate e:

Add 2-(3-bromophenyl)acetonitrile (d) (20.0g, 102mmol) at room temperature to ultra-dry tetrahydrofuran (400mL), slowly add sodium bis(trimethylsilyl)amide dropwise at -78℃ (NaHMDS) (127 mL, 251 mmol). After the dropwise addition was completed, the reaction was carried out at -78°C for 0.5 hour. Then methyl iodide (16.8 mL, 270 mmol) was slowly added dropwise and reacted at -78°C for 2 hours. After the reaction was completed, the temperature was raised to -10°C, saturated ammonium chloride (150 mL) was slowly added for quenching, and then extracted with ethyl acetate (200 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 40:1) to obtain the target intermediate 2-(3-bromophenyl)-2-methylpropionitrile (e) ( 22.5g, yield: 98%), traits: yellow oil.

Compound intermediate f:

Add 2-(3-bromophenyl)-2-methylpropionitrile (e) (29.0g, 129.5mmol) to ultra-dry tetrahydrofuran (300mL) at room temperature, slowly add borane-tetrahydrofuran ( 389 mL, 385.5 mmol). After the dripping is completed, it is heated to 70°C and reacted for 3.0 hours. After the reaction was completed, the temperature was lowered to 0°C and methanol (150 mL) was slowly added, and then concentrated hydrochloric acid (35 mL) was added dropwise, heated to 70°C, and reacted for 2.0 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, adjusted to pH 10 with a 10% aqueous sodium hydroxide solution, and then extracted three times with ethyl acetate (400 mL), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: dichloromethane: methanol = 30:1, 0.5% triethylamine) to obtain the target intermediate 2-(3-bromophenyl)-2-methylpropane -1-amine (f) (24.2 g, yield: 82%), properties: yellow oil.

LCMS: Rt: 0.742 min; MS m/z (ESI): 228.0 and 230.0 [M+H].

Compound intermediate g:

At room temperature, 2-(3-bromophenyl)-2-methylpropane-1-amine (f) (24.0g, 105.3mmol) and triethylamine (31.9g, 315.9mmol) were added to dichloromethane (320mL ), cooled to 0°C, and slowly added methyl chloroformate (14.9 g, 157.9 mmol) dropwise. After the addition is complete, react at room temperature for 1.0 hour. After the reaction was completed, it was cooled to 0°C, water (300 mL) was added for quenching, and then extracted with dichloromethane (200 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 5:1) to obtain the target intermediate methyl (2-(3-bromophenyl)-2-methylpropyl) Carbamate (g) (18.9 g, yield: 63.5%), properties: yellow oil.

LCMS: Rt: 1.827 min; MS m/z (ESI): 327.0 and 329.0 [M+H+CH ₃ CN].

Compound Intermediate 2:

At room temperature, methyl(2-(3-bromophenyl)-2-methylpropyl)carbamate (g) (18.9g, 66.1mmol) was added to trifluoromethanesulfonic acid (TfOH) (240mL In), the reaction system was heated to 100°C and stirred for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, slowly added to ice water (300 mL), and then extracted three times with dichloromethane (200 mL), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase chromatography column (eluent gradient: acetonitrile/water=34%, containing 0.1% trifluoroacetic acid) to obtain the target intermediate 6-bromo-4,4-dimethyl-3,4-bis Hydroisoquinoline-1(2H)-one (Intermediate 2) (9.1g, yield: 54.2%), properties: white solid.

LCMS: Rt: 1.505 min; MS m/z (ESI): 254.0 and 256.0 [M+H].

¹ H NMR (400MHz, CD ₃ OD): δ7.84(d,J=8.4Hz,1H), 7.59(d,J=2.0Hz,1H), 7.52(dd,J=8.4,2.0Hz,1H) , 3.28(s, 2H), 1.34(s, 6H).

Example 3, 2-(2-hydroxy-2-(1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(2-hydroxy-7-azaspiro[3.5 ]Nonane-7-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 1)

Compound 1-1:

At room temperature, 6-bromo-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one (Intermediate 2) (1.07g, 4.22mmol) was added to the ultra-dry N, In N-dimethylformamide (60 mL), under nitrogen protection, sodium hydride (0.253 g, 6.33 mmol) was added in batches at 0°C. After the addition, the reaction was stirred at room temperature for 0.5 hours, and then tert-butyl 3-(oxiran-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (Intermediate 1) ( 2.9g, 10.54mmol) was added. Heat to 40°C and react for 16.0 hours. After the reaction is complete, cool to 0°C, slowly add saturated ammonium chloride solution (60 mL) for quenching, and then extract three times with ethyl acetate (100 mL). After the organic phases are combined, they are washed twice with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by reverse phase chromatography column (eluent gradient: acetonitrile/water=62%, containing 0.1% trifluoroacetic acid) to obtain the crude product target intermediate 1-((6-bromo-4,4- Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)methyl)-1,5,10,10a-tetrahydro-3H-oxazolo[3,4- b] Isoquinolin-3-one (1-1) (2.02 g).

LCMS: Rt: 1.919 min; MS m/z (ESI): 455.0 and 457.0 [M+H].

Compound 1-2:

At room temperature, 1-((6-bromo-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinolin-2(1H)-yl)methyl)-1,5,10, 10a-Tetrahydro-3H-oxazolo[3,4-b]isoquinolin-3-one (1-1) (2.02g, 4.44mmol), [1,1'-bis(diphenylphosphine) Ferrocene] palladium dichloride (163 mg, 0.22 mmol) and potassium acetate (1.3 g, 13.32 mmol) were added to absolute ethanol (40 mL), replaced with CO three times, and heated to 70° C. to react for 16.0 hours. After the reaction was completed, it was cooled to room temperature, the reaction solution was concentrated under reduced pressure, saturated brine (100 mL) was added, and then extracted twice with ethyl acetate (100 mL), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 2:1,) to obtain the target intermediate ethyl-4,4-dimethyl-1-carbonyl-2-(( 3-carbonyl-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-1-yl)methyl)-1,2,3,4-tetrahydroiso Quinoline-6-carboxylate (1-2) (753 mg, yield: 38%).

LCMS: Rt: 1.847min; MS m/z(ESI): 449.0[M+H].

Compound 1-3:

Ethyl-4,4-dimethyl-1-carbonyl-2-((3-carbonyl-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]iso Quinolin-1-yl) methyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxylate (1-2) (750mg, 1.67mmol) was added to tetrahydrofuran (9.5mL), methanol To the mixed solution of (9.5 mL) and water (9.5 mL), sodium hydroxide (268 mg, 6.70 mmol) was added, heated to 70°C, and reacted for 16.0 hours. After the reaction was completed, the reaction system was cooled to room temperature, and Boc anhydride (1.09 g, 5.01 mmol) was added to react for 1.0 hour. After the reaction was complete, the reaction system was cooled to 0°C, the pH of the reaction solution was adjusted to 5.0 with 1N aqueous hydrochloric acid, and then extracted with ethyl acetate (50 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure . The residue was subjected to reverse phase chromatography column (eluent gradient: acetonitrile/water = 52%) to obtain the target intermediate 2-(2-(2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydro (Isoquinolin-3-yl)-2-hydroxyethyl)-4,4-dimethyl-1-carbonyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (1-3 ) (772mg, yield: 93%).

LCMS: Rt: 1.851min; MS m/z(ESI): 495[M+H].

Compound 1-4:

At room temperature, 2-(2-(2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2-hydroxyethyl)-4,4-di Methyl-1-carbonyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (1-3) (180mg, 0.3mmol), 2-(7-azabenzotriazole) -N,N,N',N'-tetramethylurea hexafluorophosphate (231mg, 0.6mmol) and N,N-diisopropylethylamine (157mg, 1.22mmol) are added to the ultra-dry N,N- To dimethylformamide (2.5 mL), 7-azaspirocyclo[3.5]non-2-ol hydrochloride (77 mg, 0.36 mmol) was added, and the reaction was carried out at room temperature for 1.0 hour. After the reaction was complete, water (25 mL) was added, and then extracted with dichloromethane (20 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 1:1) to obtain the target intermediate 3-(1-hydroxy-2-(6-(2-hydroxy-7-nitrogen) Heterosspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinolin-2(1H)-yl)ethyl)-3,4- Dihydroisoquinoline-2(1H)-tert-butyl carboxylate (1-4) (96 mg, yield: 46.4%).

LCMS: Rt: 1.68min; MS m/z(ESI): 618.3.

Compound 1:

To the intermediate 3-(1-hydroxy-2-(6-(2-hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3, 4-dihydroisoquinoline-2(1H)-yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (1-4) (50mg) dichloro Trifluoroacetic acid (TFA) (1 mL) was added to the methane (2 mL) solution, and the reaction solution was stirred at room temperature for 1 hour. After the reaction, it was concentrated under reduced pressure and purified by high performance liquid chromatography. The eluent gradient:

The final product 2-(2-hydroxy-2-(1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(2-hydroxy-7-azaspiro[3.5] Nonane-7-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 1) (11 mg, yield: 28%). HPLC showed that the compound contained two sets of peaks, and chiral HPLC analysis showed that the compound contained 4 isomers.

¹ H NMR (400MHz, DMSO-d6): δ9.31-9.72 (m, 1H), 9.04 (s, 1H), 7.89-7.93 (m, 1H), 7.36 (s, 1H), 7.21-7.31 (m ,5H),4.08-4.40(m,4H),3.85-4.07(m,2H),3.45-3.59(m,6H),3.33-3.43(m,1H),3.02-3.29(m,4H),2.15 (s,2H),1.47-1.54(m,4H),1.42(s,2H),1.29-1.32(m,6H).

HPLC: Rt: 6.62min 37.598%, Rt: 6.70min 62.402%, @214nm,

Rt: 6.62min 37.331%, Rt: 6.70min 62.669%, @254nm

LCMS: Rt: 1.28min; MS m/z(ESI): 518.4[M+H].

Example 4, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Base-6-(1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one hydrochloride and 2-((S) -2-Hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(1,7-bis Preparation of azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one hydrochloride mixture (compound 2)

Compound 2-1:

At room temperature, 2-(2-(2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2-hydroxyethyl)-4,4-di Methyl-1-carbonyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (1-3) (150mg, 0.304mmol), 2-(7-azabenzotriazole) -N,N,N',N'-tetramethylurea hexafluorophosphate (231mg, 0.608mmol) and N,N-diisopropylethylamine (157mg, 1.22mmol) are added to the ultra-dry N,N- Dimethylformamide (2.5 mL), then 1,7-diazaspiro[3.5]nonane-1-carboxylic acid tert-butyl ester (82.0 mg, 0.364 mmol) was added, and the reaction was carried out at room temperature for 1.0 hour. After the reaction was completed, saturated brine (30 mL) was added, extracted twice with ethyl acetate (30 mL), the organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 1:2) to obtain the target intermediate tert-butyl 3-(2-(6-(1-(tert-butoxycarbonyl) )-1,7-diazaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl) -1-Hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (2-1) (195 mg, yield: 91.5%).

LCMS: Rt: 1.851min; MS m/z(ESI): 703.0[M+H].

Compound 2-1-PK1 and Compound 2-1-PK2:

Add tert-butyl 3-(2-(6-(1-(tert-butoxycarbonyl)-1,7-diazaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl at room temperature -1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate ( 2-1) (195mg) is resolved by chiral chromatography column (SFC), resolution conditions:

仪器instrument	Waters SFC150Waters SFC150
流动相Mobile phase	55％EtOH in CO ₂ 55% EtOH in CO ₂
色谱柱Column	MICMIC
进样量Injection volume	3mL3mL
流速Flow rate	70g/min70g/min
循环进样时间Cycle injection time	4.5min4.5min
溶样浓度Dissolved sample concentration	140mg/40mL140mg/40mL

A pair of enantiomers 2--1-PK1 (main peak, 81 mg) was obtained; the other pair of enantiomers 2--1-PK2 (39 mg).

Chiral-HPLC: 2-1-PK1, Rt: 1.987min; 2-1-PK2, Rt: 3.642min.

Compound 2:

Trifluoroacetic acid (1 mL) was added to a dichloromethane (2 mL) solution of Intermediate 2--1-PK1 (81 mg) at 0°C, and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated under reduced pressure, and the residue was purified by HPLC (the chromatographic conditions were the same as in Example 3) to obtain the target compound 2-((R)-2-hydroxy-2-((S)-1,2 ,3,4-Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(1,7-diazaspiro[3.5]nonane-7-carbonyl)-3 ,4-Dihydroisoquinoline-1(2H)-one hydrochloride and 2-((S)-2-hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinoline) -3-yl)ethyl)-4,4-dimethyl-6-(1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1 (2H)-ketone hydrochloride mixture (compound 2) (2.31 mg, yield: 4%).

¹ H NMR (400MHz, CD ₃ OD): δ8.11-8.01 (m, 1H), 7.48 (d, J = 1.2 Hz, 1H), 7.40 (d, J = 7.6 Hz, 1H), 7.34-7.19 ( m, 4H), 5.69 (s, 1H), 4.52-4.30 (m, 3H), 4.21 (s, 1H), 4.02-3.86 (m, 2H), 3.71-3.56 (m, 4H), 3.48 (s, 1H),3.32-3.28(m,4H),3.07(d,J=6.4Hz,2H),2.49-2.91(m,2H),2.85-2.11(m,2H),1.40(d,J=7.2Hz ,6H).

HPLC: 97.881%@214nm, 97.094%@254nm

LCMS: Rt: 0.436min; MS m/z(ESI): 503.2[M+H].

