WO2019233461A1 - Tropomyosin receptor kinase inhibitor, preparation method therefor and use thereof - Google Patents

Abstract

Disclosed in the present invention are a pyrazolo[1,5-a]pyrimidine derivative having a structure of formula (I), a pharmaceutical composition comprising the compound of formula (I), and use of the compound in the preparation of a medicament for preventing or treating diseases associated with tropomyosin receptor kinases, in particular for preventing or treating cancers associated with tropomyosin receptor kinases. Each substituent in formula (I) has the same definition as that in the description.

Description

Promyosin receptor kinase inhibitor and preparation method and application thereof Technical field

The present application belongs to the technical field of medicine and specifically relates to pyrazolo [1,5-a] pyrimidine derivatives and uses thereof for the treatment of pain, cancer, and inflammation-related diseases.

This application claims the priority of Chinese patent CN201810597223.9 (application date: June 8, 2018, invention name tropomyosin receptor kinase inhibitor, preparation method and application thereof).

Background technique

Tromyosin receptor kinase (Trk) is a family of receptor tyrosine kinases that regulate the strength and plasticity of mammalian nervous system synapses. The activation of Trk receptors affects the survival and differentiation of neurons through multiple signaling pathways, and also significantly affects the function of neurons. The common ligand of the Trk receptor is neurotrophins, which play a key role in the nervous system, and the binding between two pairs is highly specific (J. Mol. Biol. 1999, 90, 149). Each neurotrophic factor has corresponding Trk receptors with different affinities. The combination causes the Trk receptor to dimerize and activate phosphorylation, causing downstream signals including RAS / MAPK / ERK, PLCγ, and PI3K / Akt pathways to activate and regulate cells. Survival and other functions (Cancers 2018, 10, 105).

Inhibitors of the Trk / neurotrophic factor pathway have been shown to be effective in many painful preclinical animal models. For example, the antibody RN-624, which antagonizes NGF and TrkA, has been shown to be effective in animal models of inflammatory and neuropathic pain (Neuroscience 1994, 62, 327; Eur. J. Neurosci. 1999, 11, 837). In addition, some literatures indicate that brain-derived neurotrophic factor (BDNF) levels and TrkB signaling increase in the dorsal root ganglia after inflammation (Brain Research 1997, 749, 358). A number of studies have shown that inhibiting the BDNF / TrkB pathway through antibodies, reducing signal transduction, can alleviate neuropathic effects and associated pain (Molecular Pain 2008, 4, 27). At present, many small molecule inhibitors of Trk kinase have been shown to be useful for treating pain (Expert Opin. Ther. Patents 2009, 19, 305).

In addition, the use of NGF antibodies or small molecule inhibitors of TrkA, B, and C to block the neurotrophic factor / Trk pathway has been shown to be effective in preclinical models of inflammatory diseases, such as inflammatory lung diseases, including asthma (Pharmacology & Therapeutics 2008, 117,52), Interstitial cystitis (The Journal of Urology 2005, 173, 1016), inflammatory bowel disease (including ulcerative colitis and Crohn's disease (Gut 2000, 46, 670), inflammatory skin diseases, etc. (Archives of Dermatological Research 2006, 298, 31), eczema and psoriasis (J. Investigative Dermatology 2004, 122, 812) and so on.

Reports in the literature also show that overexpression, activation, amplification or mutation of Trk kinase is closely related to many cancers, including neuroblastoma, ovarian cancer, glioblastoma, lung adenocarcinoma, juvenile sarcoma, colorectal cancer, fibrosarcoma, Spitzoid melanoma, thyroid cancer, intrahepatic cholangiocarcinoma, large cell neuroendocrine tumor, papillary thyroid cancer, hairy astrocytoma, head and neck squamous cell carcinoma, acute myeloid leukemia, congenital mesoderm nephroma, Ductal adenocarcinoma, gastrointestinal stromal cell carcinoma, salivary gland breast secretion carcinoma (Cancers 2018, 10, 105), etc. In preclinical models of cancer, small molecule inhibitors of Trk A, B, and C effectively inhibit tumor growth and prevent tumor metastasis (Cancer Letters 2001, 169, 107; Leukemia 2007, 1-10; Cancer Letters 2006, 232, 90; Cancer Res. 2008, 68, 346).

Summary of the Invention

The object of the present invention is to provide a pyrzolo [1,5-a] pyrimidine Trk inhibitor.

Another object of the present invention is to provide the use of the Trk inhibitor in the preparation of a medicament for preventing or treating a tropomyosin receptor kinase-related disease.

To achieve the objective of the present invention, the technical solution of the present invention is as follows:

The compound of the present invention represented by the following formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

R ^{1 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, -COR ⁵ , -SO ₂ R ⁵ or- SOR ^{5 is} optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further by one or Multiple selected from deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

Y is-(CH ₂ ) _n -or -O (CH ₂ ) _n- ;

n is selected from 0, 1, 2 or 3;

Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

R ^{3 is} selected from H, deuterium, halogen, cyano, nitro, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy;

X is selected from CH or N.

An embodiment of the present invention, which is a compound described by the general formula (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

R ^{1 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, -COR ⁵ , -SO ₂ R ⁵ or- SOR ^{5 is} optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further by one or Multiple selected from deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

R ^{3 is} selected from H, deuterium, halogen, cyano, nitro, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy;

X is selected from CH or N.

The embodiment of the present invention is characterized by:

R ^{1 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further selected from deuterium , Halogen, hydroxy, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl , C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further by one or Multiple selected from deuterium, halogen, hydroxyl, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C _3- C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

R ³ is halogen; X is selected from CH or N.

The embodiment of the present invention is characterized by:

R ^{1 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or 4-10 membered heterocyclic ring Substituted by a substituent;

R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy, or 4 -10 membered heterocyclic group substituted by a substituent;

R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl; alternatively, R ¹ , R ² and R ⁴ may independently form C ₃ -C ₈ -cycloalkyl or 4-10 membered heterocyclic group;

R ³ is halogen;

X is selected from CH or N.

The embodiment of the present invention is characterized by:

R ^{1 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -SO ₂ R ⁷ , C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl or C ₁ -C ₄ alkoxy substituted by a substituent;

R ^{7 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, C _1- C ₄ alkyl, C ₃ -C ₆ cycloalkyl or C ₁ -C ₄ alkoxy substituted by a substituent;

Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

R ³ is halogen; X is CH or N.

The embodiment of the present invention is characterized by:

R ^{1 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, hydroxyl, halogen, -SO ₂ R ⁷ or C ₁ -C ₄ Substituted with alkoxy substituents;

R ^{7 is} selected from H or C ₁ -C ₈ alkyl; R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

Alternatively, R ¹ , R ² and R ⁴ may independently form a 4-10 membered heterocyclic group;

R ³ is fluorine or chlorine; X is selected from CH or N.

The embodiment of the present invention is characterized by:

R ^{1 is} selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, optionally further Substituted with one or more substituents selected from deuterium, hydroxyl, F, Cl, -SO ₂ CH ₃ or methoxy;

R ² and R ⁴ are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclo Jiji

Alternatively, R ¹ , R ² and R ⁴ may independently form a morpholinyl group;

R ³ is fluorine or chlorine; X is selected from CH or N.

In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is selected from the following structures:

More preferably, it is selected from the following structures:

The compound of formula (I), its stereoisomer or its pharmaceutically acceptable salt according to the present invention is a novel Trk inhibitor, and thus can be used to prepare a medicament for preventing or treating a disease related to tropomyosin receptor kinase. The diseases include, but are not limited to, pain, cancer, and inflammation.

The invention further provides the use of the compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof for the prevention or treatment of cancer.

As a further preferred scheme, the cancer includes: neuroblastoma, ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, leukemia, Lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cell lymphoma, acute myeloid leukemia , Multiple myeloma, melanoma or mesothelioma, juvenile sarcoma, fibrosarcoma, large cell neuroendocrine tumor, hairy cell astrocytoma, head and neck squamous cell carcinoma, congenital mesoderm nephroma, ductal adenocarcinoma, Salivary gland breast-like secretory cancer and appendix cancer.

Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective dose of the aforementioned compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Another aspect of the present invention provides the application of the pharmaceutical composition in the preparation of a medicament for preventing or treating cancer.

Detailed description: Unless stated to the contrary, the following terms used in the specification and claims have the following meanings.

In the present invention, "C ₁ -C ₈ alkyl" refers to a straight-chain alkyl group and a branched alkyl group including 1 to 8 carbon atoms. The alkyl group refers to a saturated aliphatic hydrocarbon group, such as methyl, ethyl, n- Propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2 -Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-tris Methylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl Methyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4 -Methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl , 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2- Dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl 2-ethylpentyl, 2-methyl-3-ethylpentyl, or various branched isomers thereof, and the like.

In the present invention, "cycloalkyl" refers to a saturated monocyclic hydrocarbon substituent, and "C ₃ -C ₈ cycloalkyl" refers to a monocyclic cycloalkyl including 3 to 8 carbon atoms, for example, Restrictive examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like; "C ₃ -C ₆ cycloalkyl" refers to a single ring including 3 to 6 carbon atoms Cycloalkyl, for example: Non-limiting examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

In the present invention, "alkenyl" refers to an alkyl group as defined above, which is composed of at least two carbon atoms and at least one carbon-carbon double bond, and "C ₂ -C ₈ alkenyl" refers to a straight chain containing 2 to 8 carbons. Or containing branched alkenyl. Examples are vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, and the like.

In the present invention, "alkynyl" refers to an alkyl group as defined above composed of at least two carbon atoms and at least one carbon-carbon triple bond, and "C ₂ -C ₈ alkynyl" refers to a straight-chain or Contains branched alkynyl. For example, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl and the like.

In the present invention, "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O) r (where r is an integer of 0, Heteroatoms of 1, 2), but excluding the ring part of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. "4-10 membered heterocyclyl" refers to a cyclic group containing 4 to 10 ring atoms. Non-limiting examples of monocyclic heterocyclyls include tetrahydropyranyl, dihydropyranyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl Wait. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups. In the present invention, "alkoxy" means -O- (alkyl), wherein alkyl is as defined above. "C ₁ -C ₈ alkoxy" refers to an alkyl group having 1 to 8 carbon atoms, non-limiting embodiments include methoxy, ethoxy, propoxy, butoxy and the like.

