CN114057740B

CN114057740B - Spiropyrimidinone derivatives, method for the production thereof, pharmaceutical compositions and use thereof

Info

Publication number: CN114057740B
Application number: CN202111528169.0A
Authority: CN
Inventors: 金赟; 彭军; 吴金华
Original assignee: Shanghai Semerode Biotechnology Co ltd; Shanghai Simr Biotechnology Co ltd
Current assignee: Shanghai Semerode Biotechnology Co ltd; Shanghai Simr Biotechnology Co ltd
Priority date: 2021-12-15
Filing date: 2021-12-15
Publication date: 2024-04-02
Anticipated expiration: 2041-12-15
Also published as: WO2023109471A1; CN114057740A

Abstract

The present invention provides a compound represented by the general formula (I), its cis-trans isomer, its enantiomer, its diastereoisomer, its racemate, its solvate, its hydrate or its pharmaceutically acceptable salt or its prodrug, a process for preparing the same, a pharmaceutical composition containing the same and the compound as Lp-PLA ₂ Use of an inhibitor, wherein R ₁ ，R ₂ ，R _x ，R _y Q, m, n, p, u and A are as defined herein.

Description

Spiropyrimidinone derivatives, method for the production thereof, pharmaceutical compositions and use thereof

Technical Field

The present invention relates to novel pyrimidinone compounds, processes for their preparation, pharmaceutical compositions containing them and their use in therapyPLA ₂ Use in mediated diseases.

Background

Lipoprotein-associated phospholipase A ₂ (Lp-PLA ₂ ) Phospholipase A, which is involved in the hydrolysis of lipoprotein lipids or phospholipids ₂ An enzyme, also known as platelet activating factor acetylhydrolase (PAF-AH). Lp-PLA ₂ Moves with Low Density Lipoprotein (LDL) and rapidly cleaves oxidized phosphatidylcholine molecules resulting from oxidation of LDL. Lp-PLA ₂ Hydrolysis of the sn-2 ester of oxidized phosphatidylcholine gives the lipid mediators lysophosphatidylcholine (lysoPC) and oxidized non-esterified fatty acids (NEFA). The literature reports that lysoPC and NEFA can elicit an inflammatory response, and therefore Lp-PLA ₂ Mediating oxidative inflammatory reactions in vivo. (Zalewski A et al, arterioscler. Thromb. Vasc. Biol., 25, 5, 923-31 (2005)).

Literature (WO 96/13484, WO 96/1941, WO97/02242, WO97/12963, WO97/21675, WO97/21676, WO97/41098, WO97/41099, WO99/2442, WO00/10980, WO00/66566, WO00/66567, WO00/68208, WO01/60805, WO02/30904, WO02/30911, WO03/015786, WO03/016287, WO 03/04101712, WO03/042179, WO03/042206, WO03/042218, WO00/66 WO03/086400, WO03/87088, WO08/04886, US2008/0103156, US2008/0090851, US2008/0090852, WO08/048866, W005/003118, W006/063281, W006/063113, WO2008/141176, WO2013013503A1, WO2013014185A1, WO2014114248A1, WO2014114694A1, WO2016011930A1, JP200188847 US2008/0279846A1, US 2010/023965 A1, US 2008/0280829 A1) describe a number of Lp-PLAs ₂ Inhibitors and/or uses thereof for the treatment of disorders involving vascular endothelial dysfunction or diseases related thereto, and for the treatment of disorders related to Lp-PLA ₂ Disorders of lipid oxidation associated with activity (e.g., associated with the formation of lysophosphatidylcholine and oxidized free fatty acids) and disorders involving activated monocytes, macrophages or lymphocytes or associated with increased participation of monocytes, macrophages or lymphocytes. Examples of specific diseases include neurodegenerative diseases (e.g., alzheimer's disease, parkinson's disease, huntington's disease, vascular dementia), various neuropsychiatric diseases such as schizophrenia and autism, peripheral and brain Atherosclerosis, stroke, metabolic bone disease (e.g., bone marrow abnormality), dyslipidemia, paget's disease, type II diabetes, hypertension, angina, myocardial infarction, ischemia, reperfusion injury, metabolic syndrome, insulin resistance and hyperparathyroidism, diabetic complications (e.g., macular edema, diabetic retinopathy and post uveitis, diabetic ulcers and diabetic nephropathy), diabetic peripheral neuropathic pain, inflammatory pain, neuropathic pain, various types of cancer (e.g., prostate cancer, colon cancer, breast cancer, kidney cancer, lung cancer, ovarian cancer, and the like), macular edema, wound healing, male erectile dysfunction, rheumatoid arthritis, chronic Obstructive Pulmonary Disease (COPD), sepsis, acute and chronic inflammation, psoriasis and multiple sclerosis.

Scientific research results further prove that Lp-PLA ₂ Inhibitors may be used to treat atherosclerosis. Wilensky et al demonstrated Lp-PLA in a pig model of diabetes mellitus and hypercholesterolemia that accelerates coronary atherosclerosis ₂ The effect of inhibitors on atherosclerotic plaque components (Wilensky et al, nature Medicine,10,1015-1016 (2008)). Clinical studies have also found Lp-PLA ₂ The inhibitor stabilizes the atherosclerotic plaque in patients with atherosclerotic plaque, preventing further plaque development and rupture (Serrouys et al Circulation 118:1172-1182 (2008)).

Research shows that the Lp-PLA is high ₂ Activity is associated with the risk of high dementia, including Alzheimer's Disease (AD) and mixed dementia (Van Objen et al, annals of Neurology,59,139 (2006); fitzpatrick et al, atheroscleosis 235:384-391 (2014)). Higher levels of oxidized LDL were observed in AD patients (Kassner et al Current Alzheimer Research,5, 358-366 (2008); dildar et al Alzheimer Dis Assoc Disord,24, april-June (2010); sinem et al Current Alzheimer Research, 7, 463-469 (2010)).

Furthermore, US2008/0279846 describes Lp-PLA ₂ Inhibitors reduce blood brain barrier leakage and brain amyloid (Abeta) loading and are useful for treating diseases associated with blood brain barrier leakage, such as AbetaAlzheimer's disease and vascular dementia. In clinical studies, lp-PLA ₂ The inhibitor has a remarkable effect of preventing further deterioration of cognitive function in Alzheimer's patients (Maher-Edwards et al, alzheimer's) & Dementia：Translational Research & Clinical Interventions 1，131-140（2015））。

Neuroinflammation, including the release of a variety of cytotoxic cytokines, is a common feature of all neurodegenerative diseases, including multiple sclerosis, amyotrophic lateral sclerosis, parkinson's disease, alzheimer's disease, etc. (Perry, acta neuroskill, 120,277-286 (2010)). Lp-PLA ₂ Inhibitors reduce the release of various cytokines by inhibiting lysoPC production (Shi et al, atheroscleosis 191,54-62 (2007)). Thus, lp-PLA is inhibited ₂ Is a potential treatment method for neurodegenerative diseases (including multiple sclerosis, amyotrophic lateral sclerosis, parkinsonism and the like).

LysoPC is also involved in leukocyte activation, induction of apoptosis and mediation of vascular endothelial cell dysfunction (Wilensky et al, current Opinion in Lipidology,20, 415-420, (2009)). Thus, it is considered that Lp-PLA ₂ Inhibitors may be used to treat tissue damage associated with diabetes by reducing the production of lysoPC. High Lp-PLA ₂ Activity correlates with risk of high diabetic retinopathy onset (Siddiqui et al, diabetes, 61, 1344-1353 (2018)). Lp-PLA ₂ The inhibitor can inhibit the major pathological changes of retinopathy of diabetic rat model (Canning et al, P.N.A.S. 113, 7213-7218 (2016.) clinical studies also showed Lp-PLA ₂ The inhibitor can improve macular edema symptom and vision of diabetic retinopathy patients (Staurenghi et al, ophtalmology 122, 990-996 (2015.) these studies demonstrate Lp-PLA ₂ The inhibitor can be used for treating diabetic retinopathy.

Studies have shown that Lp-PLA in diabetic patients ₂ The activity was higher than that of normal persons (Serban et al J.cell. Mol. Med. 6:643-647, (2002); garg et al Indian J. Med. Res. 141:107-114, (2015)). Whereas, as described above, lp-PLA ₂ Is mediated by oxidative inflammatory reactions, presumably inhibiting Lp-PLA ₂ The activity can be used for treating various complications of diabetes patients caused by in vivo oxidative inflammatory reaction, such as diabetic nephropathy, diabetic peripheral neuropathy, diabetic skin ulcer, etc.

Glaucoma and age-related macular degeneration (AMD) are retinal neurodegenerative diseases. Inflammation plays an important role in the pathogenesis of glaucoma and AMD (Buschini et al, progress in Neurobiology, 95, 14-25 (2011); tezel, progress in Brain Research, vol.173, ISSN 0079-6123, chapter 28). Thus, lp-PLA ₂ Inhibitors may provide potential therapeutic applications for glaucoma and AMD.

In vivo Lp-PLA in male erectile dysfunction patients ₂ Is significantly higher than normal and is believed to be high in Lp-PLA ₂ Activity predicts early male erectile dysfunction (Otuncteur et al, androlog 47:706-710 (2015)), suggesting that inhibitors may be useful in the treatment of male erectile dysfunction.

In prostate cancer tissue with Lp-PLA ₂ High expression, reduced Lp-PLA ₂ Can reduce prostate cell canceration and promote prostate cancer cell apoptosis in vitro experiments (Vainio et al, oncostarget, 2:1176-1190 (2011)), suggesting Lp-PLA ₂ The inhibitor can be used for treating prostatic cancer.

Thus, lp-PLA ₂ The inhibitor can be used for treating or preventing Lp-PLA ₂ Various diseases associated with inhibition of activity. The invention discovers a new Lp-PLA ₂ An inhibitor.

Disclosure of Invention

An object of the present invention is to provide a compound represented by the general formula (I), a cis-trans isomer, an enantiomer, a diastereomer, a racemate, a solvate, a hydrate, or a pharmaceutically acceptable salt or ester thereof, or a prodrug thereof.

Another object of the present invention is to provide a process for producing a compound represented by the general formula (I).

Another object of the present invention is to provide a compound represented by the general formula (I) as Lp-PLA ₂ Use of an inhibitor for the manufacture of a medicament for the prevention, treatment or amelioration of a disease associated with Lp-PLA2 inhibition, such as diabetic complications, neuroinflammation-related disorders, or/and atherosclerosis, which are diabetic retinopathy/diabetic macular edema, diabetic nephropathy, diabetic neuropathy, diabetic peripheral neuropathic pain or/and diabetic foot, neuroinflammation-related disorders, such as alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis or/and parkinson's disease.

It is another object of the present invention to provide a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds of formula (I), cis-trans isomers, enantiomers, diastereomers, racemates, solvates, hydrates, or pharmaceutically acceptable salts or esters thereof, or prodrugs thereof, and a pharmaceutically acceptable carrier and/or adjuvant.

It is another object of the present invention to provide a method for preventing, treating or ameliorating a disease associated with Lp-PLA2 inhibition, comprising administering a compound of the general formula (I), its cis-trans isomer, enantiomer, diastereomer, racemate, solvate, hydrate or a pharmaceutically acceptable salt or ester thereof, or a prodrug thereof or a composition of the present invention.

In a first aspect, a compound of formula I, a cis-trans isomer thereof, an enantiomer thereof, a diastereomer thereof, a racemate thereof, a solvate thereof, a hydrate thereof or a pharmaceutically acceptable salt thereof or a prodrug thereof,

wherein the method comprises the steps of

m is 1 or 2; preferably, m is 1;

n, u is 0,1 or 2; preferably, n, u is 0 or 1;

q is-O-, -S-or-NR _a -, preferably Q is-O-;

R _a is H, C _1-6 Alkyl, C _1-3 Haloalkyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl;

R ₁ is H, halogen, cyano, amino, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Alkylamino, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, R ₁ May be substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl or 3-8 membered heteroaryl;

(R ₂ ) _p represents hydrogen on the ring by p R ₂ Substituted, each R ₂ The same or different;

p is 2,3,4,5 or 6;

R ₂ ，R _x ，R _y independently selected from the following substituents: h, halogen, hydroxy, carboxy, cyano, C _1-6 Alkyl, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl, 3-8 membered heteroaryl, -C (O) NR _b R _c ，-S(O) ₂ NR _b R _c And may be substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl or 3-8 membered heteroaryl;

R _x ，R _y Together with the carbon atoms to which they are attached, are capable of forming a 3-6 membered saturated ring which is a fully carbocyclic ring or a heterocyclic ring containing one or more atoms selected from the group consisting of N, O and S, and which may be substituted with one or more R _m Substitution;

where p are independently R ₂ Substituted on the ring, there being at least two R ₂ Attached to the same carbon atom and forming, together with the carbon atoms to which they are attached, a 3-to 6-membered saturated ring which is a fully carbocyclic ring or a heterocyclic ring containing one or more atoms selected from N, O and S, and which may be substituted with one or more R _m Substitution;

R _m is C _1-6 Alkyl, C _1-3 Haloalkyl, halogen, cyano, -OR _c ，-NR _b R _c ，C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, and,C _6-10 Aryl or 3-8 membered heteroaryl;

R _b is H, C _1-6 Alkyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl;

R _c is L, L-C (O) -, L-CH ₂ -or L-S (O) ₂ -，

Wherein L is H, C _1-6 Alkyl, C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl, L may be substituted with one or more of the following groups: halogen, hydroxy, C _1-6 Alkoxy, cyano, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl;

a is

Z is N or CR ₃ The method comprises the steps of carrying out a first treatment on the surface of the Preferably, Z is CR ₃ ；

Z' is N or CR ₄ The method comprises the steps of carrying out a first treatment on the surface of the Preferably Z' is CR ₄ ；

R ₃ , R ₄ , R ₅ , R ₆ Independently H, CN, halogen, C _1-3 Alkyl or C _1-3 A haloalkyl group;

v is N or CR ₉ Preferably, V is CR ₉ Wherein R is ₉ Is H, CN, halogen, C _1-3 Alkyl, C _1-3 Haloalkyl or-O-W;

w is a 5 or 6 membered heteroaryl or phenyl, preferably pyridinyl, pyrimidinyl, pyrazolyl or phenyl, which may be optionally substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkyl, C _1-3 Alkoxy, C _1-3 Haloalkyl and C _1-3 Haloalkoxy groups.

Detailed Description

In a preferred embodiment, the compound of formula (I) has one of the following 12 structures:

wherein the method comprises the steps of

m is 1 or 2; preferably, m is 1;

q is-O-, -S-or-NR _a -; preferred Q is-O-;

R _x ，R _y independently selected from the following substituents: h, halogen, hydroxy, carboxy, cyano, C _1-6 Alkyl, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl, 3-8 membered heteroaryl, -C (O) NR _b R _c ，-S(O) ₂ NR _b R _c And may be substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl or 3-8 membered heteroaryl;

R _x ，R _y together with the carbon atoms to which they are attached, are capable of forming a 3-6 membered saturated ring which is either fully carbocyclic or contains one or more heterocyclic rings selected from the group consisting of N, O and S atoms, and which may be substituted with one or more of the following substituents: c (C) _1-6 Alkyl, C _1-3 Haloalkyl, halogen, cyano, oxo (=o), -OR _c ，-NR _b R _c ，C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl;

u is 0 or 1, preferably u is 0, while R _x ，R _y All are H;

R ₁ is H, halogen, cyano, amino, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Alkylamino, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl, R ₁ Optionally substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl or 3-8 membered heteroaryl; preferably, R ₁ Is H, halogen, cyano, C _1-6 Alkyl or C _1-6 An alkoxy group; preferably, R ₁ Is H, halogen, cyano, C _1-3 Alkyl or C _1-3 An alkoxy group; more preferably，R ₁ Is H, fluoro, chloro, cyano, methyl, ethyl or methoxy; most preferably, R ₁ Is H or methoxy;

R _m is C _1-6 Alkyl, C _1-3 Haloalkyl, halogen, cyano, oxo (=o), -OR _c ，-NR _b R _c ，C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl;

R _b is H, C _1-6 Alkyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl;

R _c is L, L-C (O) -, L-CH ₂ -or L-S (O) ₂ -，

Wherein L is H, C _1-6 Alkyl, C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl, L optionally substituted with one or more of the following groups: halogen, hydroxy, C _1-6 Alkoxy, cyano, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl;

a is

Z is N or CR _3； Preferably, Z is CR ₃ ；

R ₃ , R ₄ , R ₅ , R ₆ Independently H, CN, halogen or C _1-3 A haloalkyl group;

w is a 5 or 6 membered heteroaryl or phenyl, preferably pyridinyl, pyrimidinyl, pyrazolyl or phenyl, which may be optionally substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkyl, C _1-3 Alkoxy, C _1-3 Haloalkyl and C _1-3 Halogenated compoundsAn alkoxy group;

in formula (I) and the above 12 formulae, preferably, A is

R ₅ , R _6, R ₇ , R ₈ , R ₉ H, F or CN independently;

or A is

R ₅ , R _6, R ₇ , R ₈ H, F or CN independently;

R ₉ is-O-W;

w is a 5-or 6-membered heteroaryl or phenyl, preferably pyridinyl, pyrimidinyl, pyrazolyl or phenyl, which may optionally be substituted by one or more substituents C _1-3 Haloalkyl, C _{1- 3} Haloalkoxy, CN, halogen and C _1-6 An alkyl group.

More preferably, A is

R ₇ , R ₈ H, F or CN independently;

R ₉ is-O-W;

w is pyridinyl, pyrimidinyl, pyrazolyl or phenyl, which may be optionally substituted with one or more substituents independently selected from the group consisting of: halogen, CN, CF ₃ 、-OCF ₃ 、CHF ₂ And CH (CH) ₃ ；

Most preferably, a is selected from the following groups:

。

in one embodiment, the compound is any one of the following compounds:

the compounds of the above formula, salts thereof (e.g., pharmaceutically acceptable salts) may exist as stereoisomers (e.g., it contains one or more asymmetric carbon atoms). The individual stereoisomers (enantiomers and diastereomers) and mixtures thereof are included within the scope of the invention.

The invention also includes various deuterated forms of the compounds of the above formula, salts thereof (e.g., pharmaceutically acceptable salts). Each available hydrogen atom attached to a carbon atom may be independently substituted with a deuterium atom. One of ordinary skill in the art will understand how to synthesize deuterated forms of the compounds of the above formula, salts thereof (e.g., pharmaceutically acceptable salts). Commercially available deuterated starting materials may be used in the preparation of the deuterated forms of the compounds of the above formula, salts thereof (e.g., pharmaceutically acceptable salts), or conventional techniques employing deuterating reagents such as lithium aluminum deuteride may be used to synthesize these compounds.

In addition to the free base or free acid forms of the compounds described herein, salt forms of the compounds are also within the scope of the invention. Salts or pharmaceutically acceptable salts of the compounds of the invention may be prepared in situ during the final isolation and purification of the compounds, or by separately reacting the purified compounds in free acid or free base form with the appropriate base or acid, respectively. For a review of suitable pharmaceutical salts see Berge et al, J.Pharm.Sci.66, 1-19,1977; P L Gould, international Journal of Pharmaceutics,33 (1986), 201-217; and Bighley et al, encyclopedia of Pharmaceutical Technology, marcel Dekker Inc, new York 1996, volume 13, page 453-497.