Example 5, 2-((R)-2-hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Base-6-(1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one hydrochloride and 2-((S) -2-Hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(1,7-bis Preparation of azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one hydrochloride mixture (compound 3)

Compound 3:

Trifluoroacetic acid (1 mL) was added to a dichloromethane (2 mL) solution of Intermediate 2--1-PK2 (39 mg) at 0°C, and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated under reduced pressure, and the residue was purified by HPLC (the chromatographic conditions were the same as in Example 3) to obtain the target compound 2-((R)-2-hydroxy-2-((R)-1,2 ,3,4-Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(1,7-diazaspiro[3.5]nonane-7-carbonyl)-3 ,4-Dihydroisoquinoline-1(2H)-one hydrochloride and 2-((S)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinoline) -3-yl)ethyl)-4,4-dimethyl-6-(1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1 (2H)-ketone hydrochloride mixture (compound 3) (6.08 mg, yield: 22%).

¹ H NMR (400MHz, CD ₃ OD): δ8.05(t,J=7.2Hz,14.8Hz,1H), 7.47(d,J=1.2Hz,1H), 7.40(d,J=7.6Hz,1H ), 7.34-7.20(m, 4H), 5.69(s, 1H), 4.42(s, 2H), 4.21(s, 1H), 4.15-4.04(m, 2H), 3.99-3.80(m, 2H), 3.72-3.43 (m, 6H), 3.27-3.18 (m, 2H), 3.14-2.95 (m, 2H), 2.50-2.35 (m, 2H), 2.30-2.12 (m, 2H), 1.39 (s, 6H) ).

HPLC: 99.405%@214nm, 99.108%@254nm

LCMS: Rt: 0.594min; MS m/z(ESI): 503.2[M+H].

Example 6, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Base-6-(6-azaspiro[2.5]octane-6-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one and 2-((S)-2-hydroxy-2 -((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(6-azaspiro[2.5]octane- Preparation of 6-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one mixture (compound 4)

Compound 4-1:

At room temperature, 2-(2-(2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2-hydroxyethyl)-4,4-di Methyl-1-carbonyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (1-3) (150mg, 0.3mmol), 2-(7-azabenzotriazole) -N,N,N',N'-tetramethylurea hexafluorophosphate (231mg, 0.6mmol) and N,N-diisopropylethylamine (157mg, 1.216mmol) are added to the ultra-dry N,N- Dimethylformamide (2.5 mL), then 6-azaspirocyclo[2.5]octane hydrochloride (54 mg, 0.36 mmol) was added, and the reaction was carried out at room temperature for 1.0 hour. After the reaction was completed, water (25 mL) was added, and then extracted with dichloromethane (200 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 1:1) to obtain the target intermediate tert-butyl 3-(2-(4,4-dimethyl-1-carbonyl) -6-(6-Azaspiro[2.5]octane-6-carbonyl)-3,4-dihydroisoquinolin-2(1H)-yl)-1-hydroxyethyl)-3,4-di Hydroisoquinoline-2(1H)-carboxylate (4-1) (140 mg, yield: 83.5%).

Compound 4--1-PK1 and 4--1-PK-2:

Add tert-butyl 3-(2-(4,4-dimethyl-1-carbonyl-6-(6-azaspiro[2.5]octane-6-carbonyl)-3,4-dihydroiso Quinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (4-1) was resolved by chiral column (resolution conditions As in Example 4), two pairs of enantiomers 4--1-PK1 (main peak) and 4--1-PK2 were obtained.

LCMS: Rt: 1.90min; MS m/z(ESI): 588.3[M+H].

Chiral-HPLC: 4-1-PK1, Rt: 1.86min.

Compound 4:

Trifluoroacetic acid (1 mL) was added to a dichloromethane (2 mL) solution of Intermediate 4--1-PK1 (112 mg), and the reaction solution was stirred at room temperature for 1 hour. After the reaction is completed, it is cooled in an ice bath, and the pH value is adjusted to 9 by adding sodium carbonate aqueous solution. Extract with ethyl acetate, wash the organic phase with saturated brine, dry and filter with anhydrous sodium sulfate, and concentrate the filtrate under reduced pressure to obtain the final product 2-((R)-2-hydroxy-2-((S)-1,2) ,3,4-Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(6-azaspiro[2.5]octane-6-carbonyl)-3,4- Dihydroisoquinolin-1(2H)-one and 2-((S)-2-hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl Yl)-4,4-dimethyl-6-(6-azaspiro[2.5]octane-6-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one mixture (compound 4) (25mg, yield: 27%).

¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.6 Hz, 1H), 7.44 (d, J = 1.2 Hz, 1H) 7.38 (dd, J = 8 Hz, 1.2 Hz, 1H), 7.10-7.16(m,3H),7.05-7.07(m,1H),4.01-4.12(m,3H),3.94(dd,J=12.8Hz,4Hz,1H), 3.80(s,2H),3.55- 3.69(m,2H),3.41(s,2H),2.81-3.02(m,3H),1.51(s,2H),1.33-1.39(m,8H),0.41(d,J=8.8Hz,4H) .

HPLC: 95.959%@214nm, 96.363%@254nm

LCMS: Rt: 0.86min; MS m/z(ESI): 488.3[M+H].

Example 7, 2-((S)-2-hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(2-hydroxy -7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 5) and 2 -((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(2-hydroxy-7-aza Preparation of spiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 6)

Compound 5-1-1:

At room temperature, 6-bromo-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one (Intermediate 2) (1.1g, 4.34mmol) was added to the ultra-dry N, To N-dimethylformamide (60mL), add sodium hydride (0.26g, 6.5mmol) in batches at 0°C. After the addition is complete, stir at room temperature for 0.5 hours, and then add tert-butyl 3-(ethylene oxide -2-yl)-3,4-dihydroisoquinoline-2(1H)carboxylate (Intermediate 1) (1.8 g, 6.54 mmol). The reaction was heated at 40°C for 16.0 hours. HPLC and LCMS showed two sets of product peaks, two pairs of enantiomers respectively. After the reaction was completed, ice water (60 mL) was slowly added for quenching, extracted twice with ethyl acetate (100 mL), the organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was added with ethyl acetate (20mL), the solid precipitated out by ultrasound, the filter cake was collected by filtration and recrystallized from ethyl acetate twice to obtain one pair of enantiomers, (1R,10aS)-1-((6-bromo -4,4-Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)methyl)-1,5,10,10a-tetrahydro-3H-oxazolo [3,4-b]isoquinolin-3-one and (1S,10aR)-1-((6-bromo-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinoline -2(1H)-yl)methyl)-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-3-one mixture (5-1-PK1 ), is the main peak of the two groups of peaks in HPLC (425 mg, yield: 23%).

LCMS:Rt:1.92min; MS m/z(ESI): 455.0 and 457.0[M+H].

Compound 5-2:

Compound 5-PK1 (425mg, 0.94mmol), [1,1'-bis(diphenylphosphine)ferrocene] palladium dichloride (40mg, 0.05mmol) and potassium acetate (196mg, 1.88) at room temperature mmol) was added to absolute ethanol (10 mL), CO was replaced 3 times, and the mixture was heated to 70° C. and reacted for 16.0 hours. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated under reduced pressure, saturated brine (50 mL) was added, and then extracted three times with ethyl acetate (50 mL). The organic phases were combined and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. Get a pair of enantiomers, 4,4-dimethyl-1-carbonyl-2-(((1R,10aS)-3-carbonyl-1,5,10,10a-tetrahydro-3H-oxazole And [3,4-b]isoquinolin-1-yl)methyl)-1,2,3,4-tetrahydroisoquinolin-6-carboxylic acid ethyl ester and 4,4-dimethyl-1 -Carbonyl-2-(((1R,10aS)-3-carbonyl-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-1-yl)methyl ) A mixture of ethyl 1,2,3,4-tetrahydroisoquinoline-6-carboxylate (5-2) (580 mg).

LCMS: Rt: 1.895min; MS m/z(ESI): 449.0[M+H].

Compound 5-3:

Compound 5-2 (580mg, 1.30mmol) was added to a mixed solution of tetrahydrofuran (7mL), methanol (7mL) and water (7mL) at room temperature, sodium hydroxide (207mg, 5.19mmol) was added, and the reaction was carried out at 70°C for 16.0 hours . After the reaction was completed, the reaction system was cooled to room temperature, and Boc anhydride (1.13 g, 5.19 mmol) was added to react for 1.0 hour. After the reaction was complete, the reaction system was cooled to 0°C, the pH of the reaction solution was adjusted to 5.0 with 1N aqueous hydrochloric acid, and then extracted with ethyl acetate (50 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure . The residue was subjected to reverse phase chromatography column (eluent gradient: acetonitrile/water=52%) to obtain the target intermediate, 2-((R)-2-((S)-2-(tert-butoxycarbonyl)-1 ,2,3,4-Tetrahydroisoquinolin-3-yl)-2-hydroxyethyl)-4,4-dimethyl-1-carbonyl-1,2,3,4-tetrahydroisoquinoline -6-carboxylic acid and 2-((S)-2-((R)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2 -Hydroxyethyl)-4,4-dimethyl-1-carbonyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid mixture (5-3) (363mg, two-step yield : 79%).

LCMS: Rt: 1.799min; MS m/z(ESI): 495.2[M+H].

Compound 5-4:

Compound 5-3 (180mg, 0.3mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (231mg, 0.6mmol) at room temperature mmol) and N,N-diisopropylethylamine (157mg, 1.216mmol) were added to ultra-dry N,N-dimethylformamide (2.5mL), and then 7-azaspiro ring [3.5] non- 2-alcohol hydrochloride (77mg, 0.36mmol), react at room temperature for 1.0 hour. After the reaction was completed, water (25 mL) was added, and the mixture was extracted three times with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 1:1) to obtain the target intermediate, tert-butyl (R)-3-((S)-1-hydroxy-2 -(6-(2-Hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H )-Yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate and tert-butyl(S)-3-((R)-1-hydroxy-2-(6- (2-Hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl) Ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate mixture (5-4) (90 mg, yield: 43.5%).

LCMS: Rt: 1.68min; MS m/z(ESI): 618.3[M+H].

Compounds 5-4-PK1 and 5-4-PK2:

The compound 5-4 was resolved by chiral chromatography column, resolution conditions:

仪器instrument	Waters SFC150Waters SFC150
流动相Mobile phase	55％EtOH in CO ₂ 55% EtOH in CO ₂
色谱柱Column	WHELKWHELK
进样量Injection volume	7mL7mL
流速Flow rate	80g/min80g/min
循环进样时间Cycle injection time	5min5min
溶样浓度Dissolved sample concentration	50mg/40mL50mg/40mL

Two single-configuration compounds are obtained. The peak 1 compound is tert-butyl (R)-3-((S)-1-hydroxy-2-(6-(2-hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4 ,4-Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (5-4-PK1), peak 2 compound is tert-butyl (S)-3-((R)-1-hydroxy-2-(6-(2-hydroxy-7-azaspiro[3.5]nonane -7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)ethyl)-3,4-dihydroisoquinoline-2 (1H)-Carboxylic acid ester (5-4-PK2).

LCMS: Rt: 1.68min; MS m/z(ESI): 618.3[M+H].

Chiral-HPLC: 5-4-PK1, Rt: 2.79min; 5-4-PK2, Rt: 3.76min.

Compound 5:

At room temperature, the compound tert-butyl (R)-3-((S)-1-hydroxy-2-(6-(2-hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4 ,4-Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate Trifluoroacetic acid (1 mL) was added to a solution of (5-4-PK1) (24 mg) in dichloromethane (2 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated under reduced pressure, and purified by high performance liquid chromatography (the chromatographic conditions were the same as in Example 3) to obtain the final product 2-((S)-2-hydroxy-2-((R)-1,2,3,4- Tetrahydroisoquinolin-3-yl)ethyl)-6-(2-hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-3,4-bis Hydroisoquinoline-1(2H)-one hydrochloride (Compound 5) (5 mg, yield: 27%).

¹ H NMR (400MHz, CD ₃ OD): δ8.03(d,J=8Hz,1H),7.43(s,1H),7.35(d,J=8Hz,1H),7.21-7.31(m,4H) ,4.21-4.50(m,4H),3.98(dd,J=14Hz,4.8Hz,1H),3.58-3.70(m,6H),3.28-3.35(m,4H),2.29-2.31(m,2H) ,1.68-1.72(m,4H),1.54(s,2H),1.39(d,J=8Hz,6H).

HPLC: 98.233%@214nm, 98.402%@254nm

LCMS: Rt: 0.74min; MS m/z(ESI): 518.5[M+H].

Compound 6:

To the compound tert-butyl (S)-3-((R)-1-hydroxy-2-(6-(2-hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4- Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (5- Trifluoroacetic acid (1 mL) was added to a dichloromethane (2 mL) solution of 4-PK2) (21 mg), and the reaction solution was stirred at room temperature for 1 hour. After the reaction is completed, it is concentrated under reduced pressure, and purified by high performance liquid chromatography (the chromatographic conditions are the same as in Example 3) to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4- Tetrahydroisoquinolin-3-yl)ethyl)-6-(2-hydroxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-3,4-bis Hydroisoquinoline-1(2H)-one hydrochloride (Compound 6) (4 mg, yield: 27%).

¹ H NMR (400MHz, CD ₃ OD): δ8.03(d,J=8Hz,1H),7.43(s,1H),7.35(d,J=8Hz,1H),7.21-7.31(m,4H) ,4.31-4.50(m,2H),4.28-4.30(m,2H),3.98(dd,J=14Hz,4.8Hz,1H),3.58-3.70(m,6H),3.28-3.32(m,4H) , 2.29-2.31 (m, 2H), 1.67-1.74 (m, 4H), 1.54 (s, 2H), 1.39 (d, J = 8Hz, 6H).

