PHARMACEUTICAL COMPOSITION WITH IMPROVED PHARMACOKINETIC PROPERTIES

Abstract

Provided in the present disclosure is a novel pharmaceutical composition, which mainly consists of an alkyl glycoside and an active ingredient as shown in formula I. It is surprisingly found in the present invention that the combined use of the alkyl glycoside can significantly and unexpectedly improve multiple PK characteristics of the active ingredient as shown in formula I, for example, an improvement in Cmax of 10 times or more, or even 52.2 times, an improvement in AUC0-inf of 5.1 times or more, or even 17.1 times, and an improvement in which Tmax is rapidly shortened to 0.17 h or less. On the basis of the excellent and improved PK characteristics, the bioavailability can be significantly enhanced and the blood drug concentration can be rapidly increased, which is expected to further reduce the administration dose and improve the medication safety, and the pharmaceutical composition is particularly suitable for acute attack treatment, alleviation and prevention of migraine and neuropathic pain indications.

Description

Pharmaceutical compositions with improved pharmacokinetic properties Technical Field

[0001] The present invention belongs to the field of medical technology, and particularly relates to a pharmaceutical composition comprising an alkyl glycoside and having improved pharmacokinetic (PK) properties and an application thereof. Background Art

[0002] Migraine is a common neurovascular disease characterized by recurrent, mostly unilateral, moderate to severe throbbing headaches that usually last 4 to 72 hours and are accompanied by symptoms such as nausea, vomiting, and sensitivity to sound and light (phonophobia and photophobia) [1]. Migraine attacks are often preceded by a period of sensory disturbances, which are called migraine with aura. One in seven migraine patients in my country experience aura symptoms [2]. The main types of aura include visual disturbances such as dark spots, bright spots, and flashes, as well as symptoms such as parosmia, dizziness, tinnitus, and difficulty speaking. Currently, more than 1 billion people worldwide are affected by migraine [3]. The rate of medical consultation for migraine patients in my country is only 52.9%, and the correct diagnosis rate by physicians is only 13.8%. In addition, there is a widespread lack of preventive treatment and excessive use of analgesics [2]. Migraine attacks can last for hours or even days and may seriously affect normal daily activities. This creates a significant demand for acute therapies that can quickly relieve headaches. In recent years, with the progress of migraine research at home and abroad, new therapeutic targets such as calcitonin gene-related peptide (CGRP) have emerged, enriching the treatment options for migraine.

[0003] Calcitonin gene-related peptide (CGRP) is a 37-amino acid polypeptide, whose N-terminal disulfide bond and amino-terminal C-terminus play an important role in activating receptors[4]. CGRP is mainly located in the c-fibers and Aδ-fibers in the trigeminal ganglion, dorsal root ganglion, and the central nervous system[5]. There are two forms of CGRP in the human body, namely α-CGRP and β-CGRP, which differ by only 1-3 amino acids in different species. α-CGRP is highly expressed in sensory neurons, while β-CGRP mainly plays a role in the enteric nervous system[6]. CGRP exerts its biological function by binding to membrane receptors. CGRP receptors are mainly composed of calcitonin receptor-like receptors (CLR) and receptor activity modifying protein 1 (RAMP1), and receptor component protein (RCP) is also required to exert signal transduction function[7]. CGRP primarily plays a role in vasodilation in the body, and studies suggest that it plays an important role in the development and progression of migraine. Clinical evidence shows that CGRP levels in the jugular vein of migraine patients increase during an attack, and intravenous injection of CGRP can cause moderate to severe headaches. The use of CGRP antagonists can relieve migraine pain and related symptoms [4].

[0004] Currently, the specific treatment modalities for acute migraine in the guidelines for diagnosis and treatment of migraine include triptans, ergotamines, ditans, and gempams [2]. Triptans are 5-HT1B / 1D receptor agonists and have been widely used in the acute treatment of migraine over the past two decades. Triptans have a high level of evidence and recommendation in the guidelines, but they also have significant cardiovascular risks, and many patients have low response rates. Ditans are 5-HT1F receptor agonists and do not have the adverse effects of vasoconstriction of triptans, but they have central nervous system depressant effects [8]. Gepandora's antagonists developed for CGRP receptors are a new treatment for migraine without cardiovascular risks or central nervous system depressant effects [7]. This provides a new option for patients who do not respond to triptans and ditans. Currently, marketed gepant-type drugs developed for CGRP receptors include AbbVie's ubrogepant and atogepant, and Pfizer's rimegepant. All three drugs are oral tablets. Migraine patients often experience symptoms such as nausea and vomiting, making oral medications less feasible. Migraine attacks also last for a long time, severely impacting daily life. Pfizer's new drug Zavegepant (vazegepant) was launched in the United States in March 2023 in the form of a nasal spray. The approved indication is the acute treatment of migraine with or without aura in adults, but it cannot be used for migraine prevention. It has the most common adverse reactions (at least 2% of ZAVZPRET-treated patients are more likely to experience them than placebo) - taste disturbance, nausea, nasal discomfort, and vomiting.

[0005] BMS disclosed and protected the compound structure of Zavegepant in patent application CN102834388B filed in 2011. BMS also disclosed a series of heterocyclic CGRP antagonists related to Zavegepant for the treatment of migraine in patents CN100558728C, CN100558428C, CN1914193A, and CN1972929A filed between 2003 and 2005.

[0006] Boehringer Ingelheim also disclosed a CGRP antagonist in CN101146799A and WO2005084672A filed in 2005-2006, respectively, hoping to use it in the treatment of pain and other related diseases.