The compounds described herein, salts (e.g., pharmaceutically acceptable salts), deuterated forms, solvates, or hydrates thereof, can exist in one or more polymorphic forms. Thus, in another aspect, the invention provides polymorphs of a compound defined herein, a salt thereof (e.g., a pharmaceutically acceptable salt), or a solvate or hydrate of a compound described herein or a salt thereof (e.g., a pharmaceutically acceptable salt).

The present invention also includes isotopically-labeled compounds and salts, which are identical to those of formula (i) above, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most frequently found in nature. Examples of isotopes that can be incorporated into compounds of the above formula or salts thereof are isotopes of hydrogen, carbon, nitrogen, deuterium, such as ³ H、 ¹¹ C、 ¹⁴ C and C ¹⁸ F these isotopically-labelled compounds of the formula above or salts thereof are useful in drug and/or substrate tissue distribution assays. For example, the number of the cells to be processed, ¹¹ c and C ¹⁸ The F isotope may be used in PET (positron emission tomography). PET can be used for brain imaging. In one embodiment, the compound of the above formula or salt thereof is non-isotopically labeled.

Thus, the compounds of the present invention include compounds of the above formula, or salts thereof, e.g., pharmaceutically acceptable salts thereof. Representative compounds of the present invention include the specific compounds.

The invention also relates to pharmaceutical compositions comprising a compound of the invention and a pharmaceutically acceptable excipient.

The invention also relates to the treatment or prevention of Lp-PLA ₂ Comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the invention described herein. The disease may be associated with: involvement of monocytes, macrophages or lymphocytes increases the formation of lysophosphatidylcholine and oxidized free fatty acids with Lp-PLA ₂ Oxidative or endothelial dysfunction of the activity-associated lipids.

The invention also provides for the inhibition of Lp-PLA ₂ Methods of treating or preventing diseases with activity. Exemplary diseases include, but are not limited to: neurodegenerative diseases (e.g., alzheimer's disease, parkinson's disease, huntington's disease, vascular dementia), various neuropsychiatric diseases such as schizophrenia and autism, peripheral and cerebral atherosclerosis, stroke, metabolic bone diseases (e.g., bone marrow abnormalities), dyslipidemia, paget's disease, type II diabetes, hypertension, angina, myocardial infarction, ischemia, reperfusion injury, metabolic syndrome, insulin resistance and hyperparathyroidism, diabetic complications (e.g., macular edema, diabetic retinopathy and post-uveitis, diabetic ulcers and diabetic nephropathy), diabetic peripheral neuropathic pain, inflammatory pain, neuropathic pain, various cancers (e.g., prostate cancer, colon cancer, breast cancer, kidney cancer, lung cancer and ovarian cancer, etc.), macular edema, wound healing, male erectile dysfunction, rheumatoid arthritis, chronic Obstructive Pulmonary Disease (COPD), sepsis, acute and chronic inflammation, psoriasis and multiple sclerosis. The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound of the invention. The present invention is not intended to be limited to any particular stage of disease (e.g., early or late).

The invention also provides methods of treating or preventing Alzheimer's disease. The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound of the invention.

The invention also provides methods of treating or preventing atherosclerosis. The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound of the invention.

The invention also provides methods of treating or preventing ocular disorders by administering the compounds of the invention. In some embodiments, the invention provides methods of treating macular edema comprising administering to a subject a therapeutically effective amount of a compound of the invention. In some embodiments, the macular edema is associated with a diabetic eye disease (e.g., diabetic macular edema or diabetic retinopathy). In one embodiment, the macular edema is associated with posterior uveitis.

The invention also provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prophylaxis of a disease as described herein.

The invention also provides a compound of the invention for use in the treatment or prophylaxis described herein.

As used herein, "and/or" refers to any and all possible combinations including one or more of the associated listed items. It will be further understood that the terms "comprises" and/or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof.

Generally, the nomenclature used herein and the laboratory procedures in organic chemistry, pharmaceutical chemistry, biology described herein are those well known and commonly employed in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Where there are multiple definitions of terms used in this application, the definitions of this section control unless otherwise indicated.

Definition of the definition

As used herein, unless otherwise indicated, the term "disease" refers to any change in the state of an organism or some organ that interrupts or interferes with the performance of a function and/or causes symptoms (such as discomfort, dysfunction, adverse stress, or even death) in a diseased person or person in contact therewith.

As used herein, unless otherwise indicated, "diabetic retinopathy" refers to the result of chronic progressive retinal microvascular leakage and obstruction caused by diabetes. "diabetic macular edema" refers to retinal thickening or hard exudation deposits due to accumulation of extracellular fluid within a range of disc diameters of the fovea caused by diabetes.

As used herein, unless otherwise indicated, "neurodegenerative disease" refers to a different class of central nervous system disorders characterized by progressive, progressive loss of nerve tissue and/or nerve tissue function. Neurodegenerative diseases are a class of neurological diseases in which neurological diseases are characterized by progressive, progressive loss of neural tissue and/or altered neural function, typically resulting from progressive, progressive loss of neural tissue. In some embodiments, the neurodegenerative diseases described herein include neurodegenerative diseases in which a defective blood brain barrier (e.g., a permeable blood brain barrier) is present. Examples of neurodegenerative diseases in which a defective blood brain barrier exists include, but are not limited to, alzheimer's disease, huntington's disease, parkinson's disease, vascular dementia, and the like.

As used herein, unless otherwise indicated, "vascular dementia" is also referred to as "multi-infarct dementia," which refers to a group of syndromes caused by different mechanisms, all of which result in vascular damage in the brain. For example, the major subtypes of vascular dementia are vascular mild cognitive impairment, multi-infarct dementia, vascular dementia due to major single infarcts (affecting the thalamus, anterior cerebral arteries, parietal lobes or cingulate gyrus), vascular dementia due to hemorrhagic lesions, small vessel diseases (including, for example, vascular dementia due to lacunar lesions and binswanger disease (Binswanger disease)), and mixed dementia.

As used herein, unless otherwise indicated, "neuropathic pain" is pain that is motivated or caused by primary damage and dysfunction of the nervous system.

As used herein, unless otherwise indicated, "inflammatory pain" is pain caused by local acute inflammation or chronic inflammation stimulating nerves.

As used herein, unless otherwise indicated, "diabetic peripheral neuropathic pain" refers to pain caused by nerve damage that is concurrent with diabetes, the nerve damage in diabetes being due at least in part to reduced blood flow and hyperglycemia.

As used herein, unless otherwise indicated, "blood brain barrier" or "BBB" is used interchangeably herein to refer to a permeable barrier present in blood vessels that pass through brain tissue that severely restricts and tightly regulates the exchange of substances between blood and brain tissue. The blood brain barrier component includes endothelial cells that form the innermost lining of all blood vessels, tight junctions between adjacent endothelial cells that are structural associates of the BBB, the basement membrane of endothelial cells, and enlarged podophy covering nearly all exposed vessel outer surfaces of nearby astrocytes.

As used herein, unless otherwise indicated, "metabolic bone disease" refers to a different type of bone disease characterized by progressive and progressive loss of bone tissue. Metabolic bone disease as described herein is a bone metabolic disease in which there is a condition of reduced diffuse bone density and/or reduced bone strength. Such diseases are characterized by histological appearance. Exemplary metabolic bone diseases include, but are not limited to, osteoporosis characterized by reduced mineral and bone matrix and osteomalacia characterized by reduced mineral but intact bone matrix.

As used herein, unless otherwise indicated, "osteopenic disease" or "osteopenia" may be used interchangeably herein, referring to conditions having reduced calcification and/or bone density, as descriptive terms used to refer to all skeletal systems in which calcification and/or reduced bone density is observed. Osteopenia also refers to a reduction in bone mass due to insufficient synthesis of osteoid (osteoid).

As used herein, unless otherwise indicated, "osteoporosis" refers to a condition in which mineral and/or bone matrix is reduced.

As used herein, unless otherwise indicated, "alkyl" is a monovalent, saturated, chain of carbon atoms of the specified number. For example, C _1-3 Alkyl refers to an alkyl group having 1 to 3 carbon atoms. C (C) _1-6 Alkyl refers to an alkyl group having 1 to 6 carbon atoms. Alkyl groups may be straight or branched. In some embodiments, branched alkyl groups may have one, two, or three branches. Exemplary alkyl groups include, but are not limited to, methyl, methylethyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl)Radical and tert-butyl).

As used herein, unless otherwise indicated, an "alkoxy" substituent is a group of formula "R-O-" wherein R is alkyl as defined above. For example, C _1-3 Alkoxy refers to such alkoxy substituents containing 1 to 3 carbons. Exemplary alkoxy substituents include, but are not limited to, methoxy, ethoxy, n-propoxy, n-butoxy, n-pentoxy, n-hexoxy, isopropoxy, isobutoxy, sec-butoxy, tert-butoxy, isopentoxy, and neopentoxy.

As used herein, unless otherwise indicated, "cycloalkyl" refers to monovalent saturated cyclic hydrocarbon groups including bridged and spiro rings, preferably having 3 to 7 ring carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or [1.1.1] propranes, as well as those specifically exemplified below.

As used herein, unless otherwise indicated, "aryl" means a hydrocarbon group containing one or more aromatic rings, such as phenyl or naphthyl, and the like.

As used herein, unless otherwise indicated, "heteroaryl" means a stable monocyclic, bicyclic, or tricyclic ring having up to 8 atoms in each ring, wherein at least one ring is aromatic and at least one ring contains 1 to 4 heteroatoms selected from O, N and S. Heteroaryl groups within the scope of this definition include, but are not limited to, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, quinazolinyl, pyrazolyl, indolyl, isoindolyl, 1H, 3H-1-oxoisoindolyl, benzotriazolyl, furanyl, thienyl, pyridomorpholinyl, pyridopiperidinyl, pyridopyrrolidinyl, benzothienyl, benzofuranyl, benzodioxan, benzodioxabenzene, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, benzoxazolyl, imidazolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinolinyl, thiazolyl, isothiazolyl, 1, 2, 3-triazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 4-thiadiazolyl, 1, 3, 5-triazinyl, 1, 2, 4, 5-tetrazinyl, tetrazolyl, xanthenyl, phenazinyl, phenothiazinyl, phenoxazinyl, azepinyl, oxazinyl and thiazinyl. Particular heteroaryl groups have a 5-or 6-membered ring, for example furyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, diazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridomorpholinyl, pyridopiperidinyl, pyridopyrrolidinyl.

As used herein, unless otherwise indicated, "heterocycle" or "heterocyclyl" refers to a cyclic hydrocarbon in which 1 to 4 carbon atoms have been replaced by heteroatoms independently selected from N, N (R), S, S (O), S (O) and O. The heterocycle may be saturated or unsaturated, but is not aromatic. Heterocyclic groups may also be those containing 1, 2 or 3 rings, including bridged and spiro ring structures. Examples of suitable heterocyclyl groups include, but are not limited to: azetidine, oxetane, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, 2-oxopyrrolidinyl, pyrrolinyl, pyranyl, dioxolanyl, piperidinyl, 2-oxopiperidinyl, pyrazolinyl, imidazolinyl, thiazolinyl, dithiodienyl, oxathiolenyl, dioxanyl, dioxazolyl, oxathiazolyl (oxathiozolyl), oxazolonyl, piperazinyl, morpholino, thiomorpholino, 3-oxomorpholino, dithianyl, trithianyl and oxazinyl.

The term "bridged ring compound" refers to one or more atoms (i.e., C, O, N or S) attached to two non-adjacent carbon or nitrogen atoms. Preferred bridged rings include, but are not limited to: one carbon atom, two carbon atoms, one nitrogen atom, two nitrogen atoms and one carbon-nitrogen group. Notably, one bridge always converts a single ring to a tricyclic ring. In bridged rings, substituents on the ring may also be present on the bridge.

The term "spiro compound" refers to a polycyclic compound in which two monocyclic rings share one carbon atom, the shared carbon atom being referred to as a spiro atom.

As used herein, unless otherwise indicated, "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). Halo refers to a halogen group: fluorine (-F), chlorine (-Cl), bromine (-Br), or iodine (-I).

As used herein, unless otherwise indicated, "Haloalkyl "is alkyl substituted with one or more halo substituents, which may be the same or different. For example, C _1-3 Haloalkyl refers to haloalkyl substituents containing 1 to 3 carbons. Exemplary haloalkyl substituents include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, 1-chloro-2-fluoroethyl, trifluoropropyl, 3-fluoropropyl, and 2-fluoroethyl.

As used herein, a ring may be spiro-fused or unilaterally-fused when two substituents on the ring, together with their interconnecting atoms, combine to form another ring, unless otherwise indicated. A spiro-fused ring system consists of two rings having only one carbon atom in common. A single-sided-fused ring system consists of two rings that share only two atoms and one bond.

As used herein, unless otherwise indicated, "optionally substituted" means that the group or ring may be unsubstituted or that the group or ring may be substituted with one or more substituents as defined herein.

As used herein, unless otherwise indicated, "4-, 5-or 6-membered saturated ring optionally containing a heteroatom selected from N or O" means a 4-, 5-or 6-membered saturated carbocyclic ring and one carbon atom ring member may optionally be replaced by a heteroatom selected from N or O, for example, cyclobutyl, cyclopentyl, cyclohexenyl, azetidinyl (aziridinyl), pyrrolidinyl, piperidinyl, oxetanyl, tetrahydrofuranyl and tetrahydro-2H-pyranyl.

As used herein, unless otherwise indicated, "treatment", "treating" or "treatment" in reference to a disease refers to: (1) reducing the disease or reducing one or more biological manifestations of the disease, (2) interfering with (a) one or more points in a biological cascade that results in or causes the disease or (b) one or more biological manifestations of the disease, (3) reducing one or more symptoms or effects associated with the disease, and/or (4) reducing the progression of the disease or one or more biological manifestations of the disease, and/or (5) reducing the severity of the disease or the likelihood of biological manifestations of the disease.

As used herein, unless otherwise indicated, "preventing" refers to the prophylactic administration of a drug to reduce the likelihood of, or delay the onset of, a disease or its biological manifestation.

As used herein, unless otherwise indicated, "subject" refers to mammalian subjects (e.g., dogs, cats, horses, cows, sheep, goats, monkeys, etc.), and in particular human subjects.

As used herein, unless otherwise indicated, "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the subject compound and exhibits minimal undesirable toxicological effects. These pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in free acid or free base form with the appropriate base or acid, respectively.

As used herein, unless otherwise indicated, the term "therapeutically effective amount" refers to an amount that results in the treatment or prevention of a disease as compared to a corresponding subject that does not receive the amount, but which is low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. The therapeutically effective amount of a compound will vary with the particular compound selected (e.g., in view of potency, efficacy, and half-life of the compound): a selected route of administration; a treated disease; severity of the disease treated; age, body type, weight and physical condition of the patient are treated: a medical history of the patient being treated; duration of treatment; the nature of concurrent therapy; the desired therapeutic effect will vary depending on factors such as the desired therapeutic effect but can still be determined in a conventional manner by one skilled in the art.

Synthesis of Compounds

Those skilled in the art will appreciate that if the substituents described herein are not compatible with the synthetic methods described herein, the substituents may be protected with suitable protecting groups that are stable under the reaction conditions. The protecting groups may be removed at appropriate points in the reaction sequence to give the desired intermediate or target compound. Suitable protecting groups and methods for protecting and deprotecting various substituents using such suitable protecting groups are well known to those skilled in the art; examples of this can be found in I.Greene and P.Wuts, protecting Groups in Chemical Synthesis (third edition), john Wiley & Sons, N.Y. (1999). In some cases, substituents that are reactive under the reaction conditions used may be specifically selected. In these cases, the reaction conditions transform the selected substituent into another substituent that can be used as an intermediate compound or another substituent that is a desired substituent in the target compound.

General scheme

General scheme general schemes provide general synthetic routes to compounds of formula 1.8, wherein R ₁ ，R ₂ ，R _x ，R _y Q, m, n, p, u, A are as defined for formula (I). The route comprises the following steps:

Step (i): generating compound (1.2) by removing Boc protecting group from compound (1.1);

step (ii) of reacting the compound (1.2) with the compound (1.3) to produce the compound (1.5);

step (iv): obtaining a compound (1.7) by ring-closing the compound (1.5);

step (v) Compounds (1.7) and HQ- (CH) ₂ ) _m -a reaction to give the final product (1.8), namely a compound of formula I;

or comprises the following steps:

step (iii): reacting compound (1.2) with compound (1.4) to form compound (1.6);

step (iv'): obtaining a compound (1.7) by ring-closing the compound (1.6);

wherein R is ₁ ，R ₂ ，R _x ，R _y Q, m, n, p, u, A are as defined herein.

Specifically, the final compound 1.8 can be prepared by the steps i, ii, iv, v, and also by the steps i, iii, iv, v. Step (i) compound 1.2 may be formed by removing the Boc protecting group from compound 1.1 in a suitable acidic reagent, such as a hydrogen chloride/1, 4-dioxane solution, at a suitable temperature, such as room temperature. Step (ii) or (iii) may be used as SN _Ar The reaction is carried out using a suitable basic reagent, such as triethylamine, and compound 1.2 is reacted with 1.3 to form compound 1.5 or compound 1.2 is reacted with 1.4 to form compound 1.6 in a suitable solvent, such as acetonitrile, at a suitable temperature, such as room temperature. Either compound 1.5 or 1.6 can be obtained as the same intermediate 1.7 by the same procedure step (iv) which can be carried out by reacting either compound 1.5 or 1.6 with a suitable reagent, such as triethylamine/methanesulfonyl chloride, or thionyl chloride, at a suitable temperature, such as 0 ^o C or room temperature, the hydroxy group is converted to mesylate or chloro, and then further reacted without purification in a basic reagent such as potassium carbonate or sodium carbonate and a suitable solvent such as acetonitrile with heating to afford compound 1.7. Step (v) reacting 1.7 with the corresponding alcohol, thiol, amine HQ- (CH) ₂ ) _m -A (Q is-O-, -S-, -NR) _a (-) in a suitable solvent such as acetonitrile under suitable base such as NaH to give the final product 1.8.

Use of the same

The compound of the invention is Lp-PLA ₂ An inhibitor. Thus, these compounds are useful in therapy, e.g., in the treatment or prevention of and Lp-PLA ₂ Comprises the use of a therapeutically effective amount of Lp-PLA ₂ Inhibitor treatment a subject in need of such treatment. Accordingly, one aspect of the invention relates to the treatment or prevention of and Lp-PLA ₂ Methods of treating diseases associated with activity. Those skilled in the art will appreciate that a particular disease or treatment thereof may involve association with Lp-PLA ₂ One or more potential mechanisms associated with activity, including one or more of the mechanisms described herein.

In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prophylaxis of any of the diseases disclosed in the following published patent applications: WO96/13484, WO96/19451, WO97/02242, WO97/12963, WO97/21675, WO97/21676, WO97/41098, WO97/41099, WO99/24420, WO00/10980, WO00/66566, WO00/66567, WO00/68208, WO01/60805, WO02/30904, WO02/30911, WO03/015786, WO03/016287, WO 03/0410112, WO03/042179, WO03/042206, WO03/042218, WO03/086400, WO03/87088, WO08/048867, US2008/0103156, US2008/0090851, US2008/0090852, WO08/048866, WO 05/118 (CA 2530816 Al), WO 06/060630813, WO 2008/176, WO2013013503A1, WO2013014185A1, WO96 A1, WO 03/5782 A1, WO 200188847 A1, WO 023592 a, WO 023595/023595, WO 023595 and WO 023595/023595.