"Halogen" means fluorine, chlorine, bromine or iodine.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or a physiologically pharmaceutically acceptable salt or prodrug thereof with other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers and excipients.形 剂。 Shape agent. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and then exerts the biological activity.

In the preparation steps of the present invention, the abbreviations of the reagents used represent:

MTBE, methyl tert-butyl ether

Sec-BuLi Sec-BuLi

THF: Tetrahydrofuran

Pd2 (dba) 3 Tris (dibenzylideneacetone) dipalladium

t-Bu3P-HBF4 Tri-n-butylphosphonium tetrafluoroborate

Hexane n-hexane

EA, ethyl acetate

DCM: Dichloromethane

DIEA N, N-diisopropylethylamine

DMF, N, N-dimethylformamide

TEA Triethylamine

(Ph3P) 2PdCl2 Dichloroditriphenylphosphonium palladium

HOBt 1-Hydroxybenzotriazole

EDCI 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 Nuclear magnetic resonance proton spectrum of the compound of Example 1;

Figure 2 Nuclear magnetic resonance proton spectrum of the compound of Example 2;

Figure 3 Nuclear magnetic resonance proton spectrum of the compound of Example 3a;

Figure 4 Nuclear magnetic resonance proton spectrum of the compound of Example 3b;

Figure 5 Nuclear magnetic resonance proton spectrum of the compound of Example 4;

Figure 6 Nuclear magnetic resonance proton spectrum of the compound of Example 5;

Figure 7 Nuclear magnetic resonance proton spectrum of the compound of Example 6;

Figure 8 Nuclear magnetic resonance proton spectrum of the compound of Example 7a;

Figure 9 Nuclear magnetic resonance proton spectrum of the compound of Example 7b;

Figure 10 Nuclear magnetic resonance proton spectrum of the compound of Example 8a;

Figure 11 Nuclear magnetic resonance proton spectrum of the compound of Example 8b;

Figure 12 Nuclear magnetic resonance proton spectrum of the compound of Example 9;

Figure 13 Nuclear magnetic resonance proton spectrum of the compound of Example 10.

Detailed ways

The present invention is described below with reference to specific examples. Those skilled in the art can understand that these examples are only used to illustrate the present invention, and they do not limit the scope of the present invention in any way.

Unless otherwise specified, the experimental methods in the following examples are conventional methods. Unless otherwise specified, the medicinal materials, reagent materials, etc. used in the following examples are all commercially available products.

Example 1

(R, 1 ³ E, 1 ⁴ E) -6-cyclopropyl-3 ⁵ -fluoro-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine -3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 1: Synthesis of 2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidine-1-carboxylic acid (R) -tert-butyl ester

At room temperature, add tert-butyl pyrrolidine-1-carboxylate (2.86mL, 16.03mmol), (-)-spartin (4.50g, 19.20mmol), and MTBE (35.0mL) to a 250mL three-necked flask, and replace with nitrogen. Three times, slowly reduce the temperature to -78 ° C. After the temperature stabilizes, add Sec-BuLi (17.01mL, 21.70mmol, 1.3N cyclohexane solution) dropwise, stir at -75 ° C for 3 hours, and slowly drop at -65 ° C. ZnCl ₂ (14.68 mL, 14.36 mmol, 1N THF solution) was cooled to about -78 ° C and stirred for 20 minutes. Warm to room temperature, and under the protection of nitrogen, 3-bromo-5-fluoro-2-methoxypyridine (3.44 g, 16.70 mmol), Pd (OAc) ₂ (0.18 g, 0.84 mmol), and t-Bu ₃ P are sequentially added. -HBF ₄ (0.28 g, 1.00 mmol), and reacted at room temperature for 16 hours. 1.50 mL of ammonia was added dropwise to the reaction system, and the mixture was stirred at room temperature for 1 hour. The reaction solution was filtered through celite, washed with MTBE (20 mL x 3), separated, and the organic phase was concentrated. Column chromatography (Hexane: EA = 20: 1) After decompression and concentration, 2.60 g of a yellow oil was obtained with a yield of 52.53%. ¹ H NMR (400MHz, Chloroform-d) δ 7.84 (d, J = 7.2 Hz, 1H), 7.09 (dd, J = 29.5, 6.1 Hz, 1H), 4.99 (dd, J = 52.8, 8.0 Hz, 1H ), 3.93 (s, 3H), 3.53 (m, 2H), 2.28 (m, 1H), 1.82 (m, 3H), 1.46 (s, 9H).

Step 2: Synthesis of (R) -5-fluoro-2-methoxy-3- (pyrrolidin-2-yl) pyridine

At room temperature, 100-mL single-necked flasks were sequentially charged with 2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidine-1-carboxylic acid (R) -tert-butyl ester (2.60 g, 8.77 mmol), Trifluoroacetic acid (7.80 mL) and dichloromethane (7.80 mL) were stirred at room temperature for 1.5 hours. The reaction of the raw materials was complete and the reaction was stopped. The reaction solution was evaporated to dryness under reduced pressure to obtain 3.94 g of a yellow oil, which was directly used for the next reaction.

Step 3: 5- (2- (5-Fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid (R) -Ethyl Synthesis

(R) -5-fluoro-2-methoxy-3- (pyrrolidin-2-yl) pyridine (1.72 g, 8.78 mmol), 5-chloropyrazolo [ 1,5-a] Pyrimidine-3-carboxylic acid ethyl ester (2.38 g, 10.54 mmol), DIEA (5.25 g, 40.65 mmol), DMF (19.30 mL), the temperature was slowly raised to 90 ° C, and the reaction was stirred for 15.5 h. After the reaction was cooled, EA (100 mL) was added to dissolve and disperse, washed with water (100 mL), and separated, and washed with saturated brine (100 mL) and saturated sodium bicarbonate aqueous solution (100 mL) in that order, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Column chromatography (PE: EA = 1: 1) gave 1.83 g of a yellow solid. MS (ESI) m / z: 386 [M + H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ 8.28 (s, 1H), 8.15 (d, J = 5.2Hz, 1H), 7.91 (s, 1H), 7.04 (d, J = 7.2Hz, 1H), 5.84 (d, J = 4.5Hz, 1H), 5.09 (m, 1H), 4.37 (m, 2H), 4.18-4.11 (m, 1H), 4.02 (s, 3H), 3.96 (m, 1H), 2.47 (m, 1H), 2.04 (m, 2H), 1.95 (m, 1H), 1.42 (m, 3H).

Step 4: 5- (2- (5-Fluoro-2-hydroxypyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidine-3-carboxylic acid (R)- Synthesis of ethyl ester

Add 5- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidine-3 to a 100 mL single-necked flask at room temperature -Carboxylic acid (R) -ethyl ester (1.83 g, 4.75 mmol), HBr (18.30 mL, 33% AcOH solution), slowly raised to 90 ° C, and stirred for 4.5 hours. EA (100 mL) was added to dissolve and disperse, washed with water (100 mL), and separated, and successively washed with a saturated solution of NaHCO ₃ (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure. Column chromatography (DCM: EtOH = 20: 1) gave 1.34 g of a yellow solid. MS (ESI) m / z: 372 [M + H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ 12.92 (br, 1H), 8.29 (s, 1H), 8.21 (d, J = 7.5Hz, 1H), 7.30 (s, 1H), 7.17 (s, 1H ), 6.93 (d, J = 4.7 Hz, 1H), 5.13 (m, 1H), 4.37 (q, J = 8.7 Hz, 2H), 4.16 (m, 1H), 3.95 (m, 1H), 2.49 (m , 1H), 2.13–2.07 (m, 2H), 1.95 (m, 1H), 1.42 (t, J = 8.8Hz, 3H).

Step 5: 5- (2- (5-Fluoro-2- (trifluoromethyl-sulfonyloxy) pyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] Synthesis of pyrimidine-3-carboxylic acid (R) -ethyl ester

At room temperature, add a 5- (2- (5-fluoro-2-hydroxypyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidine-3-carboxyl to a 100 mL single-necked flask. Acid (R) -ethyl ester (1.34 g, 3.61 mmol), DMF (13.0 mL), sonicated to dissolve completely. To the reaction system were sequentially added 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl) sulfonyl) methylsulfonamide (1.42 g, 3.97 mmol), TEA (0.44 g, 4.33 mmol), and the reaction was stirred at room temperature for 24 hours. EA (110 mL) was added to the reaction system, followed by washing with saturated NaHCO ₃ solution (100 mL), water (100 mL), and saturated brine (100 mL), drying over anhydrous sodium sulfate, evaporation to dryness under reduced pressure, and column chromatography (PE: EA = 1: 1) gave 1.60 g of a foamy white solid. MS (ESI) m / z: 504.0 [M + H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.78 (d, J = 7.8 Hz, 1H), 8.37 (s, 1H), 8.17 (s, 1H), 7.93 (d, J = 8.8 Hz, 1H) , 6.69 (d, J = 7.8 Hz, 1H), 5.37 (m, 1H), 4.08 (m, 1H), 4.04 (q, J = 7.2 Hz, 2H), 3.65 (m, 1H), 2.08 (m, 3H), 1.91 (m, 1H), 1.11 (t, J = 7.2Hz, 3H).

Step 6: Ethyl (S) -5- (2- (5-fluoro-2-vinylpyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3- Synthesis of formate

At room temperature, a 100-mL single-necked flask was charged with 5- (2- (5-fluoro-2- (trifluoromethyl-sulfonyloxy) pyridin-3-yl) pyrrolidin-1-yl) pyrazolo in order. [1,5-a] Pyrimidine-3-carboxylic acid (R) -ethyl ester (1.75 g, 3.47 mmol), tributyl vinylene (1.73 g, 5.21 mmol), lithium chloride (0.74 g, 17.38 mmol ), (Ph ₃ P) ₂ PdCl ₂ (0.24 g, 0.34 mmol) and DMF (35.0 mL), replaced with nitrogen three times, slowly warmed to 80 ° C., and stirred the reaction for 4 hours. Cooled to room temperature, washed with saturated NaHCO ₃ solution (100 mL), water (100 mL), and saturated brine (100 mL) in this order, dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and column chromatography (PE: EA = 1: 1) There was obtained 1.23 g of a white solid. MS (ESI) m / z: 382.0 [M + H] ⁺ . ¹ H NMR (400MHz, Chloroform-d) δ 8.36 (s, 1H), 8.29 (s, 1H), 8.16 (br, 1H), 7.05 (br, 2H), 6.42 (d, J = 16.8 Hz, 1H ), 5.81--5.53 (m, 2H), 5.20 (m, 1H), 4.36 (m, 2H), 4.11 (q, J = 7.2Hz, 2H), 2.58 (m, 1H), 2.07 (m, 3H) , 1.26 (t, J = 7.2 Hz, 3H).