[0007] Considering the current status of the treatment of migraine and neuropathic pain, there is still a need for novel compounds that specifically treat migraine and neuropathic headaches to meet and enrich clinical needs. The applicant of the present invention previously provided a novel CGRP antagonist compound as shown in Formula I (hereinafter referred to as "Formula I compound") in Chinese patent application CN202310479996.8 and PCT application PCT / CN2024 / 090110: Studies conducted in prior patent applications have shown that the compound of Formula I can effectively inhibit CGRP-stimulated cAMP production, thus being used as a CGRP antagonist for the prevention, alleviation, or treatment of CGRP-related diseases, particularly migraine or neuropathic pain. In particular, the compound of Formula I exhibits low IC50 values, with some specific structural compounds sharing common properties exhibiting significantly lower IC50 values, reaching pM levels. Additional experimental data demonstrates that the compound of Formula I also exhibits good mitochondrial stability, potentially leading to lower side effects and improved PK properties.

[0008] Migraine and neuralgia are acute pains, and patients often expect to obtain a drug that is fast-acting and safe to quickly control and relieve pain. Therefore, improving the PK properties of drugs, such as obtaining a shorter T max , in order to shorten the time for the drug to reach its peak and take effect quickly, and to increase Cmax and AUC to improve bioavailability, reduce drug dosage and improve drug safety, which are of particular clinical significance for migraine and pain indications.

[0009] In practice, not all active molecules exhibit high bioavailability, a rapid time to peak activity, and the resulting rapid onset of action after administration. These PK properties are related to the physicochemical and biological properties of the active drug molecule. The use of penetration enhancers is one approach to drug delivery, but the PK improvements achieved through penetration enhancement vary widely based on the active drug molecule and are often unpredictable. Furthermore, it is rare to see a single penetration enhancer significantly improve multiple PK properties simultaneously.

[0010] Alkyl glycoside is one of the optional penetration enhancers. Published Chinese patent applications CN201780081719.4, CN201780044057.3, CN200980157305.0, CN201180053436.1, and CN201980001004.2 disclose studies on the PK properties improvement of alkyl glycosides combined with different types of active pharmaceutical molecules. These results show that the PK improvement results vary greatly for different types of active pharmaceutical molecules. For example, a rough comparison shows that before and after the combined use of alkyl glycosides: maxFor some active molecule combinations, there was no change whether or not alkyl glycosides were used, while for some combinations, the effect was increased to about 3 times. For AUC, there was no change whether or not alkyl glycosides were used in combination with various active molecules, or there was a slight increase to about 1.5 times after the combination of alkyl glycosides. max Some active molecules combined with alkyl glycosides can extend the duration of drug release by about 2 times, while other combinations can shorten it. Therefore, it is difficult to predict the effect of combining active pharmaceutical molecules with alkyl glycosides on PK properties.

[0011] Based on the above, further optimizing and improving the PK properties of CGRP antagonists, such as the compound of formula I described in Chinese patent application CN202310479996.8 and PCT application PCT / CN2024 / 090110, and compounds with similar activities in the prior art such as zavegepant, rimegepant, ubrogepant, atogepant, telcagepant, olcegepant, etc., to meet the special clinical needs of migraine and neuralgia, still has important practical significance.

[0012] Summary of the Invention

[0013] The primary purpose of the present invention is to improve the PK properties of CGRP antagonists, such as the compound of formula I described in Chinese patent application CN202310479996.8 and PCT application PCT / CN2024 / 090110.

[0014] In subsequent studies of the compound of Formula I, the inventors of this application unexpectedly discovered that the combination of the compound of Formula I with an alkyl glycoside surprisingly significantly and / or simultaneously improves multiple PK properties, including Cmax, AUC, and Tmax. These significant improvements, especially simultaneous significant improvements, in PK properties are of particular clinical significance and value for the emergency treatment, relief, and prevention of migraine and neuralgic headaches, for which the compound of Formula I is indicated.

[0015] Based on the above primary objectives and corresponding unexpected discoveries, the present invention first provides a pharmaceutical composition comprising a. an active ingredient and b. an alkyl glycoside, wherein the active ingredient is preferably a compound of Formula I (including its isomers, pharmaceutically acceptable salts, or solvates) referred to herein. As demonstrated by the experimental results provided in the Examples of this application, the combination of the compound of Formula I with the alkyl glycoside can significantly improve multiple PK properties of the active ingredient. For example, compared to a pharmaceutical composition without the addition of b. an alkyl glycoside, the active ingredient C can be provided at a concentration of more than 10 times, more than 30 times, 10 to 52.2 times, or 30 to 52.2 times higher. max value, and / or provide 5.1 times or more, 8.6 times or more, 5.1 times to 17.1 times, or 8.6 times to 17.1 times the AUC of the active ingredient 0-inf value, and / or optionally provide a T shortened to 0.17h or less max Value. C max The value of the substantial increase and T max The time to peak effect was reduced from 1 hour to less than 0.17 hours (about 10 minutes), shortening the time to peak effect by 83.3%. This means that the drug can quickly reach the peak effect after administration and treat, relieve or prevent acute attacks of migraine and neuralgia. max Value and AUC 0-inf The substantial increase in the value can help further reduce the dosage, achieve low-dose and high-efficiency effects, and reduce the risk of drug safety.

[0016] In the pharmaceutical composition of the present invention, the weight ratio of the active ingredient to the pharmaceutical composition and the content of the active ingredient in the pharmaceutical composition can be selected as needed. Generally, the weight ratio of the active ingredient can be in the range of 0.05% to 20%, and the content of the active ingredient can be 0.001 mg to 1000 mg.