In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment of an eye disease. The ocular diseases for which the present invention is applicable may be related to the disruption of the blood-retinal barrier (iBRB). Exemplary ocular diseases involve diabetic ocular diseases including macular edema, diabetic retinopathy, posterior uveitis, retinal vein occlusion, and the like. Further ocular diseases include, but are not limited to, central retinal vein occlusion, branch retinal vein occlusion, ibm (post-cataract and post-operative), retinitis pigmentosa, blepharitis, bird gun-like retinochoroidal lesions, epiretinal membranes, choroidal tumors, cystic macular edema, paracentesis telangiectasia, traction maculopathy, vitreomacular traction syndrome, retinal detachment, retinitis optica, idiopathic macular edema, and the like. Using Lp-PLA ₂ More details of inhibitors for the treatment of ocular diseases are provided in WO2012/080497, which is incorporated herein by reference.

Furthermore, some embodiments of the invention provide the use of a compound of the invention in the manufacture of a medicament for treating or preventing diabetic macular edema in a subject. In some embodiments, the invention provides the use of a compound of the invention for treating diabetic macular edema in a subject.

In certain embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for treating or preventing macular edema or a subject at risk of having macular edema. In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for treating a subject suffering from or at risk of suffering from macular edema. In another embodiment, the macular edema is associated with a diabetic eye disease, such as diabetic macular edema or diabetic retinopathy. In another embodiment, the macular edema is associated with posterior uveitis.

In certain embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prevention of glaucoma or macular degeneration. In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment of glaucoma or macular degeneration.

In one embodiment, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prevention of a disease associated with disruption of the blood-retinal barrier in a subject in need of such treatment. In one embodiment, the invention provides the use of a compound of the invention in the manufacture of a medicament for treating a disease associated with disruption of the blood-retinal barrier in a subject in need of such treatment.

In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prevention of any disorder involving endothelial dysfunction, e.g., atherosclerosis (e.g., peripheral vascular atherosclerosis and cerebrovascular atherosclerosis), diabetes, hypertension, angina pectoris, ischemia, and post-reperfusion disorders.

In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prophylaxis of any disease states which involve lipid oxidation associated with enzymatic activity, for example, other conditions in addition to conditions such as atherosclerosis and diabetes, for example, rheumatoid arthritis, stroke, inflammatory conditions of the brain (e.g., alzheimer's disease), various neuropsychiatric conditions (e.g., schizophrenia, autism), myocardial infarction, ischemia, reperfusion injury, sepsis, and acute and chronic inflammation.

In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for reducing the chance of a cardiovascular event (e.g., a heart attack, myocardial infarction, or stroke) in a patient suffering from coronary heart disease.

In some embodiments, the invention provides the use of a compound of the invention in the manufacture of a medicament for the treatment or prevention of a disease involving activated monocytes, macrophages or lymphocytes, as all of these cell types express Lp-PLA ₂ Including diseases involving activated macrophages (e.g., M1, dendritic and/or other macrophages that develop oxidative stress). Exemplary conditions include, but are not limited to, cow's skin, rheumatoid arthritis, wound healing, chronic Obstructive Pulmonary Disease (COPD), liver cirrhosis, atopic dermatitis, emphysema, chronic pancreatitis, chronic gastritis, aortic aneurysm, atherosclerosis, multiple sclerosis, alzheimer's disease, and autoimmune diseases such as lupus.

In other embodiments, the invention provides the use of a compound of the invention in the preparation of a medicament for the primary or secondary prevention of an acute coronary event (e.g., caused by atherosclerosis); adjuvant therapy to prevent restenosis; or delay the progression of diabetes or hypertensive renal insufficiency. Prevention includes treatment of subjects at risk for such conditions.

In some embodiments, the invention provides methods of treating or preventing a neurological disorder associated with abnormal Blood Brain Barrier (BBB) function, inflammation, and/or microglial activation in a subject in need of such treatment. In some embodiments, the invention provides methods of treating or preventing a neurological disorder associated with abnormal Blood Brain Barrier (BBB) function, inflammation, and/or microglial activation in a subject in need of such treatment. The method comprises administering to the subject a therapeutically effective amount of a compound of the invention. In another embodiment, the abnormal BBB is a permeable BBB. In another embodiment, the disease is a neurodegenerative disease. Examples of such neurodegenerative diseases are, but are not limited to, vascular dementia, alzheimer's disease, parkinson's disease and Huntington's disease. In one embodiment, the invention provides a method of treating or preventing a disease associated with leakage of the Blood Brain Barrier (BBB) in a subject. In some embodiments, the invention provides methods of treating a disease associated with leakage of the Blood Brain Barrier (BBB) in a subject. Exemplary diseases include, but are not limited to, cerebral hemorrhage and cerebral amyloid angiopathy. In one embodiment, the neurodegenerative disease is alzheimer's disease. In a specific embodiment, the neurodegenerative disease is vascular dementia. In one embodiment, the neurodegenerative disease is Multiple Sclerosis (MS).

In one embodiment, the compounds of the invention are useful for treating or preventing a neurodegenerative disease in a subject. The method comprises administering a compound of the invention (e.g., in the form of a pharmaceutical composition comprising a compound of the invention) to a subject in need of such treatment. In one embodiment, the compounds of the invention are useful for treating neurodegenerative diseases in a subject. Exemplary neurodegenerative diseases include, but are not limited to, alzheimer's disease, vascular dementia, parkinson's disease, and Huntington's disease. In a specific embodiment, the neurodegenerative disease of the invention is associated with an abnormal blood brain barrier. In one embodiment, the agent is administered to inhibit Lp-PLA ₂ The subject of the active agent is a human.

In one embodiment, the invention provides a method of treating or preventing a subject suffering from, or at risk of suffering from vascular dementia. The method comprises administering a compound of the invention (e.g., a pharmaceutical composition comprising a therapeutically effective amount of a compound of the invention) to a subject. In one embodiment, the invention provides a method of treating a subject suffering from or at risk of suffering from vascular dementia. In a specific embodiment, the vascular dementia is associated with Alzheimer's disease.

In certain embodiments, the invention relates to methods of treating or preventing metabolic bone disease by administering to a subject in need of such treatment a therapeutically effective amount of a compound of the invention. In some embodiments, the invention relates to methods of treating metabolic bone disease by administering to a subject in need of such treatment a therapeutically effective amount of a compound of the invention. Exemplary metabolic bone diseases include diseases associated with loss of bone mass and bone density, including but not limited to osteoporosis and osteopenia. Exemplary osteoporosis and osteopenic disorders include, but are not limited to, bone marrow abnormalities, dyslipidemia, paget's disease, type II diabetes, metabolic syndrome, insulin resistance, hyperparathyroidism and related disorders. In another embodiment, the subject in need of such treatment is a human.

It is believed that methods of preventing osteoporosis and/or osteopenia described herein may be inhibited by Lp-PLA ₂ Expression and/or inhibition of Lp-PLA ₂ Is effective in improving protein activity. Thus, some embodiments of the invention provide for inhibition of Lp-PLA by blocking enzymatic activity ₂ Is a method of (2). In another embodiment, there is provided a method of reducing and/or down-regulating Lp-PLA by ₂ Expression of RNA thereby inhibiting Lp-PLA ₂ Is a method of (2). In another embodiment, preventing and/or reducing bone loss and I or bone density loss results in preventing or reducing symptoms associated with metabolic bone diseases such as osteoporosis and/or osteopenia.

In particular embodiments, the method further comprises administering to a subject in need of treatment an additional therapeutic agent for treating metabolic bone disease. For example, when the metabolic bone disease is osteoporosis, other therapeutic agents may be used, such as biphosphates (e.g., alendronate, ibandronate, risedronate, calcitonin, raloxifene), selective estrogen modulators (SERMs), estrogen therapy, hormone replacement therapy (ET/HRT), and teriparatum.

In one embodiment, systemic inflammatory diseases such as juvenile rheumatoid arthritis, inflammatory bowel disease, kawasaki disease, multiple sclerosis, sarcoidosis, polyarteritis, psoriatic arthritis, reactive arthritis, systemic lupus erythematosus, valsalva-primordial syndrome, lyme disease, behcet's disease, ankylosing spondylitis, chronic granulomatous disease, start-stop inflammation (enthesitis) may be the root cause of posterior uveitis affecting the retina, and it may lead to macular edema. The present invention relates to methods of treating or preventing post-uveitis or any of these systemic inflammatory conditions by administering a therapeutically effective amount of a compound of the invention. In one embodiment, the invention provides a method of treating post-uveitis or any of these systemic inflammatory conditions by administering a therapeutically effective amount of a compound of the invention.

With Lp-PLA ₂ Treatment and/or prevention of activity-related disorders may be achieved using the compounds of the invention in monotherapy or in dual or multiple combination therapies. For example, the compounds of the invention may be used in combination with an antihyperlipidemic agent, an antiatherosclerotic agent, an antidiabetic agent, an antianginal agent, an antiinflammatory agent or an antihypertensive agent or an agent for reducing lipoprotein (a) (Lp (a)) to treat or prevent the diseases described herein. Examples of such agents include, but are not limited to, cholesterol synthesis inhibitors, such as statins: antioxidants, such as probucol; insulin sensitizers; calcium channel antagonists and anti-inflammatory agents, such as non-steroidal anti-inflammatory drugs (NSAIDs). Agents for reducing Lp (a) include phosphoramidates as described in WO 97/02037, WO 98/28310, WO 98/28311 and WO 98/28312. In one embodiment, the compounds of the present invention may be used in combination with one or more statins. Statins are well known cholesterol lowering agents, including atorvastatin, simvastatin, pravastatin, cerivastatin, fluvastatin, lovastatin and rosuvastatin. In some embodiments, the compounds of the present invention may be used with antidiabetic agents or insulin sensitizers. In one embodiment, the compounds of the invention may be used with pparγ activators, such as GI262570 (GlaxoSmithKline) and glitazone (glitazone) compounds, such as rosiglitazone, troglitazone and pioglitazone. The agent may be administered, for example, in a therapeutically effective amount known in the art or in an amount that is smaller or greater than the amount known in the art to provide effective treatment.

Combination therapy involves administration of therapeutic agents together in separate dosage forms or in a single dosage form. Combination therapy may include simultaneous or separate administration of therapeutic agents, which may be administered substantially simultaneously or substantially separately. Typically, combination therapy includes administering each agent such that a therapeutically effective amount of each agent is present in the body of the subject for at least a portion of the overlap.

Application method

The therapeutically effective amount of the compounds of the present invention will depend on a number of factors, including for example the age and weight of the intended recipient, the precise condition to be treated and its severity, the nature of the formulation and the route of administration, and will ultimately depend on the judgment of the prescribing physician. However, a therapeutically effective amount of a compound of the invention for treating the disorders described herein will typically range from 0.1 to 100 milligrams per kg of body weight of the recipient per day, more typically from 1 to 10 milligrams per kg of body weight per day. Thus, for example, for a 70kg adult mammal, the actual amount per day is typically 70 to 700 milligrams, and may be administered as a single dose per day or as multiple sub-doses per day, such as two, three, four, five, or six doses per day. Or the administration may be intermittent, for example, once every other day, once a week, or once a month. It is contemplated that similar dosages may be applicable to treat other conditions as described above.

The pharmaceutical compositions of the present invention may comprise one or more compounds of the present invention. In some embodiments, the pharmaceutical composition may comprise more than one compound of the invention. For example, in some embodiments, the pharmaceutical composition may comprise two or more compounds of the present invention. In addition, the pharmaceutical composition may optionally further comprise one or more additional pharmaceutically active compounds.

As used herein, "pharmaceutically acceptable excipient" refers to a pharmaceutically acceptable raw material, component, or carrier that participates in imparting morphology or consistency to the pharmaceutical composition. When mixed, each excipient is compatible with the other ingredients of the pharmaceutical composition, thereby avoiding interactions that significantly reduce the efficacy of the compounds of the invention when administered to a subject, as well as interactions that would result in pharmaceutically unacceptable pharmaceutical ingredients.

The compounds of the invention and one or more pharmaceutically acceptable excipients may be formulated into a dosage form suitable for administration to a subject by a desired route of administration. For example, dosage forms include those suitable for the following routes of administration: (1) Oral administration (including buccal or sublingual), such as tablets, capsules, caplets, pills, troches, powders, syrups, brewages, suspensions, solutions, emulsions, sachets and cachets; (2) Parenteral administration (including subcutaneous, intramuscular, intravenous or intradermal), for example, sterile solutions, suspensions and powders for reconstitution; (3) transdermal administration, such as transdermal patches; (4) rectal administration, such as suppositories; (5) Nasal inhalation, such as dry powders, aerosols, suspensions and solutions; and (6) topical administration (including buccal, sublingual or transdermal), such as creams, ointments, lotions, solutions, pastes, sprays, foams and gels. Such compositions may be prepared by any method known in the pharmaceutical arts, for example by combining a compound of the formula described above with a carrier or excipient.

Pharmaceutical compositions suitable for oral administration may be presented as discrete units, such as capsules or tablets; powder or granules; solutions or suspensions in aqueous or non-aqueous liquid form; edible foam (foams or whishes), or oil-in-water liquid emulsions or water-in-oil liquid emulsions.

Suitable pharmaceutically acceptable excipients may vary depending upon the particular dosage form selected. Furthermore, suitable pharmaceutically acceptable excipients may be selected according to the particular function they perform in the composition. For example, some pharmaceutically acceptable excipients may be selected for their ability to promote the production of a uniform dosage form. Some pharmaceutically acceptable excipients may be selected for their ability to promote the production of stable dosage forms. Some pharmaceutically acceptable excipients may be selected for their ability to facilitate the delivery or transport of one or more compounds of the invention from one organ or part of the body to another organ or part of the body upon administration to a subject. Some pharmaceutically acceptable excipients may be selected for their ability to increase patient compliance.

Suitable pharmaceutically acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, taste masking agents, colorants, anti-caking agents, humectants, chelating agents, plasticizers, tackifiers, antioxidants, preservatives, stabilizers, surfactants, and buffers. Those skilled in the art will appreciate that some pharmaceutically acceptable excipients may provide more than one function and may provide other functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

The skilled artisan is well within the knowledge and skill of the art to be able to select the appropriate amount of a suitable pharmaceutically acceptable excipient for use in the present invention. Furthermore, many sources are available to the skilled person that describe pharmaceutically acceptable excipients and can be used to select the appropriate pharmaceutically acceptable excipient. Examples include Remington pharmaceutical sciences (Remington's Pharmaceutical Sciences, mack publishing company), pharmaceutical additives manuals (The Hand book of Pharmaceutical Additives, gower publishing company limited), and pharmaceutical excipients manuals (Handbook of Pharmaceutical Excipients, american pharmaceutical association and pharmaceutical publishing company).

The pharmaceutical compositions of the present invention are prepared using techniques and methods known to those skilled in the art. Some methods commonly used in the art are described in Remington pharmaceutical sciences (Mack publishing).

In one aspect, the invention relates to a solid oral dosage form, such as a tablet or capsule, comprising a therapeutically effective amount of a compound of the invention and a release agent or filler. Suitable diluents and fillers include lactose, dextrose, glucose, mannitol, sorbitol, starches (e.g., corn starch, potato starch and pregelatinized starch), cellulose and its derivatives (e.g., microcrystalline cellulose), calcium sulfate and dibasic calcium phosphate. The oral solid dosage form may also include a binder. Suitable binders include starches (e.g., corn starch, potato starch, and pregelatinized starch), gelatin, acacia, sodium alginate, alginic acid, xanthan gum, guar gum, povidone, and cellulose and derivatives thereof (e.g., microcrystalline cellulose). The oral solid dosage form may also include a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmellose (croscarmellose), alginic acid and sodium carboxymethylcellulose. The oral solid dosage form may also contain a lubricant. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate and talc.

In a specific embodiment, the invention relates to a pharmaceutical composition comprising 0.01mg to 1000mg of one or more compounds of the above formula as described herein or a pharmaceutically acceptable salt thereof and 0.01g to 5g of one or more pharmaceutically acceptable excipients.

Example preparation

Intermediate 1

(S) -2- (((tert-Butoxycarbonyl) amino) -6- (dimethyl (oxo) -lambda) ⁶ -sulfinyl) -5-oxohexanoic acid methyl ester

At room temperature, 1- (tertiary butyl) 2-methyl groupS) 5-Oxopyrrolidine-1, 2-dicarboxylic acid ester (49 g,0.2 mol), potassium t-butoxide (34 g,0.3 mol) was added to dimethyl sulfoxide (500 mL), stirred at room temperature for 1 hour, and trimethylsulfoxide iodide (75 g,0.34 mol) was added, and stirred at room temperature for 3 hours. The reaction was poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography to give the title compound (41 g, 61%). MS: M/z [ M+H ]] ⁺ =336。

1- (tert-butyl) 2-methyl [ ]S) -5-Oxopiperidine-1, 2-dicarboxylic acid ester

At room temperatureS) -2- (((tert-Butoxycarbonyl) amino) -6- (dimethyl (oxo) -lambda) ⁶ -sulfinyl) -5-oxohexanoic acid methyl ester (4.1 g,12 mmol) and Ir [ COD ]] ₂ Cl (41 mg,0.6 mmol) was added to 1, 2-dichloroethane (50 mL), under argon, 90 ^o C stirring for 2 days. Concentrating the reaction solution and performing column chromatographyPurification gave the title compound (1.3 g, 33%). MS: M/z [ M+H ] ] ⁺ =258。

1- (tert-butyl) 2-methyl [ ]S) -5-methylenepiperidine-1, 2-dicarboxylic acid ester

At room temperature, 1- (tertiary butyl) 2-methyl groupS) 5-Oxopiperidine-1, 2-dicarboxylic acid ester (500 mg,1.9 mmol) was added to anhydrous tetrahydrofuran (50 mL), and the mixture was cooled to 0 under argon atmosphere ^o C, n-hexane solution (2.5 M,1 mL,2.5 mmol) of n-butyllithium was added dropwise, 0 ^o C stirring for 1 hour. Methyl triphenylphosphine bromide (500 mg,1.9 mmol) was added to the reaction solution and stirred at room temperature overnight. The reaction was quenched with water, extracted with dichloromethane, the organic phase concentrated and purified by column chromatography to give the compound (250 mg, 50%). MS: M/z [ M+H ]] ⁺ =256。

1-benzyl 2-methyl [ ]S) -5-methylenepiperidine-1, 2-dicarboxylic acid ester

At room temperature, 1- (tertiary butyl) 2-methyl groupS) 5-Methylpiperidine-1, 2-dicarboxylic acid ester (250 mg,1.0 mmol) and trifluoroacetic acid (228 mg,2.0 mmol) were added to dichloromethane (5 mL), and stirred at room temperature for 1 hour. The reaction solution was concentrated, and to the residue were added water (5 mL), potassium carbonate (414 mg,3.0 mmol) and benzyl chloroformate (170 mg,1.0 mmol) in this order, followed by stirring overnight at room temperature. The reaction was poured into water, extracted with dichloromethane, and the organic phase was concentrated to give the title compound (110 mg, 38%) by preparative thin layer chromatography. MS: M/z [ M+H ] ] ⁺ =290。

5-benzyl 6-methyl [ ]S) -5-azaspiro [ 2.5 ]]Octane-5, 6-dicarboxylic acid ester

Adding diethyl zinc in n-hexane solution (1 M,7.6 mL,7.6 mmol) into dichloromethane (7 mL) at room temperature, and cooling to 0 under argon protection ^o C, trifluoroacetic acid (560. Mu.L, 7.6 mmol) was added to the reaction solution, 0 ^o C stirring for 1 hour. Diiodomethane (2.0 g,7.6 mmol) was added to the reaction solution, 0 ^o C stirring for 1 hour. 1-benzyl 2-methyl radicalS) 5-Methylenepiperidine-1, 2-dicarboxylic acid ester (1.1 g,3.8 mmol) was added to the reaction solution, and stirred overnight at room temperature. The reaction solution was diluted with methylene chloride, poured into a saturated aqueous sodium hydrogencarbonate solution, and stirred at room temperature for 0.5 hours. Filtration, extraction of the filtrate with dichloromethane, concentration of the organic phase and purification by column chromatography gave the title compound (300 mg, 27%). MS: M/z [ M+H ]] ⁺ =304。