Step 7: Ethyl (S) -5- (2- (2- (2- (cyclopropylamino) ethyl) -5-fluoropyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1 Synthesis of 5,5-a] pyrimidine-3-formate

Ethyl (S) -5- (2- (5-fluoro-2-vinylpyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5- a] Pyrimidine-3-formate (1.23g, 3.22mmol), cyclopropylamine (5.52g, 96.75mmol), glacial acetic acid (5.81g, 96.75mmol) and ethanol (12.30mL), protected by nitrogen, slowly warmed to 75 The reaction was carried out under reflux at 2 ° C for 20.5 hours. The reaction solution was cooled to room temperature, and the pH was adjusted to neutral with a saturated sodium carbonate aqueous solution, and extracted with EA (150 mL). The organic phase was dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and column chromatography (DCM: EtOH = 25: 1) gave a yellow color. 378.2 mg of solid, 605.4 mg of recovered material. The recovered raw materials were further subjected to the above operation, and combined to obtain 507.2 mg of a yellow solid. MS (ESI) m / z: 439.0 [M + H] ⁺ .

Step 8: ethyl (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5-fluoropyridin-3-yl ) Synthesis of pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylate

Add ethyl (S) -5- (2- (2- (2- (cyclopropylamino) ethyl) -5-fluoropyridin-3-yl) pyrrolidin-1-yl to a 10 mL single-necked flask at room temperature ) Synthesis of pyrazolo [1,5-a] pyrimidine-3-carboxylic acid ester (0.20g, 0.45mmol), tetrahydrofuran (4.0mL), dissolve by ultrasound, reduce the temperature to 0 ° C, add N-Boc-O-pair Tosyl-hydroxylamine (131.0 mg, 0.45 mmol) and N-methylmorpholine (22.5 mg, 0.23 mmol) were returned to room temperature and the reaction was stirred overnight. Diluted with dichloromethane (40mL), washed with saturated aqueous sodium bicarbonate solution (300mL), extracted with dichloromethane (40mL x 3), dried over anhydrous Na ₂ SO ₄ , evaporated to dryness under reduced pressure, and column chromatography (PE: EA = 1 : 1) 100.0 mg of a yellow solid was obtained. MS (ESI) m / z: 554.0 [M + H] ⁺ .

Step 9: (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5-fluoropyridin-3-yl) pyrrole Synthesis of Alkyl-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid

Add ethyl (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5 to a 10 mL single-necked flask at room temperature -Fluoropyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid ester (85.0mg, 0.15mmol) and ethanol (3.0mL), stir until all is dissolved After that, lithium hydroxide monohydrate (38.6 mg, 0.92 mmol, dissolved in 0.5 mL of water) was added, the temperature was slowly raised to 70 ° C, and the reaction was stirred for 15 hours. The reaction solvent was evaporated to dryness under reduced pressure at 40 ° C to obtain a crude product as a yellow solid. MS (ESI) m / z: 526.0 [M + H] ⁺ .

Step 10: (R, 1 ³ E, 1 ⁴ E) -6-cyclopropyl-3 ⁵ -fluoro-6,7-diazepine-1 (5,3) -pyrazolo [1,5- a) Synthesis of pyrimidine-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctadecane-8-one

At room temperature, the crude product from the previous step was added to a 10 mL single-necked flask, and then dichloromethane (1.0 mL) and trifluoroacetic acid (1.0 mL) were added, and the mixture was dispersed by ultrasonication, and the reaction was stirred for 2 hours. The reaction solvent was evaporated to dryness under reduced pressure at 40 ° C. to obtain a yellow oil. The oil was pumped for 4 hours at room temperature to obtain a pale yellow crude product 1. MS (ESI) m / z: 426.0 [M + H] ⁺ .

At room temperature, put the above crude product 1 into a 10 mL single-necked flask, and then add HOBt (103.7 mg, 0.77 mmol), EDCI (310.7 mg, 0.77 mmol) and DMF (4.0 mL), disperse by ultrasound, and add TEA (310.7 mg, 3.07) mmol), protected by nitrogen, and reacted at room temperature overnight. Evaporate to dryness under reduced pressure and purify by reverse column chromatography (0.1% formic acid in water / acetonitrile, 95% to 0) to obtain 11.0 mg of a white solid. MS (ESI) m / z: 408.0 [M + H] ⁺ . ¹ H NMR (300 MHz, Chloroform-d) δ 9.73 (br, 1H), 8.33 (d, J = 7.7 Hz, 1H), 8.31 (d, J = 2.5 Hz, 1H), 8.29 (s, 1H), 7.08 (dd, J = 9.4, 2.8 Hz, 1H), 6.33 (d, J = 7.7 Hz, 1H), 5.58 (t, J = 7.4 Hz, 1H), 4.01-3.65 (m, 6H), 2.94 (dd , J = 16.4, 8.6 Hz, 1H), 2.62 (dt, J = 13.3, 6.7 Hz, 1H), 2.43 (dt, J = 12.2, 6.0 Hz, 1H), 2.24 (dt, J = 13.9, 7.5 Hz, 1H), 1.96 (dt, J = 13.7, 6.7 Hz, 1H), 1.17 (dd, J = 10.3, 5.6 Hz, 1H), 0.87 (dd, J = 10.3, 4.0 Hz, 1H), 0.51 (d, J = 6.3 Hz, 2H).

Example 2

(1 ³ E, 1 ⁴ E, 2 ² R) -6-cyclopropylamino-3 ⁵ -fluoro-5-methyl-6,7-diazalide-1 (5,3) -pyrazolo [ 1,5-a] pyrimidine-3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one

(1 ³ E, 1 ⁴ E, 2 ² R) -6-cyclopropylamino-3 ⁵ -fluoro-5-methyl-6,7-diazalide-1 (5,3) -pyrazolo [ The method for preparing 1,5-a] pyrimidin-3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one is similar to that in Example 1.

¹ H NMR (300 MHz, Chloroform-d) δ 8.33 (d, J = 9.0 Hz, 1 H), 8.31 (d, J = 6.0 Hz 1 H), 8.24 (s, 1 H), 7.21 (dd, J = 9.0, 3.0Hz, 0.5H), 7.11 (dd, J = 9.0, 3.0Hz, 0.5H), 6.33 (d, J = 7.6Hz, 1H), 5.55 (t, J = 6.0Hz, 0.5H), 5.48 (t , J = 6.0 Hz, 0.5H), 4.17 (m, 1H), 3.97 (m, 1H), 3.78 (m, 1H), 3.64 (m, 1H), 3.41 (m, 1H), 2.57 (ddq, J = 19.1, 9.9, 8.6 Hz, 1H), 2.40 (dd, J = 13.1, 6.7 Hz, 1H), 2.23 (m, 1H), 2.01 (m, 1H), 1.88 (m, 6.3 Hz, 1H), 1.49 (d, J = 6.9 Hz, 1.5H), 1.16 (m, 1.5H), 1.00 (m, 1H), 0.88 (m, 1H), 0.70 (m, 1H), 0.43 (m, 1H). MS (ESI) m / z: 422.2 [M + H] ⁺ .

Example 3

(1 ³ E, 1 ⁴ E, 2 ² R, 5R) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazalide-1 (5,3) -pyrazole And [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one and (1 ³ E, 1 ⁴ E, 2 ² R, 5S ) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine-3 (3, 2) -Pyridin-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 1: Synthesis of (1 ³ E, 1 ⁴ E, 2 ² R) -6-cyclopropylamino-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 ( 5,3) -pyrazolo [1,5-a] pyrimidin-3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 2: (1 ³ E, 1 ⁴ E, 2 ² R, 5R) -6-Cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 (5,3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one and (1 ³ E, 1 ⁴ E, 2 ² R, 5S) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine-3 Preparation of (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

Take (1 ³ E, 1 ⁴ E, 2 ² R) -6-cyclopropylamino-3 ⁵ -fluoro-5-methyl-6,7-diazalide-1 (5,3) -pyrazolo [1,5-a] Pyrimidine-3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one (200 mg, 0.47 mmol) Example 3a: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazalide-1 (5, 3) -pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one (28.3mg, yield 28.3%) and Example 3b: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 (5 , 3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one (35.6mg, yield 35.6%) .

Separation preparation method:

Instrument: waters SFC200

Column: ChiralPak OD, 250 × 30mm I.D., 5μm

Mobile phase: A is CO ₂ ; B is MEOH (0.1% NH ₃ H ₂ 0)

Wavelength: 254nm

Column temperature: 38 ° C

Flow rate: 60g / min

Single injection volume: 5mL

Single injection concentration: 10mg / mL

Number of injections: 4

Solvent: MeOH

Back pressure: 100bar

Gradient condition: B 40%

Cycle time: 8min

Collection conditions: concentrated under reduced pressure at 40 ° C

Separation detection method:

Instrument: waters UPCC

Column: ChiralPak OD, 2.1 × 150mm I.D., 3μm

Mobile phase: A is CO ₂ ; B is MEOH (0.1% DEA)

Wavelength: 254nm

Column temperature: 40 ° C

Flow rate: 1mL / min

Injection volume: 2 μL

Injection concentration: 2mg / mL

Solvent: MeOH

Back pressure: 1500psi

Gradient conditions: B 5% -40%

Running time: 10min

Retention time: 4.471min, 4.788min

Example 3a: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 (5 , 3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one, with a retention time of 4.471 min. MS (ESI) m / z: 422.2 [M + H] ^{+ .1} H NMR (400MHz, CDCl ₃ ) δ8.63 (br, 1H), 8.31 (dd, J = 23.1,15.6Hz, 3H), 7.16– 7.07 (m, 1H), 6.33 (d, J = 7.6 Hz, 1H), 5.49 (t, J = 17.1 Hz, 1H), 4.16 (t, J = 8.0 Hz, 1H), 4.01-3.92 (m, 1H ), 3.81 (m, 1H), 3.46 (dd, J = 33.0, 21.6 Hz, 2H), 3.18 (m, 1H), 2.60 (m, 1H), 2.39 (m, 1H), 2.24 (m, 1H) , 1.89 (d, J = 11.6 Hz, 1H), 1.29-1.21 (d, J = 7.6 Hz, 3H), 1.07-0.93 (m, 2H), 0.74-0.66 (m, 1H), 0.51-0.42 (m , 1H).