[0017] In the pharmaceutical composition of the present invention, the alkyl glycoside is mainly composed of a hydrophobic alkyl carbon chain and a hydrophilic oligosaccharide covalently coupled. In some embodiments, the covalently coupled linking group includes a glycosidic bond, a thioglycosidic bond or an amide bond. In other embodiments, the hydrophobic alkyl carbon chain is selected from C5 to C 16 Alkyl chain, C 10 ~C 16 Alkyl chain, C 11 ~C 14 Alkyl chain, C 11 Alkyl chain, C 12 Alkyl chain, C 13 Alkyl chain or C 14In other embodiments, the hydrophilic oligosaccharide comprises a monosaccharide or a disaccharide, wherein the monosaccharide comprises glucose and the disaccharide comprises maltose, trehalose or sucrose. In other embodiments, the alkyl glycoside is undecyl maltoside, dodecyl maltoside, tridecyl maltoside or tetradecyl maltoside.

[0018] In some more specific embodiments of the present invention, the weight ratio of the alkyl glycoside to the pharmaceutical composition can be 0.05% to 20%, 0.05% to 10%, 0.05% to 5%, 0.1% to 5%, 0.1% to 0.5%, 0.1% to 0.3%, 0.05%, 0.2% or 5%. All types of alkyl glycosides can be selected and used with reference to this dosage, especially when the alkyl glycoside is specifically selected as undecyl maltoside, dodecyl maltoside, tridecyl maltoside or tetradecyl maltoside, the weight ratio of the alkyl glycoside to the pharmaceutical composition can be 0.05% to 20%, 0.05% to 10%, 0.05% to 5%, 0.1% to 5%, 0.1% to 0.5%, 0.1% to 0.3%, 0.05%, 0.2% or 5%.

[0019] The pharmaceutical compositions provided herein can be formulated for any route of administration, including but not limited to nasal, oral, transdermal, or transmucosal administration. Nasal administration is particularly preferred, including nasal drops, nasal sprays, nasal aerosols, nasal gels, and nasal creams. Oral administration often fails to achieve rapid onset of action due to drug disintegration, dissolution, and absorption after entering the gastrointestinal tract, especially for drugs that are affected by food. Injections offer rapid onset of action, but patients are often unable to self-administer the drug, requiring professional assistance in hospitals or clinics with medical facilities. Migraine and neuralgic headaches are acute pain symptoms, and nasal administration can avoid the drawbacks of oral administration, such as first-pass effects and slow onset of action, as well as the poor compliance and medical requirements of injections. This makes it particularly suitable for administering the compound of Formula I. Furthermore, the present invention demonstrates that nasal administration of the compound in combination with an alkyl glycoside exhibits particularly significant and surprising improvements in PK properties, making it particularly valuable for the acute treatment of migraine and neuralgic headaches.

[0020] In some specific embodiments, the nasal administration form can be a single dose form, a double dose form or a multiple dose form. For a nasal spray dosage form, the delivery volume of the nasal spray can be 50ul-200ul / spray. In some more specific embodiments, the pharmaceutical composition can be in the form of a solution, and further in the form of an aqueous solution.

[0021] In some embodiments, the pharmaceutical composition can be in sterile or non-sterile form.

[0022] The present invention also provides other optional technical solutions, in which the provided pharmaceutical compositions can be added with or without other types of pharmaceutical excipients as needed, including but not limited to one or a combination of two or more of buffers, osmotic pressure regulators, wetting agents, thickeners, pH regulators, metal chelators and preservatives.

[0023] In addition to the surprising and significant PK improvement effect of the combination of alkyl glycosides and compounds of formula I, the inventors also unexpectedly discovered that a variety of CGRP antagonists that are on the market or under development, similar to compounds of formula I, especially gepant-type CGRP antagonists such as zavegepant, also have a certain PK improvement effect after nasal administration in combination with alkyl glycosides, with Cmax increased by 2.7 times and AUC increased by 5.4 times. This effect is completely unexpected because according to the records of CN202180077330.9, after oral administration of zavegepant in combination with alkyl glycosides, Cmax increased by 2.7 times and AUC increased by 5.4 times. max , AUC had no significant changes, T max On the contrary, it was extended.

[0024] Based on this discovery, another object of the present invention is to provide a nasal preparation comprising a CGRP antagonist and an alkyl glycoside, wherein the CGRP antagonist is a gepant-type CGRP antagonist drug, for example, including but not limited to zavegepant, rimegepant, ubrogepant, atogepant, telcagepant, olcegepant or any compound of formula I described in the present invention.

[0025] In some specific embodiments, the alkyl glycoside in the provided nasal formulation is mainly composed of a hydrophobic alkyl carbon chain and a hydrophilic oligosaccharide covalently coupled. In some embodiments, the covalently coupled linking group includes a glycosidic bond, a thioglycosidic bond, or an amide bond. In some embodiments, the hydrophobic alkyl carbon chain is selected from C5 to C 16 Alkyl chain, C 10 ~C 16 Alkyl chain, C 11 ~C 14 Alkyl chain, C 11 Alkyl chain, C 12 Alkyl chain, C 13 Alkyl chain or C 14Alkyl chain. In some embodiments, the hydrophilic oligosaccharide includes a monosaccharide or a disaccharide, the monosaccharide includes glucose, and the disaccharide includes maltose, trehalose or sucrose. In some embodiments, the alkyl glycoside is undecyl maltoside, dodecyl maltoside, tridecyl maltoside or tetradecyl maltoside. In some embodiments, the weight ratio of the alkyl glycoside to the pharmaceutical composition can be 0.05% to 20%, 0.05% to 10%, 0.05% to 5%, 0.1% to 5%, 0.1% to 0.5%, 0.1% to 0.3%, 0.05%, 0.2% or 5%.

[0026] In some specific embodiments, the present invention also provides the use of the aforementioned pharmaceutical composition or nasal formulation of the present invention as a CGRP antagonist.