(S) -5-azaspiro [ 2.5 ]]Octane-6-carboxylic acid methyl ester

At room temperature, 5-benzyl 6-methyl radicalS) -5-azaspiro [ 2.5 ]]Octane-5, 6-dicarboxylic acid ester (300 mg,1.0 mmol) and palladium on carbon (10%, 30 mg) were added to methanol (10 mL) and hydrogenated overnight at room temperature under normal pressure. The reaction solution was filtered, and the filtrate was concentrated to give the crude title compound which was used directly in the next reaction. MS: M/z [ M+H ]] ⁺ =170。

(S) - (5-azaspiro [ 2.5 ]]Octane-6-yl) methanol

At room temperature, will S) -5-azaspiro [2.5 ]]Octane-6-carboxylic acid methyl ester (190 mg,1.1 mmol) was added to anhydrous tetrahydrofuran (2 mL), lithium aluminum hydride (45 mg,1.1 mmol) was added with stirring, and the mixture was stirred at room temperature for 2 hours. The reaction solution was quenched with sodium sulfate decahydrate, stirred at room temperature for 30 minutes, filtered, and the cake was rinsed with tetrahydrofuran, and the filtrate was concentrated to give the title compound (100 mg,2 steps yield: 64%). MS: M/z [ M+H ]] ⁺ =142。

(S) - (5- (2, 6-di)Chloropyrimidin-4-yl) -5-azaspiro [2.5]Octane-6-yl) methanol

At room temperature, willS) - (5-azaspiro [2.5 ]]Octane-6-yl) methanol (100 mg,0.7 mmol) and sodium carbonate solid (150 mg,1.4 mmol) were added to acetonitrile (2 mL), cooled to 0 under argon ^o C, 2, 4, 6-trichloropyrimidine (130 mg,0.7 mmol) was added and stirred overnight at room temperature. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (100 mg, 50%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =288。

(S) -3 '-chloro-8', 9', 9a', 10 '-tetrahydro-1'H, 6'H-spiro [ cyclopropane-1, 7' -pyridine [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

At room temperature, willS) - (5- (2, 6-dichloropyrimidin-4-yl) -5-azaspiro [2.5 ]]Octane-6-yl) methanol (50 mg,0.2 mmol) and thionyl chloride (250 mg,0.2 mmol) were added to dichloromethane (2 mL), and stirred at room temperature overnight. The reaction solution was concentrated, and to the residue were added successively potassium carbonate solid (50 mg,0.3 mmol) and acetonitrile (2 mL), 80 ^o C stirring overnight. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (20 mg, 45%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =252。

Intermediate 2

1-benzyl-5-oxopyrrolidine-2-carboxylic acid methyl ester

Methyl 5-oxopyrrolidine-2-carboxylate (4.0 g,28.0 mmol) and sodium hydride (60%, 2.24 g,56.0 mmol) were added to anhydrous tetrahydrofuran (50 mL) at room temperature and warmed to 60℃under argon ^o C benzyl bromide (5.7 g,33.6 mmol), 60 ^o C stirring for 2 hours. The reaction solution was cooled to room temperature, then poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=3/1) to give the title compound (4.2 g, 64%). MS: M/z [ M+H ]] ⁺ =234。

1-benzyl-5- (hydroxymethyl) pyrrolidin-2-one

1-benzyl-5-oxopyrrolidine-2-carboxylic acid methyl ester (4.2 g,18.02 mmol) was added to anhydrous tetrahydrofuran (40 mL) at room temperature, lithium borohydride (800 mg,36.0 mmol) was added with stirring, and stirred at room temperature for 1 hour. The reaction solution was quenched with sodium sulfate decahydrate and stirred at room temperature for 30 minutes. Filtration and concentration of the filtrate gave crude title compound (3.6 g, 97%). MS: M/z [ M+H ]] ⁺ =206。

1-benzyl-5- (((tert-butyldimethylsilyl) oxy) methyl) pyrrolidin-2-one

1-benzyl-5- (hydroxymethyl) pyrrolidin-2-one (3.6 g,14.4 mmol) was dissolved in dichloromethane (50 mL) at room temperature, imidazole (2.9 g,43.2 mmol) and t-butyldimethylchlorosilane (3.2 g,21.6 mmol) were added and stirred at room temperature for 30 minutes. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (3.6 g, 78%). MS: M/z [ M+H ]] ⁺ =320。

4-benzyl 5- (((tert-butyldimethylsilyl) oxy) methyl) -4-azaspiro [ 2.4 ] heptane

1-benzyl-5- (((t-butyl)Dimethylsilanyloxy) methyl-pyrrolidin-2-one (2.0 g,6.27 mmol) was added to anhydrous tetrahydrofuran (30 mL) and cooled to 0 under argon protection ^o C, tetraisopropyl titanate (4.5 mL,15.67 mmol) and ethyl magnesium bromide in tetrahydrofuran (2 M,15 mL,30 mmol) were added with stirring at room temperature overnight. The reaction solution was quenched with water, the reaction solution was filtered, and the filtrate was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=20/1) to give the title compound (1.3 g, 63%). MS: M/z [ M+H ]] ⁺ =332。

(4-benzyl-4-azaspiro [ 2.4 ] heptan-5-yl) methanol

Under the protection of argon, 4-benzyl 5- (((tert-butyldimethylsilyl) oxy) methyl) -4-azaspiro [ 2.4 ]Heptane (3.1 g,9.36 mmol) was added to a tetrahydrofuran solution (1 M,10.8 mmol,10.8 mL) of anhydrous tetrahydrofuran (40 mL) and tetrabutylammonium fluoride and stirred at room temperature for 3 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=15/1 to 10/1) to give the title compound (800 mg, 94%). MS: M/z [ M+H ]] ⁺ =218。

(4-azaspiro [ 2.4 ] heptane-5-yl) methanol

(4-benzyl-4-azaspiro [ 2.4 ] at room temperature]Heptan-5-yl) methanol (700 mg,3.22 mmol) and palladium on carbon (10%, 105 mg) were added to methanol (25 mL), 50 ^o C, hydrogenation for 3 days at normal pressure. The reaction solution was filtered, and the filtrate was concentrated to give the crude title compound (1.2 g, 256%). MS: M/z [ M+H ]] ⁺ =128。

(4- (2, 6-dichloropyridin-4-yl) -4-azaspiro [ 2.4 ] heptan-5-yl) methanol

2, 4, 6-trichloropyrimidine (582 mg,3.2 mmol) was added to acetonitrile (20 mL) at room temperature and cooled to 0 under argon ^o C. Sodium carbonate solid (510 mg,4.8 mmol) and (4-azaspiro [ 2.4 ] were added sequentially with stirring]The crude product of heptan-5-yl) methanol (600 mg,4.7 mmol) was stirred at room temperature for 2 hours. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (160 mg, 12%) by preparative thin layer chromatography (petroleum ether/ethyl acetate=5/1). MS: M/z [ M+H ] ] ⁺ =274。

3 '-chloro-7', 8', 8a', 9 '-tetrahydro-1'H-spiro [ cyclopropane-1, 6' -pyrrolo [1', 2': 3, 4]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

(4- (2, 6-dichloropyridin-4-yl) -4-azaspiro [ 2.4 ] at room temperature]Heptan-5-yl) methanol (120 mg,0.52 mmol) was added to dichloromethane (8 mL), and thionyl chloride (120 mg,1.04 mmol) was added dropwise at room temperature and stirred at room temperature for 10 minutes. The reaction solution was concentrated, and potassium carbonate (290 mg,2.08 mmol) and acetonitrile (12 mL), 85 were added to the residue in this order ^o C stirring overnight. The reaction was filtered and the filtrate was concentrated to give the title compound (80 mg, 65%) by preparative thin layer chromatography (dichloromethane/methanol=20/1). MS: M/z [ M+H ]] ⁺ =238。

Intermediate 3

1, 1-bis (iodomethyl) cyclopropane

Triphenylphosphine (26.0 g,99.0 mmol) and imidazole (6.5 g,96.0 mmol) were added to dichloromethane (30 mL) at room temperature, and cooled to 0 under argon ^o C, iodine (25.0 g,99.0 mmol) was added, 0 ^o C stirring for 1 hour. A solution of 1, 1-cyclopropane-dimethanol (5.0 g,48.0 mmol) in methylene chloride (20) mL) was added to the reaction solution, and the mixture was stirred at this temperature for 3 hours. The reaction solution was poured into brine (15%), extracted with ethyl acetate, the organic phase was washed with saturated sodium sulfite solution, the organic phase was concentrated, petroleum ether (200 mL) and ethyl acetate (10 mL) were added to the residue in this order, stirred at room temperature for 30 minutes, filtered, and the filtrate was concentrated to give the title compound (12 g, 75%). ¹ H NMR(400 MHz, CDCl ₃ ) d 3.34(br. s., 4 H), 1.02(br. s., 4 H)。

5- (tert-butyl) 6-ethyl 5-azaspiro [ 2.4 ] heptane-5, 6-dicarboxylic acid ester

Sodium hydride (60%, 3.0 g,75.0 mmol) was added to N, N-dimethylformamide (40 mL) at room temperature, and the temperature was lowered to 0 under the protection of argon gas ^o C, 1-bis (iodomethyl) cyclopropane (9.6 g,28.9 mmol) and ethyl (t-butoxycarbonyl) glycinate (6.2 g,30.5 mmol) in N, N-dimethylformamide (40 mL), 0 ^o C stirred for 1.5 hours. Acetic acid (2.5. 2.5 mL), 0 ^o C stirring for 2 hours. The reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (4.8 g, 62%). MS: M/z [ M+H-Boc ]] ⁺ =170。

(5-azaspiro [ 2.4 ] heptane-6-yl) methanolic hydrochloride

At room temperature, 5- (tert-butyl) 6-ethyl 5-azaspiro [ 2.4 ]]Heptane-5, 6-dicarboxylic acid ester (540 mg,2.0 mmol) was added to anhydrous tetrahydrofuran (6 mL), lithium borohydride (88 mg,4.0 mmol) was added with stirring, and stirred overnight at room temperature. The reaction solution was quenched with sodium sulfate decahydrate, stirred at room temperature for 0.5 hour, filtered, and the filtrate was concentrated, and the residue was added to dichloromethane (3 mL) and ethyl acetate solution of hydrogen chloride (4 m,6 mL). Stirring was carried out at room temperature for 30 minutes, and the reaction was concentrated to give the crude title compound (350 mg, 107%). MS: M/z [ M+H ] ] ⁺ =128。

(5- (2, 6-dichloropyrimidin-4-yl) -5-azaspiro [ 2.4 ] heptane-6-yl) methanol

2, 4, 6-trichloropyrimidine (760 mg,4.18 mmol) was added to acetonitrile (12 mL) at room temperature and cooled to 0 under argon ^o C, sodium carbonate solid (680 mg,6.42 mmol) and (5-azaspiro [ 2.4 ] were added sequentially with stirring]Heptan-6-yl) methanol hydrochloride (350 mg,2.14 mmol) was stirred at room temperature for 1 hour. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (140 mg, 19%) by preparative thin layer chromatography (petroleum ether/ethyl acetate=4/1). MS: M/z [ M+H ]] ⁺ =273。

3 '-chloro-8 a', 9 '-dihydro-1'H, 6'H, 8'H-spiro [ cyclopropane-1, 7' -pyrrolo [1', 2': 3, 4]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

(5- (2, 6-dichloropyrimidin-4-yl) -5-azaspiro [ 2.4 ] at room temperature]Heptan-6-yl) methanol (140 mg,0.51 mmol) was added to dichloromethane (6 mL), and thionyl chloride (184 mg,1.53 mmol) was added dropwise with stirring and stirred at room temperature for 10 minutes. The reaction solution was concentrated, and to the residue were added successively potassium carbonate solid (345 mg,2.5 mmol) and acetonitrile (8 mL), 85 ^o C stirring overnight. The reaction was filtered and the filtrate was concentrated to give the title compound (80 mg, 66%) by preparative thin layer chromatography (dichloromethane/methanol=20/1). MS: M/z [ M+H ]] ⁺ =238。

Intermediate 4

5-hydroxy-6-oxa-7-azaspiro [ 3.5 ] nonyl-7-ene-8-carboxylic acid ethyl ester

Cyclobutanecarboxaldehyde (1 g,11.9 mmol) and pyrrolidine (1.1 g,15.5 mm) were reacted at room temperatureol) was added to anhydrous toluene (12 mL), 4A molecular sieves (0.1 g) were added with stirring, and stirred at room temperature for 2 hours. Anhydrous tetrahydrofuran (12, mL) was added to the reaction solution, followed by addition of ethyl 3-bromo-2- (hydroxyimino) propionate (2.6 g,12.6 mmol,J. Org. Chem.1982, 47, 2147), stirring at room temperature for 0.5 hours. Triethylamine (1.3 g,12.7 mmol) was added to the reaction solution, and stirred at room temperature overnight. The reaction solution was concentrated, purified by column chromatography (petroleum ether/ethyl acetate=5/1), and purified by preparative thin layer chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (510 mg, 20%). ¹ H NMR(400 MHz, DMSO-d ₆ )d 7.44 - 7.14(m, 1 H), 5.26(br. s., 1 H), 4.21(q, J = 6.8 Hz, 2 H), 2.56(m, 1 H), 2.32(m, 1 H), 2.09 - 1.95(m, 1 H), 1.95 - 1.66(m, 4 H), 1.59(d, J = 10.3 Hz, 1 H), 1.25(t, J = 7.1 Hz, 3 H); MS: m/z [M+H] ⁺ =214。

6-azaspiro [ 3.4 ] octane-7-carboxylic acid ethyl ester

At room temperature, 5-hydroxy-6-oxa-7-azaspiro [ 3.5 ]]Ethyl nonyl-7-ene-8-carboxylate (510 mg,2.4 mmol) was added to ethanol (30 mL), raney-Ni catalyst (510 mg) was added under argon atmosphere and hydrogenated overnight at room temperature under normal pressure. Filtration and concentration of the filtrate gave the crude title compound (460 mg, 102%). ¹ H NMR(400 MHz, DMSO-d ₆ )d 4.06(d, J = 6.4 Hz, 2 H), 3.63(br. s., 1 H), 2.86(br. s., 1 H), 2.76(br. s., 1 H), 2.04(d, J = 8.3 Hz, 1 H), 1.96 - 1.67(m, 7 H), 1.23 - 1.11(m, 3 H); MS: m/z [M+H] ⁺ =184。

(6-azaspiro [ 3.4 ] octyl-7-yl) methanol

At room temperature, 6-azaspiro [ 3.4 ]]Crude octane-7-carboxylic acid ethyl ester (360 mg,1.97 mmol) was added to anhydrous tetrahydrofuran (10 mL) and methanol (1) mL), lithium borohydride (220 mg,5.8 mmol), 50 ^o C stirring for 2 hours. The reaction solution was cooled to 0 ^o And C, quenching the reaction solution by using sodium sulfate decahydrate. The mixture was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated to give the crude title compound (290 mg, 105%). MS: M/z [ M+H ]] ⁺ =142。

(6- (6-chloro-2-methoxypyrimidin-4-yl) -6-azaspiro [ 3.4 ] octyl-7-yl) methanol

At room temperature, (6-azaspiro [ 3.4 ]]Octyl-7-yl) methanol (290 mg,2.06 mmol) and 4, 6-dichloro-2-methoxypyrimidine (358 mg,2 mmol) were dissolved in isopropanol (20 mL), and sodium carbonate (848 mg,8 mmol), 80 was added with stirring ^o C stirred for 3 hours. The reaction solution was cooled to room temperature, the reaction mixture was diluted with dichloromethane, filtered through celite, and the filtrate was concentrated to give the title compound (160 mg, 24%) by preparative thin layer chromatography (petroleum ether/ethyl acetate=2/1). ¹ H NMR(400 MHz, CDCl ₃ )d 6.05(br. s., 1 H), 5.55(br. s., 1 H), 4.29(br. s., 1 H), 3.93(s, 3 H), 3.78 - 3.60(m, 2 H), 3.53 - 3.42(m, 1 H), 3.39(br. s., 1 H), 2.17 - 2.10(m, 1 H), 2.10 - 1.85(m, 7 H)； MS: m/z [M+H] ⁺ =284。

3 '-chloro-8 a', 9 '-dihydro-1'H, 6'H, 8'H-spiro [ cyclobutane-1, 7' -pyrrolo [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

(6- (6-chloro-2-methoxypyrimidin-4-yl) -6-azaspiro [ 3.4 ] at room temperature]Octyl-7-yl) methanol (140 mg, 0.49 mmol) and thionyl chloride (176 mg,1.48 mmol) were added to dichloromethane (3 mL) and stirred overnight at room temperature. The reaction solution was concentrated and dissolved in water (2 mL), 0 ^o Sodium hydroxide solid (78 mg,1.96 mmol), 0 was added at C ^o C stirred for 0.5 hours. Reaction liquidPoured into water, extracted with dichloromethane and the organic phase concentrated to afford the title compound (47 mg, 38%) as a white solid by preparative thin layer chromatography (dichloromethane/methanol=40/1). ¹ H NMR(400 MHz, CDCl ₃ )d 5.60(s, 1 H), 4.22(d, J = 7.8 Hz, 2 H), 4.09 - 3.97(m, 1 H), 3.47(d, J = 10.8 Hz, 1 H), 3.23(d, J = 10.8 Hz, 1 H), 2.37 - 2.27(m, 1 H), 2.18 - 1.88(m, 7 H); MS: m/z [M+H] ⁺ =252。

Intermediate 5

2- (((tert-Butyldimethylsilyl) oxy) methyl) -3-alkenylpiperidine-1-carboxylic acid tert-butyl ester

Methyl triphenylphosphine bromide (407 mg,1.14 mmol) was added to anhydrous tetrahydrofuran (10 mL) at room temperature, and cooled to 0 under argon ^o C, n-hexane solution (2.5 M,0.456 mL,1.14 mmL) of n-butyllithium was added dropwise, 0 ^o C stirred for 0.5 hours. Tert-butyl 2- (((tert-butyldimethylsilyl) oxy) methyl) -3-oxopiperidine-1-carboxylate (300 mg,0.87 mmol,Bioorganic and Medicinal Chemistry Letters2017, 27, 2210-2215) was slowly dropped into the reaction solution, and stirred at room temperature overnight. The reaction solution was cooled to 0 ^o C, quenched by addition of saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phase concentrated to give the title compound (92 mg, 29%) by preparative thin layer chromatography (dichloromethane/methanol=100/1). MS: M/z [ M+H-Boc ]] ⁺ =242。