Example 3b: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -6-cyclopropyl-3 ⁵ -fluoro-5-methyl-6,7-diazepine-1 (5 , 3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one, retention time 4.788min. MS (ESI) m / z: 422.2 [M + H] ⁺ . ¹ H NMR (400MHz, CDCl ₃ ) δ9.11 (br, 1H), 8.42–8.22 (m, 3H), 7.20 (d, J = 9.3 Hz, 1H), 6.39–6.25 (d, J = 7.6 Hz, 1H), 5.56 (t, J = 7.2 Hz, 1H), 4.18 (t, J = 8.0 Hz, 1H), 3.98 (s, 1H), 3.86 (d, J = 24.8 Hz, 2H), 2.80 (d, J = 16.3 Hz, 1H), 2.53 (m, 1H), 2.42 (m, 1H), 2.30 (m, 1H), 2.22 (m, 1H ), 1.88 (d, J = 11.6 Hz, 1H), 1.57-1.42 (d, J = 7.6 Hz, 3H), 1.02 (m, 1H), 0.93-0.85 (m, 1H), 0.44 (m, 2H) .

Example 4

(R, 1 ³ E, 1 ⁴ E) -3 ⁵ -fluoro-6-methyl-6,7-diazalide-1 (5,3) -pyrazolo [1,5-a] pyrimidine- Synthesis of 3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 1: Ethyl (S) -5- (2- (5-fluoro-2- (2- (methylamino) ethyl) pyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1 Synthesis of 5,5-a] pyrimidine-3-formate

At room temperature, add ethyl (S, E) -5- (2- (5-fluoro-2- (prop-1-en-1-yl) pyridin-3-yl) pyrrolidine- 1-yl) pyrazolo [1,5-a] pyrimidine-3-carboxylic acid ester (1.00 g, 2.62 mmol), 30% methylamine ethanol solution (30 mL) and glacial acetic acid (10 mL), vacuumed and replaced The temperature was slowly raised to 75 ° C. under nitrogen, and the reaction was refluxed for 19 hours to stop the reaction. The reaction solution was cooled to room temperature, EA was added, and a saturated aqueous solution of sodium carbonate was added. The aqueous phase was back-extracted with EA. The organic phases were combined, dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and column chromatography (DCM / EtOH system) to obtain a yellow solid. 544.3mg, yield 45.74%. MS (ESI) m / z: 413.3 [M + H] ⁺ .

Step 2: Ethyl (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-methylhydrazino) ethyl) -5-fluoropyridin-3-yl) Synthesis of pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylate

At room temperature, add ethyl (S) -5- (2- (5-fluoro-2- (2- (methylamino) ethyl) pyridin-3-yl) pyrrolidine-1- ) Pyrazolo [1,5-a] pyrimidine-3-carboxylic acid ester (439.0 mg, 1.04 mmol), tetrahydrofuran (9.00 mL) and N-methylmorpholine (158.0 mg, 1.56 mmol), cooled to 0 ℃, N-Boc-O p-toluenesulfonyl-hydroxylamine (898.3mg, 3.12mmol) was added in batches. After the addition was completed, the reaction was returned to room temperature for 19 hours. The reaction was stopped, EA was added, and a saturated sodium carbonate aqueous solution was washed. The phases were extracted with EA, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , evaporated to dryness under reduced pressure, and column chromatography (PE / EA system) to obtain 68.2 mg of product. MS (ESI) m / z: 528.3 [M + H] ⁺ .

Step 3: (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-methylhydrazino) ethyl) -5-fluoropyridin-3-yl) pyrrolidine Synthesis of 1-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid

At room temperature, add ethyl (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-methylhydrazyl) ethyl) -5 to a 25 mL single-necked flask -Fluoropyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid ester (62.8mg, 0.13mmol) and ethanol (2.00mL), after all dissolved Then, an aqueous solution (0.5 mL) of lithium hydroxide monohydrate (32.5 mg, 0.77 mmol) was added, the temperature was gradually raised to 70 ° C., and the reaction was stirred for 6 hours, and the reaction was completed. The reaction solution was cooled to room temperature and evaporated to dryness under reduced pressure to obtain crude product 1 as a yellow solid. MS (ESI) m / z: 500.1 [M + H] ⁺ .

Step 4: (R, 1 ³ E, 1 ⁴ E) -3 ⁵ -Fluoro-6-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a Synthesis of] pyrimidine-3 (3,2) -azabenzene-2 (1,2) -pyrrolidine cyclooctane-8-one

At room temperature, the above crude product 1 was added to a 25 mL single-necked flask, and then dichloromethane (2.00 mL) and trifluoroacetic acid (2.00 mL) were added, and the solution was sonicated. The mixture was stirred for 2 hours at room temperature, and the reaction was stopped. The reaction system was evaporated to dryness under reduced pressure to obtain a slightly yellow crude product 2. MS (ESI) m / z: 400.5 [M + H] ⁺ .

At room temperature, put the above crude product 2 into a 25 mL single-necked flask, and then add HOBt (87.3 mg, 0.64 mmol), EDCI (123.9 mg, 0.64 mmol), DMF (3.00 mL) and TEA (261.6 mg, 2.58 mmol) Evacuate, replace with nitrogen, and react overnight at room temperature. The reaction is complete. The reaction system was evaporated to dryness under reduced pressure, and column chromatography (0.1% formic acid aqueous solution / acetonitrile system) yielded 16.5 mg of the product. MS (ESI) m / z: 382.4 [M + H] ^{+ .1} H NMR (300MHz, Chloroform-d) δ8.35 (d, J = 9.4, 1H), 8.34 (d, J = 2.8, 1H), 8.31 (s, 1H), 7.10 (dd, J = 9.4, 2.8 Hz, 1H), 6.34 (d, J = 7.6 Hz, 1H), 5.54 (t, J = 7.5 Hz, 1H), 4.12 (dd, J = 15.0, 6.2 Hz, 1H), 3.99 (dd, J = 7.9, 7.2 Hz, 1H), 3.86 (dd, J = 8.7, 7.8 Hz, 1H), 3.72–3.69 (m, 1H), 3.67–3.65 ( m, 1H), 2.87--2.79 (m, 1H), 2.77 (s, 3H), 2.66--2.56 (m, 1H), 2.47--2.41 (m, 1H), 2.30--2.23 (m, 1H), 1.97-- 1.90 (m, 1H).

Example 5

(R, 1 ³ E, 1 ⁴ E) -6-ethyl-3 ⁵ -fluoro-6,7-diazalide-1 (5,3) -pyrazolo [1,5-a] pyrimidine- 3 (3,2) -Azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 1: Ethyl (S) -5- (2- (2- (2- (ethylamino) ethyl) -5-fluoropyridin-3-yl) pyrrolidin-1-yl) pyrazolo [ Synthesis of 1,5-a] pyrimidine-3-formate

At room temperature, add ethyl (S) -5- (2- (5-fluoro-2-vinylpyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5 to a 100 mL single-necked bottle. -a] pyrimidine-3-formate (1.52g, 3.98mmol), 30% ethylamine ethanol solution (40.80mL) and glacial acetic acid (13.60mL), evacuate, replace nitrogen, slowly warm to 75 ° C, reflux reaction. Stir for 23.5 hours to stop the reaction. The reaction solution was cooled to room temperature, EA was added, and a saturated sodium carbonate aqueous solution was added for washing. The aqueous phase was back-extracted with EA. The organic phases were combined, dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and column chromatography (DCM / EtOH system). 610 mg of solid with a yield of 33.66%. MS (ESI) m / z: 427.5 [M + H] ⁺ .

Step 2: Ethyl (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-ethylhydrazino) ethyl) -5-fluoropyridin-3-yl ) Synthesis of pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylate

At room temperature, add ethyl (S) -5- (2- (2- (2- (ethylamino) ethyl) -5-fluoropyridin-3-yl) pyrrolidine-1 to a 50 mL single-necked flask. -Yl) pyrazolo [1,5-a] pyrimidine-3-carboxylate (753.7 mg, 1.77 mmol), tetrahydrofuran (15 mL) and N-methylmorpholine (178.8 mg, 1.77 mmol), cooled to 0 ℃, N-Boc-O p-toluenesulfonyl-hydroxylamine (2.54g, 8.84mmol) was added in portions. After the addition was completed, the reaction was returned to room temperature for 17 hours. The reaction was stopped, EA was added, and a saturated sodium carbonate aqueous solution was washed. The phases were extracted again with EA, the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , evaporated to dryness under reduced pressure, and column chromatography (PE / EA system) to obtain 101.6 mg of product. MS (ESI) m / z: 542.6 [M + H] ⁺ .

Step 3: (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-ethylhydrazino) ethyl) -5-fluoropyridin-3-yl) pyrrolidine Synthesis of 1-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid

At room temperature, add ethyl (S) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-ethylhydrazyl) ethyl) -5 to a 25 mL single-necked flask -Fluoropyridin-3-yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid ester (101.6 mg, 0.19 mmol) and ethanol (4.00 mL) until all are dissolved Then, an aqueous solution (0.8 mL) of lithium hydroxide monohydrate (47.2 mg, 1.13 mmol) was added, and the temperature was gradually raised to 70 ° C, followed by stirring for 5 hours, and the reaction was completed. The reaction system was cooled to room temperature and evaporated to dryness under reduced pressure to obtain crude product 1 as a yellow solid. MS (ESI) m / z: 514.5 [M + H] ⁺ .