[0027] In some specific embodiments, the present invention provides the use of a pharmaceutical composition or nasal formulation of the present invention in the preparation of a medicament for preventing, treating, or alleviating CGRP-mediated diseases, including migraine and neuropathic pain. Accordingly, the present invention also provides a method for preventing, treating, or alleviating migraine, neuropathic pain, or CGRP-mediated diseases, comprising administering the aforementioned pharmaceutical composition or nasal formulation of the present invention to a subject in need thereof.

[0028] Detailed Description of the Invention

[0029] The compounds of Formula I used as active ingredients in the present invention include the compounds of Formula I described in Chinese patent application CN202310479996.8 and PCT application PCT / CN2024 / 090110, in particular the compounds described in the specific embodiments and examples therein. The entire contents of the above-mentioned Chinese patent application CN202310479996.8 and PCT application PCT / CN2024 / 090110 are hereby incorporated into this application as a part thereof. Even though they have been incorporated in their entirety, for the sake of clarity and completeness, this application also provides a detailed description of the compounds of Formula I through the following structural descriptions and the preparation, characterization, and activity tests of the specific examples below.

[0030] Compounds of formula I:

[0031] In one embodiment, the structure of the compound of formula I of the present invention (including its isomers, pharmaceutically acceptable salts or solvates) is defined as follows:

[0032] in,

[0033] X 1 、X 2 Each independently is CH or N;

[0034] X3 、X 4 are each independently C, CH or N; alternatively, X 3 、X 4 The optional value is N at different times;

[0035] X 5 is C, CH, CH2, NH or N;

[0036] Y 1 is CH2, O or NH;

[0037] Y 2 O or S;

[0038] R 2 is independently at each occurrence -H, -(C1-C6)alkyl or -(C3-C8)cycloalkyl, n is 0, 1, 2, 3, 4, 5 or 6,

[0039] Or, two R 2 The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected to R 2 The attached ring forms a spiro ring structure, a fused ring structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S;

[0040] Or, -(R 2 ) n The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 2 ) n The attached ring forms a spiro ring structure, a fused ring structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S;

[0041] R 3 is independently at each occurrence -H, -(C1-C6)alkyl or -(C3-C8)cycloalkyl, o is 0, 1, 2, 3, 4, 5 or 6,

[0042] Or, two R 3 The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected to R 3 The attached ring forms a spiro ring structure, a fused ring structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S;

[0043] Or, -(R 3 ) o The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 3 ) oThe attached ring forms a spiro ring structure, a fused ring structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S;

[0044] A is the structure shown in the following formula II:

[0045] The G chain group and the two carbon atoms to which it is attached together form a 5- to 7-membered aliphatic ring or aliphatic heterocyclic ring, wherein the heteroatoms constituting the ring are -O-, -S- or

[0046] R 1 is independently at each occurrence -H, -(C1-C6)alkyl or -(C3-C8)cycloalkyl, m is 0, 1, 2, 3, 4, 5 or 6,

[0047] Or, two R 1 The atoms to which it is attached together form a 3-8 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected to R 1 The attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S,

[0048] Or, -(R 1 ) m The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 1 ) m The attached ring constitutes a spiro ring structure, a fused ring structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S.

[0049] In some embodiments, the compound of formula I of the present invention is a chiral structure represented by formula I-2 or formula I-3:

[0050] Among them, X 1 、X 2 、X 3 、X 4 、X 5 、Y 1 、Y 2 ,A,R 2 、R 3 , n, o are as defined above.

[0051] In some embodiments, the A group in the compound of Formula I, Formula I-2, or Formula I-3 is further selected as follows:

[0052] Selection of the G chain group in A: If the G chain group and the two carbon atoms to which it is attached together form a 5- or 6-membered aliphatic ring, the G chain group may optionally have 0 or 1 double bonds, and the rest are single bonds; if the G chain group and the two carbon atoms to which it is attached together form a 7-membered aliphatic ring, the G chain group may optionally have 0, 1 or 2 double bonds, and the rest are single bonds; if the G chain group and the two carbon atoms to which it is attached together form a 5-membered aliphatic heterocyclic ring, the G chain group may optionally have single bonds; if the G chain group and the two carbon atoms to which it is attached together form a 6- or 7-membered aliphatic heterocyclic ring, the G chain group may optionally have 0 or 1 double bonds, and the rest are single bonds. The G chain group in A can be further specifically selected from:

[0053] Among the further specific choices of the G chain group, there are some examples The symmetric groups and for example Considering the asymmetry of the groups attached to the two connection sites of the G chain group, when connecting the attached asymmetric groups, the specific selection structure of the asymmetric group of each G chain group represents two different specific connection methods, such as the asymmetric group Represents two situations As an example, when the attached asymmetric group is an oxopyridine structure When the G chain group is still asymmetric As an example, the two cases represented are connected to the oxopyridine structure in the following ways:

[0054] R in A 1 Group selection: R 1 Each occurrence is independently -H, -(C1-C3)alkyl or -(C3-C6)cycloalkyl, m is 0, 1, 2 or 3, or two R 1 The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected to R 1 The attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S, or -(R 1 ) m The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 1 ) m The attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, and the heteroatom in the aliphatic heterocycle is O, N or S. 1 The group can further select: R 1 is independently -H, methyl, ethyl or cyclopropane at each occurrence, m is 0, 1 or 2, or, two R 1The atoms to which it is attached together form cyclopropane, cyclobutane or cyclopentane, and the cyclopropane, cyclobutane or cyclopentane and R 1 The attached ring forms a spiro ring structure, a fused ring structure or a bridged ring structure, or, -(R 1 ) m The atoms to which it is attached together form a cyclopropane, cyclobutane or cyclopentane, which is in combination with -(R 1 ) m The attached rings form a spiro ring structure, a fused ring structure or a bridged ring structure.