(5-azaspiro [ 2.5 ] octyl-4-yl) methanolic hydrochloride

A solution of tert-butyl 2- (((tert-butyldimethylsilyl) oxy) methyl) -3-methylpiperidine-1-carboxylate (82 mg,0.24 mmol) and diethyl zinc in n-hexane (1 M,2.4 mL,2.4 mmL) was added to the di-phase at room temperatureCooling to 0 in chloromethane (20 mL) under the protection of argon ^o Diiodomethane (1.3 g,4.8 mmol) was added dropwise and the mixture was stirred overnight at room temperature after completion of the dropwise addition. The reaction solution was cooled to 0 ^o The reaction mixture was quenched with sodium sulfate decahydrate, filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, methanol (2 mL) and ethyl acetate solution of hydrogen chloride (4 m,1 mL) were added sequentially to the residue, and stirred at room temperature for 5 hours. The reaction solution was concentrated to give the crude title compound (90 mg, 211%). MS: M/z [ M+H ]] ⁺ =142。

(5- (6-chloro-2-methoxypyrimidin-4-yl) -5-azaspiro [ 2.5 ] octyl-4-yl) methanol

At room temperature, (5-azaspiro [ 2.5 ]]Octyl-4-yl) methanol hydrochloride crude (90 mg,0.51 mmol), 4, 6-dichloro-2-methoxypyrimidine (90 mg,0.39 mmol) and sodium carbonate solid (127 mg,1.2 mmol) were added to isopropanol (5 mL), 80 ^o C stirring for 2 days. The reaction solution was cooled to room temperature, poured into water, extracted with dichloromethane, and the organic phase was concentrated to give the title compound (20 mg, 14%) by preparative thin layer chromatography (petroleum ether/ethyl acetate=2/1). ¹ H NMR(400 MHz, CDCl ₃ )d 6.24(s, 1 H), 4.79 - 4.50(m, 2 H), 4.01(br. s., 1 H), 3.88(s, 3 H), 3.81(d, J = 7.3 Hz, 1 H), 3.14(d, J = 7.3 Hz, 1 H), 2.19 - 1.93(m, 2 H), 1.74(br. s., 1 H), 1.71 - 1.59(m, 1 H), 0.88 - 0.76(m, 1 H), 0.68 - 0.49(m, 1 H), 0.47 - 0.36(m, 1 H), 0.21(br. s., 1 H); MS: m/z [M+H] ⁺ =284。

3 '-chloro-7', 8', 9a', 10 '-tetrahydro-1'H, 6'H-spiro [1, 9' -pyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

(5- (6-chloro-2-methoxypyrimidin-4-yl) -5-azaspiro [ 2.5 ] at room temperature]Octyl-4-yl) methanol (20 mg,0.07 mmol) and thionyl chloride (26 mg,0.22 mmol) were added to dichloromethane (3 mL) and stirred at room temperature overnight. The reaction mixture was concentrated, water (2 mL) was added to the residue, and the mixture was cooled to 0 ^o C, sodium hydroxide solid (11 mg,0.28 mmol) was added with stirring. 0 ^o C stirred for 0.5 hours. The reaction was poured into water, extracted with dichloromethane and the organic phase concentrated to give the crude title compound (24 mg, 113%). MS: M/z [ M+H ]] ⁺ =252。

Intermediate 6

3-azaspiro [ 5.5 ] undecane-3-carboxylic acid tert-butyl ester

At room temperature, 3-azaspiro [ 5.5 ]]Undecane (900 mg,5.87 mmol), di-tert-butyl dicarbonate (1.92 g,8.81 mmol) and sodium carbonate (1.87 g,17.61 mmol) were added sequentially to dichloromethane (10 mL) and stirred at room temperature for 5 hours. The reaction solution was poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography to give the title compound (1.62 g, 94%). MS: M/z [ M+H-tBu] ⁺ =198。

2-formyl-3-azaspiro [ 5.5 ] undecane-3-carboxylic acid tert-butyl ester

3-azaspiro [ 5.5 ] under argon protection ]Tert-butyl undecane-3-carboxylate (1 g,3.95 mmol) and tetramethyl ethylenediamine (458 mg,3.95 mmol) were added to anhydrous tetrahydrofuran (10 mL), and cooled to-60 under argon ^o C, sec-butyllithium in n-hexane (1.3 M,3.03 mL,3.95 mmol) was added dropwise thereto, 50 ^o C ~ -20 ^o C stirred for 0.5 hours. Cooling the reaction solution to-60 deg.f ^o C, N-dimethylformamide (432 mg,5.93 mmol), -50 ^o C is stirred for 1 hour, and is stirred for 1 hour at room temperature. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (563 mg, 51%). MS/z [ M+H ]tBu] ⁺ =226。

2- (hydroxymethyl) -3-azaspiro [ 5.5 ] undecane-3-carboxylic acid tert-butyl ester

At room temperature, 2-formyl-3-azaspiro [ 5.5 ]]Tert-butyl undecane-3-carboxylate (460 mg,1.64 mmol) was added to ethanol (5 mL), sodium borohydride (124 mg,3.27 mmol) was added with stirring, and stirred at room temperature for 2 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (430 mg, 93%). MS: M/z [ M+H-tBu] ⁺ =228。

(3- (2, 6-dichloropyrimidin-4-yl) -3-azaspiro [ 5.5 ] undecan-2-yl) methanol

At room temperature, 2- (hydroxymethyl) -3-azaspiro [ 5.5 ] ]Tert-butyl undecane-3-carboxylate (430 mg,1.52 mmol) was added to dichloromethane (3 mL), and an ethyl acetate solution of hydrogen chloride (4M, 1.9 mL) was added under stirring and stirred at room temperature for 2 hours. The reaction solution was concentrated, acetonitrile (5 mL) and sodium carbonate (483 mg,4.56 mmol) were successively added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated to give the title compound (410 mg, 8% by preparative thin layer chromatography). MS: M/z [ M+H ]] ⁺ =330。

3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclohexane-1, 8' -pyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

(3- (2, 6-dichloropyrimidin-4-yl) -3-azaspiro [ 5.5 ] at room temperature]Undec-2-yl) methanol (360 mg,1.09 mmol) was added to dichloromethaneTo alkane (5 mL), thionyl chloride (389 mg,3.27 mmol) was added with stirring and stirred at room temperature for 1 hour. The reaction solution was concentrated, acetonitrile (5 mL) and potassium carbonate (451 mg,3.1 mmol) were successively added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by preparative thin layer chromatography to give the title compound (220 mg, 60%). MS: M/z [ M+H ]] ⁺ =294。

Intermediate 7

1, 1-dichloro-2-oxo-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, tert-butyl 4-methylenepiperidine-1-carboxylate (49 g,248.4 mmol) and zinc copper reagent (49 g) were added sequentially to anhydrous ethylene glycol dimethyl ether (500 mL), and the temperature was controlled to 30 ^o C-40 ^o And C, dropwise adding trichloroacetyl chloride (49.4 g,273.2 mmol), and stirring at room temperature for 3 hours. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by column chromatography to give the title compound (38.6 g, 50%). MS: M/z [ M+H-tBu] ⁺ =252。

2-oxo-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

Sequentially adding 1, 1-dichloro-2-oxo-7-aza-spiro [ 3.5 ] at room temperature]Tert-butyl nonane-7-carboxylate (37 g,120.1 mmol) was added to saturated aqueous ammonium chloride (200 mL) and methanol (200 mL), cooled to 0 under argon ^o Zinc powder (37 g,565.8 mmol) was added in portions and stirred overnight at room temperature. The reaction was filtered, the filter cake was rinsed with ethyl acetate, the organic phase was concentrated and purified by column chromatography to give the title compound (24.2 g, 84%). MS: M/z [ M+H-tBu] ⁺ =184。

7-azaspiro [ 3.5 ] nonanes

At room temperature, 2-oxo-7-azaspiro [ 3.5 ]]Tert-butyl nonane-7-carboxylate (5 g,20.92 mmol) was added to polyethylene glycol (50 mL), hydrazine hydrate (7.32 g,146.44 mmol) and potassium hydroxide solid (7.03 g,125.52 mmol), 200 g, were added with stirring ^o C stirring for 4 hours. The reaction was cooled to room temperature, poured into water, extracted with diethyl ether, and the organic phase was concentrated to give the title compound (3.32 g, 126%). MS: M/z [ M+H ]] ⁺ =126。

7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

7-azaspiro [ 3.5 ] at room temperature]Nonane (2.8 g,22.4 mmol) was added to dichloromethane (50 mL), di-tert-butyl dicarbonate (7.32 g,33.6 mmol) and potassium carbonate (9.27 g,67.3 mmol) were added sequentially with stirring and stirred overnight at room temperature. The reaction solution was poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography to give the title compound (4.5 g, 89%). MS: M/z [ M+H-tBu] ⁺ =170。

6-formyl-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

Tetramethyl ethylenediamine (1.03 g,8.89 mmol) and 7-azaspiro [ 3.5 ] were sequentially added at room temperature]Tert-butyl nonane-7-carboxylate (2 g,8.89 mmol) was added to anhydrous tetrahydrofuran (20 mL), and the mixture was cooled to-60℃under argon ^o C, dropwise adding sec-butyllithium in n-hexane (1.3 M,6.84 mL,8.89 mmol) under stirring, wherein the addition is completed to-20 ^o C stirring for 30 minutes. Cooling the reaction solution to-60 deg.f ^o C, N-dimethylformamide (973 mg,13.3 mmol) was added dropwise, and the addition was completed to-60 ^o C is stirred for 1 hour and 2 hours at room temperature. Pouring the reaction solution into saturated ammonium chloride aqueous solution, extracting with ethyl acetate, and collecting the extract The organic phase was concentrated and purified by column chromatography to give the title compound (0.83 g, 37%). MS: M/z [ M+H-tBu] ⁺ =198。

6- (hydroxymethyl) -7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, 6-formyl-7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (1.63 g,6.4 mmol) was added to absolute ethanol (10 mL), sodium borohydride (487 mg,12.8 mmol) was added with stirring, and stirred at room temperature for 2 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (1.43 g, 87%). MS: M/z [ M+H-tBu] ⁺ =200。

(7- (6-chloro-2-methoxypyrimidin-4-yl) -7-azaspiro [ 3.5 ] nonan-6-yl) methanol

At room temperature, 6- (hydroxymethyl) -7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (1.43 g,6.4 mmol) was added to dichloromethane (100 mL), and an ethyl acetate solution of hydrogen chloride (4M, 27.8 mL) was added under stirring and stirred at room temperature for 1 hour. The reaction solution was concentrated, acetonitrile (10 mL), 4, 6-dichloro-2-methoxypyrimidine (2.29 g,12.8 mmol) and sodium carbonate (2.32 g, 21.87 mmol) were sequentially added to the residue, 85 ^o C stirred for 16 hours. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by column chromatography to give the title compound (0.783 g, 35%). MS: M/z [ M+H ] ] ⁺ =298。

3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

At the room temperature, the temperature of the mixture is higher than the room temperature,(6-chloro-2-methoxypyrimidin-4-yl) -7-azaspiro [ 3.5 ]]Nonan-6-yl) methanol (100 mg,0.33 mmol) was added to dichloromethane (5 mL), and thionyl chloride (118 mg,0.99 mmol) was added under stirring and stirred at room temperature for 1 hour. The reaction solution was concentrated, acetonitrile (5 mL) and potassium carbonate (182 mg,1.32 mmol) were successively added to the residue, 85 ^o C stirring overnight. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (38 mg, 43%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =266。

Intermediate 8

8-azaspiro [ 4.5 ] decane-8-carboxylic acid tert-butyl ester

At room temperature, 8-azaspiro [ 4.5 ]]Decane hydrochloride (1 g,7.18 mmol) was added to dichloromethane (10 mL), di-tert-butyl dicarbonate (2.35 g,10.77 mmol) and potassium carbonate (2.98 g,21.54 mmol) were added sequentially with stirring, and stirred overnight at room temperature. The reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (1.41 g, 82%). MS: M/z [ M+H-tBu] ⁺ =184。

7-formyl-8-azaspiro [ 4.5 ] decane-8-carboxylic acid tert-butyl ester

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Tetramethyl ethylenediamine (0.65 g,5.56 mmol) and 8-azaspiro [ 4.5 ] were sequentially added at room temperature ]Tert-butyl decane-8-carboxylate (1.33 g,5.56 mmol) was added to anhydrous tetrahydrofuran (10 mL), and the mixture was cooled to-60℃under argon ^o C, dropwise adding sec-butyllithium in n-hexane (1.3 M,5.56 mL,7.23 mmol) under stirring, wherein the addition is completed to-20 ^o C stirring for 30 minutes. Cooling the reaction solution to-60 deg.f ^o C, N-dimethylformamide (610 mg,8.34 mmol), -60 ^o C is stirred for 1 hour and 2 hours at room temperature. Pouring the reaction solution into saturated ammonium chloride aqueous solution, extracting with ethyl acetate, concentrating the organic phase,purification by column chromatography gave the title compound (0.132 g, 9%). MS: M/z [ M+H-tBu] ⁺ =212。

7- (hydroxymethyl) -8-azaspiro [ 4.5 ] decane-8-carboxylic acid tert-butyl ester

7-formyl-8-azaspiro [ 4.5 ] at room temperature]Decane-8-carboxylic acid tert-butyl ester (132 mg,0.49 mmol) was added to anhydrous methanol (10 mL), sodium borohydride (92.68 mg,2.45 mmol) was added with stirring, and stirred at room temperature for 3 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (130 mg, 98%). MS: M/z [ M+H-tBu] ⁺ =214。

(8- (6-chloro-2-methoxypyrimidin-4-yl) -8-azaspiro [ 4.5 ] decan-7-yl) methanol

At room temperature, 6- (hydroxymethyl) -7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (51 mg,0.19 mmol) was added to dichloromethane (3 mL), and an ethyl acetate solution of hydrogen chloride (4M, 0.25 mL) was added under stirring and stirred at room temperature for 3 hours. The reaction solution was concentrated, acetonitrile (10 mL), 4, 6-dichloro-2-methoxypyrimidine (68 mg,0.38 mmol) and sodium carbonate (165 mg,1.56 mmol) were sequentially added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated to give the title compound (55 mg, 93%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =312。

3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclopentane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

At room temperature(8- (6-chloro-2-methoxypyrimidin-4-yl) -8-azaspiro [ 4.5)]Decane-7-yl) methanol (55 mg,0.18 mmol) was added to dichloromethane (5 mL), and thionyl chloride (64 mg,0.54 mmol) was added with stirring, and stirred at room temperature for 1 hour. The reaction solution was concentrated, and water (5 mL) and an aqueous sodium hydroxide solution (15%, 186, mg) were added in this order to the residue, followed by stirring at room temperature for 0.5 hours. The reaction solution was extracted with dichloromethane, and the organic phase was concentrated to give the title compound (28 mg, 72%). MS: M/z [ M+H ]] ⁺ =280。

Intermediate 8

2-hydroxy-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, 2-oxo-7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (5 g,20.89 mmol) was added to methanol (50 mL), sodium borohydride (2.37 g,62.67 mmol) was added with stirring, and the mixture was stirred at room temperature for 5 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (4.93 g, 98%). MS: M/z [ M+H-tBu ] ⁺ =186。

2- ((tert-Butyldimethylsilyl) oxy) -7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

Sequentially adding 2-hydroxy-7-aza spiro [ 3.5 ] at room temperature]Nonane-7-carboxylic acid tert-butyl ester (4.83 g,20.01 mmol) and imidazole (2.72 g,40.02 mmol) were added to dichloromethane (50 mL), tert-butyldimethylchlorosilane (4.52 g,30.01 mmol) was added with stirring, and stirred at room temperature for 3 hours. The reaction solution was poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography to give the title compound (6.8 g, 96%). MS: M/z [ M+H-tBu] ⁺ =300。

2- ((tert-Butyldimethylsilyl) oxy) -6-formyl-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

Tetramethyl ethylenediamine (0.69 g,5.91 mmol) and 2- ((tert-butyldimethylsilyl) oxy) -7-azaspiro [ 3.5 ] at room temperature]Tert-butyl nonane-7-carboxylate (2.1 g,5.91 mmol) was added to anhydrous tetrahydrofuran (15 mL), and the mixture was cooled to-60℃under argon ^o C, dropwise adding sec-butyllithium in n-hexane (1.3 M,4.55 mL,5.91 mmol) under stirring, wherein the addition is completed to-20 ^o C stirring for 30 minutes. Cooling the reaction solution to-60 deg.f ^o C, dropwise adding N, N-dimethylformamide (650 mg,8.87 mmol), and ending the dropwise adding to-60 ^o C is stirred for 1 hour and 2 hours at room temperature. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (2.2 g, 97%). MS: M/z [ M+H-tBu ] ⁺ =328。

2- (tert-Butyldimethylsilyl) oxy) -6- (hydroxymethyl) -7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, 2- ((tert-butyldimethylsilyl) oxy) -6-formyl-7-azaspiro [ 3.5]Nonane-7-carboxylic acid tert-butyl ester (310 mg,0.81 mmol) was added to anhydrous methanol (3 mL), sodium borohydride (91 mg,2.43 mmol) was added under stirring, and stirred at room temperature for 3 hours, the reaction was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (298 mg, 95%). MS: M/z [ M+H-tBu] ⁺ =330。

(2- ((tert-Butyldimethylsilyl) oxy) -7- (6-chloro-2-methoxypyrimidin-4-yl) -7-azaspiro [ 3.5 ] nonan-6-yl) methanol

At room temperature, 2- (tert-butyl) di-Methylsilyl) oxy) -6- (hydroxymethyl) -7-azaspiro [ 3.5]Nonane-7-carboxylic acid tert-butyl ester (296 mg,0.77 mmol) was added to dichloromethane (10 mL) and trifluoroacetic acid (1 mL), and stirred at room temperature for 1 hour. Potassium carbonate (320 mg,2.31 mmol) was added thereto, and the mixture was stirred at room temperature for 0.5 hours. The reaction solution was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated. Acetonitrile (5 mL), 4, 6-dichloro-2-methoxypyrimidine (196 mg,1.06 mmol) and sodium carbonate (169 mg,1.59 mmol) were sequentially added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated to give the title compound (55 mg, 17%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =428。

3- ((tert-butyldimethylsilyl) oxy) -3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

(2- ((tert-Butyldimethylsilyl) oxy) -7- (6-chloro-2-methoxypyrimidin-4-yl) -7-azaspiro [ 3.5 ] at room temperature]Non-6-yl) methanol (55 mg,0.13 mmol) and triethylamine (66 mg,0.26 mmol) were added to dichloromethane (5 mL), and methanesulfonic anhydride (45 mg,0.26 mmol) was added with stirring and stirred at room temperature for 3 hours. The reaction solution was poured into water, dichloromethane was extracted, the organic phase was concentrated, and water (5 mL) and an aqueous sodium hydroxide solution (15%, 123 mg) were sequentially added to the residue, followed by stirring at room temperature for 0.5 hours. The reaction solution was extracted with dichloromethane, and the organic phase was concentrated to give the title compound (18 mg, 41%). MS: M/z [ M+H ]] ⁺ =396。

3- ((tert-butyldimethylsilyl) oxy) -3'- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ] c]Pyrimidine]-1' -one

3- ((tert-Butyldimethylsilyl) oxy) -3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]1' -Ketone (17 mg,0.043 mmol), (3-fluoro-4- (2-trifluoromethyl) pyridin-4-yl) phenyl) methanol (37 mg,0.13 mmol) and cesium carbonate (42 mg,0.13 mmol) were added to toluene (5 mL), 110 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by preparative thin layer chromatography to give the title compound (23 mg, 83%). MS: M/z [ M+H ]] ⁺ =647。