Step 4: (R, 1 ³ E, 1 ⁴ E) -3 ⁵ -Fluoro-6-ethyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a Synthesis of] pyrimidine-3 (3,2) -azabenzene-2 (1,2) -pyrrolidine cyclooctane-8-one

At room temperature, the above crude product 1 was added to a 25 mL single-necked flask, and then dichloromethane (3.00 mL) and trifluoroacetic acid (3.00 mL) were added, and the solution was sonicated, and stirred at room temperature for 1.5 hours. The reaction system was evaporated to dryness under reduced pressure to obtain a slightly yellow crude product 2. MS (ESI) m / z: 414.4 [M + H] ⁺ .

At room temperature, put the crude product 2 into a 25 mL single-necked flask, and add HOBt (126.7 mg, 0.94 mmol), EDCI (180.0 mg, 0.94 mmol), DMF (6.00 mL), and TEA (380.0 mg, 3.75 mmol). Evacuate, replace with nitrogen, and react overnight at room temperature. The reaction is complete. The reaction system was evaporated to dryness under reduced pressure, and column chromatography (0.1% formic acid aqueous solution / acetonitrile system) yielded 14.4 mg of the product. MS (ESI) m / z: 396.4 [M + H] ⁺ .HPLC: 97.87%. ¹ H NMR (300 MHz, Chloroform-d) δ 9.42 (br, 1H), 8.41–8.31 (m, 3H), 7.10 (dd, J = 8.4, 2.8 Hz, 1H), 6.36 (d, J = 7.9 Hz, 1H), 5.57 (t, J = 7.5 Hz, 1H), 4.04-3.95 (m, 2H), 3.92-3.83 ( m, 2H), 3.78--3.70 (m, 1H), 3.63 (d, J = 17.8Hz, 1H), 2.98 (q, J = 8.4Hz, 2H), 2.66-2.57 (m, 1H), 2.29-2.18 (m, 2H), 1.26--1.20 (t, J = 6.0 Hz, 3H).

Example 6

(R, 1 ³ E, 1 ⁴ E) -3 ⁵ -fluoro-6-isopropyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine -3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

(R, 1 ³ E, 1 ⁴ E) -3 ⁵ -fluoro-6-isopropyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine The method for preparing -3 (3,2) -pyridine-2 (1,2) -pyrrolidinecycloocta-8-one is similar to that in Example 1.

¹ H NMR (400MHz, CDCl ₃ ) δ 9.41 (br, 1H), 8.33 (d, J = 8.0 Hz, 1H), 8.32 (s, 2H), 7.09 (dd, J = 9.4, 2.3 Hz, 1H) , 6.34 (d, J = 7.9 Hz, 1H), 5.56 (t, J = 7.3 Hz, 1H), 4.20-4.14 (m, 1H), 4.00 (q, J = 8.0 Hz, 1H), 3.88-3.82 ( m, 1H), 3.65 (dd, J = 15.4, 4.0 Hz, 1H), 3.56 (d, J = 16.2 Hz, 1H), 3.09 (p, J = 6.2 Hz, 1H), 2.86 (dd, J = 15.1 , 10.1Hz, 1H), 2.61 (dd, J = 13.4, 6.8Hz, 1H), 2.44 (dt, J = 12.7, 6.3Hz, 1H), 2.25 (dt, J = 13.7, 7.3Hz, 1H), 1.94 (dd, J = 13.5, 6.8 Hz, 1H), 1.29 (d, J = 6.3 Hz, 3H), 1.22 (d, J = 6.3 Hz, 3H). MS (ESI) m / z: 410.2 [M + H] ⁺ .

Example 7

(1 ³ E, 1 ⁴ E, 2 ² R, 5R) -3 ⁵ -fluoro-5,6-dimethyl-6,7-diazalide-1 (5,3) -pyrazolo [1 , 5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one and (1 ³ E, 1 ⁴ E, 2 ² R, 5S) -3 ⁵ -fluoro-5,6-dimethyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine-3 (3,2) -pyridine-2 (1,2) -Pyrrolidine cyclooctane-8-one

Step 1: Synthesis of (1 ³ E, 1 ⁴ E, 2 ² R) -5,6-dimethyl-3 ⁵ -fluoro-6,7-diazepine-1 (5,3 ) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 2: (1 ³ E, 1 ⁴ E, 2 ² R, 5R) -3 ⁵ -Fluoro-5,6-dimethyl-6,7-diazepine-1 (5,3) -pyrazole And [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one and (1 ³ E, 1 ⁴ E, 2 ² R, 5S ) -3 ⁵ -Fluoro-5,6-dimethyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine-3 (3,2)- Preparation of pyridin-2 (1,2) -pyrrolidinecyclooctane-8-one

Take (1 ³ E, 1 ⁴ E, 2 ² R) -5,6-dimethyl-3 ⁵ -fluoro-6,7-diazalide-1 (5,3) -pyrazolo [1, 5-a] Pyrimidine-3 (3,2) -azabenzene-2 (1,2) -pyrrolidinecyclooctane-8-one (260 mg, 0.65 mmol) was prepared by column separation to obtain Example 7a: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-5,6-dimethyl-6,7-diazalide-1 (5,3) -pyrazolo [1 , 5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one (23 mg, yield 17.7%) and Example 7b: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-5,6-dimethyl-6,7-diazalide-1 (5,3) -pyrazolo [1,5- a] Pyrimidine-3 (3,2) -pyridine-2 (1,2) -pyrrolidine cyclooctane-8-one (56 mg, yield 43.1%).

Separation preparation method:

Instrument: Qingbohua LC2000 High Performance Liquid Chromatography

Column: YMC Triart C18 250 * 20mm 10um

Mobile phase: A: acetonitrile; mobile phase B: 0.1% trifluoroacetic acid aqueous solution

Wavelength: 230nm

Column temperature: 35 ° C

Flow rate: 10ml / min

Solvent: acetonitrile

Preparation conditions: 0min A25B75 25min A25B75 30min A26B74 35min A28B72

Collection conditions: concentrated under reduced pressure at 40 ° C

Separation detection method:

Instrument: Agilent 1260 Infinity II

Column: Xtimate C18 4.6 * 50mm 3um

Mobile phase: A is pure acetonitrile; B is 0.1% trifluoroacetic acid water

Wavelength: 230nm

Column temperature: 35 ° C

Flow rate: 1mL / min

Injection volume: 20 μL

Retention time: 2.371min, 2.844min

Solvent: acetonitrile

Test conditions: B 10% -76%

Example 7a: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-5,6-dimethyl-6,7-diazepine-1 (5,3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one, retention time 2.371 min. MS (ESI) m / z: 396.2 [M + H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.55 (br, 1H), 8.82 (d, J = 7.7Hz, 1H), 8.37 ( m, 1H), 8.11 (s, 1H), 7.54 (d, J = 9.8Hz, 1H), 6.72 (d, J = 7.8Hz, 1H), 5.40 (t, J = 6.8Hz, 1H), 4.20 ( m, 1H), 4.08 (q, J = 7.5 Hz, 1H), 3.84 (m, 1H), 3.53-3.42 (m, 1H), 3.23 (dd, J = 14.5, 10.6 Hz, 1H), 3.04 (s , 3H), 2.62 (dt, J = 12.6, 6.5 Hz, 1H), 2.24 (dt, J = 11.7, 5.9 Hz, 1H), 2.08 (dq, J = 12.9, 7.4 Hz, 1H), 1.83-1.72 ( m, 1H), 1.31 (d, J = 6.9Hz, 3H).

Example 7b: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-5,6-dimethyl-6,7-diazepine-1 (5,3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one, retention time 2.844min. MS (ESI) m / z: 396.2 [M + H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.64 (br, 1H), 8.80 (d, J = 7.7Hz, 1H), 8.42 ( d, J = 2.6 Hz, 1H), 8.13 (s, 1H), 7.59 (d, J = 12.6 Hz, 1H), 6.69 (d, J = 7.8 Hz, 1H), 5.61 (t, J = 7.1 Hz, 1H), 4.14 (d, J = 7.2Hz, 1H), 3.96 (s, 1H), 3.91 (d, J = 5.0Hz, 1H), 3.88--3.75 (m, 2H), 2.65 (s, 3H), 2.59 (s, 1H), 2.31 (dq, J = 11.7, 5.7, 5.1 Hz, 1H), 2.14–1.99 (m, 1H), 1.80 (dq, J = 13.5, 7.1 Hz, 1H), 1.46 (d, J = 6.9Hz, 3H).

Example 8

(1 ³ E, 1 ⁴ E, 2 ² R, 5R) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one and (1 ³ E, 1 ⁴ E, 2 ² R, 5R) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine-3 (3,2) -Pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 1: Synthesize (1 ³ E, 1 ⁴ E, 2 ² R) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5 , 3) -pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

Step 2: (1 ³ E, 1 ⁴ E, 2 ² R, 5R) -3 ⁵ -Fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5,3)- Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one and (1 ³ E, 1 ⁴ E, 2 ² R , 5S) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine-3 (3 Preparation of 2,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

Take (1 ³ E, 1 ⁴ E, 2 ² R) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazalide-1 (5,3) -pyrazolo [ 1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidine cyclooctane-8-one (320 mg, 0.78 mmol) was prepared by column separation to obtain Example 8a: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctadecane-8-one (26 mg, yield 16.2%) and Example 8b: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazalide-1 (5,3) -pyrazolo [ 1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidine cyclooctane-8-one (53mg, yield

33.1%).