[0055] According to an embodiment of the present invention, the group A is preferably a structure represented by the following formula II-1, II-2 or II-3:

[0056] In formula II-1, II-2 and II-3, X 6 -O-, -S- or

[0057] Two places in formula II-2 All refer to single bonds, or any one of them is a double bond and the other is a single bond;

[0058] Three places in formula II-3 All refer to single bonds, or any one of them is a double bond and the other two are single bonds.

[0059] When the A group is preferably a structure represented by formula II-1, II-2 or II-3, the compound containing an oxopyridoaliphatic ring or aliphatic heterocyclic structure provided by the present invention has better CGRP antagonistic activity, lower side reactions, and / or better PK metabolic effects.

[0060] According to the specific embodiment of the present invention, compared with Y 2 is O, when Y 2 When S, X 6 Compounds with -S- generally have better CGRP antagonist activity. 2 is S, and X 6 For -S-.

[0061] According to an embodiment of the present invention, the A group may also preferably be a structure shown in the following formula II-4 or II-5:

[0062] In formula II-4 or II-5, X 6 -O-, -S- or

[0063] In formula II-4 Refers to single or double bonds;

[0064] Two places in formula II-5 All refer to single bonds, or any one of them is a double bond and the other is a single bond.

[0065] As demonstrated in the specific embodiments of the present invention, the heteroatom X in A is usually 6 As the position gradually moves away from the -NH- group in A, the CGRP antagonist activity of the compound tends to decrease. 6 When the ring is large, such as a 6-membered ring or a 7-membered ring, even if the heteroatom X 6 Located at the position shown in II-4 or II-5, the CGRP antagonist activity of the compound is still within an acceptable range.

[0066] According to an embodiment of the present invention, when the G chain group in A and the two carbon atoms to which it is attached together form an aliphatic ring, the aliphatic ring is preferably unsubstituted.

[0067] In one embodiment, the G chain group in A and the two carbon atoms to which it is attached together form a 5-7 membered aliphatic ring, and m is 0.

[0068] According to an embodiment of the present invention, the A group can be selected as follows:

[0069] The A group is particularly preferably selected from

[0070] Based on the particularly preferred selection of the A group at this time, the compounds containing an oxopyridoaliphatic ring or aliphatic heterocyclic structure provided by the present invention have more excellent CGRP antagonistic activity, lower side reactions, and / or better PK metabolic effects.

[0071] According to an embodiment of the present invention, the A group can be selected as follows:

[0072] The A group is particularly preferably selected from Based on the particularly preferred selection of the A group at this time, the compounds containing an oxopyridoaliphatic ring or aliphatic heterocyclic structure provided by the present invention have more excellent CGRP antagonistic activity, lower side reactions, and / or better PK metabolic effects.

[0073] In other embodiments, the present invention can be used for R 2 The group is further selected as: R 2 is independently -H, -(C1-C3)alkyl or -(C3-C6)cycloalkyl at each occurrence, and n is 0, 1, 2 or 3; or, two R 2 The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected to R 2The attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, and the two R 2 The provided ring atoms are all C atoms; or, -(R 2 ) n The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 2 ) n The attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, wherein -(R 2 ) n The provided ring atoms are all C atoms. 2 The group can further be selected as: R 2 is independently -H, methyl, ethyl, cyclopropyl or cyclobutyl at each occurrence, and n is 0, 1 or 2; or, two R 2 The atoms to which it is attached together form a 3-5 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a 3-5 membered aliphatic ring or aliphatic heterocyclic ring with R 2 The attached ring forms a spiro ring structure, and the two R 2 The provided ring atoms are all C atoms; or, -(R 2 ) n The atoms to which it is attached together form a 3-5 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 2 ) n The attached ring forms a spiro ring structure, wherein -(R 2 ) n The provided ring atoms are all C atoms.

[0074] In other embodiments, the present invention can be used for R 3 The group is further selected as: R 3 is independently -H, -(C1-C3)alkyl or -(C3-C6)cycloalkyl at each occurrence, and n is 0, 1, 2 or 3; or, two R 3 The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected to R 3 The attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, and the two R 3 The provided ring atoms are all C atoms; or, -(R 3 ) o The atoms to which it is attached together form a 3-6 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 3 ) oThe attached ring forms a spiro ring structure, a cyclic structure or a bridged ring structure, wherein -(R 3 ) o The provided ring atoms are all C atoms. 3 The group can further be selected as: R 3 is independently -H, -(C1-C3)alkyl or -(C3-C6)cycloalkyl at each occurrence, and n is 0, 1, 2 or 3; or, two R 3 The atoms to which it is attached together form a 3-5 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a 3-5 membered aliphatic ring or aliphatic heterocyclic ring with R 3 The attached ring forms a spiro ring structure, and the two R 3 The provided ring atoms are all C atoms; or, -(R 3 ) o The atoms to which it is attached together form a 3-5 membered aliphatic ring or aliphatic heterocyclic ring, wherein the aliphatic ring or aliphatic heterocyclic ring is optionally connected with -(R 3 ) o The attached ring forms a spiro ring structure, wherein -(R 3 ) o The provided ring atoms are all C atoms.

[0075] In other embodiments, the compound of formula I of the present invention is more preferably a structure shown in formula I-1:

[0076] In Formula Ⅰ-1, X 1 、X 2 、X 3 、X 4 、X 5 、Y 1 、Y 2 , A may be optionally defined as any of the above descriptions of the present invention. In Formula I-1, R 2 、R 3 is independently at each occurrence -H, -CH3 or absent.

[0077] In other embodiments, more than one H in the compounds of formula I or more preferably in the compounds of formula I-1 of the present invention is substituted with D, particularly wherein the G chain group or R 1 One or more H in is replaced by D.

[0078] In some embodiments, R in the compound of formula I or more preferably in the compound of formula I-1, formula I-2 or formula I-3 of the present invention is 3 More than one H in the D is replaced, and further selection is made with X 5 Connected R 3 It is -CD3.