Intermediate 9

2-methoxy-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, 2-oxo-7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (5 g,20.89 mmol) was added to methanol (50 mL), sodium borohydride (2.37 g,62.67 mmol) was added with stirring, and stirred at room temperature for 5 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, the organic phase was concentrated, acetonitrile (30 mL) and methyl iodide (8.96 g,62.67 mmol) were sequentially added to the residue, sodium hydride (60%, 2.51 g,62.67 mmol) was added with stirring, and the mixture was stirred at room temperature for 3 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (1.9 g, 36%). MS: M/z [ M+H-tBu ] ⁺ =200。

6-formyl-2-methoxy-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

Tetramethyl ethylenediamine (1.3 g,11.16 mmol) and 2-methoxy-7-azaspiro [ 3.5 ] were sequentially added at room temperature]Tert-butyl nonane-7-carboxylate (1.9 g,7.44 mmol) was added to anhydrous tetrahydrofuran (20 mL), and the temperature was lowered under argon atmosphereTo-60 ^o C, dropwise adding sec-butyllithium in n-hexane (1.3 M,8.58 mL,11.16 mmol) under stirring, wherein the addition is completed to-20 ^o C stirring for 30 minutes. Cooling the reaction solution to-60 deg.f ^o C, dropwise adding N, N-dimethylformamide (1.63 g,22.32 mmol), and ending the dropwise adding to-60 ^o C is stirred for 1 hour and 2 hours at room temperature. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (0.57 g, 27%). MS: M/z [ M+H-tBu] ⁺ =228。

6- (hydroxymethyl) -2-methoxy-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, 6-formyl-2-methoxy-7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (380 mg,1.34 mmol) was added to anhydrous methanol (5 mL), sodium borohydride (150 mg,4.02 mmol) was added under stirring, and stirred at room temperature for 3 hours, the reaction was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (365 mg, 95%). MS: M/z [ M+H-tBu ] ⁺ =230。

(7- (6-chloro-2-methoxypyrimidin-4-yl) -2-methoxy-7-azaspiro [ 3.5 ] nonan-6-yl) methanol

At room temperature, 6- (hydroxymethyl) -2-methoxy-7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (365 mg,1.28 mmol) was added to dichloromethane (2 mL), and an ethyl acetate solution of hydrogen chloride (4M, 2 mL) was added under stirring and stirred at room temperature for 1 hour. The reaction solution was concentrated, acetonitrile (10 mL), 4, 6-dichloro-2-methoxypyrimidine (344 mg,1.92 mmol) and sodium carbonate (385 mg,3.63 mmol) were sequentially added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated to give the title compound (310 mg, 74%) by preparative thin layer chromatography. MS: m/z [M+H] ⁺ =328。

3 '-chloro-3-methoxy-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

(7- (6-chloro-2-methoxypyrimidin-4-yl) -2-methoxy-7-azaspiro [ 3.5 ] at room temperature]Nonan-6-yl) methanol (270 mg,0.82 mmol) was added to dichloromethane (5 mL), and thionyl chloride (118 mg,0.99 mmol) was added under stirring and stirred at room temperature for 1 hour. The reaction solution was concentrated, acetonitrile (5 mL) and potassium carbonate solid (298 mg,2.16 mmol) were successively added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by preparative thin layer chromatography to give the title compound (198 mg, 93%). MS: M/z [ M+H ]] ⁺ =296。

Intermediate 10

1, 4-dioxa-10-azadispiro [ 4.1.5 ⁷ . 1 ⁵ ]Tridecane-10-carboxylic acid tert-butyl ester

At room temperature, 2-oxo-7-azaspiro [ 3.5 ]]Nonane-7-carboxylic acid tert-butyl ester (5 g,20.89 mmol), ethylene glycol (1.94 g,31.34 mmol) and pyridine 4-methylbenzenesulfonate (1.05 g,4.18 mmol) were added sequentially to toluene (50 mL), 120 ^o C stirring overnight. The reaction solution was concentrated and purified by column chromatography to give the title compound (3.6 g, 61%). MS: M/z [ M+H-tBu] ⁺ =228。

9-formyl-1, 4-dioxa-10-azadispiro [ 4.1.5 ⁷ . 1 ⁵ ]Tridecane-10-carboxylic acid tert-butyl ester

Tetramethyl ethylenediamine (2.15 g,18.52 mmol) and 1, 4-dioxa-10-azadispiro [ 4.1.5 ] were sequentially added at room temperature ⁷ . 1 ⁵ ]Tert-butyl tridecane-10-carboxylate (3.5 g,12.35 mmol) was added to anhydrous tetrahydrofuran (40 mL), and cooled to-60℃under argon ^o C, dropwise adding sec-butyllithium in n-hexane (1.3 M,14.25 mL,18.52 mmol) under stirring, wherein the addition is completed to-20 ^o C stirring for 30 minutes. Cooling the reaction solution to-60 deg.f ^o C, dropwise adding N, N-dimethylformamide (2.71 g,37.05 mmol), and ending the dropwise adding to-60 ^o C is stirred for 1 hour and 2 hours at room temperature. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (1.23 g, 32%). MS: M/z [ M+H-tBu] ⁺ =256。

9- (hydroxymethyl) -1, 4-dioxa-10-azadispiro [ 4.1.5 ⁷ . 1 ⁵ ]Tridecane-10-carboxylic acid tert-butyl ester

At room temperature, 9-formyl-1, 4-dioxa-10-azadispiro [ 4.1.5 ⁷ . 1 ⁵ ]Tridecane-10-carboxylic acid tert-butyl ester

(1.13 g,3.63 mmol) was added to anhydrous methanol (10 mL), sodium borohydride (270 mg,7.26 mmol) was added under stirring, and stirred at room temperature for 3 hours, the reaction was quenched with saturated aqueous ammonium chloride, extracted with dichloromethane, and the organic phase was concentrated to give the title compound (1.1 g, 97%). MS: M/z [ M+H-tBu] ⁺ =258。

7- (6-chloro-2-methoxypyrimidin-4-yl) -6- (hydroxymethyl) -7-azaspiro [ 3.5 ] non-2-one

At room temperature, 9- (hydroxymethyl) -1, 4-dioxa-10-azadispiro [ 4.1.5 ⁷ . 1 ⁵ ]Tert-butyl tridecane-10-carboxylate (1.1 g,3.51 mmol) was added to dichloromethane (10 mL)A solution of hydrogen chloride in ethyl acetate (4M, 0.32 mL) was added under stirring, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, acetonitrile (10 mL), 4, 6-dichloro-2-methoxypyrimidine (942 mg,5.27 mmol) and sodium carbonate (2.07 g, 19.5 mmol) were sequentially added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, and the filtrate was concentrated to give the title compound (180 mg, 8%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =312。

3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1', 3-diketones

7- (6-chloro-2-methoxypyrimidin-4-yl) -6- (hydroxymethyl) -7-azaspiro [ 3.5 ] at room temperature]Nonan-2-one (58 mg,0.19 mmol) was added to dichloromethane (5 mL), and thionyl chloride (67.8 mg,0.57 mmol) was added with stirring and stirred at room temperature for 3 hours. The reaction solution was concentrated, acetonitrile (3 mL) and potassium carbonate solid (62 mg,0.45 mmol) were successively added to the residue, 85 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by preparative thin layer chromatography to give the title compound (37 mg, 88%). MS: M/z [ M+H ]] ⁺ =280。

3'- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1', 3-diketones

3' -chloro-3-methoxy-6 ', 7', 9a ', 10' -tetrahydro-1 ' is sequentially added at room temperature 'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine ]-1' -Ketone (33 mg,0.12 mmol), (3-fluoro-4- (2-trifluoromethyl) pyridin-4-yl) phenyl) methanol (51 mg,0.18 mmol) and cesium carbonate (78)mg,0.24 mmol) was added to toluene (3 mL), 120 ^o C stirring overnight. The reaction was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and purified by preparative thin layer chromatography to give the title compound (41 mg, 64%). MS: M/z [ M+H ]] ⁺ =531。

Intermediate 11

2-methoxy-6-vinyl-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

At room temperature, 6-formyl-2-methoxy-7-azaspiro [ 3.5 ]]Tert-butyl nonane-7-carboxylate (2.4 g,8.47 mmol) was added to anhydrous tetrahydrofuran (80 mL), methyltriphenylphosphorus bromide (3.63 g,10.16 mmol) was added under stirring, and the temperature was reduced to 0 under argon ^o Sodium hydride (60%, 1.02 g,25.41 mmol) was added in portions, and the mixture was stirred at room temperature for 3 hours after the addition. The reaction solution was quenched with saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (700 mg, 29%). MS: M/z [ M+H-tBu] ⁺ =226。

6- (2-hydroxyethyl) -2-methoxy-7-azaspiro [ 3.5 ] nonane-7-carboxylic acid tert-butyl ester

2-methoxy-6-vinyl-7-azaspiro [ 3.5 ] at room temperature]Nonane-7-carboxylic acid tert-butyl ester (700 mg,2.49 mmol) was added to 9-borobicyclo [ 3.3.1 ] ]The mixture was stirred overnight at room temperature in a tetrahydrofuran solution of nonane (0.5 M,20 mL,9.96 mmol). Adding water (2 mL), sodium hydroxide aqueous solution (3M, 10 mL) and hydrogen peroxide (37%, 10 mL) into the reaction solution in sequence, the reaction solution 50 ^o C stirring for 2 hours. The reaction solution was cooled to room temperature, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=2/1) to give the title compound (750 mg, 100%). MS: M/z [ M+H-tBu] ⁺ =244。

2- (2-methoxy-7-azaspiro [ 3.5 ] nonan-6-yl) ethyl-1-ol hydrochloride

At room temperature, 6- (2-hydroxyethyl) -2-methoxy-7-azaspiro [ 3.5 ]]Tert-butyl nonane-7-carboxylate (750 mg,2.50 mmol) was added to a solution of hydrogen chloride in 1, 4-dioxane (10 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated to give the crude title compound (590 mg, 100%). MS: M/z [ M+H ]] ⁺ =200。

2- (7- (2, 6-dichloropyrimidin-4-yl) -2-methoxy-7-azaspiro [ 3.5 ] nonan-6-yl) ethyl-1-ol

At room temperature, 2- (2-methoxy-7-azaspiro [ 3.5 ]]Nonan-6-yl) ethyl-1-ol hydrochloride (570 mg,2.86 mmol) was added to acetonitrile (10 mL), 4, 6-dichloro-2-methoxypyrimidine (512 mg,2.86 mmol) and sodium carbonate solid (909 mg,8.58 mmol) were added sequentially with stirring, and the reaction stirred at room temperature overnight. The reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (dichloromethane/methanol=50/1) to give the title compound (390 mg, 40%). MS: M/z [ M+H ] ] ⁺ =342。

7- (6-chloro-2-methoxypyrimidin-4-yl) -6- (2-chloroethyl) -2-methoxy-7-azaspiro [ 3.5 ] nonane

2- (7- (2, 6-dichloropyrimidin-4-yl) -2-methoxy-7-azaspiro [ 3.5 ] at room temperature]Nonan-6-yl) ethan-1-ol (350 mg,1.02 mmol) was added to dichloromethane (20 mL) and cooled to 0 ^o Thionyl chloride (394 mg,3.06 mmol) was added thereto and stirred at reflux for 2 hours. The reaction solution was concentrated to give the title compound (350 mg, 95%). MS: M/z [ M+H ]] ⁺ =360。

2 '-chloro-3-methoxy-6', 7', 7a', 8', 10', 11 '-hexahydro-4'HSpiro [ cyclobutane-1, 9' -pyrido [1, 2 ]c]Pyrimido [1, 6 ]a]Pyrimidine]-4' -one

7- (6-chloro-2-methoxypyrimidin-4-yl) -6- (2-chloroethyl) -2-methoxy-7-azaspiro [ 3.5 ] under the protection of argon]Nonane (350 mg,0.97 mmol) and potassium carbonate (402 mg,2.91 mmol) were added to acetonitrile (10 mL), 85 ^o C was stirred overnight, the reaction solution was filtered, the organic phase was concentrated, and purified by column chromatography (dichloromethane/methanol=100/1 to 50/1) to give the title compound (200 mg, 66%). MS: M/z [ M+H ]] ⁺ =310。

Intermediate 12

6-Oxopiperidine-2-carboxylic acid

6-oxo-1, 6-dihydropyridine-2-carboxylic acid (10 g,72 mmol) and palladium on carbon (10%, 1 g) were added to methanol (100 mL) at room temperature and hydrogenated overnight at room temperature under normal pressure. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (5 g, 50%). LC-MS: M/z [ M+H ] ] ⁺ =144。

6-Oxopiperidylmethyl-2-carboxylic acid methyl ester

Under the protection of argon, the methanol (10 mL) is cooled to 0 ^o Thionyl chloride (830 mg,7.7 mmol) was added dropwise thereto with stirring, and the mixture was stirred at room temperature for 10 minutes. 6-Oxopiperidine-2-carboxylic acid (1 g,7 mmol) was added to the reaction solution and stirred overnight at room temperature. The reaction solution was concentrated, toluene (10 mL) and triethylamine (1.4 g,14 mmol) were successively added to the residue, and stirred at room temperature for 0.5 hour, the reaction solution was filtered, and the filtrate was concentrated to give the title compound (850 mg, 77%). MS: m/z [M+H] ⁺ =158。

1-benzyl-6- ((benzyloxy) methyl) piperidin-2-one

Methyl 6-oxopiperidylmethyl-2-carboxylate (850 mg,5.4 mmol) was added to methanol (10 mL) at room temperature, sodium borohydride (307 mg,8.1 mmol) was slowly added with stirring, and the mixture was stirred at room temperature for 1 hour. The reaction solution was quenched with sodium sulfate decahydrate, filtered, and the filtrate was concentrated. Adding the residue into dimethyl sulfoxide (20 mL), and cooling to 0 under argon ^o Sodium hydride (3.6 g,21 mmol) was slowly added with stirring and stirred at room temperature for 30 minutes. Benzyl bromide (3.6 g,21 mmol) was added to the reaction and stirred overnight at room temperature. The reaction solution was quenched with water, poured into water, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (900 mg, 54%). MS: M/z [ M+H ] ] ⁺ =310。

4-benzyl-5- ((benzyloxy) methyl) -4-azaspiro [ 2.5 ] octane

Cooling anhydrous tetrahydrofuran (60 mL) to-40 deg.f under argon protection ^o C, a tetrahydrofuran solution (1M,20 mL,19.4 mmol) of ethyl magnesium bromide and tetraisopropyl titanate (2 g,7.2 mmol) were added in this order with stirring, and stirred at room temperature for 5 minutes. A solution of 1-benzyl-6- ((benzyloxy) methyl) piperidin-2-one (2.0 g,6.5 mmol) in anhydrous tetrahydrofuran (1 mL) was added to the reaction solution, which was stirred under reflux overnight. The reaction solution was quenched with 10% aqueous sodium hydroxide, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography to give the title compound (1.1 g, 53%). MS: M/z [ M+H ]] ⁺ =322。

(4-azaspiro [ 2.5 ] oct-5-yl) methanolic hydrochloride

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4-benzyl-5- ((benzyloxy) methyl) -4-azaspiro [ 2.5 ] at room temperature]Octane (200 mg,0.6 mmol) and palladium on carbon (10%, 30 mg) were added to an ethyl acetate solution of hydrogen chloride (4M, 5 mL) and methanol (5 mL), 40 ^o C, hydrogenation overnight at normal pressure. The reaction solution was filtered, and the filtrate was concentrated to give the crude title compound (200 mg, 182%). MS: M/z [ M+H ]] ⁺ =142。

(4- (2, 6-dichloropyrimidin-4-yl) -4-azaspiro [ 2.5 ] octyl-5-yl) methanol

At room temperature, (4-azaspiro [ 2.5 ] was added sequentially]Octane-5-yl) methanol hydrochloride (100 mg,0.6 mmol), 2, 4, 6-trichloropyrimidine (146 mg,0.8 mmol) and sodium carbonate solid (149 mg,1.4 mmol) were added to acetonitrile (2 mL) and stirred overnight at room temperature. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (100 mg, 62%) by preparative thin layer chromatography. MS: M/z [ M+H ] ] ⁺ =288。

3 '-chloro-8', 9', 9a', 10 '-tetrahydro-1'H, 7'H-spiro [ cyclopropane-1, 6' -pyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

(4- (2, 6-dichloropyrimidin-4-yl) -4-azaspiro [ 2.5 ] at room temperature]Octyl-5-yl) methanol (100 mg,0.3 mmol) and thionyl chloride (47 mg,0.4 mmol) were added to dichloromethane (2 mL) and stirred overnight at room temperature. The reaction solution was concentrated, and to the residue were added successively potassium carbonate solid (83 mg,0.6 mmol) and acetonitrile (5 mL), 80 ^o C stirring overnight. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated to give the title compound (80 mg, 100%) by preparative chromatography. MS: M/z [ M+H ]] ⁺ =252。

Intermediate 13

4-Methylenepiperidine-1-carboxylic acid benzyl ester

Methyl triphenylphosphine bromide (10 g,27.8 mmol) was added to anhydrous tetrahydrofuran (30 mL) at room temperature, and cooled to 0 under argon ^o C, temperature control 0 ^o Tetrahydrofuran solution (2.5 M,11 mL,27.8 mmol) of n-butyllithium was added dropwise at a rate of 0 or below ^o C stirring for 1 hour. Temperature control 0 ^o Next, a solution of benzyl 4-oxopiperidine-1-carboxylate (5 g,21.4 mmol) in anhydrous tetrahydrofuran (20) mL) was added dropwise to the reaction solution, 0 ^o C was stirred at room temperature overnight after 1 hour. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (3.8 g, 76%). MS: M/z [ M+H ] ] ⁺ =232。

6-azaspiro [ 2.5 ] octane-6-carboxylic acid benzyl ester

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Cooling dichloromethane (60 mL) to 0 under argon protection ^o Adding diethyl zinc in n-hexane solution (1 M,60 mL,60 mmol) under stirring, and controlling temperature to 0 ^o Trifluoroacetic acid (6.8 g,60 mmol) and 0.g were added dropwise below C ^o C stirred for 0.5 hours. Diiodomethane (16 g,60 mmol) was added to the reaction solution and stirred at room temperature for 30 minutes. Benzyl 4-methylenepiperidine-1-carboxylate (7 g,30 mmol) was added to the reaction solution and reacted overnight at room temperature. The reaction solution was poured into a saturated aqueous sodium hydrogencarbonate solution, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography to give the title compound (6.1 g, 82%). MS: M/z [ M+H ]] ⁺ =246。

6-azaspiro [ 2.5 ] octane hydrochloride

At room temperature, 6-azaspiro [ 2.5 ]]Octane-6-carboxylic acid benzyl ester (8.3 g, 33.9)mmol) and palladium on carbon (10%, 830 mg) were added to methanol (30 mL) and hydrogenated overnight at room temperature and pressure. The reaction solution was filtered, a 1, 4-dioxane solution (4M, 30 mL) of hydrogen chloride was added to the filtrate, the mixture was stirred at room temperature for 30 minutes, the reaction solution was concentrated, tetrahydrofuran (15 mL) was added to the residue, and the mixture was stirred at room temperature for 30 minutes. Filtration and drying of the filter cake gave the title compound (4.3 g, 86%). MS: M/z [ M+H ]] ⁺ =112。

6-azaspiro [ 2.5 ] octane-6-carboxylic acid tert-butyl ester

At room temperature, 6-azaspiro [ 2.5 ] was added sequentially]Octane hydrochloride (4.27 g,29 mmol), triethylamine (6.2 g,61 mmol) and di-tert-butyl dicarbonate (7 g,32 mmol) were added to dichloromethane (50 mL) and stirred overnight at room temperature. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with methylene chloride, and the organic phase was concentrated and purified by column chromatography to give the title compound (5.3 g, 87%). MS: M/z [ M+H ]] ⁺ =212。