HPLC: 99.168%@214nm, 99.232%@254nm

LCMS: Rt: 0.74min; MS m/z(ESI): 518.5[M+H].

Example 8, 2-((S)-2-hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Base-6-(2,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 7) and 2- ((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(2 Preparation of ,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 8)

Compound 7-1:

Compound 5-3 (140mg, 0.3mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (231mg, 0.6mmol) at room temperature mmol) and N,N-diisopropylethylamine (157mg, 1.216mmol) were added to ultra-dry N,N-dimethylformamide (2.5mL), added 2,7-diazaspiro[3.5]non Tert-butyl alkane-2-carboxylate (77 mg, 0.36 mmol) was reacted at room temperature for 1.0 hour. After the reaction was completed, water (25 mL) was added, and the mixture was extracted three times with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 1:1) to obtain the target intermediate, tert-butyl (R)-3-((S)-2-(6- (2-(tert-Butoxycarbonyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-dihydroisoquine Lin-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate and tert-butyl(S)-3-((R)- 2-(6-(2-(tert-butoxycarbonyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4 -Dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate mixture (7-1) (166mg, Yield: 83.4%).

LCMS: Rt: 1.88min; MS m/z(ESI): 703.4[M+H].

Compound 7-1-PK1 and 7-1-PK2:

Compound 7-1 was resolved by chiral resolution (the resolution conditions were the same as those in Example 7), and two compounds with a single configuration were obtained. The compound of peak 1 was tert-butyl (R)-3-((S)-2-( 6-(2-(tert-Butoxycarbonyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-carbonyl-3,4-di Hydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (7-1-PK1) (45mg), peak 2 The compound is tert-butyl (S)-3-((R)-2-(6-(2-(tert-butoxycarbonyl)-2,7-diazaspiro[3.5]nonane-7- (Carbonyl)-4,4-Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline- 2(1H)-carboxylate (7-1-PK2) (107 mg).

LCMS: Rt: 1.88min; MS m/z(ESI): 703.4[M+H].

Chiral-HPLC: 7-1-PK1, Rt: 3.13min; 7-1-PK2, Rt: 4.05min.

Compound 7:

To the compound tert-butyl (R)-3-((S)-2-(6-(2-(tert-butoxycarbonyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl )-4,4-Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2 Trifluoroacetic acid (1 mL) was added to a solution of (1H)-carboxylate (7-1-PK1) (24 mg) in dichloromethane (2 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction is completed, it is concentrated under reduced pressure, and purified by high performance liquid chromatography (chromatographic conditions are the same as in Example 3) to obtain the final product 2-((S)-2-hydroxy-2-((R)-1,2,3,4- Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(2,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydro Isoquinoline-1(2H)-one hydrochloride (Compound 7) (5 mg, yield: 27%).

¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.6 Hz, 1H), 7.45 (s, 1H), 7.37 (d, J = 7.6 Hz, 1H), 7.21-7.31 (m, 4H),4.35-4.50(m,3H),3.92-4.01(m,5H),3.57-3.73(m,6H),3.30-3.36(m,4H),1.87-1.99(m,4H),1.39- 1.41(d,J=7.6Hz,6H).

HPLC: 99.704%@214nm, 99.917%@254nm

LCMS: Rt: 0.42min; MS m/z(ESI): 503.3[M+H].

Compound 8:

To the compound tert-butyl (S)-3-((R)-2-(6-(2-(tert-butoxycarbonyl)-2,7-diazaspiro[3.5]nonane-7-carbonyl )-4,4-Dimethyl-1-carbonyl-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2 Trifluoroacetic acid (1 mL) was added to a solution of (1H)-carboxylate (7-1-PK2) (21 mg) in dichloromethane (2 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction, it was concentrated under reduced pressure and purified by high performance liquid chromatography (chromatographic conditions were the same as in Example 3) to obtain the final product 2-((R)-2-hydroxy-2-((S)-1,2,3,4- Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(2,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydro Isoquinolin-1(2H)-one hydrochloride (Compound 8) (4 mg, yield: 27%).

HPLC: 96.311%@214nm, 99.784%@254nm

LCMS: Rt: 0.52min; MS m/z(ESI): 503.3[M+H].

Example 9, 2-((S)-2-hydroxy-2-((R)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Base-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one hydrochloride (compound 9) and 2 -((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-( Preparation of 2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 10)

Compound 9-1:

Compound 5-3 (135mg, 0.3mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (231mg, 0.6mmol) at room temperature mmol) and N,N-diisopropylethylamine (157mg, 1.216mmol) were added to ultra-dry N,N-dimethylformamide (2.5mL), and 7-azaspiro ring [3.5]non-2 was added -Ketone hydrochloride (77mg, 0.36mmol), react at room temperature for 1.0 hour. After the reaction was completed, water (25 mL) was added, and then extracted with dichloromethane (20 mL) three times. The organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase chromatography column (eluent gradient: petroleum ether: ethyl acetate = 1:1) to obtain the target intermediate, tert-butyl (R)-3-((S)-2-(4, 4-Dimethyl-1-carbonyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-2(1H)-yl) -1-Hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate and tert-butyl(S)-3-((R)-2-(4,4-dimethyl Group-1-carbonyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxy Ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate mixture (9-1) (153 mg, yield: 91.0%).

LCMS: Rt: 1.54min; MS m/z(ESI): 616.3[M+H].

Compound 9-1-PK1 and 9-1-PK2:

The compound 9-1 was subjected to chiral resolution (the resolution conditions were the same as in Example 4), and two compounds with a single configuration were obtained. Among them, the compound of peak 1 was tert-butyl (R)-3-((S)-2-( 4,4-Dimethyl-1-carbonyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-2(1H)- Yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (9-1-PK1)(24mg), peak 2 compound is tert-butyl(S)- 3-((R)-2-(4,4-Dimethyl-1-carbonyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-di Hydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (9-1-PK2) (88 mg).

LCMS: Rt: 1.54min; MS m/z(ESI): 616.3[M+H].

Chiral-HPLC: 9-1-PK1, Rt: 3.12min; 9-1-PK2, Rt: 3.99min.

Compound 9:

To the intermediate tert-butyl (R)-3-((S)-2-(4,4-dimethyl-1-carbonyl-6-(2-carbonyl-7-azaspiro[3.5]nonane- 7-carbonyl)-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (9 Trifluoroacetic acid (1 mL) was added to a dichloromethane (2 mL) solution of -1-PK1) (24 mg), and the reaction solution was stirred at room temperature for 1 hour. After the reaction is completed, it is concentrated under reduced pressure, and purified by high performance liquid chromatography (chromatographic conditions are the same as in Example 3) to obtain the final product 2-((S)-2-hydroxy-2-((R)-1,2,3,4- Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-di Hydroisoquinoline-1(2H)-one hydrochloride (Compound 9) (5 mg, yield: 27%).

¹ H NMR (400MHz, DMSO-d6): δ9.90 (d, J = 8Hz, 1H), 9.08 (d, J = 8 Hz, 1H), 7.92 (d, J = 7.6 Hz, 1H), 7.40 (s ,1H),7.33(dd,J=8Hz,1.2Hz,1H),7.21-7.28(m,4H),5.89(s,1H),4.32-4.38(m,2H),4.21-4.24(m,1H) ), 3.97-4.02(m,2H),3.57-3.60(m,6H),3.17-3.37(m,5H),2.84-2.89(m,4H),1.64-1.76(m,2H),1.32(d ,J=12.8Hz,6H).

HPLC: 98.504%@214nm, 98.008%@254nm

LCMS: Rt: 1.28min; MS m/z(ESI): 516.3[M+H].

Compound 10:

To the compound tert-butyl (S)-3-((R)-2-(4,4-dimethyl-1-carbonyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7 -Carbonyl)-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (9- Trifluoroacetic acid (1 mL) was added to a dichloromethane (2 mL) solution of 1-PK2) (21 mg), and the reaction solution was stirred at room temperature for 1 hour. After the reaction, it was concentrated under reduced pressure and purified by high performance liquid chromatography (chromatographic conditions were the same as in Example 3) to obtain the final product 2-((R)-2-hydroxy-2-((S)-1,2,3,4- Tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-6-(2-carbonyl-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-di Hydroisoquinoline-1(2H)-one hydrochloride (Compound 10) (4 mg, yield: 27%).

HPLC: 98.682%@214nm, 98.990%@254nm

LCMS: Rt: 1.28min; MS m/z(ESI): 516.3[M+H].

Example 10, 2-((R)-2-((S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2-hydroxyl (Ethyl)-4,4-Dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (Intermediate 3) synthesis

Compound h:

Disperse methyltriphenylphosphonium bromide (238g, 0.67mol) in anhydrous tetrahydrofuran (1.5L), cool to -70℃ under nitrogen protection, and slowly add sodium bis(trimethylsilyl)amide (334mL) dropwise , 0.67mol), control the temperature to be lower than -50°C, slowly rise to room temperature after the addition is completed, and stir for 2 hours. Re-cool to -70℃, slowly add tert-butyl(S)-3-formyl-3,4-dihydroisoquinoline-2(1H)-carboxylate (c) in tetrahydrofuran solution dropwise, control the temperature to be low At -50°C, slowly rise to room temperature overnight after the addition is complete. After the reaction was detected by TLC, it was cooled to 0°C, quenched by adding saturated ammonium chloride solution, 1N aqueous hydrochloric acid was slowly added to adjust the pH to 3-4, and ethyl acetate (500 mL) was added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was recrystallized by adding a mixed solvent (ethyl acetate/petroleum ether = 1/4), filtered to remove the precipitated triphenylphosphine oxide, and the filtrate was silica gel column chromatography (ethyl acetate/petroleum ether = 1/8) to obtain the target product (S)-3-vinyl-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (h) (85 g, 98%).

Chiral-HPLC:Rt:1.883

Compound i:

Dissolve (S)-3-vinyl-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (h) (25.9g, 0.1mol) in ethyl acetate/acetonitrile (500mL/ 500mL) solution, cool to 0°C, add an aqueous solution of sodium periodate (32.1g, 0.15mol) and ruthenium trichloride hydrate (1.6g, 7.7mmol) within 10 minutes, and stir the reaction solution at 0°C for 10 minutes minute. After the reaction is detected by TLC, it is quenched by adding saturated sodium thiosulfate solution (150mL), stirred for 30 minutes and then separated into layers. The organic phase is washed with saturated brine (200mL X 2), dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure The residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether = 1/2-1/1) to obtain the product (S)-3-((S)-1,2-dihydroxyethyl Yl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (i) (less polar product, 14g, yield 48%). Increase the polarity of the eluent (ethyl acetate/petroleum ether=2/1-1/0) to obtain the product (S)-3-((R)-1,2-dihydroxyethyl)-3,4 -Dihydroisoquinoline-2(1H)-tert-butyl carboxylate (i-1) (more polar product, 8g, yield 28%).

Compound j:

Add (S)-3-((S)-1,2-dihydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (i) (14g, 0.047mol ) Was dissolved in dichloromethane (150mL), triethylamine (9.90mL, 0.072mol) was added, and p-toluenesulfonyl chloride (10.0g, 0.052mol) was added in batches under stirring. The reaction solution was stirred at 40°C overnight . The reaction solution was cooled to room temperature, washed with saturated brine (100 mL × 2), and the organic phase was dried with anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (ethyl acetate/petroleum ether = 1/4) to obtain the target product (S)-3-((S)-1-hydroxy-2-(toluenesulfonyl (Oxo)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (j) (12.6 g, yield 59%).

Compound k:

Add (S)-3-((S)-1-hydroxy-2-(tosyloxy)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester ( j) (12.6g, 28.2mmol) was dissolved in N,N-dimethylformamide (150mL), sodium hydride (1.70g, 42.3mmol) was added in batches under nitrogen protection, and the reaction solution was stirred at 40°C for 1 hour . TLC detected that the reaction was complete. Cooled to 0℃, quenched with saturated brine dropwise, the reaction solution was directly purified by reversed-phase column (water/acetonitrile=50/10), the solution obtained by column chromatography was extracted with ethyl acetate (200mL X 3), the organic phase After drying with sodium sulfate and filtering, the filtrate was concentrated under reduced pressure to obtain intermediate (S)-3-((S)-oxiran-2-yl)-3,4-dihydroisoquinoline-2(1H)- Tert-butyl carboxylate (k) (6.5 g, yield 72%).

Compound m:

At room temperature, 6-bromo-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one (Intermediate 2) (1.07g, 4.22mmol) was added to the ultra-dry N, In N-dimethylformamide (40mL), under nitrogen protection, sodium hydride (0.253g, 6.33mmol) was added in batches at 0°C. After the addition, the reaction was stirred at 40°C for 2 hours, and then (S)-3-( (S)-Ethylene oxide-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (k) (2.9 g, 10.54 mmol) was added. Heat to 40°C and react for 16.0 hours. After the reaction was completed, the reaction solution was cooled to 0°C, slowly poured into saturated ammonium chloride solution (200mL) for quenching, filtered, and the filter cake was slurried with methanol to obtain the crude product (1R,10aS)-1-((6-bromo-4) ,4-Dimethyl-1-oxo-3,4-dihydroisoquinoline-2(1H)-yl)methyl)-1,5,10,10a-tetrahydro-3H-oxazolo[ 3,4-b]isoquinolin-3-one (m) (350 mg, yield 18%).

LCMS: Rt: 1.738 min; MS m/z (ESI): 455.1, 457.1 [M+H].

Diffusion method (dichloromethane-methyl tert-butyl ether system) was used to obtain a single crystal sample of compound m. The obtained sample was subjected to X-ray crystal diffraction analysis at the Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences to determine the absolute structure of compound m. type. The test results are shown in Table 1 and Figure 1.