[0079] In some embodiments, the H on the pyrazole ring of the indazolyl group in the compounds of Formula I or, more preferably, the compounds of Formula I-1, Formula I-2, or Formula I-3 of the present invention is substituted with D.

[0080] In some embodiments, the compound of formula I of the present invention can be specifically selected from:

[0081] In other embodiments, the compound of formula I of the present invention can also be specifically selected from:

[0082] In the present invention, when a later technical solution further defines a previous technical solution, it may only further define some of the technical features. In this case, the undefined technical features may be optionally defined in the previous technical solution or anywhere in the present invention.

[0083] definition

[0084] Unless stated to the contrary or otherwise defined, the following terms used in the specification and claims have the following meanings.

[0085] "Alkyl" refers to an aliphatic hydrocarbon group, including a saturated hydrocarbon group. An alkyl group may be a straight-chain or branched chain alkyl group. For example, -(C1-C6)alkyl or -(C1-C3)alkyl. -(C1-C6)alkyl refers to an alkyl group having 1 to 6 carbon atoms, such as 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, and n-hexyl.

[0086] "Ring" refers to any covalently closed structure, including, for example, a carbocyclic ring (e.g., an aryl group, a cycloalkyl group), a heterocyclic group (e.g., a heteroaryl group, a heterocycloalkyl group), an aromatic group (e.g., an aryl group, a heteroaryl group), a non-aromatic group (e.g., a cycloalkyl group, a heterocycloalkyl group, etc.). The ring can be a monocyclic ring or a polycyclic ring. Typical polycyclic rings generally include bicyclic and tricyclic rings. The rings of the present invention generally have 3-20 ring atoms, for example, 3 ring atoms, 4 ring atoms, 5 ring atoms, 6 ring atoms, 7 ring atoms, 8 ring atoms, 9 ring atoms, 10 ring atoms, 11 ring atoms, 12 ring atoms, 13 ring atoms, 14 ring atoms, 15 ring atoms, 16 ring atoms, 17 ring atoms, 18 ring atoms, 19 ring atoms or 20 ring atoms.

[0087] An "aliphatic ring" is a non-aromatic, saturated or unsaturated cyclic hydrocarbon whose skeletal atoms are all carbon atoms. In the present invention, an aliphatic ring typically has 3 to 8, 3 to 6, or 5 to 7 ring carbon atoms and may also be referred to as a 3- to 8-membered aliphatic ring, a 3- to 6-membered aliphatic ring, or a 5- to 7-membered aliphatic ring. Typical examples of aliphatic rings include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclopropene, cyclobutene, cyclopentene, cyclohexene, and cycloheptene.

[0088] "Cycloalkyl" refers to a saturated cyclic hydrocarbon substituent, preferably cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. The cycloalkyl group may be substituted or unsubstituted.

[0089] "Aliphatic heterocycle", also referred to as "aliphatic heterocycle" in the present invention, is a non-aromatic heterocycle formed by replacing one or more of the carbon atoms in the skeleton of the "aliphatic ring" with heteroatoms. The replaced heteroatoms can usually be selected from O, S, N, etc.

[0090] "Membered" refers to the number of atoms that make up the ring. Typical five-membered rings include cyclopentyl, pyrrole, imidazole, thiazole, furan, and thiophene. Typical six-membered rings include cyclohexane, pyridine, pyran, pyrazine, thiopyran, pyridazine, pyrimidine, and benzene. A ring containing heteroatoms among its backbone atoms is considered a heterocycle.

[0091] "Spirocycle" or "spirocyclic structure" refers to a structure formed by two rings sharing a backbone ring atom, for example wait.

[0092] "Bridged ring" or "bridged ring structure" refers to a structure formed by two rings sharing two skeleton ring atoms, and the two shared skeleton ring atoms are not adjacent, for example wait.

[0093] "Parallel ring" or "parallel ring structure" refers to a structure formed by two rings sharing two adjacent skeleton ring atoms, for example wait.

[0094] "Oxo" refers to the replacement of a hydrogen on a carbon with =0.

[0095] "Substituted" means that one or more hydrogen atoms, preferably up to 5 (e.g., 1, 2, 3, 4, 5), more preferably 1 to 3 hydrogen atoms, in a group can be replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) without undue effort which substitutions are possible or impossible. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated bond.

[0096] "Antagonist" refers to a drug that has no intrinsic activity but can block the effects mediated by receptor agonists after binding to the receptor.

[0097] "Optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes that the event or circumstance occurs or does not occur.

[0098] The term "substituted or unsubstituted" herein refers to any group that is monosubstituted or polysubstituted by a specified substituent to the extent that such monosubstituted or polysubstituted (including multiple substitutions on the same moiety) is chemically permitted, and each substituent can be located at any available position on the group and can be attached through any available atom on the substituent. "Any available position" refers to any position on the group that is chemically accessible by methods known in the art or methods taught herein and does not produce an overly unstable molecule. When there are two or more substituents on any group, each substituent is defined independently of any other substituent and can therefore be the same or different.

[0099] The term "compounds of the present invention" as used herein is intended to encompass compounds of the general formula (I) as defined herein or any preferred or specific embodiment thereof (including compounds of formula (I-1) and example compounds), their stereoisomers, pharmaceutically acceptable salts, tautomers or solvates.

[0100] The term "pharmaceutically acceptable" as used herein refers to molecular entities and compositions that are or are approvable by relevant agencies in various countries, or listed in generally recognized pharmacopeias for use in animals, and more particularly in humans, or that do not produce adverse, allergic or other untoward reactions when administered in appropriate amounts to animals, such as humans.