5-formyl-6-azaspiro [ 2.5 ] octane-6-carboxylic acid tert-butyl ester

At room temperature, 6-azaspiro [ 2.5 ]]Tert-butyl octane-6-carboxylate (400 mg,1.9 mmol) was added to anhydrous tetrahydrofuran (5 mL), and the mixture was cooled to-78 under argon gas atmosphere ^o C, controlling the temperature to-50 ^o Under C, tetramethyl ethylenediamine (660 mg,5.7 mmol) and sec-butyllithium in n-hexane (1.3 M,2.2 mL 2.9 mmol) were successively added dropwise, 30 ^o C stirring for 15 minutes. Cooling to-78 deg.f ^o C, N-dimethylformamide (416 mg,5.7 mmol) was added dropwise thereto, and the mixture was stirred at room temperature overnight. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to give the title compound (110 mg, 24%). MS: M/z [ M+H ]] ⁺ =240。

5- (hydroxymethyl) -6-azaspiro [ 2.5 ] octane-6-carboxylic acid tert-butyl ester

At room temperature, 5-formyl-6-azaspiro [ 2.5 ] ]Tert-butyl octane-6-carboxylate (110 mg,0.46 mmol) was added to absolute ethanol (5 mL), and sodium borohydride solid (17 mg,0.46 mmol) was added under stirring and stirred at room temperature for 2 hours. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (110 mg, 100%). MS: M/z [ M+H ]] ⁺ =242。

(6- (2, 6-dichloropyrimidin-4-yl) -6-azaspiro [ 2.5 ] octyl-5-yl) methanol

At room temperature, 5- (hydroxymethyl) -6-azaspiro [ 2.5 ]]Tert-butyl octane-6-carboxylate (360 mg,1.5 mmol) was added to a solution of hydrogen chloride in ethyl acetate (4M, 5 mL) and dichloromethane (5 mL), and stirred at room temperature for 1 hour. The reaction solution was concentrated, and to the residue were added sodium carbonate solid (318 mg,3.0 mmol), 2,4, 6-trichloropyrimidine (366 mg,3.0 mmol) and acetonitrile (5 mL) in this order, followed by stirring overnight at room temperature. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (260 mg, 60%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =288。

3 '-chloro-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclopropane-1, 8' -pyridinyl [1', 2': 3, 4]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

(6- (2, 6-dichloropyrimidin-4-yl) -6-azaspiro [ 2.5 ] at room temperature]Octyl-5-yl) methanol (160 mg,0.6 mmol) and thionyl chloride (1 mL) were added to dichloromethane (1 mL) and stirred overnight at room temperature. The reaction solution was concentrated, and to the residue were added successively potassium carbonate solid (230 mg,1.7 mmol) and acetonitrile (5 mL), followed by reflux Stir overnight. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated to give the title compound (80 mg, 50%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =252。

Intermediate 14

8-formyl-3-oxa-9-azaspiro [ 5.5 ] undecane-9-carboxylic acid tert-butyl ester

3-oxa-9-azaspiro [ 5.5 ] at room temperature]Tert-butyl undecane-9-carboxylate (750 mg,2.94 mmol) and tetramethyl ethylenediamine (0.51 g,4.41 mmol) were added to tetrahydrofuran (10 mL), and the mixture was cooled to-60℃under argon ^o C, sec-butyllithium in n-hexane (1.3 M,4.5 mL,5.88 mmol) was added dropwise thereto, 50 ^o C was stirred for 1 hour, N-dimethylformamide (0.32 g,4.41 mmol), -50 ^o C is stirred for 1 hour, and is stirred for 1 hour at room temperature. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (340 mg, 41%). MS: M/z [ M+H-Boc ]] ⁺ =184。

8- (hydroxymethyl) -3-oxa-9-azaspiro [ 5.5 ] undecane-9-carboxylic acid tert-butyl ester

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8-formyl-3-oxa-9-azaspiro [ 5.5 ] at room temperature]Undecane-9-carboxylic acid tert-butyl ester (340 mg,1.2 mmol) was added to tetrahydrofuran (10 mL), sodium borohydride (40 mg,1.2 mmol) was added with stirring, and stirred at room temperature for 0.5 h. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (340 mg, 99%). MS: M/z [ M+H-Boc ] ] ⁺ =186。

(9- (6-chloro-2-methoxypyrimidin-4-yl) -3-oxa-9-azaspiro [ 5.5 ] undecan-8-yl) methanol

8- (hydroxymethyl) -3-oxa-9-azaspiro [ 5.5 ] at room temperature]Undecane-9-carboxylic acid tert-butyl ester (340 mg,1.2 mmol) was added to trifluoroacetic acid (1 mL) and dichloromethane (4 mL) and stirred at room temperature for 5 min. The pH of the reaction system was adjusted to 8-9 with sodium carbonate solids. The reaction solution was concentrated, and 4, 6-dichloro-2-methoxypyrimidine (0.32 g,1.78 mmol), sodium carbonate (0.63 g,5.95 mmol) and acetonitrile (30 mL) were sequentially added to the residue, 85 ^o C stirring overnight. The reaction solution was filtered, and the filtrate was concentrated and purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (210 mg, 54%). MS: M/z [ M+H ]] ⁺ =328。

3 '-chloro-2, 3, 5, 6, 6', 7', 9a', 10 '-octahydro-1'H, 9'H-spiro [ pyran-4, 8' -pyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

(9- (6-chloro-2-methoxypyrimidin-4-yl) -3-oxa-9-azaspiro [ 5.5 ] at room temperature]Undecan-8-yl) methanol (141 mg,0.43 mmol) and triethylamine (0.13 g,1.29 mmol) were added to dichloromethane (12 mL), methanesulfonic anhydride (74 mg,0.65 mmol) was added with stirring, and stirred at room temperature for 20 minutes. The reaction solution was adjusted to pH 8-9 with sodium carbonate solids. The reaction solution was concentrated, and water (8. 8 mL) and potassium carbonate (0.29 g,2.1 mmol) were successively added to the residue. Stirring for 30 minutes at room temperature. Dichloromethane extraction and concentration of the organic phase gave the title compound (60 mg, 47%) by preparative thin layer chromatography (dichloromethane/methanol=20/1). MS: M/z [ M+H ] ] ⁺ =296。

Intermediate 15

1- (tert-butyl) 4-ethyl 4-methylpiperidine-1, 4, 4-tricarboxylic acid

Adding 1- (tert-butyl) 4-ethylpiperidine-1, 4-dicarboxylic acid ester (45.0 g,175.1 mmol) into anhydrous tetrahydrofuran (180 mL) at room temperature, and cooling to-60 under argon gas ^o C, adding lithium diisopropylamide in tetrahydrofuran/n-heptane (2 M,131 mL,262 mmol) with stirring, 60 ^o C stirred for 0.5 hour, methyl chloroformate (18.2 g,192.6 mmol) was added, 60 ^o C stirring for 30 minutes and stirring for two hours at room temperature. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=5/1) to give the title compound (41 g, 91%). MS: M/z [ M+H-Boc ]] ⁺ =216。

4, 4-bis (hydroxymethyl) piperidine-1-carboxylic acid tert-butyl ester

1- (tert-butyl) 4-ethyl 4-methylpiperidine-1, 4, 4-tricarboxylic acid (10.0 g,31.7 mmol) was added to tetrahydrofuran (50 mL) and methanol (50 mL) at room temperature, and the reaction was warmed to 50 ^o C, temperature control 50 ^o C-60 ^o Sodium borohydride (6.0 g,158.5 mmol), 60 is added in portions ^o C stirring for 1 hour. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the crude title compound (8 g, 103%). MS: M/z [ M+H-Boc ] ] ⁺ =146。

4, 4-bis (((t-butyldimethylsilyl) oxy) methyl) piperidine-1-carboxylic acid tert-butyl ester

Tert-butyl 4, 4-bis (hydroxymethyl) piperidine-1-carboxylate (45.0 g,183 mmol) and imidazole (37.9 g,549 mmol) were added sequentially to dichloromethane (100 mL) at room temperature, tert-butyldimethylchlorosilane (55.2 g,366 mmol) was added with stirring, and stirred at room temperature for 2 hours. Pouring the reaction solution into water, extracting with dichloromethane, concentrating the organic phase, and performing column chromatographyPurification (petroleum ether/ethyl acetate=10/1) afforded the title compound (66 g, 77%). MS: M/z [ M+H-Boc ]] ⁺ =373。

4, 4-bis (((tert-butyldimethylsilyl) oxy) methyl) -2-formylpiperidine-1-carboxylic acid tert-butyl ester

At room temperature, 4-bis (((tert-butyldimethylsilyl) oxy) methyl) piperidine-1-carboxylic acid tert-butyl ester (20.0 g,42.28 mmol) and tetramethyl ethylenediamine (7.35 g,64.42 mmol) are added to anhydrous tetrahydrofuran (120 mL), and cooled to-60 under argon protection ^o C, controlling the temperature to-50 ^o Below C, sec-butyllithium in cyclohexane (1.3 M,71.5 mL,93.0 mmol) was added dropwise, -50 ^o Stirring C for 1 hr, adding N, N-dimethylformamide (4.56 g,63.42 mmol), -50 ^o C stirring 1 h, stirring 2 hours at room temperature. The reaction solution was poured into saturated aqueous ammonium chloride, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (12 g, 57%). MS: M/z [ M+H-Boc ] ] ⁺ =402。

(4, 4-bis (((tert-butyldimethylsilyl) oxy) methyl) piperidin-2-yl) methanol

Tert-butyl 4, 4-bis (((tert-butyldimethylsilyl) oxy) methyl) -2-formylpiperidine-1-carboxylate (4.0 g,8.0 mmol) was added to a dichloromethane solution of trifluoroacetic acid (10%, 40, mL) at room temperature and stirred at room temperature for 5 minutes. The reaction solution was adjusted to 8-9 with potassium carbonate solid, filtered, and the filtrate was concentrated. The residue was added to methanol (20 mL), sodium borohydride (907 mg,24.0 mmol) was added with stirring, and the mixture was stirred at room temperature for 1 hour. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the crude title compound (4 g, 124%). MS: M/z [ M+H ]] ⁺ =404。

(4, 4-bis (((tert-butyldimethylsilyl) oxy) methyl) -1- (6-chloro-2-methoxypyrimidin-4-yl) piperidin-2-yl) methanol

Crude (4, 4-bis (((tert-butyldimethylsilyl) oxy) methyl) piperidin-2-yl) methanol (3.0 g,7.44 mmol), 4, 6-dichloro-2-methoxypyrimidine (1.32 g,7.44 mmol) and sodium carbonate solid (1.58 g,14.88 mmol) were added to acetonitrile (30 mL) at room temperature, 90 ^o C stirring overnight. Filtration, concentration of the filtrate, and column chromatography purification (petroleum ether/ethyl acetate=10/1 to 5/1) gave the title compound (1 g, 25%). MS: M/z [ M+H ] ] ⁺ =546。

8, 8-bis (((tert-butyldimethylsilyl) oxy) methyl) -3-chloro-6, 7, 8, 9, 9a, 10-hexahydro-1HPyrido [1', 2': 3, 4]Imidazole [1, 2 ]c]Pyrimidin-1-one

(4, 4-bis (((t-butyldimethylsilyl) oxy) methyl) -1- (6-chloro-2-methoxypyrimidin-4-yl) piperidin-2-yl) methanol (1.0 g,2.0 mmol) was added to dichloromethane (20 mL) at room temperature, triethylamine (303 mg,3.0 mmol) and methanesulfonyl chloride (273 mg,2.4 mmol) were added sequentially to the reaction solution under stirring, and stirring was carried out at room temperature for 20 minutes. The reaction solution was concentrated, water (16 mL) was added to the residue, and the pH of the system was adjusted to 9-10 with potassium carbonate solid under stirring, and stirred at room temperature for 30 minutes. Dichloromethane extraction, concentration of the organic phase, and column chromatography purification (dichloromethane/methanol=20/1) gave the title compound (450 mg, 48%). MS: M/z [ M+H ]] ⁺ =514。

8, 8-bis (((tert-butyldimethylsilyl) oxy) methyl) -3- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6, 7, 8, 9, 9a, 10-hexahydro-1HPyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidin-1-one

8, 8-bis (((tert-butyldimethylsilyl) oxy) methyl) -3-chloro-6, 7, 8, 9, 9a, 10-hexahydro-1, sequentially at room temperatureHPyrido [1', 2': 3, 4 ]Imidazole [1, 2 ]c]Pyrimidin-1-one (450 mg,0.88 mmol), (3-fluoro-4- [ (2- (trifluoromethyl) pyridin-4-yl) oxy)]Phenyl) methanol (0.28 g,0.97 mmol) and cesium carbonate (0.86 g,2.64 mmol) were added to toluene (20 mL), 110 ^o C stirring overnight. The reaction solution was concentrated, poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography (dichloromethane/methanol=20/1) to give the title compound (670 mg, 99%). MS: M/z [ M+H ]] ⁺ =765。

3- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -8, 8-bis (hydroxymethyl) -6, 7, 8, 9, 9a, 10-hexahydro-1HPyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidin-1-one

At room temperature, 8-bis (((tert-butyldimethylsilyl) oxy) methyl) -3- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6, 7, 8, 9, 9a, 10-hexahydro-1HPyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidin-1-one (670 mg,0.88 mmol) was added to anhydrous tetrahydrofuran (10 mL), and a tetrahydrofuran solution (1 M,2.2 mL,2.2 mmol) of tetrabutylammonium fluoride was added with stirring, followed by stirring at room temperature for 1 hour. The reaction solution was poured into a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated and purified by column chromatography (dichloromethane/methanol=20/1 to 10/1) to give the title compound (350 mg, 74%). MS: M/z [ M+H ] ] ⁺ =537。

(3- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -8- (hydroxymethyl) -1-oxo-6, 7, 8, 9, 9a, 10-hexahydro-1HPyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidin-8-yl) methanesulfonic acid methyl ester

3- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -8, 8-bis (hydroxymethyl) -6, 7, 8, 9, 9a, 10-hexahydro-1HPyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine-1-one (100 mg,0.19 mmol) was added to dichloromethane (10 mL), and the temperature was lowered to 0 under argon atmosphere ^o C, adding a solution of triethylamine (23.07 mg,0.23 mmol) and methanesulfonyl chloride (13.06 mg,0.11 mmol) in dichloromethane (1 mL) sequentially under stirring, and stirring at room temperature for 15 minutes. The reaction solution was poured into water, extracted with dichloromethane, and the organic phase was concentrated to prepare a thin layer chromatography (dichloromethane/methanol=10/1) to give the title compound (60 mg, 51%). MS: M/z [ M+H ]] ⁺ =615。

Intermediate 16

5-formyl-2- (3- (trifluoromethyl) phenoxy) benzonitrile

2-fluoro-5-formylbenzonitrile (15.0 g,0.1 mol), 3- (trifluoromethyl) phenol (16.0 g,0.1 mol) and potassium carbonate (13.8 g,0.1 mol) were added to DMF (100 mL), 105 at room temperature ^o C was stirred for 8 hours, the reaction solution was poured into water, extracted with dichloromethane, the organic phase was concentrated, ethanol (50 mL) was added to the residue, stirred at room temperature for 30 minutes, filtered, and the filter cake was concentrated to dryness to give the title compound (22.1 g, 76%). ¹ H NMR(400 MHz, DMSO-d ₆ )d 9.96(s, 1 H), 8.49(s, 1 H), 8.14(br.s, 1 H), 7.88(d, 2 H), 7.47(d, 2 H), 7.19(d, 1 H)。

5- (hydroxymethyl) -2- (3- (trifluoromethyl) phenoxy) benzonitrile

At room temperature, 5-formylThe base-2- (3- (trifluoromethyl) phenoxy) benzonitrile (24.3 g,83.44 mmol) was added to methanol (250 mL), sodium borohydride (4.86 g,127.9 mmol) was added in portions and reacted at room temperature for 0.5 hours. The reaction solution was poured into water, extracted with methylene chloride, and the organic phase was concentrated to give the title compound (24.2 g, 99%). ¹ H NMR(400 MHz, DMSO-d ₆ )d 7.80(s, 1 H), 7.73 - 7.53(m, 3 H), 7.52 - 7.27(m, 2 H), 7.09(br.s, 1 H), 5.41(s, 1 H), 4.51(s, 2 H)。

Referring to the table below, the following intermediates 17 to 23 were prepared with reference to the preparation method of intermediate 16, using the compounds described in the column "raw materials" as raw materials.

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Intermediate 24

4- (benzyloxy) -2- (trifluoromethoxy) pyridine

4- (benzyloxy) pyridin-2-ol (1.91 g,9.48 mmol) and 1-trifluoromethyl-1, 2-phenyliodides-3 [ ]H) Ketone (1 g,3.16 mmol) was added to nitromethane (25 mL) and stirred overnight at 100 ℃. The reaction solution was concentrated, and the residue was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the title compound (530 mg, 47%). MS: M/z [ M+H ]] ⁺ =270。

2- (trifluoromethoxy) pyridin-4-ol

4- (benzyloxy) -2- (trifluoromethoxy) pyridine (530 mg,1.97 mmol) and Pd/C (10%, 125 mg) were added to methanol (20 mL) and stirred overnight under hydrogen at 60℃and 3 bar pressure. Filtering the reaction solution, and filtering the filtrateConcentration gave the crude title compound (330 mg, 94%). MS: M/z [ M+H ] ] ⁺ =180。

(3, 5-difluoro-4- ((2- (trifluoromethoxy) pyridin-4-yl) oxy) phenyl) methanol

3, 4, 5-trifluorobenzaldehyde (294.4 mg,1.84 mol), 2- (trifluoromethoxy) pyridin-4-ol (330 mg,1.84 mol) and potassium carbonate (330.6 mg,2.39 mol) were added to DMF (10 mL), stirred at 120℃for 2 hours, the reaction solution was poured into water, extracted with ethyl acetate, the organic phase was concentrated to give a crude 3, 5-difluoro-4- ((2- (trifluoromethoxy) pyridin-4-yl) oxy) benzaldehyde, which was added to ethanol (250.0 mL), sodium borohydride (69.61 mg,1.84 mmol) was added and stirred at room temperature for 1 hour. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was dried and concentrated to give the title compound (400 mg, 68%). ¹ H NMR(400 MHz，DMSO-d ₆ )d 8.30(d，J = 5.9 Hz，1 H)，7.29(d，J = 9.3 Hz，2 H)，7.07(d，J = 3.9 Hz，1 H)，6.95(s，1 H)，5.56(br. s.，1 H)，4.56(d，J = 4.9 Hz，2 H); MS: m/z [M+H] ⁺ =322。

Intermediate 25

4- (4-chloro-3- (trifluoromethyl) phenoxy) benzaldehyde

2-chloro-5-hydroxytrifluorotoluene (15 g,76.53 mmol), 4-fluorobenzaldehyde (9.50 g,76.53 mmol) and potassium carbonate (21.12 g,153.06 mmol) were added to DMF (150 mL), stirred overnight at 120℃and the reaction mixture was poured into water, extracted with ethyl acetate, the organic phase was concentrated, and purified by column chromatography (petroleum ether/ethyl acetate=50/1 to 5/1) to give the title compound (14.6 g, 64%). MS: M/z [ M+H ]] ⁺ =301。