Table 1 Sample and crystal data and structure refinement of compound m

From the X-ray crystal diffraction results, it can be confirmed that the absolute configuration of compound m is as follows:

Compound n:

Add (1R,10aS)-1-((6-bromo-4,4-dimethyl-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)methyl) at room temperature -1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-3-one (m) (1.84g, 4mmol), [1,1'-bis(two Phenylphosphine) ferrocene] dichloride palladium dichloromethane complex (170mg, 0.2mmol) and potassium acetate (1.3g, 12mmol) were added to absolute ethanol (40mL), CO replaced 3 times, heated to React at 70°C for 3 hours. After the reaction was completed, it was cooled to room temperature, the reaction solution was concentrated under reduced pressure, saturated brine (100 mL) was added, and then extracted twice with ethyl acetate (100 mL), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase chromatography column (eluent gradient: petroleum ether/ethyl acetate = 1/1) to obtain 4,4-dimethyl-1-oxo-2-(((1R,10aS)- 3-oxo-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-1-yl)methyl)-1,2,3,4-tetrahydro Ethyl isoquinoline-6-carboxylate (n) (crude product 1.3 g, yield: 72%).

LCMS: Rt: 1.892min; MS m/z(ESI): 449.2[M+H].

Intermediate 3:

4,4-Dimethyl-1-oxo-2-(((1R,10aS)-3-oxo-1,5,10,10a-tetrahydro-3H-oxazolo[3, 4-b]isoquinolin-1-yl)methyl)-1,2,3,4-tetrahydroisoquinolin-6-carboxylic acid ethyl ester (n) (750mg, 1.67mmol) was added to tetrahydrofuran (9.5 mL), methanol (9.5 mL) and water (9.5 mL) mixed solvent, then sodium hydroxide (268 mg, 6.70 mmol) was added, heated to 70°C, and reacted for 16.0 hours. The reaction system was cooled to room temperature, Boc anhydride (430 mg, 2 mmol) was added, and the reaction was carried out at room temperature for 1.0 hour. After the reaction was complete, the reaction system was cooled to 0°C, the pH of the reaction solution was adjusted to 5.0 with 1N aqueous hydrochloric acid, and extracted with ethyl acetate (50 mL) three times, the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase chromatography (eluent: 52% acetonitrile in water) to obtain the target product 2-((R)-2-((S)-2-(tert-butoxycarbonyl)-1,2,3 ,4-Tetrahydroisoquinolin-3-yl)-2-hydroxyethyl)-4,4-dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-6- Carboxylic acid (Intermediate 3) (430 mg, yield: 87%).

LCMS: Rt: 1.093min; MS m/z(ESI): 495.4[M+H].

Example 11, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(2-methyl Oxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 12) Preparation

Compound 12-2:

Add tert-butyl 2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (12-1) (1.0g, 4.18mmol) to methanol (10.0mL) at room temperature, at 0°C Sodium borohydride (474mg, 12.5mmol) was added and reacted at room temperature for 1.5 hours. The reaction was quenched by adding aqueous ammonium chloride solution (20.0 mL), and extracted twice with ethyl acetate (50.0 mL). The organic phases were combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (petroleum ether: ethyl acetate = 1:1) to obtain the target intermediate 2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (12- 2) (970mg, yield: 96%).

LCMS:Rt:1.446min; MS m/z(ESI):142.1[M+H-Boc].

Compound 12-3:

Add 2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (12-2) (300mg, 1.245mmol) to ultra-dry N,N-dimethylformamide at room temperature (3.0 mL), then sodium hydride (200 mg, 4.98 mmol) was added to react at room temperature for 0.5 hours, methyl iodide (1.06 mg, 7.47 mmol) was added, and the reaction was continued for 1.5 hours. After the reaction was completed, water (50 mL) was added, and then it was extracted 3 times with ethyl acetate (40 mL). The organic phases were combined, washed 3 times with saturated brine (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (petroleum ether: ethyl acetate = 5:1) to obtain the target intermediate 2-methoxy-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester ( 12-3) (313 mg, yield: 85%).

Compound 12-4:

Add tert-butyl 2-methoxy-7-azaspiro[3.5]nonane-7-carboxylate (12-3) (313 mg, 1.23 mmol) to the 4N hydrochloric acid/dioxane solution at room temperature ( 3.0mL), react at room temperature for 1.5 hours, and concentrate the reaction solution under reduced pressure to obtain the intermediate 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) (200mg, yield: 85%) ).

LCMS:Rt:0.571min; MS m/z(ESI): 156.5[M+H].

Compound 12-5:

Add 2-((R)-2-((S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2-hydroxyethyl at room temperature Yl)-4,4-dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (Intermediate 3) (500mg, 1.01mmol), 2-(7 -Azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (570mg, 1.5mmol) and N,N-diisopropylethylamine (520mg, 4mmol) Add to ultra-dry N,N-dimethylformamide (10 mL). Then, 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) (230 mg, 1.2 mmol) was added, and the reaction was carried out at room temperature for 1.5 hours. After the reaction was completed, saturated brine (50 mL) was added, and the mixture was extracted twice with ethyl acetate (20 mL). The organic phases were combined and washed with saturated brine (50 mL) three times, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent: petroleum ether/ethyl acetate = 1/4) to obtain the target intermediate (S)-3-((R)-1-hydroxy-2-(6-( 2-Methoxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-oxo-3,4-dihydroisoquinoline-2(1H)- (Yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (12-5) (560 mg, yield: 88%).

LCMS: Rt: 1.734min; MS m/z(ESI): 632.4[M+H].

Compound 12:

(S)-3-((R)-1-hydroxy-2-(6-(2-methoxy-7-azaspiro[3.5]nonane-7-carbonyl)-4,4- Dimethyl-1-oxo-3,4-dihydroisoquinoline-2(1H)-yl)ethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (12-5) (560mg, 0.88mmol) was dissolved in 4N hydrochloric acid/dioxane solution (5mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction is complete, it is concentrated under reduced pressure, and the residue is purified by high performance liquid chromatography (eluent gradient as shown in the table below):

Obtain the target compound 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(2-methoxy Yl-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 12) ( 361mg, yield: 73%). ¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.9 Hz, 1H), 7.43 (s, 1H), 7.36 (d, J = 7.9 Hz, 1H), 7.33-7.20 (m, 4H), 4.49-4.28 (m, 3H), 3.99-3.95 (m, 2H), 3.70-3.56 (m, 6H), 3.30-3.24 (m, 4H), 3.21 (s, 3H), 2.26-2.24 ( m,2H),1.72-1.54(m,6H),1.38(d,J=7.6Hz,6H).

LCMS: Rt: 1.370min; MS m/z(ESI): 532.4[M+H].

Example 12, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of 6-(6-azaspiro[2.5]octane-6-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 11)

Referring to the synthesis method of Example 11, 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) was replaced with 6-azaspirocyclo[2.5]octane hydrochloride, Obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl -6-(6-Azaspiro[2.5]octane-6-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 11).

¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.9Hz, 1H), 7.45 (d, J = 1.6Hz, 1H), 7.38 (dd, J = 7.9Hz, 1.6Hz, 1H ),7.33-7.18(m,4H),4.51-4.36(m,2H),4.35-4.27(m,1H),3.97(dd,J=14.1Hz,4.8Hz,1H),3.81-3.79(m, 2H), 3.67-3.56 (m, 6H), 3.40 (s, 2H), 3.31-3.27 (m, 2H), 1.51-1.49 (m, 2H), 1.39 (d, J = 7.7 Hz, 6H), 0.45 -0.37(m,4H).

LCMS: Rt: 1.040min; MS m/z(ESI): 488.4[M+H].

Example 13, 7-(2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4 -Dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carbonyl)-7-azaspiro[3.5]nonane-2-carbonitrile (compound 13)

Compound 13-2:

Dissolve 2-cyano-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (13-1) (600mg, 2.4mmol) in dichloromethane (2mL) at room temperature, and slowly add trifluoro Acetic acid (1 mL), and the reaction solution was stirred at room temperature for 1 hour. After the completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain intermediate 7-azaspiro[3.5]nonane-2-carbonitrile trifluoroacetate (13-2) (crude product, 590 mg, yield: 100%). Used directly in the next step.

Compound 13-3:

Add 2-((R)-2-((S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2-hydroxyethyl at room temperature Yl)-4,4-dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (Intermediate 3) (990mg, 2mmol), 2-(7- Azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.14g, 3mmol) and N,N-diisopropylethylamine (520mg, 4mmol) were added To ultra-dry N,N-dimethylformamide (10 mL). Then add 7-azaspiro[3.5]nonane-2-carbonitrile trifluoroacetate (13-2) (590mg, 2.4mmol) and N, N-diisopropylethylamine (520mg, 4mmol) The mixture was reacted at room temperature for 1.5 hours. After the reaction was completed, saturated brine (50 mL) was added, and the mixture was extracted twice with ethyl acetate (20 mL). The organic phases were combined and washed with saturated brine three times, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent: petroleum ether/ethyl acetate = 1/4) to obtain the target intermediate (S)-3-((R)-2-(6-(2-cyano -7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)-1- Hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (13-3) (890 mg, yield: 71%).

LCMS: Rt: 1.632min; MS m/z(ESI): 627.3[M+H].

Compound 13:

(S)-3-((R)-2-(6-(2-cyano-7-azaspiro[3.5]nonane-7-carbonyl)-4,4-dimethyl-1 at room temperature -Oxo-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester ( 13-3) (890 mg, 1.42 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (2 mL) was slowly added, and the reaction solution was stirred at room temperature for 1 hour. After the reaction is complete, cool to 0°C, dilute with water, slowly add saturated sodium carbonate aqueous solution to adjust the pH to 10 in an ice bath, extract with ethyl acetate (50mL 2), combine the organic phases, and wash with saturated brine (50mL X 3), dried with anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was lyophilized after adding a small amount of water to obtain the target compound 7-(2-((R)-2-hydroxy-2-((S)) -1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline- 6-carbonyl)-7-azaspiro[3.5]nonane-2-carbonitrile (Compound 13) (567 mg, yield: 75%).

¹ H NMR (400MHz, CD ₃ OD): δ8.02 (d, J = 7.9Hz, 1H), 7.42 (d, J = 1.5Hz, 1H), 7.35 (dd, J = 7.9Hz, 1.5Hz, 1H ),7.14-7.10(m,3H),7.05-7.03(m,1H),4.11-3.97(m,3H),3.97-3.88(m,1H),3.69-3.54(m,5H),3.31-3.29 (m,3H),2.93-2.91(m,3H),2.36-2.33(m,2H),2.20-2.18(m,2H),1.77-1.74(m,2H),1.65-1.61(m,2H) ,1.37(d,J=2.1Hz,6H).

LCMS: Rt: 1.318min; MS m/z(ESI): 527.3[M+H].

Example 14, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of 6-(2-azaspiro[4.5]decane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 15)

Referring to the synthesis method of Example 11, 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) was replaced with 2-azaspirocyclo[4.5]decane hydrochloride, Obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl -6-(2-Azaspiro[4.5]decane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 15).

¹ H NMR (400MHz, CD ₃ OD): δ8.06-8.02 (m, 1H), 7.54 (s, 1H), 7.53-7.45 (m, 1H), 7.31-7.21 (m, 4H), 4.49-4.31 (m, 3H), 4.02-3.95 (m, 1H), 3.68-3.56 (m, 5H), 3.49-3.43 (m, 2H), 3.30-3.22 (m, 3H), 1.86-1.76 (m, 2H) ,1.58-1.49(m,6H),1.47-1.30(m,10H).

LCMS: Rt: 1.465 min; MS m/z (ESI): 516.2 [M+H].

Example 15, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of 6-(7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 16)

With reference to the synthesis method of Example 11, 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) was replaced with 7-azaspiro[3.5]nonane hydrochloride to obtain The target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl- 6-(7-Azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 16).

¹ H NMR (400MHz, CD ₃ OD): δ 8.03 (d, J = 8.0 Hz, 1H), 7.42 (d, J = 1.6 Hz, 1H), 7.36-7.34 (m, 1H), 7.31-7.21 ( m, 4H), 4.45-4.29 (m, 3H), 4.00-3.95 (m, 1H), 3.67-3.59 (m, 6H), 3.30-3.27 (m, 4H), 1.95-1.82 (m, 6H), 1.70(s,2H),1.56(s,2H),1.39(d,J=7.2Hz,6H).

LCMS: Rt: 1.455min; MS m/z(ESI): 502.2[M+H].

Example 16, 6-(4,4-difluoro-6-azaspiro[2.5]octane-6-carbonyl)-2-((R)-2-hydroxy-2-((S)-1, 2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 17 ) Preparation

Referring to the synthesis method of Example 11, replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 4,4-difluoro-6-azaspiro[2.5] Octane hydrochloride to obtain the target product 6-(4,4-difluoro-6-azaspiro[2.5]octane-6-carbonyl)-2-((R)-2-hydroxy-2-(( S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one salt Acid salt (compound 17).

¹ H NMR (400MHz, CD ₃ OD): δ8.06(d,J=7.6Hz,1H),7.48(s,1H),7.40(dd,J=7.6Hz,1.6Hz,1H),7.31-7.25 (m,3H),7.22(d,J=7.2Hz,1H),4.50-4.37(m,2H),4.33-4.29(m,1H),4.05-3.96(m,2H),3.88(s,1H) ), 3.68-3.50 (m, 5H), 3.48 (s, 1H), 3.28-3.25 (m, 2H), 1.73-1.63 (m, 2H), 1.39 (d, J = 4.4 Hz, 6H), 0.95- 0.90(m,2H),0.57(s,2H).

LCMS: Rt: 1.455 min; MS m/z (ESI): 524.1 [M+H].