[0101] The term "pharmaceutically acceptable salt" as used herein refers to a salt of a compound of the present invention that is pharmaceutically acceptable and possesses the desired pharmacological activity of the parent compound. Specifically, such salts are non-toxic and can be inorganic acid addition salts or organic acid addition salts and base addition salts.

[0102] It should be understood that when selecting the various groups in the compound structure of the present invention, the groups that are connected, coordinated or influenced by each other should be selected accordingly under the premise of complying with the chemical valence rules.

[0103] According to the contents of the present invention, in accordance with common technical knowledge and means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0104] Preparation of compounds of formula I

[0105] To further illustrate the present invention, the following examples provide a detailed description of the compound of formula I, its preparation method, application, pharmaceutical composition, and activity test provided by the present invention.

[0106] The following abbreviations or terms have the following meanings: K2CO3 represents potassium carbonate; H2O represents water; DMF represents N,N-dimethylformamide; KF represents potassium fluoride; DIPEA or DIEA represents N,N-diisopropylethylamine; n-BuLi represents n-butyllithium; THF represents tetrahydrofuran; TFA represents trifluoroacetic acid; HCl / Dioxane represents a solution of hydrogen chloride in dioxane; HCl represents a solution of hydrogen chloride; DMSO represents dimethyl sulfoxide; CHI or MeI represents iodomethane; H2 represents hydrogen; Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium; t-BuOK represents potassium tert-butoxide; NMP represents N-methylpyrrolidone; MeOH represents methanol; EtOH represents ethanol; Pd(PPh3)2Cl2 represents bis(triphenylphosphinepalladium dichloride); DIBAL-H represents diisobutylaluminum hydride; TEA represents triethylamine; DCM represents dichloromethane; DCE represents 1,2-dichloroethane; dioxane represents 1,4-dioxane; NBS represents N-bromosuccinimide; NaH represents sodium hydride; LiHMDS represents lithium bis(trimethylsilylamide); NaBH4 represents sodium borohydride; Pd(dppf)Cl2 represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; Zn represents zinc; Ruphos represents 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl; I2 represents iodine; NaSH represents sodium hydrosulfide; CuI represents cuprous iodide; Pd / C represents palladium on carbon; AcSH represents thioacetic acid; DIAD represents diisopropyl azodicarboxylate; PPh3 represents triphenylphosphine; CH3CN or MeCN represents acetonitrile; PtO2 represents platinum dioxide; NaOMe represents sodium methoxide; KHMDS represents potassium bis(trimethylsilyl)amide;DEAD represents diethyl azodicarboxylate; MWI represents microwave-induced reaction; AcCl represents acetyl chloride; NaOH represents sodium hydroxide; TBTU represents 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate; DMAP represents 4-dimethylaminopyridine; Ms2O represents methanesulfonic anhydride; LiOH represents lithium hydroxide; B(OCH3)3 represents trimethyl borate; Cs2CO3 represents cesium carbonate; TCDI represents N,N'-thiocarbonyldiimidazole; TosCl represents p-toluenesulfonyl chloride; and DSC represents N,N'-disuccinimidyl carbonate. LDA represents lithium diisopropylamide; XantPhos represents 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; LAH represents lithium aluminum tetrahydride; DPPA represents diphenylphosphoryl azide; DMC represents dimethyl carbonate; TMSCN represents trimethylsilyl cyanide; s-BuLi represents sec-butyllithium; PyBOP represents 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate; DBU represents 1,8-diazabicyclo[5.4.0]undec-7-ene; DMSu represents dimethyl succinate; Pd(dppf)Cl2·DCM represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex.

[0107] In addition, in the following preparation examples and examples, when the structural formula of a compound does not match the chemical nomenclature, the structural formula of the compound shall prevail.

[0108] The following first provides exemplary methods for synthesizing intermediate compounds of the compound of Formula I of the present invention. The starting materials and other additives used in the synthesis of the provided intermediate compounds can be obtained through common commercial sources or by combining conventional chemical reaction synthesis methods with methods reported in prior art literature. Some intermediate compounds can also be obtained directly through common commercial sources or by combining conventional chemical reaction synthesis methods with methods reported in prior art literature.

[0109] Intermediate Preparation Example 1: Preparation of 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropane-1-ol (can be used in Intermediate Preparation Example 2)

[0110] Step 1: Preparation of 4-(2-chloro-6-methoxypyridin-3-yl)isoxazole

[0111] Under a nitrogen atmosphere, 3-bromo-2-chloro-6-methoxypyridine (10.0 g) and potassium carbonate (12.4 g) were dissolved in 1,4-dioxane (100 mL) and water (10 mL). 4-Isoxazoleboronic acid (9.7 g) was then dissolved in 1,4-dioxane (100 mL) and placed in a 150 mL constant pressure dropping funnel. 1,1-bis(diphenylphosphino)diphenylferric palladium chloride (1.63 g) was added. The reaction mixture was brought to 90°C and 4-Isoxazoleboronic acid was slowly added dropwise over 3 hours. The reaction mixture was allowed to react for 2 hours after the addition was complete. TLC indicated that the reaction was complete. The reaction mixture was brought to room temperature, 500 mL of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 9.0 g of the title compound.

[0112] MS (ESI) m / z (M+H) + =211.0.

[0113] Step 2: Preparation of 2-(2-chloro-6-methoxypyridin-3-yl)acetonitrile

[0114] Dissolve 4-(2-chloro-6-methoxypyridin-3-yl)isoxazole (9.0 g) in methanol (100 mL) and water (10 mL), add potassium fluoride (12.4 g), and place the system in a thick-walled reaction flask. Keep sealed at 100°C for 6 hours. TLC indicates the reaction is complete. The mixture is brought to room temperature and concentrated to remove the methanol. 500 mL of water is added and extracted three times with ethyl acetate. The organic phases are combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 7.1 g of the title compound.