Separation preparation method:

Instrument: Qingbohua LC2000 High Performance Liquid Chromatography

Column: YMC Triart C18 250 * 20mm 10um

Mobile phase: A: acetonitrile; mobile phase B: 0.05% trifluoroacetic acid aqueous solution

Wavelength: 230nm

Column temperature: 35 ° C

Flow rate: 10ml / min

Solvent: acetonitrile

Preparation conditions: 0min A33B67 25min A33B67 30min A35B65

Collection conditions: concentrated under reduced pressure at 40 ° C

Separation detection method:

Instrument: Agilent 1260 Infinity II

Column: Xtimate C18 4.6 * 50mm 3um

Mobile phase: A is pure acetonitrile; B is 0.1% trifluoroacetic acid water

Wavelength: 230nm

Column temperature: 35 ° C

Flow rate: 1mL / min

Injection volume: 20 μL

Retention time: 1.898min, 2.369min

Solvent: acetonitrile

Test conditions: B 10% -72%

Example 8a: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5, 3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one, with a retention time of 1.898min. MS (ESI) m / z: 410.1 [M + H] ^{+ .1} H NMR (400MHz, DMSO-d ₆ ) δ9.55 (br, 1H), 8.82 (d, J = 7.7Hz, 1H), 8.37 ( m, 1H), 8.11 (s, 1H), 7.54 (d, J = 9.8Hz, 1H), 6.72 (d, J = 7.8Hz, 1H), 5.40 (t, J = 6.8Hz, 1H), 4.20 ( m, 1H), 4.08 (q, J = 7.5 Hz, 1H), 3.84 (m, 1H), 3.53-3.42 (m, 1H), 3.23 (dd, J = 14.5, 10.6 Hz, 1H), 3.04 (s , 3H), 2.62 (dt, J = 12.6, 6.5 Hz, 1H), 2.24 (dt, J = 11.7, 5.9 Hz, 1H), 2.08 (dq, J = 12.9, 7.4 Hz, 1H), 1.83-1.72 ( m, 1H), 1.04 (m, 6H).

Example 8b: (1 ³ E, 1 ⁴ E, 2 ² R, 5R or S) -3 ⁵ -fluoro-6-ethyl-5-methyl-6,7-diazepine-1 (5, 3) -Pyrazolo [1,5-a] pyrimidin-3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one, retention time: 2.369min. MS (ESI) m / z: 410.1 [M + H] ^{+ .1} H NMR (400MHz, DMSO-d ₆ ) δ9.02 (br, 1H), 8.76 (d, J = 7.7Hz, 1H), 8.39 ( d, J = 2.5 Hz, 1H), 8.06 (s, 1H), 7.56 (dd, J = 10.1, 2.4 Hz, 1H), 6.66 (d, J = 7.8 Hz, 1H), 5.59 (s, 1H), 4.22--3.94 (m, 1H), 3.90--3.73 (m, 2H), 3.68--3.59 (m, 1H), 2.90--2.74 (m, 1H), 2.67 (p, J = 8.3, 7.5Hz, 1H), 2.53 (m, 1H), 2.48–2.40 (m, 1H), 2.30 (dq, J = 11, 6.6Hz, 1H), 2.07 (dt, J = 13.2, 7.2Hz, 1H), 1.77 (td, J = 12.9, 6.3Hz, 1H), 1.44 (d, J = 6.9Hz, 3H), 0.98 (t, J = 6.9Hz, 3H).

Example 9

(R, 1 ³ E, 1 ⁴ E) -3 ⁵ -fluoro-6-isobutyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine -3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclooctane-8-one

(R, 1 ³ E, 1 ⁴ E) -3 ⁵ -fluoro-6-isobutyl-6,7-diazepine-1 (5,3) -pyrazolo [1,5-a] pyrimidine The method for preparing -3 (3,2) -pyridine-2 (1,2) -pyrrolidinecycloocta-8-one is similar to that in Example 1.

¹ H NMR (400MHz, CDCl ₃ ) δ 9.29 (br, 1H), 8.30 (dd, J = 9.5, 7.2 Hz, 3H), 7.07 (dd, J = 9.4, 2.8 Hz, 1H), 6.32 (d, J = 7.7Hz, 1H), 5.57-5.53 (t, J = 7.3Hz, 1H), 3.99-3.91 (m, 2H), 3.87-3.80 (m, 2H), 3.63 (dt, J = 16.0, 3.5Hz , 1H), 2.86-2.76 (m, 2H), 2.65 (dt, J = 13.3, 6.8Hz, 1H), 2.54 (dd, J = 11, 8.4Hz, 1H), 2.43 (dt, J = 12.6, 6.2 Hz, 1H), 2.25 (dt, J = 13.5, 7.5 Hz, 1H), 1.97-1.88 (dt, J = 14.2, 7.2 Hz, 1H), 1.82 (dt, J = 13.6, 6.6 Hz, 1H), 1.12 (d, J = 6.5Hz, 3H), 0.96 (d, J = 6.7Hz, 3H) .MS (ESI) m / z: 424.2 [M + H] ⁺ .

Example 10

(R, 1 ³ E, 1 ⁴ E) -6-cyclopropyl-3 ⁵ -fluoro-6,7-diaza-1 (5,3) -pyrazoline [1,5-a] pyrimidine- Synthesis of 3 (3,2) -pyridine-2 (1,2) -pyrrolidinecyclotridecane-8-one

Step 1: 2- (2-bromo-4-fluorophenyl) ethanol

Dissolve 2- (2-bromo-4-fluorophenyl) acetic acid (46.63g, 200.00mmol) in 500mL of THF, add borane tetrahydrofuran solution (1.0M, 300mL) at 0 ° C, and terminate after 18h at 16 ° C . The reaction solution was quenched with 1N dilute hydrochloric acid, extracted with EA, and the organic phase was washed with saturated brine, dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to give 42.57 g of a yellow oil with a yield of 97%.

Step 2: 2-bromo-4-fluoro-1- (2- (4-methoxybenzyloxy) ethyl) benzene

At room temperature, add NaH (4.10 g, 102.00 mmol) and 200 mL of THF to a four-necked flask, and add 2- (2-bromo-4-fluorophenyl) ethanol (14.90 g, 102.00 mmol) at 0 ° C at 0 ° C. The reaction was continued for 0.5 h. Tetrabutylammonium iodide (36.28 g, 96.00 mmol) and p-methoxybenzyl chloride (12.78 g, 82.00 mmol) were added. The reaction was terminated at 50 ° C for 4 h. The reaction solution was quenched with 1N dilute hydrochloric acid, extracted with EA, and the organic phase was washed with saturated brine, dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to give 15.37 g of a colorless oil with a yield of 67%.

Step 3: 5-fluoro-2- (2- (4-methoxybenzyloxy) ethyl) benzaldehyde

At room temperature, 2-bromo-4-fluoro-1- (2- (4-methoxybenzyloxy) ethyl) benzene (15.37 g, 45.50 mmol) was dissolved in THF (500 mL) at -78 ° C. Under the protection of N ₂ , sec-butyllithium (70 mL) was added, stirred at -78 ° C for 1 h, and then DMF (18.17 g, 230.00 mmol) was added. The temperature was controlled at -78 ° C and the reaction was terminated for 0.5 h. The reaction was quenched with a saturated ammonium chloride solution. EA was added for extraction, washed with water, dried over anhydrous MgSO ₄ , filtered, concentrated, and purified by column chromatography (EA / PE system) to obtain 8.01 g of a yellow oil with a yield of 62%.

Step 4: (R, E) -N- (5-fluoro-2- (4-methoxybenzyloxy) ethyl) benzylidene) -2-tert-butyl-2-sulfonimide

At room temperature, 5-fluoro-2- (2- (4-methoxybenzyloxy) ethyl) benzaldehyde (8.00 g, 28.00 mmol), R-tert-butylsulfinamide ( 3.56 g, 30.00 mmol), Cs ₂ CO ₃ (6.33 g, 19.00 mmol) and DCM 100 mL, protected by N ₂ , terminated after 16 h at 25 ° C. The reaction solution was poured into water, and the layers were separated. The organic phase was washed with saturated brine, dried over anhydrous MgSO ₄ , filtered, and concentrated to obtain 10.88 g of a yellow oil with a yield of 100%.

Step 5: N-((R) -3-((1,3-dioxan-2-yl) -1- (5-fluoro-2- (2- (4-methoxybenzyloxy) ethyl ) Phenyl) propyl) -2-tert-butyl-2-sulfonylimide

At room temperature, Mg powder (0.87g, 36.00mmol), 1-bromo-3,3-dimethoxypropane (7.10g, 36.00mmol), and THF (100mL) were added to the four-necked flask in this order. ₂ , N ₂ protection, spot heating to initiate the reaction, reaction at 25 ° C for 2 h. (R, E) -N- (5-fluoro-2- (4-methoxybenzyloxy) ethyl) methylene) -2-tert-butyl-2-sulfonimide was added dropwise under an ice bath (10.88g, 28.00mmol) in THF (40mL). The reaction was terminated after rising to 25 ° C after dropping. The reaction was quenched by adding saturated NH ₄ Cl solution under ice bath. The layers were separated. The aqueous phase was extracted with EA and the organic phases were combined. , Dried over anhydrous MgSO ₄ , filtered, and concentrated to obtain 14.20 g of white solid in 100% yield.

Step 6: (R) -2- (2- (3,4-Dihydro-2H-pyrrole-2-yl) -4-fluoro) phenylethanol

At room temperature, (S) -N-((R) -3-((1,3-dioxan-2-yl) -1- (5-fluoro-2- (2- ( 4-methoxybenzyloxy) ethyl) phenyl) propyl) -2-tert-butyl-2-sulfonimide (14.20 g, 28.00 mmol) and H ₂ O (25 mL), dripped on ice TFA (100 mL) was added, and the reaction was stirred for 20 min after dripping, and then raised to 25 ° C. for 22 h to terminate the reaction. The reaction solution was directly used in the next step.

Step 7: (R) -2- (4-fluoro-2- (pyrrolidin-2-yl) phenylethanol

At room temperature, add (R) -2- (2- (2,3,4-dihydro-2H-pyrrole-2-yl) -4-fluoro) phenylethanol (previous reaction solution) to the single-necked flask in sequence, ice salt Triethylsilane (9.77g, 84.00mmol) was added in batches in the bath, and the temperature was raised to 17 ° C for 1 hour. The reaction was terminated. The reaction was quenched by adding 2N HCl aqueous solution, adjusting the pH of the system to 2-3, washing with EA, and 1N NaOH aqueous solution. Adjust the pH to 9-10, extract with DCM, dry with anhydrous Na ₂ SO ₄ , filter, and concentrate to obtain 3.80 g of light yellow oil. The total yield in two steps is 65%.