Example 17, 6-(1,1-difluoro-6-azaspiro[2.5]octane-6-carbonyl)-2-((R)-2-hydroxy-2-((S)-1, 2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 18 ) Preparation

With reference to the synthesis method of Example 11, 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) was replaced with 1,1-difluoro-6-azaspiro[2.5 ]Octane hydrochloride to obtain the target product 6-(1,1-difluoro-6-azaspiro[2.5]octane-6-carbonyl)-2-((R)-2-hydroxy-2-( (S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one Hydrochloride (Compound 18).

¹ H NMR (400MHz, CD ₃ OD): δ 8.05 (d, J = 8.0 Hz, 1H), 7.48 (d, J = 1.6 Hz, 1H), 7.41-7.39 (m, 1H), 7.31-7.21 ( m, 4H), 4.49-4.41 (m, 2H), 4.37-4.31 (m, 1H), 4.00-3.95 (m, 1H), 3.80 (s, 2H), 3.67-3.57 (m, 4H), 3.31- 3.30 (m, 2H), 3.29-3.27 (m, 2H), 1.71-1.68 (m, 4H), 1.40-1.39 (m, 6H), 1.26 (t, J = 8.4 Hz, 2H).

LCMS: Rt: 1.409min; MS m/z(ESI): 524.3[M+H].

Example 18, 6-(1,1-difluoro-5-azaspiro[2.4]heptane-5-carbonyl)-2-((R)-2-hydroxy-2-((S)-1, 2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 19 ) Preparation

With reference to the synthesis method of Example 11, 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) was replaced with 1,1-difluoro-5-azaspiro[2.4 ]Heptane hydrochloride to obtain the target product 6-(1,1-difluoro-5-azaspiro[2.4]heptane-5-carbonyl)-2-((R)-2-hydroxy-2-( (S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one Hydrochloride (Compound 19).

¹ H NMR (400MHz, CD ₃ OD): δ 8.04 (d, J = 8.0 Hz, 1H), 7.61-7.54 (m, 1H), 7.52-7.47 (m, 1H), 7.28-7.18 (m, 4H) ), 4.43-4.27 (m, 3H), 4.00-3.95 (m, 1H), 3.87-3.71 (m, 2H), 3.67-3.47 (m, 6H), 3.25-3.22 (m, 2H), 2.25-2.00 (m,2H),1.58-1.43(m,2H),1.39(d,J=6.8Hz,6H).

LCMS:Rt:1.060min; MS m/z(ESI):510.2[M+H].

Example 19, 6-(6,6-difluoro-2-azaspiro[3.3]heptane-2-carbonyl)-2-((R)-2-hydroxy-2-((S)-1, 2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 20 ) Preparation

Referring to the synthesis method of Example 13, replace 2-cyano-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (13-1) with 6,6-difluoro-2-aza Spiro[3.3]heptane-2-carboxylic acid tert-butyl ester to obtain the target product 6-(6,6-difluoro-2-azaspiro[3.3]heptane-2-carbonyl)-2-((R) -2-Hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl-3,4-dihydroisoquine Lin-1(2H)-one hydrochloride (Compound 20).

¹ H NMR (400MHz, CD ₃ OD): δ8.03 (d, J = 8.0 Hz, 1H), 7.68 (d, J = 1.6 Hz, 1H), 7.58 (dd, J = 8.0 Hz, 1.6 Hz, 1H) ), 7.31-7.25 (m, 3H), 7.22 (d, J = 7.2 Hz, 1H), 4.50-4.34 (m, 4H), 4.33-4.29 (m, 1H), 4.26 (s, 2H), 3.98 ( dd,J=14.0Hz,4.8Hz,1H), 3.67-3.57(m,4H), 3.29-3.27(m,2H), 2.83(t,J=11.6Hz,4H), 1.40(d,J=8.4 Hz, 6H).

LCMS: Rt: 1.381min; MS m/z(ESI): 510.2[M+H].

Example 20, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of yl-6-(3-azaspiro[5.5]undecane-3-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 21)

Referring to the synthesis method of Example 11, replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 3-azaspiro[5.5]undecane hydrochloride, Obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl -6-(3-Azaspiro[5.5]undecane-3-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (Compound 21).

¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.6 Hz, 1H), 7.43 (d, J = 1.2 Hz, 1H), 7.37-7.35 (m, 1H), 7.31-7.21 ( m, 4H), 4.49-4.34 (m, 3H), 4.01-3.98 (m, 1H), 3.74-3.56 (m, 6H), 3.34-3.33 (m, 2H), 3.30-3.27 (m, 2H), 1.47-1.28(m,20H).

LCMS: Rt: 1.523min; MS m/z(ESI): 530.5[M+H].

Example 21, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl 6-(2-oxo-6-azaspiro[3.4]octane-6-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one hydrochloride (compound 22) preparation

Referring to the synthesis method of Example 13, the 2-cyano-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (13-1) was replaced with 2-oxo-6-azaspiro[3.5] 3.4] tert-butyl octane-6-carboxylate to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinoline-3- Yl)ethyl)-4,4-dimethyl-6-(2-oxo-6-azaspiro[3.4]octane-6-carbonyl)-3,4-dihydroisoquinoline-1( 2H)-keto hydrochloride (compound 22).

¹ H NMR (400MHz, DMSO-d ₆ ): δ9.71 (s, 1H), 9.03-9.01 (m, 1H), 7.91 (d, J = 8.0 Hz, 1H), 7.52 (d, J = 4.8 Hz ,1H),7.47(d,J=8.0Hz,1H),7.27-7.23(m,4H),4.55-4.49(m,3H),4.25-4.21(m,1H),3.98(dd,J=14.0 Hz, 6.0Hz, 1H), 3.68 (s, 1H), 3.59-3.33 (m, 7H), 3.25-3.16 (m, 3H), 3.08-2.93 (m, 3H), 2.12-2.03 (m, 2H) ,1.32(d,J=11.2Hz,6H).

LCMS: Rt: 1.217min; MS m/z(ESI): 502.4[M+H].

Example 22, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of 6-(2-oxa-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one (Compound 34)

Referring to the synthesis method of Example 13, the 2-cyano-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (13-1) was replaced with 2-oxa-7-azaspiro[3.5] 3.5] tert-butyl nonane-7-carboxylate to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinoline-3- Yl)ethyl)-4,4-dimethyl-6-(2-oxa-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1( 2H)-ketone (Compound 34).

¹ H NMR (400MHz, CD ₃ OD): δ 8.03 (d, J = 8.0 Hz, 1H), 7.42 (d, J = 1.2 Hz, 1H), 7.37-7.35 (m, 1H), 7.14-7.09 ( m, 3H), 7.05-7.03 (m, 1H), 4.49-4.45 (m, 4H), 4.09-4.01 (m, 3H), 4.01-3.91 (m, 1H), 3.69-3.64 (m, 3H), 3.61-3.54 (m, 2H), 3.34-3.30 (m, 2H), 2.97-2.89 (m, 3H), 1.98 (s, 2H), 1.85 (s, 2H), 1.37 (d, J = 1.6Hz, 6H).

LCMS: Rt: 0.834min; MS m/z(ESI): 504.4[M+H].

Example 23, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of 6-(1-oxa-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one (Compound 35)

Referring to the synthesis method of Example 11, replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 1-oxa-7-azaspiro[3.5]nonane Hemioxalate to obtain the intermediate (S)-3-((R)-2-(4,4-dimethyl-1-oxo-6-(1-oxa-7-azaspiro[3.5 ]Nonane-7-carbonyl)-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxy Tert-butyl ester (35-1).

Compound 35:

To (S)-3-((R)-2-(4,4-dimethyl-1-oxo-6-(1-oxa-7-azaspiro[3.5]nonane-7-carbonyl )-3,4-dihydroisoquinoline-2(1H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (35- 1) Add zinc bromide (69.0 mg, 0.307 mmol) to a solution of (35.0 mg, 0.061 mmol) in dichloromethane (2 mL) and react at 30°C for 3.0 hours. After the reaction was completed, an aqueous sodium bicarbonate solution (10 mL) was added, and it was extracted three times with ethyl acetate (20 mL). The organic phases were combined, washed once with saturated brine (30 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to high performance liquid chromatography (eluent gradient:

) Purification to obtain the target compound 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4- Dimethyl-6-(1-oxa-7-azaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one (Compound 35) (9.17 mg, yield: 30%). ¹ H NMR (400MHz, DMSO-d6): δ7.93-7.92 (m, 1H), 7.39 (s, 1H), 7.34 (d, J = 8.0 Hz, 1H), 7.09-7.07 (m, 3H), 7.01(d,J=4.8Hz,1H),5.49-5.31(m,1H),4.92(d,J=5.6Hz,1H),4.46-4.45(m,1H),4.04(s,1H),3.95 -3.92(m,2H),3.86-3.70(m,3H),3.57-3.40(m,5H),2.81-2.67(m,3H),2.42-2.22(m,2H),2.21-2.08(m, 5H),1.30-1.23(m,6H).

LCMS: Rt: 1.221min; MS m/z(ESI): 504.4[M+H].

Example 24, 3,3-difluoro-2'-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl )-6'-(2-Methoxy-7-azaspiro[3.5]nonane-7-carbonyl)-2',3'-dihydro-1'hydro-spiro[cyclobutane-1, Preparation of 4'-isoquinoline]-1'-one hydrochloride (Compound 37)

Compound 37-2:

Add 2-(3-bromophenyl)acetonitrile (37-1) (30g, 153mmol) to N,N-dimethylformamide (600mL) at 0°C, then add sodium hydride (60%) in batches (14g, 352mmol), react for 0.5 hour. Then 1,3-dibromo-2,2-dimethoxypropane (60 g, 229 mmol) was added dropwise, and the temperature was raised to 60° C. to react for 16 hours. After the reaction was completed, the reaction solution was cooled to room temperature, slowly added to ice water (1L), extracted with ethyl acetate (1L) three times, the organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate and filtered, and the filtrate was reduced Pressure concentration. The crude product was purified by normal phase chromatography column (eluent gradient: petroleum ether/ethyl acetate = 10/1) to obtain the target intermediate 1-(3-bromophenyl)-3,3-dimethoxycyclobutane -1-carbonitrile (37-2) (16 g, yield: 35.3%).

Compound 37-3:

Add 1-(3-bromophenyl)-3,3-dimethoxycyclobutane-1-carbonitrile (37-2) (16g, 54mmol) to tetrahydrofuran (160mL) and water (80mL) at 0°C ), then add concentrated hydrochloric acid (80 mL) dropwise, and react at room temperature for 16 hours. After the completion of the reaction, the reaction system was cooled to 0°C, the pH of the reaction solution was adjusted to 8.0 with potassium carbonate, extracted three times with ethyl acetate (200 mL), the organic phase was washed three times with saturated brine, dried and filtered with anhydrous sodium sulfate, and the filtrate Concentrated under reduced pressure to obtain the target intermediate 1-(3-bromophenyl)-3-oxocyclobutane-1-carbonitrile (37-3) (10 g, yield: 74.0%), and its properties: yellow liquid. ¹ H NMR (400MHz, DMSO-d ₆ ): δ7.82 (t, J = 1.9Hz, 1H), 7.66-7.59 (m, 2H), 7.45 (t, J = 7.9Hz, 1H), 4.12-4.04 (m, 2H), 3.97-3.89 (m, 2H).

Compound 37-4:

Add 1-(3-bromophenyl)-3-oxocyclobutane-1-carbonitrile (37-3) (10g, 40mmol) to ultra-dry dichloromethane (40mL) at 0°C, and then add double (2-Methoxyethyl) aminosulfur trifluoride (BAST) (18.4 g, 84 mmol), heated to 30° C. under the protection of nitrogen, and reacted for 48 hours. After the reaction was completed, the reaction system was reduced to 0°C, slowly poured into an ice-water mixture (500 mL), adjusted to pH 8 with solid sodium bicarbonate, extracted three times with dichloromethane (200 mL), and the organic phase was washed with saturated brine Three times, dry and filter with anhydrous sodium sulfate, and concentrate the filtrate under reduced pressure. The crude product was purified by normal phase chromatography column (eluent gradient: petroleum ether/ethyl acetate = 10/1) to obtain the target intermediate 1-(3-bromophenyl)-3,3-difluorocyclobutane-1 -Formonitrile (37-4) (5.89 g, yield: 54.1%).

¹ H NMR (400MHz, DMSO-d ₆ ): δ 7.76 (t, J = 1.8 Hz, 1H), 7.63 (ddd, J = 7.9, 1.8, 1.0 Hz, 1H), 7.54 (ddd, J = 7.8, 1.8,1.0Hz,1H),7.45(t,J=7.9Hz,1H),3.58-3.52(m,2H),3.49-3.36(m,2H).

Compound 37-5:

Add 1-(3-bromophenyl)-3,3-difluorocyclobutane-1-carbonitrile (37-4) (6.29g, 23.1mmol) to ultra-dry tetrahydrofuran (100mL) at 0°C, nitrogen Protect, add 1N borane tetrahydrofuran solution (69.3mL, 69.3mmol) dropwise, react at room temperature for 1 hour, and then heat to 70°C for 3 hours. The reaction system was cooled to 0° C., methanol (30 mL) and concentrated hydrochloric acid (5.9 mL, 71 mmol) were added dropwise, and the reaction was heated to 70° C. for 3 hours. After the completion of the reaction, the reaction solution was concentrated, water (200 mL) and ethyl acetate (100 mL) were added to the crude product, the pH was adjusted to 8 with solid sodium carbonate, extracted three times with ethyl acetate (200 mL), and washed with saturated sodium chloride solution Three times, dry and filter with anhydrous sodium sulfate, and concentrate the filtrate under reduced pressure to obtain the target intermediate (1-(3-bromophenyl)-3,3-difluorocyclobutyl)methylamine (37-5) (crude product, 6.6) g, yield: 100%).