1-chloro-2- (trifluoromethyl) -4- (4-vinylphenoxy) benzene

Adding 4- (4-chloro-3- (trifluoromethyl) phenoxy) benzaldehyde (14.6 g,48.67 mmol) and methyltriphenyl phosphorus bromide (19.12 g,53.53 mmol) into THF (150 mL), and cooling to 0 under argon gas ^o Sodium hydride (60%, 9.73 g,243.35 mmol) was slowly added with stirring and stirred overnight at room temperature. The reaction solution was quenched with water, poured into ice water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=50/1 to 5/1) to give the title compound (13.6 g, 94%). MS: M/z [ M+H ]] ⁺ =299。

2- (4- (4-chloro-3- (trifluoromethyl) phenoxy) phenyl) ethyl-1-ol

1-chloro-2- (trifluoromethyl) -4- (4-vinylphenoxy) benzene (13.6 g,45.48 mmol) was dissolved in anhydrous tetrahydrofuran (150 mL) at room temperature and cooled to 0 ^o C addition of 9-boron bicyclo [ 3.3.1 ]]Tetrahydrofuran solution of nonane (0.5 M,136 mL,68.00 mmol) was stirred overnight at room temperature. To the reaction mixture were added water (14, 14 mL), aqueous sodium hydroxide (3M, 70 mL) and hydrogen peroxide (30%, 70% mL) in this order, and the mixture was stirred at 50℃for 2 hours. The reaction solution was quenched with saturated aqueous sodium thiosulfate, poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by column chromatography (dichloromethane/methanol=30/1) to give the title compound (10.6 g, 74%). MS: M/z [ M+H ] ] ⁺ =317。

Intermediate 26

4-bromo-2- (difluoromethyl) pyridine

4-bromopyridine-formaldehyde (10 g,53.76 mmol) was added to dichloromethane (100 mL), and the temperature was reduced to 0 ^o C. Stirring to obtain diethylaminosulfur trifluoride(17.3 g,107.52 mmol) the reaction mixture was dropped and stirred at room temperature overnight. The reaction solution was slowly dropped into water, extracted with dichloromethane, and the organic phase was concentrated to give the crude title compound which was directly used in the next reaction. MS: M/z [ M+H ]] ⁺ =208。

2- (difluoromethyl) -4-methoxypyridine

4-bromo-2- (difluoromethyl) pyridine (11.18 g,53.76 mmol), sodium methoxide (5.81 g,107.52 mmol) was added to methanol (100 mL). 90 ^o C stirring overnight. The reaction solution was cooled to room temperature and poured into water (300 mL), followed by extraction with ethyl acetate. The organic phase was concentrated to give crude title compound which was used directly in the next reaction. MS: M/z [ M+H ]] ⁺ =160。

2- (difluoromethyl) pyridin-4-ols

2- (difluoromethyl) -4-methoxypyridine (9.54 g,59.95 mmol) was added to hydrobromic acid (40% aqueous, 58 mL). 90 ^o C stirring for 2 days. The reaction solution was concentrated, the residue was diluted with water, sodium hydrogencarbonate solid was added with stirring until no bubble was generated in the system, ethyl acetate was extracted, and the organic phase was concentrated and purified by column chromatography (petroleum ether/ethyl acetate=10/1 to 1/1) to give the title compound (2.5. 2.5 g, three steps total yield 26%). MS: M/z [ M+H ] ] ⁺ =146。

4- ((2- (difluoromethyl) pyridin-4-yl) oxy) -3-fluorobenzaldehyde

3, 4-difluorobenzaldehyde (350 mg,2.46 mmol), 2- (difluoromethyl) pyridin-4-ol (357 mg,2.46 mmol) and potassium carbonate solid (1019 mg,7.38 mmol) were added to N, N-dimethylformamide (5 mL),120 ^o c stirred for 3 hours. The reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (510 mg, 78%) by preparative thin layer chromatography. MS: M/z [ M+H ]] ⁺ =268。

(4- ((2- (difluoromethyl) pyridin-4-yl) oxy) -3-fluorophenyl) methanol

4- ((2- (difluoromethyl) pyridin-4-yl) oxy) -3-fluorobenzaldehyde (505 mg,1.89 mmol) was added to absolute ethanol (5 mL), naBH was added with stirring ₄₍ 140 mg,3.78 mmol), stirred at room temperature for 2 hours. The reaction solution was poured into water, extracted with dichloromethane, and the organic phase was concentrated to give the title compound (381 mg, 75%). MS: M/z [ M+H ]] ⁺ =271。

Examples

Example 1

Method A

3'- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclopropane-1, 8' -pyrido [1', 2': 3,4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

3' -chloro-6 ', 7', 9a ', 10' -tetrahydro-1 ' is reacted at room temperature 'H, 9'H-spiro [ cyclopropane-1, 8' -pyridinyl [1', 2': 3,4 ]Imidazole [1, 2 ]c]Pyrimidine]1' -Ketone (30 mg,0.12 mmol), (3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) phenyl) methanol (50 mg,0.17 mmol) and sodium hydride (7 mg,0.18 mmol) were added to acetonitrile (5 mL) and stirred at room temperature for 1 hour. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (9 mg, 15%) by preparative thin layer chromatography. ¹ H NMR(400 MHz, CDCl ₃ )d 8.56(d, J = 5.4 Hz, 1 H), 7.32(d, J = 11.2 Hz, 1 H), 7.28 - 7.14(m, 3 H), 6.95(d, J = 3.4 Hz, 1 H), 5.43(s, 2 H), 5.04(s, 1 H), 4.35 - 4.13(m, 1 H), 4.01 - 3.84(m, 1 H), 3.67(dd, J = 7.3, 11.2 Hz, 1 H), 3.53(dd, J = 3.9, 12.7 Hz, 1 H), 3.22 - 3.07(m, 1 H), 2.09 - 1.92(m, 2 H), 1.11(d, J = 12.7 Hz, 1 H), 0.88(d, J = 12.7 Hz, 1 H), 0.53 - 0.38(m, 4 H); MS: m/z [M+H] ⁺ =503。

Example 2

Method B

3'- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -8', 9', 9a', 10 '-tetrahydro-1'H, 7'H-spiro [ cyclopropane-1, 6' -pyridine [1', 2': 3, 4]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

3' -chloro-8 ', 9', 9a ', 10' -tetrahydro-1 ' is reacted at room temperature 'H, 7'H-spiro [ cyclopropane-1, 6' -pyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -Ketone (80 mg,0.3 mmol), (3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) phenyl) methanol (86 mg,0.3 mmol) and cesium carbonate solid (195 mg,0.6 mmol) were added to toluene (10 mL), 100 ^o C stirring overnight. The reaction solution was filtered, and the filtrate was concentrated to give the title compound (50 mg, 33%) by preparative thin layer chromatography. ¹ H NMR(400 MHz, CDCl ₃ )d 8.55(d, J = 5.4 Hz, 1 H), 7.34(d, J = 10.8 Hz, 1 H), 7.27 - 7.25(m, 1 H), 7.23 - 7.12(m, 2 H), 6.95(d, J = 5.4 Hz, 1 H), 5.41(s, 2 H), 5.35(s, 1 H), 3.94 - 3.84(m, 3 H), 2.04(t, J = 12.5 Hz, 1 H), 1.85(br. s., 1 H), 1.79 - 1.71(m, 1 H), 1.53(br. s., 1 H), 1.41 - 1.29(m, 1 H), 1.20 - 1.13(m, 1 H), 0.96 - 0.84(m, 2 H), 0.78 - 0.68(m, 1 H), 0.58 - 0.48(m, 1 H); MS: m/z [M+H] ⁺ =503。

Example 3

Method C

3'- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a', 10 '-tetrahydro-1' H, 9 'H-spiro [ oxetane-3, 8' -pioglitazonePyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidine]-1' -one

(3- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -8- (hydroxymethyl) -1-oxo-6, 7, 8, 9, 9a, 10-hexahydro-1) at room temperatureHPyrido [1', 2': 3, 4]Imidazo [1, 2 ]c]Pyrimidin-8-yl) methanesulfonic acid methyl ester (60 mg,0.098 mmol) was added to anhydrous tetrahydrofuran (20 mL), sodium hydride (60%, 19.6 mg,0.49 mmol), 50 ^o C stirred for 24 hours. The reaction solution was quenched with saturated aqueous ammonium chloride, poured into water, extracted with ethyl acetate, and the organic phase was concentrated to give the title compound (15 mg, 30%) by preparative thin layer chromatography (dichloromethane/methanol=10/1). ¹ H NMR(400 MHz, CDCl ₃ )d 8.58(d, J = 5.4 Hz, 1 H), 7.33(d, J = 11.2 Hz, 1 H), 7.24(d, J = 9.3 Hz, 2 H), 7.22 - 7.16(m, 1 H), 6.96(d, J = 5.4 Hz, 1 H), 5.46(s, 2 H), 5.05(s, 1 H), 4.69 - 4.52(m, 2 H), 4.43(s, 2 H), 4.33 - 4.17(m, 1 H), 3.74(d, J = 7.3 Hz, 2 H), 3.62 - 3.49(m, 1 H), 3.15 - 2.93(m, 1 H), 2.44(d, J = 12.2 Hz, 1 H), 2.26(d, J = 12.7 Hz, 1 H), 1.74(br. s., 1 H), 1.64 - 1.56(m, 1 H); MS: m/z [M+H] ⁺ =519。

Example 4

Method D

3'- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -3-hydroxy-6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

3- ((tert-Butyldimethylsilanyloxy) -3' - ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a ', 10' -tetrahydro-1 ' at room temperature 'H, 9'H-spirocyclic ring[ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]1' -Ketone (16 mg,0.026 mmol) was added to anhydrous tetrahydrofuran (2 mL), tetrabutylammonium fluoride (20.39 mg,0.078 mmol) was added with stirring, and stirred at room temperature for 5 hours. The reaction was poured into water, extracted with dichloromethane, and the organic phase was concentrated and purified by preparative thin layer chromatography to give the title compound (5.5 mg, 40%). ¹ H NMR(400 MHz, CDCl ₃ )d 8.57(d, J = 5.4 Hz, 1 H), 7.32(d, J = 10.8 Hz, 1 H), 7.25 - 7.14(m, 3 H), 6.95(d, J = 4.4 Hz, 1 H), 5.42(s, 2 H), 5.02(s, 1 H), 4.48 - 4.33(m, 1 H), 4.26 - 4.15(m, 1 H), 3.82(br. s., 1 H), 3.72 - 3.63(m, 1 H), 3.42(d, J = 10.3 Hz, 1 H), 3.15 - 2.99(m, 1 H), 2.44(d, J = 5.4 Hz, 1 H), 2.25(d, J = 4.9 Hz, 1 H), 2.04(br. s., 1 H), 1.92(d, J = 13.2 Hz, 1 H), 1.72 - 1.62(m, 4 H); MS: m/z [M+H] ⁺ =533。

Example 5

Method E

3-amino-3 '- ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a', 10 '-tetrahydro-1'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]-1' -one

3' - ((3-fluoro-4- ((2- (trifluoromethyl) pyridin-4-yl) oxy) benzyl) oxy) -6', 7', 9a ', 10' -tetrahydro-1 ' at room temperature 'H, 9'H-spiro [ cyclobutane-1, 8' -pyridine [1', 2': 3, 4 ]]Imidazole [1, 2 ]c]Pyrimidine]1', 3-Dione (33 mg,0.12 mmol) and methylamine hydrochloride (13 mg,0.2 mmol) were added to methanol (3 mL), sodium cyanoborohydride (21 mg,0.33 mmol) was added with stirring, and stirred overnight at room temperature. The reaction was quenched with water, extracted with dichloromethane, and the organic phase concentrated to give the title compound (9 mg, 25%) which was purified by preparative thin layer chromatography. ¹ H NMR(400 MHz, CDCl ₃ )d 8.58 (d, J = 5.4 Hz, 1 H), 7.33 (d, J = 10.8 Hz, 1 H), 7.27 - 7.25 (m, 1 H), 7.23 - 7.15 (m, 2 H), 6.97 (br. s., 1 H), 5.42 (br. s., 2 H), 5.05 (s, 1 H), 4.29 - 4.19 (m, 1 H), 3.83 (br. s., 1 H), 3.76 - 3.61 (m, 2 H), 3.51 (d, J = 12.2 Hz, 1 H), 3.16 - 2.97 (m, 1 H), 2.58 (br. s., 3 H), 2.52 - 2.34 (m, 3 H), 2.30 - 2.16 (m, 2 H), 2.12 - 1.97 (m, 2 H), 1.67 - 1.59 (m, 1 H); MS: m/z [M+H] ⁺ =546。

Examples 6-34 listed in the following table were prepared by procedures analogous to those described in examples 1-5 starting from the corresponding intermediates:

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biological testing and data

The compound of the invention is Lp-PLA ₂ Inhibitors useful for the treatment and prevention of Lp-PLA ₂ Mediated diseases. The biological activity of the compounds of the invention can be used to determine the compound as Lp-PLA ₂ Any suitable test for the activity of the inhibitor, as well as tissue and in vivo models.

The biological activity data for each compound is reported as an average of at least one experiment or multiple experiments. It is to be understood that the data described herein may vary reasonably depending on the particular conditions and methods used by the person performing the experiment.

Lipoprotein-related phospholipase A2 (Lp-PLA) ₂ ) Human plasma assay.

Human plasma assays utilize thioester analogs of PAF (phosphatidylcholine), where hydrolysis results in the formation of phospholipids containing free sulfhydryl groups. The amount of thiol groups was continuously determined by reaction with CPM (7-diethylamino-3- (4' -maleimidophenyl) -4-methylcoumarin), a maleimide with increased fluorescence after Michael addition of thiol groups. The assay can detect Lp-PLA in human plasma ₂ Is, for example, through Lp-PLA ₂ Specific inhibition of the inhibitor was determined.

The Thio-PAF assay was performed as a quenched 75. Mu.L assay. Compound source plates were prepared by preparing 1:3 (volume) serial dilutions of the individual compounds in pure DMSO on 96-well microtiter plates. 3. Mu.L of the compound on the source plate was transferred by a Raini multichannel pipette into a 96-well microplate to which 57. Mu.L of assay buffer had been added in advance, and the compound on the source plate was 20-fold diluted. The test buffer contained a mixture of 50mM HEPES,pH 7.4, 150mM NaCl, 1mM CHAPS. 20-fold dilution of the compound was performed by a Rainin multichannel pipette Transfer 1 μl into a 96 well Greiner 655076 (black) microplate with pre-added aliquots and thawed 40 μl of pooled human plasma. The plates were mixed well by shaking in an ELISA plate shaker for 20 seconds. After 30 minutes of pre-incubation at room temperature, 10. Mu.L of substrate solution containing 2.5mM 2-thio-PAF [ from ethanol stock ] in assay buffer consisting of 50mM HEPES,pH 7.4, 150mM NaCl, 1mM CHAPS was added to a 96-well Greiner 655076 (black) microplate by means of a Rainin multichannel pipette]32. Mu.M CPM [ from DMSO mother liquor ]]And 3.2mM NEM (N-ethylmaleimide) [ New preparation in DMSO per experiment ]]. After 2 min, the reaction was quenched with 25 μl of 5% aqueous trifluoroacetic acid (TFA). Plates were centrifuged at 2000rpm for 1 min. A Biotek Synergy H1 (H1 MF) microplate reader was used with an ex:380/em:485 plate. IC using GraphPad Prism 6.0 and Excel ₅₀ Data, curves and QC analysis.

Examples determination of Activity

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Claims

1. A compound of the formula:

wherein the method comprises the steps of

m is 1 or 2;

u is independently 0 or 1;

q is-O-;

R ₁ is H, halogen, cyano, amino, C _1-6 Alkyl, C _1-3 Alkoxy, C _1-3 Alkylamino, C _1-3 A haloalkyl group;

R _x ，R _y independently selected from the following substituents: h, halogen, hydroxy, carboxy, cyano, C _1-6 Alkyl, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl, 3-8 membered heteroaryl, -C(O)NR _b R _c ，-S(O) ₂ NR _b R _c And optionally substituted with one or more of the following substituents: halogen, cyano, C _1-6 Alkoxy, C _3-8 Cycloalkyl, 3-8 membered heterocyclyl or 3-8 membered heteroaryl;

R _m is C _1-6 Alkyl, C _1-3 Haloalkyl, halogen, cyano, -OR _c ，-NR _b R _c ，C _3-6 Cycloalkyl, 3-8 membered heterocyclyl, C _6-10 Aryl or 3-8 membered heteroaryl;

R _b is H, C _1-6 Alkyl, C _3-8 Cycloalkyl or 3-8 membered heterocyclyl;

R _c is L, L-C (O) -, L-CH ₂ -or L-S (O) ₂ -，

a is

Z is N or CR ₃ ；

Z' is N or CR _4；

R ₃ ,R ₄ ,R ₅ ,R ₆ Independently H, CN, halogen, C _1-3 Alkyl or C _1-3 A haloalkyl group;

v is N or CR _9， Wherein R is ₉ Is H, CN, halogen, C _1-3 Alkyl, C _1-3 Haloalkyl or-O-W; w is a 5-or 6-membered heteroaryl or phenyl group, which may optionally be substituted with one or more of the following substituents Substitution: halogen, cyano, C _1-6 Alkyl, C _1-3 Alkoxy, C _1-3 Haloalkyl and C _1-3 Haloalkoxy groups.

2. The compound of claim 1, wherein u is 0, r _x ,R _y H.

3. The compound of claim 1 or 2, wherein R ₁ Is H, halogen, C _1-6 Alkyl or C _1-6 An alkoxy group.

4. The compound of claim 1 or 2, wherein m is 1.

5. The compound of claim l or 2, wherein a is

R ₅ ,R ₆ ,R ₇ ,R ₈ ,R ₉ H, F or CN, independently.

6. The compound of claim l or 2, or a pharmaceutically acceptable salt thereof, wherein a is

R ₅ ,R ₆ ,R ₇ ,R ₈ H, F or CN independently;

R ₉ is-O-W;

w is a 5-or 6-membered heteroaryl or phenyl group, which may optionally be substituted by one or more substituents C _1-3 Haloalkyl, C _1-3 Haloalkoxy, CN, halogen and C _1-6 An alkyl group.

7. The compound of claim l or 2, wherein a is

R ₇ ,R ₈ H, F or CN independently;

R ₉ is-O-W;

w is pyridinyl, pyrimidinyl, pyrazolyl or phenyl optionally substituted with one or more substituents independently selected from the group consisting of: halogen, CN, CF ₃ 、-OCF ₃ 、CHF ₂ And CH (CH) ₃ 。

8. The compound of claim 1 or 2, wherein R _m Is H, hydroxy, methoxy or methylamino.

9. The compound of claim i or 2, wherein a is selected from the group consisting of:

10. The compound of claim 1 having one of the following structures:

11. a composition comprising a compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

12. The compound according to any one of claims 1 to 10 or the composition according to claim 11 for use in the manufacture of a medicament for the treatment, prevention or amelioration of Lp-PLA ₂ Use in medicine for inhibiting related diseases.

13. A process for preparing a compound according to any one of claims 1 to 10, the process comprising the steps of:

step (iv): obtaining a compound (1.7) by ring-closing the compound (1.5);

or comprises the following steps:

step (iii): reacting compound (1.2) with compound (1.4) to form compound (1.6);

step (iv'): obtaining a compound (1.7) by ring-closing the compound (1.6);

Step (v) Compounds (1.7) and HQ- (CH) ₂ ) _m -a reaction to give the final product (1.8), the final product (1.8) being a compound of formula:

wherein R is ₁ ，R _x ，R _y ，R _m Q, m, u, A are as defined in any of claims 1 to 10.