Step 8: (R) -5- (2- (5-Fluoro-2- (2-hydroxyethyl) phenyl) pyrrole-1-yl) pyrazoline [1,5-a] pyrimidine-3-carboxylic acid Ethyl ester

At room temperature, (R) -2- (4-fluoro-2- (pyrrolidin-2-yl) phenylethanol (3.80 g, 18.00 mmol), 5-chloropyrazolo [1,5 -A] pyrimidine-3-carboxylate (4.10g, 18.00mmol), TEA ( 3.64g, 36.00mmol) and EtOH (50mL), evacuated substituted N _2, N ₂ protection, 24 ℃ reaction was terminated after 16h. rotary Ethanol was removed, DCM and water were added, and the mixture was stirred and separated. The aqueous phase was removed. The organic phase was washed with saturated brine, washed with saturated NaHCO ₃ aqueous solution, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated, and purified by column chromatography (EA / PE. System) to obtain 3.56 g of a yellow foamy solid with a yield of 50%.

Step 9: (R) -5- (2- (5-Fluoro-2- (2-methylsulfonyl) ethyl) phenyl) pyrrole-1-yl) pyrazoline [1,5-a] pyrimidine- Ethyl 3-formate

At room temperature, (R) -5- (2- (5-fluoro-2- (2-hydroxyethyl) phenyl) pyrrole-1-yl) pyrazoline [1,5-a ] Pyrimidine-3-carboxylic acid ethyl ester (2.00 g, 5.02 mmol), TEA (1.52 g, 15.06 mmol) and 50 mL of DCM. Methanesulfonyl chloride (1.16 g, 10.04 mmol) was added under an ice bath, and the reaction temperature was naturally raised to 26 ° C for 2 h. After termination. The reaction solution was poured into water, stirred, extracted, and separated. The organic phase was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to obtain a yellow liquid. The crude product was directly used in the next step with a yield of 100%.

Step 10: (R) -5- (2- (2- (2 (Cyclopropylamino) ethyl) -5-fluorophenyl) pyrrole-1-yl) pyrazoline [1,5-a] pyrimidine Ethyl-3-formate

(R) -5- (2- (5-fluoro-2- (2-methanesulfonyl) ethyl) phenyl) pyrrole-1-yl) pyrazoline [1, 5-a] Pyrimidine-3-carboxylic acid ethyl ester (crude from the previous step, 5.02 mmol), cyclopropylamine (0.86 g, 15.06 mmol), and DMF (30 mL) under N ₂ protection, the reaction was stirred at 80 ° C. for 3 h and terminated. The reaction solution was poured into water, extracted with EA, and the EA phase was washed with 1N dilute hydrochloric acid. The aqueous phase was added with sodium carbonate to adjust pH = 12, added with EA extraction, washed with saturated brine, dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to obtain Light yellow liquid 1.69 g, yield 77%.

Step 11: (R) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5-fluorophenyl) pyrrolidine-1 Synthesis of Ethyl) pyrazolo [1,5-a] pyrimidine-3-carboxylic acid ethyl ester

(R) -5- (2- (2- (2- (2 (cyclopropylamino) ethyl) -5-fluorophenyl) pyrrole-1-yl) pyrazoline [1] , 5-a] Pyrimidine-3-carboxylic acid ethyl ester (1.67 g, 3.82 mmol), N-tert-butoxycarbonyl-3- (4-cyanophenyl) dumazine (1.25 g, 4.97 mmol), and DMF (25 mL) The reaction was stopped at 21 ° C for 16h. EA was added, washed with water, and the aqueous phase was back-extracted with EA. The organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated, and purified by column chromatography (PE / EA system) to obtain 1.02 g of a yellow oily solid with a yield of 68%. .

Step 12: (R) -5- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5-fluorophenyl) pyrrolidine-1 -Yl) Synthesis of pyrazolo [1,5-a] pyrimidin-3-carboxylic acid

(R) -5- (2- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5-fluorobenzene (Yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid ethyl ester (0.70 g, 1.34 mmol) and ethanol (10 mL), stirred until completely dissolved, then added sodium hydroxide ( 0.32 g, 8.04 mmol) of H ₂ O (5 mL) solution, heated to 70 ° C. for 16 h and terminated, cooled to room temperature, concentrated to remove most ethanol, added DCM and H ₂ O, 1N HCl to adjust the pH to 3-4, The layers were stirred and separated, and the aqueous phase was extracted with DCM. The organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to obtain 0.63 g of a white solid with a yield of 95%.

Step 13: (R) -5- (2- (2- (2- (2- (cyclopropylhydrazinyl) ethyl) -5-fluorophenyl) pyrrolidin-1-yl) pyrazolo [1, 5-a] pyrimidine-3-carboxylic acid

(R) -5- (2- (2- (2- (2- (2- (tert-butylcarbonyl) -1-cyclopropylhydrazino) ethyl) -5-fluorobenzene Yl) pyrrolidin-1-yl) pyrazolo [1,5-a] pyrimidin-3-carboxylic acid (0.63 g, 1.20 mmol) and HCl in ethanol (10 mL), stirred until completely dissolved, and reacted at 32 ° C for 1 h Then it was terminated and concentrated to dryness under reduced pressure to obtain 0.60 g of a slightly yellow solid with a yield of 100%.

Step 14: (R, ¹³ E, ¹⁴ E) -6-cyclopropyl-3 ⁵ -fluoro-6,7-diaza-1 (5,3) -pyrazoline [1,5-a] pyrimidine -3 (3,2) -phenyl-2 (1,2) -pyrrolidinecyclotridecane-8-one

At room temperature, four-necked flask were successively added TBTU (0.68g, 2.12mmol), DMAP (0.034g, 0.30mmol), DMF (6mL) and DCM (30mL), evacuated and replaced with N _2, N ₂ protection, DIEA (1.09 g, 8.46 mmol) was added. (R) -5- (2- (2- (2- (1- (Cyclopropylhydrazinyl) ethyl) -5-fluorophenyl) pyrrolidin-1-yl) pyrazolo [1,5- a] Pyrimidine-3-carboxylic acid (0.60g, 1.41mmol) was dissolved in a mixed solution of DMF (6mL) and DCM (6mL) and divided into five equal parts, and an equal portion of the above solution was added after every 1h at 32 ° C. After completion of the reaction, the reaction was terminated at 32 ° C. for 1 h, and then concentrated and dried under reduced pressure, and purified by column chromatography (DCM / CH ₃ OH system) to obtain 0.22 g of an off-white solid with a yield of 42%. ¹ H NMR (400MHz, Chloroform-d) δ 9.85 (br, 1H), 8.30 (d, J = 9.3 Hz, 2H), 7.20 (m, 1H), 6.90 (t, J = 8.0 Hz, 1H), 6.78 (d, J = 10.2 Hz, 1H), 6.33 (d, J = 7.7 Hz, 1H), 5.67 (t, J = 6.6 Hz, 1H), 3.99 (dt, J = 14.0, 7.9 Hz, 2H), 3.90--3.72 (m, 1H), 3.65--3.50 (m, 2H), 2.94--2.88 (m, 1H), 2.67--2.50 (m, 2H), 2.50--2.33 (m, 1H), 2.21 (dt, J = 13.3, 7.0 Hz, 1H), 1.93 (dt, J = 12.7, 6.5 Hz, 1H), 1.14 (m, 1H), 0.90 (d, J = 10.2 Hz, 1H), 0.54 (m, 2H) .MS (ESI) m / z: 407 [M + H] ⁺ .

Example 11

Trk kinase inhibitory activity test

This test uses of TrkB, inhibition of isotope γ- ³³ p-ATP assays test compound kinase TrkA, TrkC, and the obtained compound of the half inhibitory concentration of inhibitory activity IC _50, the reported Trk inhibitors LOXO-101 As a positive control, LOXO-101 was purchased from Shanghai Sinok Medical Technology Co., Ltd., batch number: SCT0142170801.

Basal reaction buffer

20mM Hepes (pH 7.5), 10mM MgCl ₂ , 1mM EGTA, 0.02% Brij35, 0.02mg / ml BSA, 0.1mM Na3VO4, 2mM DTT, 1% DMSO.

2. Compound formulation

The compound was dissolved to a specific concentration with 100% DMSO, and then it was gradient diluted to a different concentration of the test sample (DMSO solution) using an automatic sampling device.

3. Reaction steps

3.1 Dilute the reaction substrate with the basic reaction buffer;

3.2 Add the kinase to the substrate solution and mix gently;

3.3 Use an automatic sample loading system to add 100% DMSO-diluted compounds of different concentrations to the kinase solution and incubate for 20 min at room temperature;

3.4 Add ³³ P-ATP (10 μM, 10 μCi / μl) at room temperature to start the kinase reaction, and react for 2 h.

4. Detection

The reaction solution was subjected to ion exchange filtration system to remove unreacted ATP and ADP plasma generated by the reaction, and then measured the ³³ P isotope radiation in the substrate.

5. Data processing

The inhibitory effect of different concentrations of compounds on kinase activity was calculated based on the amount of kinase added to the inhibitor system at different concentrations, and the IC ₅₀ was inhibited by graphpad prism fitting.

The biochemical activity of the compounds of the present invention was determined through the above tests. The measured IC ₅₀ values are shown in Table 1:

Table 1 Trk kinase inhibitory activity test results

Note: "-" in the table means not tested

Conclusion: The compound of the present invention has better Trk kinase inhibitory activity than the positive drug.

Example 12

tel-NTRK1-BaF3, BaF3-LMNA-NTRK1 cell growth inhibition test

In this test, CellTiter-Glo cell proliferation fluorescence detection method was used to test the compound's inhibitory effect on the growth of tel-NTRK1-BaF3 and BaF3-LMNA-NTRK1 transgenic cells, and the half-growth inhibitory concentration GI _{50 of the} compound on the cell was obtained. The Trk inhibitor LOXO-101 was used as a positive control, and LOXO-101 was purchased from Haoyuan Chemexpress Co., Ltd ..

1. Experimental equipment and supplies

I) PreceDo target equivalent gene stable cell line library;

II) CellTiter-Glo cell proliferation fluorescence detection reagent (Promega, USA);

III) 96-well special plate for drug screening (Corning, Rochester, NY);

IV) Test compounds.