LCMS: Rt: 0.780min; MS m/z(ESI): 276.0, 278.0[M+H].

Compound 37-6:

Add triphosgene (BTC) (3.4g, 11.6mmol) to ultra-dry dichloromethane (100mL) at 0℃, then add (1-(3-bromophenyl)-3,3-difluorocyclobutyl) ) Methylamine (37-5) (6.4 g, 23.1 mmol) and triethylamine (2.3 g, 23.1 mmol). After reacting for 0.5 hours, methanol (30 mL) and triethylamine (11.6 g, 115.5 mmol) were added dropwise. After the reaction is complete, add water (50mL) to the reaction system to separate the liquids, extract 3 times with dichloromethane (70mL), wash 3 times with saturated sodium chloride solution, dry and filter with anhydrous sodium sulfate, and concentrate the filtrate under reduced pressure. Purification by phase chromatography column (eluent gradient: petroleum ether/ethyl acetate = 10/1) to obtain the target intermediate ((1-(3-bromophenyl)-3,3-difluorocyclobutyl)methyl ) Methyl carbamate (37-6) (5.2 g, yield: 67.3%).

LCMS: Rt: 1.530min; MS m/z(ESI): 334.1, 336.1 [M+H].

Compound 37-7:

Add ((1-(3-bromophenyl)-3,3-difluorocyclobutyl)methyl)methyl carbamate (37-6) (4.7g, 14mmol) to Eaton's reagent (75mL) at room temperature Under nitrogen protection, react at 125°C for 3 hours. The reaction solution was cooled to room temperature, slowly poured into crushed ice, extracted 3 times with ethyl acetate (200 mL), washed 3 times with saturated sodium chloride solution, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: petroleum ether/ethyl acetate = 1/1) to obtain the target intermediate 6'-bromo-3,3-difluoro-2',3'-dihydro -1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (37-7) (1 g, yield: 23.6%). LCMS: Rt: 1.291 min; MS m/z (ESI): 302.1, 304.1 [M+H].

Compound 37-8:

Add 6'-bromo-3,3-difluoro-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-1'-one (37 -7) (950mg, 3.14mmol), (S)-3-((S)-oxirane-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl Ester (k) (1 g, 3.77 mmol) and cesium carbonate (2.04 g, 6.29 mmol) were added to N-methylpyrrolidone (20 mL), protected by argon, and reacted at 85°C for 16 hours. After the reaction is completed, it is directly purified by reversed-phase chromatography column (eluent gradient: acetonitrile/pure water = 2/1) to obtain the target intermediate (1R,10aS)-1-((6'-bromo-3,3-di Fluoro-1'-carbonyl-1'H-spiro[cyclobutane-1,4'-isoquinoline]-2'(3'H)-yl)methyl)-1,5,10,10a-tetra Hydrogen-3H-oxazolo[3,4-b]isoquinolin-3-one (37-8) (1.3 g, yield: 82.3%).

LCMS: Rt: 1.898min; MS m/z(ESI): 503.0, 505.0 [M+H].

Compound 37-9:

(1R,10aS)-1-((6'-Bromo-3,3-difluoro-1'-carbonyl-1'H-spiro[cyclobutane-1,4'-isoquinoline]- 2'(3'H)-yl)methyl)-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-3-one(37-8)( 600mg, 1.19mmol), [1,1'-bis(diphenylphosphine)ferrocene] palladium dichloride (44mg, 0.06mmol) and potassium acetate (350mg, 3.57mmol) were added to absolute ethanol (18.0mL In ), the CO is replaced 3 times, and the reaction is heated to 70°C for 2.0 hours. After the reaction was completed, it was cooled to room temperature and concentrated under reduced pressure. The residue was purified by normal phase chromatography column (eluent gradient: petroleum ether/ethyl acetate = 5/1) to obtain the target intermediate 3,3-difluoro-1'-carbonyl-2'-(((1R, 10aS)-3-carbonyl-1,5,10,10a-tetrahydro-3H-oxazolo[3,4-b]isoquinolin-1-yl)methyl)-2',3'-dihydro -1'H-spiro[cyclobutane-1,4'-isoquinoline]-6'-carboxylic acid ethyl ester (37-9) (453 mg, yield: 76%).

LCMS: Rt: 1.883min; MS m/z(ESI): 497.1[M+H].

Compound 37-10:

3,3-Difluoro-1'-carbonyl-2'-(((1R,10aS)-3-carbonyl-1,5,10,10a-tetrahydro-3H-oxazolo[3,4 -b]isoquinolin-1-yl)methyl)-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-6'-carboxylic acid ethyl Ester (37-9) (453 mg, 0.91 mmol) was added to a mixed solution of methanol (5.0 mL), tetrahydrofuran (5.0 mL) and water (5.0 mL), sodium hydroxide (146 mg, 3.65 mmol) was added, and the mixture was heated to 70 ℃, react for 16 hours. After the reaction was completed, the reaction system was cooled to room temperature, di-tert-butyl dicarbonate (O(Boc) ₂ ) (595 mg, 2.73 mmol) was added, and the reaction was carried out at room temperature for 3.5 hours. The reaction solution was reduced to 0°C, adjusted to pH 5.0 with 1N aqueous hydrochloric acid, and then extracted three times with ethyl acetate (50 mL), the organic phase was dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. Obtain the target intermediate 2'-((R)-2-((S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2 -Hydroxyethyl)-3,3-difluoro-1'-oxo-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-6' -Carboxylic acid (37-10) (350 mg, yield: 71%).

LCMS: Rt: 1.857min; MS m/z(ESI): 543.1[M+H].

Compound 37-11:

2'-((R)-2-((S)-2-(tert-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinolin-3-yl)-2- Hydroxyethyl)-3,3-difluoro-1'-oxo-2',3'-dihydro-1'H-spiro[cyclobutane-1,4'-isoquinoline]-6'- Carboxylic acid (37-10) (70mg, 0.13mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (98mg, 0.26 mmol) and N,N-diethylethylamine (52.2mg, 0.52mmol) were added to the ultra-dry N,N-dimethylformamide (5.0mL), and then 2-methoxy-7-aza Spiro[3.5]nonane hydrochloride (12-4) (29.6mg, 0.16mmol) was reacted at room temperature for 1.5 hours. After the reaction was completed, saturated brine (50 mL) was added, extracted twice with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified with a normal phase chromatography column (eluent gradient: ethyl acetate = 100%) to obtain the target intermediate (S)-3-((R)-2-(3,3-difluoro-6'- (2-Methoxy-7-azaspiro[3.5]nonane-7-carbonyl)-1'-oxo-1'H-spiro[cyclobutane-1,4'-isoquinoline]-2 '(3'H)-yl)-1-hydroxyethyl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (37-11) (65mg, yield: 74% ).

LCMS: Rt: 1.994min; MS m/z(ESI): 680.4[M+H].

Compound 37:

To (S)-3-((R)-2-(3,3-difluoro-6'-(2-methoxy-7-azaspiro[3.5]nonane-7-carbonyl at 0℃ )-1'-oxo-1'H-spiro[cyclobutane-1,4'-isoquinoline]-2'(3'H)-yl)-1-hydroxyethyl)-3,4- Trifluoroacetic acid (1 mL) was added to a solution of dihydroisoquinoline-2(1H)-tert-butyl carboxylate (37-11) in dichloromethane (2 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (eluent gradient in reference to Example 11) to obtain the target compound 3,3-difluoro-2'-((R)-2-hydroxy- 2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6'-(2-methoxy-7-azaspiro[3.5]nonane- 7-carbonyl)-2',3'-dihydro-1'hydro-spiro[cyclobutane-1,4'-isoquinoline]-1'-one hydrochloride (compound 37) (24.28mg, Yield: 41%).

¹ H NMR (400MHz, CD ₃ OD): δ8.08 (d, J = 8Hz, 1H), 7.57 (s, 1H), 7.43 (d, J = 8Hz, 1H), 7.31-7.20 (m, 4H) ,4.44-4.37(m,2H),4.30-4.28(m,1H),3.98-3.90(m,4H),3.70-3.65(m,3H), 3.60-3.58(m,1H),3.33-3.32( m,1H),3.29-3.25(m,3H),3.21(s,3H),2.99-2.90(m,4H),2.26-2.25(m,2H),1.72-1.69(m,4H),1.55( s, 2H).

LCMS: Rt: 0.974 min; MS m/z (ESI): 580.2 [M+H].

Example 25, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of phenyl-6-(2,6-diazaspiro[3.3]heptane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 39)

Referring to the synthesis method of Example 11, replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 2,6-diazaspiro[3.3]heptane-2 -Carboxylic acid tert-butyl ester hemioxalate to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl) Ethyl)-4,4-dimethyl-6-(2,6-diazaspiro[3.3]heptane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one Hydrochloride (Compound 39).

¹ H NMR (400MHz, CD ₃ OD): δ8.07-8.03 (m, 1H), 7.67 (s, 1H), 7.58 (d, J = 8.0 Hz, 1H), 7.31-7.21 (m, 4H), 5.56(s,2H),4.49-4.29(m,10H),4.03-3.96(m,1H),3.68-3.57(m,4H),3.29-3.25(m,1H),1.41(d,J=7.2 Hz, 6H).

LCMS: Rt: 0.356min; MS m/z(ESI): 475.3[M+H].

Example 26, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of yl-6-(2,7-diazaspiro[3.5]nonane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 40)

Referring to the synthesis method of Example 11, the 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) was replaced with 2,7-diazaspiro[3.5]nonane-7 -Carboxylic acid tert-butyl ester hydrochloride to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl Yl)-4,4-dimethyl-6-(2,7-diazaspiro[3.5]nonane-2-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one salt Acid salt (compound 40).

¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.6Hz, 1H), 7.70 (s, 1H), 7.62 (d, J = 7.6Hz, 1H), 7.31-7.21 (m, 4H), 4.49-4.34 (m, 3H), 4.18 (s, 2H), 4.01-3.97 (m, 3H), 3.68-3.57 (m, 4H), 3.30-3.29 (m, 2H), 3.27-3.19 ( m, 4H), 2.07 (t, J = 5.6 Hz, 4H), 1.40 (d, J = 8.4 Hz, 6H).

LCMS: Rt: 1.102min; MS m/z(ESI): 503.4[M+H].

Example 27, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(9-hydroxyl -3-azaspiro[5.5]undecane-3-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 51) preparation

Compound 51-2:

Add 9-oxo-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (51-1) (400mg, 1.5mmol) to methanol (10.0mL) at room temperature, at 0°C Sodium borohydride (171mg, 4.5mmol) was added and reacted at room temperature for 1.5 hours. The reaction was quenched by adding aqueous ammonium chloride solution (10.0 mL), and extracted twice with ethyl acetate (50.0 mL). The organic phases were combined, washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by normal phase chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (51-2) (305mg, yield: 76%).

LCMS:Rt:1.570min; MS m/z(ESI):270.0[M+H].

Compound 51-3:

Add 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (51-2) (305mg) to 4N hydrochloric acid/dioxane solution (5.0mL) at room temperature, The reaction was carried out at room temperature for 1.5 hours, and the reaction solution was concentrated under reduced pressure to obtain the crude intermediate 9-hydroxy-3-azaspiro[5.5]undecane hydrochloride (51-3) (155 mg, yield: 66%).

For the rest of the steps, refer to the synthesis method of Example 11, and replace 2-methoxy-7-azaspiro [3.5] nonane hydrochloride (12-4) with 9-hydroxy-3-azaspiro [5.5] ten Monoalkane hydrochloride (51-3) to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl) Ethyl)-6-(9-hydroxy-3-azaspiro[5.5]undecane-3-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinoline-1(2H) -Ketone hydrochloride (Compound 51).

¹ H NMR (400MHz, CD ₃ OD): δ8.03 (d, J = 8.0 Hz, 1H), 7.44 (s, 1H), 7.36 (d, J = 8.0 Hz, 1H), 7.31-7.25 (m, 3H),7.23-7.21(m,1H),4.50-4.37(m,2H),4.34-4.30(m,1H),3.98(dd,J=14.0Hz,4.8Hz,1H),3.75-3.71(m ,2H),3.67-3.57(m,5H),3.35-3.33(m,2H),3.28-3.23(m,2H),1.76-1.74(m,4H),1.65-1.62(m,1H),1.49 -1.46(m,3H),1.39(d,J=7.6Hz,6H),1.32-1.24(m,4H).

LCMS: Rt: 1.370min; MS m/z(ESI): 546.1[M+H].

Example 28, 3-(2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4 -Dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carbonyl)-3-azaspiro[5.5]undecane-9-one hydrochloride (compound 52 ) Preparation

Referring to the synthesis method of Example 13, replace 2-cyano-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (13-1) with 9-oxo-3-azaspiro[ 5.5] Undecane-3-carboxylic acid tert-butyl ester to obtain the target product 3-(2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquine (Aline-3-yl) ethyl)-4,4-dimethyl-1-oxo-1,2,3,4-tetrahydroisoquinoline-6-carbonyl)-3-azaspiro[5.5] Undecane-9-one hydrochloride (Compound 52).

¹ H NMR (400MHz, DMSO-d ₆ ): δ9.52-9.46(m,1H), 9.02-8.95(m,1H), 7.92(d,J=8.0Hz,1H), 7.40(s,1H) ,7.34(dd,J=8.0Hz,1.2Hz,1H),7.29-7.23(m,4H),5.88(s,1H),4.40-4.23(m,3H),3.97(dd,J=13.6Hz, 6.0Hz, 1H), 3.65-3.55 (m, 5H), 3.41-3.31 (m, 1H), 3.27 (s, 2H), 3.18-3.16 (m, 2H), 2.27-2.24 (m, 4H), 1.75 -1.71(m,4H),1.61(s,2H),1.45(s,2H),1.32(d,J=10.4Hz,6H).LCMS:Rt:1.387min; MS m/z(ESI):544.1 [M+H].

Example 29, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of phenyl-6-(2,7-diazaspiro[4.4]nonane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 53)

Referring to the synthesis method of Example 11, replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 2,7-diazaspiro[4.4]nonane-2 -Tert-butyl carboxylate hemioxalate to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl) Ethyl)-4,4-dimethyl-6-(2,7-diazaspiro[4.4]nonane-2-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one Hydrochloride (Compound 53).

¹ H NMR (400MHz, CD ₃ OD): δ8.04 (d, J = 7.6 Hz, 1H), 7.60-7.51 (m, 2H), 7.31-7.21 (m, 4H), 4.49-4.34 (m, 3H) ), 4.01-3.98 (m, 1H), 3.75-3.58 (m, 7H), 3.52-3.40 (m, 2H), 3.30-3.26 (m, 1H), 2.66 (s, 4H), 2.18-2.03 (m ,4H),1.40(d,J=8.4Hz,6H).

LCMS: Rt: 0.385min; MS m/z(ESI): 503.4[M+H].

Example 30, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Preparation of 6-(8-oxa-2-azaspiro[4.5]decane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one (Compound 54)

Referring to the synthesis method of Example 11, replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 8-oxa-2-azaspiro[4.5]decane , The target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl Group-6-(8-oxa-2-azaspiro[4.5]decane-2-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one (Compound 54).

¹ H NMR (400MHz, CD ₃ OD): δ8.08-8.00 (m, 1H), 7.55 (s, 1H), 7.48 (d, J = 7.2Hz, 1H), 7.30-7.21 (m, 4H), 4.51-4.35 (m, 2H), 4.32-4.31 (m, 1H), 4.00-3.96 (m, 1H), 3.77-3.64 (m, 9H), 3.61-3.48 (m, 3H), 3.29-3.12 (m ,2H),1.96(t,J=7.2Hz,1H),1.85(t,J=6.8Hz,1H),1.67-1.64(m,2H),1.58-1.50(m,2H),1.40(d, J=7.2Hz, 6H).

LCMS: Rt: 1.226min; MS m/z(ESI): 518.4[M+H].

Example 31, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl 6-(2-oxo-1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinoline-1(2H)-one (compound 55) preparation

Referring to the synthesis method of Example 13, replace 2-cyano-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (13-1) with 2-oxo-1,7-diazepine Heterospiro[3.5]nonane-7-carboxylic acid tert-butyl ester to obtain the target compound 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinoline -3-yl)ethyl)-4,4-dimethyl-6-(2-oxo-1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydro Isoquinolin-1(2H)-one (Compound 55).

¹ H NMR (400MHz, DMSO-d ₆ ): δ8.36 (s, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.38 (s, 1H), 7.33 (d, J = 8.0 Hz, 1H ), 7.09(s,3H),7.02-7.00(m,1H), 4.92(d,J=5.2Hz,1H),3.96-3.80(m,4H),3.63-3.40(m,6H), 3.29- 3.19(m,1H),2.81-2.65(m,5H),2.33-2.23(m,1H),1.80-1.61(m,4H),1.30(d,J=2.0Hz,6H).

LCMS: Rt: 1.130min; MS m/z(ESI): 517.4[M+H].

Example 32, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-4,4-dimethyl 6-(1-methyl-1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-dihydroisoquinolin-1(2H)-one (compound 56) preparation

Compound 56-2:

Dissolve 1,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (56-1) (230mg, 1.0mmol) in methanol (10.0mL) at room temperature, and add glacial acetic acid (0.2mL) Then, sodium cyanoborohydride (320 mg, 5.0 mmol) and aqueous formaldehyde solution (1.0 mL) were sequentially added. The reaction was stirred at room temperature for 2 hours. After the reaction was completed, the pH of the reaction solution was adjusted to 9.0 with saturated sodium carbonate aqueous solution, and extracted twice with dichloromethane (30 mL). The organic phases were combined, washed twice with saturated brine (30 mL), dried and filtered with anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain the target crude product 1-methyl-1,7-diazaspiro[3.5]nonane-7 -Tert-butyl carboxylate (56-2) (210 mg, yield: 79%).

LCMS: Rt: 0.496min; MS m/z(ESI): 241.1[M+H].

Compound 56-3:

At room temperature, the crude product 1-methyl-1,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (56-2) (100mg, 0.42mmol) was dissolved in dichloromethane (4.0mL ) Solution, then add trifluoroacetic acid (2.0 mL), and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated under reduced pressure to obtain the target crude product 1-methyl-1,7-diazaspiro[3.5]nonane trifluoroacetate (56-3) (121 mg, yield: 86%).

For the remaining steps, refer to the synthesis method of Example 13, and replace 7-azaspiro[3.5]nonane-2-carbonitrile trifluoroacetate (13-2) with 1-methyl-1,7-diazepine Spiro[3.5]nonane trifluoroacetate (56-3) to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquine (Alkolin-3-yl)ethyl)-4,4-dimethyl-6-(1-methyl-1,7-diazaspiro[3.5]nonane-7-carbonyl)-3,4-di Hydroisoquinolin-1(2H)-one (Compound 56).

¹ H NMR (400MHz, CD ₃ OD): δ8.03 (d, J = 8.0Hz, 1H), 7.45 (d, J = 1.6Hz, 1H), 7.38 (dd, J = 8.0Hz, 1.6Hz, 1H) ),7.14-7.10(m,3H),7.06-7.04(m,1H),4.64-4.56(m,1H),4.09-3.98(m,3H),3.93(dd,J=14.0Hz,3.6Hz, 1H), 3.69-3.54 (m, 4H), 3.18-3.12 (m, 3H), 2.98-2.85 (m, 4H), 2.25 (s, 3H), 2.17-1.91 (m, 3H), 1.83-1.57 ( m,3H),1.38(d,J=2.0Hz,6H).

LCMS: Rt: 0.640min; MS m/z(ESI): 517.3[M+H].

Example 33, 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinolin-3-yl)ethyl)-6-(9-methyl Oxy-3-azaspiro[5.5]undecane-3-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinolin-1(2H)-one hydrochloride (compound 57 ) Preparation

Compound 57-2:

Dissolve 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (51-2) (540mg, 2mmol) in N,N-dimethylformamide (10mL) and cool At 0°C, sodium hydride (320mg, 8mmol) was added in batches, and then reacted at 0°C for 1.0 hour. Add methyl iodide (568mg, 4mmol) and stir overnight at room temperature. After the reaction was complete, it was quenched by adding saturated ammonium chloride solution, diluted with saturated brine (50 mL), extracted with ethyl acetate (20 mL) twice, the organic phase was washed twice with saturated brine, dried and filtered with anhydrous sodium sulfate, and the filtrate was reduced The target intermediate 9-methoxy-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (57-2) (crude product, 560 mg, yield: 100%) was obtained by pressure concentration.

Compound 57-3:

Dissolve 9-methoxy-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (57-2) (560mg, 2mmol) in dioxane (1mL) and cool to 0℃ , Add 4N hydrochloric acid/dioxane solution (4mL), and react at 0°C for 1.0 hour. After the completion of the reaction, the reaction solution was concentrated under reduced pressure, the residue was slurried with methyl tert-butyl ether, and filtered to obtain the target intermediate 9-methoxy-3-azaspiro[5.5]undecane hydrochloride (57-3 ) (192 mg, yield: 44%).

LCMS: Rt: 0.415min; MS m/z(ESI): 184.2[M+H].

For the rest of the steps, refer to the synthesis method of Example 11, and replace 2-methoxy-7-azaspiro[3.5]nonane hydrochloride (12-4) with 9-methoxy-3-azaspiro[5.5 ] Undecane hydrochloride (57-3) to obtain the target product 2-((R)-2-hydroxy-2-((S)-1,2,3,4-tetrahydroisoquinoline-3- Yl)ethyl)-6-(9-methoxy-3-azaspiro[5.5]undecane-3-carbonyl)-4,4-dimethyl-3,4-dihydroisoquinoline- 1(2H)-keto hydrochloride (Compound 57).

¹ H NMR (400MHz, CD ₃ OD): δ8.03(d,J=8.0Hz,1H),7.44(s,1H),7.36(d,J=8.0Hz,1H),7.34-7.16(m, 4H), 4.54-4.35 (m, 2H), 4.32 (s, 1H), 4.04-3.90 (m, 1H), 3.80-3.52 (m, 6H), 3.43-3.32 (m, 4H), 3.30-3.18 ( m,4H),1.88-1.65(m,4H),1.61(s,1H),1.57-1.34(m,11H),1.34-1.19(m,2H).LCMS:Rt:1.381min; MS m/z (ESI): 560.4 [M+H].

With reference to the above-mentioned synthesis methods of Examples 11 and 13, the following compounds have also been synthesized in this application.

Test example 1: PRMT5 enzymatic activity inhibition experiment

Materials: PRMT5/MEP50 protein was purchased from BPS bioscience company (U.S.); Histone H4 Peptide substrate was purchased from Shenggong Bioengineering (Shanghai) Co., Ltd.; Anti-Histone H4 (symmetric di methyl R3) antibody-ChIP Grade was purchased from Ai Bokang (U.S.); S-(5-Adenosyl)-L-methionine chloride dihydrochloride was purchased from Sigma (U.S.); 384-well plates, AlphaScreen Streptavidin Donor beads and AlphaScreen Protein A Acceptor beads were purchased from PerkinElmer Instruments Limited company (United States).

Enzyme activity detection: Use Echo to drive the compound into a 384-well plate to make the final concentration of the compound 0-1000 nM (initial concentration 1000 nM, 3-fold dilution, 10 points), and DMSO content 0.5%. Add 10μL of 2X PRMT5/MEP50 solution to each well and incubate at room temperature for 30 minutes. Add 10μL of 2X PRMT5/MEP50 substrate solution to each well to start the reaction, and incubate at room temperature for 60 minutes. Prepare 6X detection reagent containing AlphaScreen Protein A Acceptor beads and Histone H4 (symmetric dimethyl R3) antibody, add 5 μL to each well, and incubate for 60 minutes at room temperature. Prepare 6X detection reagent containing AlphaScreen Streptavidin Donor beads, add 5 μL to each well, and incubate at room temperature for 60 minutes. Envision detection signal value. The test results are shown in Table 2.

Test Example 2: Inhibitory activity of the compound on tumor cell proliferation

Materials and cells: Z-138 cells were purchased from ATCC (U.S.); IMDM medium and penicillin-streptomycin were purchased from Sigma (U.S.); horse serum was purchased from Hyclone (U.S.); 96-well plates were purchased from Corning ( United States); Cell-Titer Glo reagent was purchased from Promega (United States).

Cell culture: Z-138 cells were cultured in IMDM medium containing 10% horse serum + 1% penicillin-streptomycin at 37°C and 5% CO ₂ . Cells in the logarithmic growth phase can be used in experiments.

Cell proliferation activity detection: Cell-Titer Glo reagent was used to detect the proliferation inhibitory activity of the compound on Z-138 cells. Adjust the cell concentration, inoculate a 96-well plate (500/well) with 180 μL per well, and equilibrate for 10-15 minutes _{at 37°C and 5% CO 2.} Add 20 μL of the cell culture medium containing the compound to each well to make the final concentration of the compound reach 0-300 nM (initial concentration 300 nM, 3-fold dilution, 10 points), and the DMSO content is 0.1%. The cell plate was incubated at 37°C and 5% CO ₂ for 8 days. On the fourth day, the medium was changed: 100 μL of supernatant was slowly aspirated, and 100 μL of fresh culture medium containing the compound was added to keep the compound concentration unchanged. Cell-Titer Glo reagent was used to detect cell viability.

The test results are shown in Table 2.

Test Example 3: The compound's inhibitory activity experiment on Z-138 cells SDMA

Materials and cells: Z-138 cells were purchased from ATCC (U.S.); IMDM medium and penicillin-streptomycin were purchased from Sigma (U.S.); Horse serum was purchased from Hyclone (U.S.); Hoechst antibody was purchased from Invitrogen (U.S.) ); Alexa Fluor 488 goat anti-rabbit IgG antibody was purchased from Santa Company; Anti-dimethyl-Arginine symmetric (SYM11) antibody was purchased from Merck Company (USA).

Immunofluorescence detection: Use immunofluorescence to detect the effect of compounds on SDMA in Z-138 cells. Adjust the cell concentration to 1*10 ⁵ /mL, inoculate a 384-well plate (4000/well) with 40 μL per well, and equilibrate for 10-15 minutes _{at 37°C and 5% CO 2.} Using Echo, the compound was injected into a 384-well plate, so that the final concentration of the compound was 0-300 nM (initial concentration 300 nM, 3-fold dilution, 10 points), and the DMSO content was 0.1%. The cell plate was incubated at 37°C and 5% CO ₂ for 2 days. Add 40μL 8% paraformaldehyde to each well and incubate for 30 minutes at room temperature. Discard the supernatant, wash the plate with PBS, add 40 μL 0.5% PBST to each well, and incubate at room temperature for 60 minutes. Discard the supernatant, wash the plate with 0.05% PBST, add 40 μL of blocking solution to each well, and incubate for 60 minutes at room temperature. Discard the supernatant, add 20μL of primary antibody to each well, overnight at 4°C. Discard the supernatant, wash the plate with 0.05% PBST, add 20 μL of secondary antibody to each well, and incubate for 60 minutes at room temperature. The supernatant was discarded, the plate was washed with 0.05% PBST, and the fluorescence intensity was detected by ImageXpress Nano. The test results are shown in Table 2.