2. Preparation of compound medicine board

The test compound was dissolved in DMSO to prepare a 10 mM stock solution, and prepared at a 3-fold dilution to 10 mM, 3.333 mM, 1.111 mM, 0.370 mM, 0.123 mM, 0.041 mM, 0.014 mM, 0.005 mM, 0.002 mM and stored in 0.5 ml for sterilization. In a dorf tube (Corning, USA), the same volume of DMSO solvent was used as a blank control at the same time, a total of 10 concentration gradients, and the drug plate was stored under vacuum at -20 ° C.

3. Cell culture conditions

tel-NTRK1-BaF _3, BaF3-LMNA-NTRK1 cell line was cultured with RPMI 1640 (Corning, NY, USA) + 10% fetal bovine serum (Gibico, Invitrogen, USA). Cells were cultured for two generations after resuscitation and tested.

4. Testing and data processing

Take logarithmic growth phase cells (2000-2500 cells / well) and inoculate 12 × 8 96-well white opaque cell culture plates (Corning 3570, NY, USA) with a volume of 100 μL per well, and add drug to the cell plate (0.1 μL) / Well), the final concentration of the compound is 10 μM, 3.3 μM, 1.1 μM, 0.37 μM, 0.12 μM, 0.04 μM, 0.014 μM, 0.005 μM, 0.002 μM (0.4 uL of the drug solution is added to the 400 μl cell mixture, and then Add 100ul per well), incubate for 72 hours at 37 ° C, 5% CO ₂ incubator, add 20 μL CellTiter-Glo cell proliferation fluorescence detection reagent, let stand for 10 minutes, and read with Envision Plate-Reader.

5. Experimental verification

There were vehicle groups (only DMSO added) in the drug plate as negative controls.

6. Results

The Envision Plate-reader reads the corresponding fluorescence RLU per well. The raw data of the test compound RLU _Drug and the DMSO control group RLU _DMSO are normalized:

Cell Viability% = (RLU _Drug / RLU _DMSO ) * 100%

Graph Pad Prism version 6.0 was used to perform a non-linear regression curve fitting on the cell inhibition rate at a single concentration of the test compound to obtain the GI ₅₀ value.

7. Basic reaction buffer

20mM Hepes (pH 7.5), 10mM MgCl ₂ , 1mM EGTA, 0.02% Brij35, 0.02mg / ml BSA, 0.1 mM Na3VO4, 2mM DTT, 1% DMSO.

The biochemical activity of the compounds of the present invention was determined through the above tests. The measured GI ₅₀ values are shown in Table 2:

Table 2 Trk fusion mutant cell inhibitory activity test results

Note: "-" in the table means not tested.

Conclusion: Compared with the positive drug, the compound of the present invention has better Trk fusion mutant cell growth inhibitory activity.

Example 13

Metabolic stability test of compounds on human liver microsomes

The total volume of the incubation system is 250 μL. 50 mmol / L PBS buffer (pH = 7.4) is used to prepare various liver microsomal incubation solutions with a protein concentration of 0.5 mg / mL. Before the incubation, 100 μmol / L of the test compound 2.5 Mixing μL with 197.5 μL of the above incubation solution, pre-incubating in a 37 ° C water bath for 5 min, and adding 50 μL of the same reduced coenzyme Ⅱ solution (5 mmol / L) which was pre-incubated for 5 min, (the content of liver microsomal protein in the reaction system 0.5g / L, final concentration of the test compound is 1 μmol / L), incubate with shaking in a 37 ° C water bath, and take them out at 0, 5, 15, 30, 60min, and immediately add the internal standard Terfenadine (positive ion internal standard) , 25 ng / mL) and Tolbutamide (negative ion internal standard, 50 ng / mL) positive and negative internal standard mixed methanol solution 600 μL to stop the reaction. The incubation solution was shaken for 2 minutes, centrifuged (4 ° C, 16000 r / min) for 10 minutes, and the supernatant was taken for LC-MS / MS detection to quantitatively analyze the remaining amount of the parent drug. (DMSO <0.1%).

The concentration of the compound at 0 min incubation was taken as 100%, and the concentrations at other incubation time points were converted to the percentage remaining amount. The natural logarithm of the percentage remaining amount at each time point was linearly regression to the incubation time, and the slope k was calculated. According to the formula , T _1/2 = -0.693 / k calculated in vitro half-life. Clearance in liver microsomes (CLint (μL / min / mg protein) = Ln (2) * 1000 / T _1/2 (min) / Protein Conc (mg / ml)).

The test stability data of the compounds of the present invention on human liver microsomes are shown in Table 3:

Table 3 Test results of human liver microsomal metabolic stability

Note: The Trk inhibitor LOXO-195 is prepared with reference to the method of patent WO2011146336.

Conclusion: Compared with the control compound, the compound of the present invention has better metabolic stability of human liver microsomes and has better drugability than the positive control drug.

Claims (14)

1. A compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

   

   R ^{1 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, -COR ⁵ , -SO ₂ R ⁵ or- SOR ^{5 is} optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

   R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further by one or Multiple selected from deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

   R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

   R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   Y is-(CH ₂ ) _n -or -O (CH ₂ ) _n- ;

   n is selected from 0, 1, 2 or 3;

   Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

   R ^{3 is} selected from H, deuterium, halogen, cyano, nitro, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy;

   X is selected from CH or N.
2. The compound of general formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof according to claim 1, which is a compound of general formula (II), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof. The salt:

   

   R ^{1 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, -COR ⁵ , -SO ₂ R ⁵ or- SOR ^{5 is} optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

   R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further by one or Multiple selected from deuterium, halogen, hydroxyl, cyano, nitro, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

   R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

   R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

   R ^{3 is} selected from H, deuterium, halogen, cyano, nitro, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or C ₁ -C ₈ alkoxy;

   X is selected from CH or N.
3. The compound according to claim 2, characterized in that:

   R ^{1 is} selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further selected from deuterium , Halogen, hydroxy, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl , C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

   R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₂ -C ₈ alkenyl or C ₂ -C ₈ alkynyl, optionally further by one or Multiple selected from deuterium, halogen, hydroxyl, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C _3- C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or a 4-10 membered heterocyclic substituent;

   R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

   R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

   R ³ is halogen;

   X is selected from CH or N.
4. The compound according to claim 3, wherein:

   R ^{1 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁷ , -COR ⁷ , -SO ₂ R ⁷ , -SOR ⁷ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy or 4-10 membered heterocyclic ring Substituted by a substituent;

   R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -NR ⁶ R ⁸ , -NR ⁶ COR ⁵ , -COR ⁵ , -SO ₂ R ⁵ , -SOR ⁵ , C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, C ₁ -C ₈ alkoxy, or 4 -10 membered heterocyclic group substituted by a substituent;

   R ⁵ and R ⁷ are each independently selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₈ cycloalkyl, or -NR ⁸ ;

   R ⁶ and R ⁸ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

   R ³ is halogen;

   X is CH or N.
5. The compound according to claim 4, characterized in that:

   R ^{1 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, -SO ₂ R ⁷ , C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl or C ₁ -C ₄ alkoxy substituted by a substituent;

   R ^{7 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, halogen, hydroxyl, cyano, C _1- C ₄ alkyl, C ₃ -C ₆ cycloalkyl or C ₁ -C ₄ alkoxy substituted by a substituent;

   Alternatively, R ¹ , R ² and R ⁴ may independently form a C ₃ -C ₈ cycloalkyl group or a 4-10 membered heterocyclic group;

   R ³ is halogen;

   X is selected from CH or N.
6. The compound according to claim 5, characterized in that:

   R ^{1 is} selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl, optionally further selected from one or more of deuterium, hydroxyl, halogen, -SO ₂ R ⁷ or C ₁ -C ₄ Substituted with alkoxy substituents;

   R ^{7 is} selected from H or C ₁ -C ₈ alkyl;

   R ² and R ⁴ are each independently selected from H, C ₁ -C ₈ alkyl or C ₃ -C ₈ cycloalkyl;

   Alternatively, R ¹ , R ² and R ⁴ may independently form a 4-10 membered heterocyclic group;

   R ^{3 is} selected from fluorine or chlorine;

   X is selected from CH or N.
7. The compound according to claim 6, characterized in that:

   R ^{1 is} selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, optionally further Substituted with one or more substituents selected from deuterium, hydroxyl, F, Cl, -SO ₂ CH ₃ or methoxy;

   R ² and R ⁴ are each independently selected from H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclo Jiji

   Alternatively, R ¹ , R ² and R ⁴ may independently form a morpholinyl group;

   R ^{3 is} selected from fluorine or chlorine;

   X is selected from CH or N.
8. The compound, its stereoisomer or its pharmaceutically acceptable salt is selected from the following structures:

   

   
9. The compound, its stereoisomer or its pharmaceutically acceptable salt is selected from the following structures:

   
10. A pharmaceutical composition comprising the compound according to any one of claims 1 to 9, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
11. The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral solution, inhalant, ointment, suppository or Patch.
12. The compound according to any one of claims 1 to 9, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claims 10 to 11 for the prevention or treatment of tropomyosin Application of Receptor Kinase Activity to Medicines Mediating Diseases.
13. The compound according to any one of claims 1 to 9, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claims 10 to 11 for the preparation or prevention of pain, inflammation, and cancer Application in medicine.
14. The use according to claim 13, characterized in that the cancer is selected from the group consisting of neuroblastoma, ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanin Tumor, prostate cancer, leukemia, lymphoma, non-Hodgkin's lymphoma, gastric cancer, lung cancer, hepatocellular carcinoma, gastrointestinal stromal tumor, thyroid cancer, bile duct cancer, endometrial cancer, kidney cancer, anaplastic large cells Lymphoma, acute myeloid leukemia, multiple myeloma, melanoma or mesothelioma, juvenile sarcoma, fibrosarcoma, large cell neuroendocrine tumor, hairy cell astrocytoma, head and neck squamous cell carcinoma, congenital mesoderm Renal tumor, ductal adenocarcinoma, salivary gland mammary gland secretion carcinoma, and appendic carcinoma.

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