Table 2

Note: NA stands for not tested.

Test Example 4: Liver Cell Metabolic Stability Test

Experimental materials: human liver cells were purchased from Biopredic; mouse liver cells were purchased from BioIVT; acetonitrile and methanol were purchased from Merck; AOPI stains were purchased from Nexcelom; dexamethasone was purchased from NIFDC; DMSO was purchased from Beijing Solei Bao Technology Co., Ltd.; DPBS (10x), GlutaMAX ^TM -1 (100x) and human recombinant insulin were purchased from Gibco by Life Technologies; fetal bovine serum was purchased from Corning; formic acid was purchased from DIKMAPURE; Isotonic Percoll was purchased from GE Healthcare; Alprazolam was purchased from Supelco; caffeine was purchased from ChromaDex.inc; HEPES, tolbutamide and Williams' Medium E were purchased from Sigma.

Experiment preparation:

Prepare a high-concentration stock solution of the test substance powder with DMSO, and dilute it with acetonitrile to a working solution of 100μM before use. The final concentration of the test substance is 1μM.

The specific preparation information of the hepatocyte resuscitation solution is shown in Table 3 below. Mix 49.5mL Williams'E Medium and 0.5mL GlutaMAX as an incubation solution. Place the hepatocyte resuscitation solution and incubation solution in a 37°C water bath for at least 15 minutes before use. Take a tube of ultra-low-temperature preserved liver cells to ensure that the liver cells are still in a low-temperature frozen state before resuscitation. The hepatocytes were quickly placed in a 37°C water bath and gently shaken until all ice crystals were dispersed, sprayed with 70% ethanol and transferred to a biological safety cabinet. Pour the contents of the hepatocyte tubule into a centrifuge tube containing 50 mL of resuscitation medium, and centrifuge it at 100 g for 10 minutes. After centrifugation, aspirate the recovery medium and add sufficient incubation medium to obtain ^{a cell suspension with a cell density of about 1.5×10 6} cells/mL. Use Cellometer Vision to count liver cells and determine the density of viable cells. The survival rate of liver cells must be greater than 75%. Use the incubation medium to dilute the hepatocyte suspension to a viable cell density of 0.5×10 ⁶ viable cells/mL.

Table 3 Preparation of Hepatocyte Resuscitation Solution

experimental method:

Transfer 247.5 μL of suspension or culture medium of live cells (human hepatocytes or mouse hepatocytes) to a 96-well deep well plate, and place the deep well plate in a vortex incubator for 10 minutes. All samples were incubated in double parallel. Add 2.5 μL 100 μM test substance to each well to start the reaction, and put the deep-well plate back on the vortex of the incubator. Incubate the samples at 0, 15, 30, 60, 90, and 120 minutes, respectively, take 25 μL of the suspension, and add 125 μL of acetonitrile containing internal standard (100 nM alprazolam, 200 nM caffeine, 100 nM tolbutamide) to stop the reaction. Vortex for 10 minutes, centrifuge at 3220g and 4°C for 30 minutes. After centrifugation, transfer 100 μL of supernatant to the sample plate, add 150 μL of pure water to mix, and use for LC-MS/MS analysis.

All data calculations are performed by Microsoft Excel software. Detect the peak area by extracting the ion spectrum. By linearly fitting the natural logarithm of the elimination percentage of the parent drug to time, the in vitro half-life (t _1/2 ) of the parent drug is detected.

The in vitro half-life (t _1/2 ) is calculated by the slope:

in vitro t _1/2 = 0.693/k

The experimental results are shown in Table 4.

Table 4 Test results of liver cell metabolic stability

化合物Compound	人t _1/2(min) Person t _1/2 (min)	鼠t _1/2(min) T _1/2 (min)
1010	98.2498.24	36.3636.36
1111	50.8050.80	20.7720.77
1212	89.5189.51	25.2225.22
1313	186.98186.98	66.0666.06
1616	52.2552.25	19.9819.98
1818	67.0367.03	24.6824.68
3535	889.57889.57	71.5371.53
5454	419.95419.95	128.87128.87
5757	90.6390.63	76.4476.44

Test Example 5: Mouse Pharmacokinetic Experiment

Experimental materials: CB17-SCID mice were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.; DMSO, HP-β-CD (hydroxypropyl-β-cyclodextrin), MC (methyl cellulose), acetonitrile were purchased From Merck (USA).

Experimental method: 6 female CB17-SCID mice (20-30g, 4-6 weeks) were randomly divided into 2 groups with 3 mice in each group. The first group was given compound 12 by tail vein injection at a dose of 2 mg/kg and the vehicle was an aqueous solution of 5% DMSO + 95% 10% HP-β-CD. The second group was given compound 12 orally at a dose of 10 mg/kg and the vehicle was 0.5 %MC aqueous solution. Feed and water normally before the animal experiment. Mice in each group were subjected to intravenous blood sampling at 0.083 (intravenous injection group only), 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h before and after administration. The collected whole blood samples were placed in a K ₂ EDTA anticoagulation tube, and after centrifugation for 5 min (4000 rpm, 4° C.), plasma was taken for testing.

Take 10μL of mouse plasma sample, add 150μL of acetonitrile solvent (containing internal standard compound) to precipitate the protein, vortex for 0.5min, centrifuge (4700rpm, 4℃) for 15min, and use the supernatant with 0.05% (v/v) FA Dilute 2 times with water, and inject 3μL into the LC-MS/MS system (AB Sciex Triple Quad 6500+) for quantitative detection. The CB17-SCID mouse plasma standard curve (linear range: 0.5-1000ng/mL) and quality control samples were accompanied when determining the sample concentration. For the 10x diluted sample, take 2μL of the sample and add 18μL of blank plasma. After vortexing for 0.5min, add 300μL of acetonitrile solvent (containing internal standard compound) to precipitate the protein. The rest of the processing steps are the same as those without dilution.

According to the same operation described above, the difference is that compound 12 is replaced with compound 13, and the pharmacokinetic experiment is performed.

The pharmacokinetic test results are shown in Table 5.

Table 5 Mouse pharmacokinetic test results

Test Example 6: In vivo drug efficacy experiment in mice

Experimental materials: Z138 cells were purchased from ATCC; IMDM culture medium, penicillin and 0.25% pancreatin-EDTA were purchased from Gibco; horse serum was purchased from Hyclone.

Animal information: CB17-SCID mice, female, 5-6 weeks old, weighing about 13-20 grams, the animals were purchased from Shanghai Lingchang Biological Technology Co., Ltd. The mice were kept in an SPF-level environment, and each cage was independent When sending and exhausting air, all animals have free access to standard certified commercial laboratory food and drinking water.

experimental method:

Cell culture: Human mantle cell lymphoma Z-138 cell line is cultured in vitro, and the culture conditions are IMDM (cell culture medium) with 10% horse serum, 1% penicillin solution, 37°C, 5% CO ₂ incubator. Use 0.25% pancreatin-EDTA digestion solution twice a week for routine digestion and passage. When the cell saturation is 85%-90% and the number reaches the requirement, the cells are collected and counted.

Cell inoculation: 0.1ml/(containing 1×10 ⁷ ) Z-138 cell suspension (PBS: Matrigel=1:1) was subcutaneously inoculated on the right back of each mouse. On the 18th day after inoculation, when the average tumor volume reached about 130 mm ³ , the drug was administered in groups according to the tumor volume and animal body weight using a random stratified grouping method. PBS is a phosphate buffered saline solution without calcium and magnesium ions, and Matrigel is matrigel.

Administration: The dose of compound 12 is 25 mg/kg, PO, twice a day (BID) x 3 weeks, or 50 mg/kg, PO, once a day (QD) x 3 weeks. The dosage of compound 13 is 50 mg/kg, PO, twice a day (BID) x 3 weeks. Each group has 6 mice.

Tumor measurement and experimental indicators:

The tumor diameter was measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5a x b ² , a and b represent the long diameter and short diameter of the tumor, respectively. The body weight of the mice was measured twice a week.

The anti-tumor efficacy of the compound is evaluated by the tumor growth inhibition rate TGI (%):

TGI(%)=[(1-(Average tumor volume at the end of a certain treatment group-average tumor volume at the beginning of the treatment group)/(Average tumor volume at the end of the solvent control group-when the solvent control group starts Average tumor volume)] x 100%.

The experimental results are shown in Table 6, Figure 2 and Figure 3. No mice became ill or died during the experiment. In the mouse subcutaneous xenograft tumor model Z-138, compound 12 at 25 mg/kg twice a day and compound 13 at 50 mg/kg twice a day have a significant inhibitory effect on tumor growth and shrink tumors. The effect shows good anti-tumor efficacy. Compound 12 and compound 13 did not significantly affect the body weight of the mice at the tried doses, nor did they cause any death of the mice, and the mice could tolerate them.

Table 6 Tumor volume of Z-138 subcutaneous tumor model

The above are only preferred embodiments of the present invention, and do not limit the present invention in any form. Any person familiar with the profession, within the scope of the technical solution of the present invention, according to the technical essence of the present invention, Any simple modifications, equivalent replacements and improvements made to the above embodiments still fall within the protection scope of the technical solution of the present invention.

Claims

The compound of formula (I) or a pharmaceutically acceptable salt thereof,

among them,

A is a 5-14 membered spirocyclic group optionally substituted by R 6;

R 1 and R 2 are independently selected from H, C 1-4 alkyl, halogen, C 1-4 alkoxy, or

R 1 , R 2 and the C atom to which they are connected together form a C 3-8 cycloalkyl or 3-8 membered heterocycloalkyl, the cycloalkyl or heterocycloalkyl is optionally substituted by halogen;

R 3 , R 4 , and R 5 are independently selected from H, halogen and C 1-4 alkyl;

m is 1, 2, 3 or 4;

R 6 is independently selected from halogen, hydroxyl, cyano, amino, C 1-3 alkylamino, di(C 1-3 alkyl)amino, oxo, C 1-4 alkyl, C 1-4 alkoxy Group, C 1-4 alkoxycarbonyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl.
The compound of claim 1, wherein A is C 6-13 spirocycloalkyl or 6-13 membered spiroheterocycloalkyl, and the spirocycloalkyl or spiroheterocycloalkyl is optionally substituted by R 6 .
The compound of claim 1, wherein A is optionally substituted by R 6
among them

n, n', p, and q are independently selected from 1, 2, 3, and 4, and n+n'+p+q≤10;

W is selected from CH or N;

X and Y are independently selected from CH 2 , NH or O;

Z is selected from CH 2 , NH, O or a bond.
The compound of claim 3, wherein n, n', p, and q are independently selected from 1, 2, and 3, and n+n'+p+q≤10.
The compound of claim 4, wherein n and n'are independently selected from 1, 2 and 3, and p and q are independently selected from 1 and 2.
The compound of any one of claims 3-5, wherein W is N.
The compound of any one of claims 1-6, wherein A is selected from those optionally substituted by R 6
The compound of claim 7, wherein A is selected from optionally substituted by R 6
The compound of any one of claims 1-8, wherein R 6 is independently selected from halogen, hydroxyl, cyano, amino, C 1-3 alkylamino, oxo, C 1-4 alkyl, C 1 -4 alkoxy, C 1-4 alkoxycarbonyl and C 3-6 cycloalkyl.
The compound of claim 9, wherein R 6 is independently selected from halogen, hydroxy, cyano, amino, oxo, C 1-4 alkyl, C 1-4 alkoxy and C 1-4 alkoxy Carbonyl.
The compound of claim 10, wherein R 6 is independently selected from fluorine, hydroxyl, cyano, amino, oxo, methyl, methoxy, and methoxycarbonyl.
The compound of any one of claims 1-11, wherein A is selected from
The compound of claim 12, wherein A is selected from
The compound of any one of claims 1-13, wherein R 1 and R 2 are independently selected from H, C 1-4 alkyl, or R 1 , R 2 and the C atom to which they are connected together form C 3-6 Cycloalkyl, the C 3-6 cycloalkyl is optionally substituted with halogen.
The compound of claim 14, wherein R 1 and R 2 are independently selected from H, methyl, ethyl, or R 1 , R 2 and the C atom to which they are connected together form a cyclopropyl or cyclobutyl, said Cyclopropyl or cyclobutyl is optionally substituted with fluorine.
The compound of claim 15, wherein R 1 and R 2 are methyl groups.
The compound of any one of claims 1-16, wherein R 3 , R 4 , and R 5 are independently selected from H, F, Cl, methyl, and ethyl.
The compound of claim 17, wherein R 3 , R 4 , and R 5 are H.
The compound of any one of claims 1-18, wherein m is 1 or 2.
The compound of claim 19, wherein m is 1.
The compound of any one of claims 1-20, wherein the compound of formula (I) is selected from the compound of formula (II)
The compound of any one of claims 1-20, wherein the compound of formula (I) is selected from the compound of formula (III)
The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, selected from the following compounds or a pharmaceutically acceptable salt thereof:
The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, selected from the following compounds or a pharmaceutically acceptable salt thereof:
A pharmaceutical composition comprising the compound described in any one of claims 1-24 of the present application or a pharmaceutically acceptable salt thereof.
Use of the compound of any one of claims 1-24 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 25 in the preparation of a medicament for the prevention or treatment of PRMT5-mediated diseases.
A method for treating a disease mediated by PRMT5 in a mammal, comprising administering a therapeutically effective amount of the compound according to any one of claims 1-24 or a pharmaceutically acceptable salt thereof or claim 25 to a mammal in need of such treatment The pharmaceutical composition.