[0115] MS (ESI) m / z (M+H) + =183.0.

[0116] Step 3: Preparation of 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropionitrile

[0117] Under a nitrogen atmosphere, 2-(2-chloro-6-methoxypyridin-3-yl)acetonitrile (7.1 g) was dissolved in anhydrous tetrahydrofuran (80 mL). The system was cooled to -78°C and lithium bistrimethylsilylamide (1 M, 117.0 mL) was slowly added. The reaction was allowed to react for 10 minutes. Iodomethane (12.7 g) was added and the reaction continued for 20 minutes. TLC indicated the reaction was complete. The reaction system was brought to room temperature and 200 mL of saturated aqueous ammonium chloride was added. The product was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 8.1 g of crude product.

[0118] MS (ESI) m / z (M+H) + =211.1.

[0119] Step 4: Preparation of 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropanal

[0120] Under a nitrogen atmosphere, 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropionitrile (8.1 g) was dissolved in anhydrous tetrahydrofuran (80 mL). The system was cooled to -78°C and diisobutylaluminum hydride (1 M, 64.3 mL) was slowly added. The reaction was allowed to react for 2 hours. TLC indicated that the reaction was complete. The reaction system was warmed to 0°C and diluted with tetrahydrofuran (200 mL). Water (3.9 mL), 10% aqueous sodium hydroxide solution (3.9 mL), and water (9.64 mL) were added sequentially. The mixture was brought to room temperature and stirred for 10 minutes. An appropriate amount of anhydrous sodium sulfate was added and stirring continued for 20 minutes. The mixture was filtered and the filtrate was concentrated. The crude product was purified by column chromatography to obtain 5.0 g of the title compound.

[0121] MS (ESI) m / z (M+H) + =214.1.

[0122] Step 5: Preparation of 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropan-1-ol

[0123] Dissolve 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropanal (5.0 g) in methanol (50 mL), add sodium borohydride (446.0 mg), and react at room temperature for 10 minutes. TLC indicates the reaction is complete. The reaction system is concentrated, and the crude product is purified by column chromatography to obtain 4.0 g of the title compound.

[0124] MS (ESI) m / z (M+H) + =216.1.

[0125] Intermediate Preparation Example 2: Preparation of 5-bromo-6-methoxy-3,3-dimethyl-2,3-dihydrofuran[2,3-b]pyridine (can be used in Example 1)

[0126] Step 1: Preparation of 6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridine

[0127] Dissolve 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropan-1-ol (1.0 g) in anhydrous tetrahydrofuran (10 mL) and slowly add the mixture to a suspension of sodium hydride (220.8 mg) in anhydrous tetrahydrofuran (10 mL). Heat the mixture to 60°C and react for 2 hours. TLC indicates the reaction is complete. Add the reaction system dropwise to 50 mL of saturated aqueous ammonium chloride in an ice-water bath and extract three times with ethyl acetate. Combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 516 mg of the title compound.

[0128] MS (ESI) m / z (M+H) + =180.1.

[0129] Step 2: Preparation of 5-bromo-6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridine

[0130] Dissolve 6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridine (510.0 mg) in anhydrous N,N-dimethylformamide (5 mL), add N-bromosuccinimide (558 mg), and allow to react at room temperature for 20 minutes. TLC indicates completion of the reaction. Add 30 mL of water to the reaction system, extract three times with ethyl acetate, combine the organic phases, backwash twice with purified water and once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 610.0 mg of the title compound.

[0131] MS (ESI) m / z (M+H) + =258.0,260.0.

[0132] Intermediate Preparation Example 3: Preparation of (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)propionic acid methyl ester trifluoroacetate (can be used in Example 1)

[0133] Step 1: Preparation of (S)-(2-((tert-Butoxycarbonyl)amino)-3-methoxy-3-oxypropyl)zinc(II) iodide

[0134] Under a nitrogen atmosphere, activated zinc powder (2.37 g) was added to a 100 mL three-necked flask. Iodine (926.37 mg) was dissolved in anhydrous N,N-dimethylformamide and added dropwise to the flask. The system was then reacted at 60°C for 15 minutes. Methyl (S)-2-((tert-butoxycarbonyl)amino)-3-iodopropionate (6.00 g) was dissolved in anhydrous N,N-dimethylformamide (35 mL) and slowly added dropwise to the reaction system. The reaction was continued at 70°C for 2.5 hours. After the reaction was completed, the reaction system was brought to room temperature and the resulting solution was directly used in the next reaction.

[0135] Step 2: Preparation of methyl (R)-2-((tert-butoxycarbonyl)amino)-3-(7-methyl-1H-indazol-5-yl)propionate

[0136] Under a nitrogen atmosphere, 5-bromo-7-methyl-1H-indazole (2.26 g), tris(dibenzylideneacetone)dipalladium (489.52 mg), and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (499.29 mg) were dissolved in anhydrous N,N-dimethylformamide (5 mL). The solution from the first step was added to the reaction system, and the mixture was heated at 70°C for 3 hours. After completion of the reaction, the reaction system was filtered through celite, and the filtrate was collected. 30 mL of water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by silica gel column chromatography to obtain 2.1 g of the title compound.

[0137] MS (ESI) m / z (M+H) + =334.1.

[0138] Step 3: Preparation of (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)propionic acid methyl ester trifluoroacetate

[0139] Methyl (R)-2-((tert-Butoxycarbonyl)amino)-3-(7-methyl-1H-indazol-5-yl)propanoate (1 g) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (10 mL) and allowed to react at room temperature for 1 hour. After completion of the reaction, the reaction system was concentrated to obtain the crude product, which was used directly in the next reaction without purification.

[0140] MS (ESI) m / z (M+H) + =234.1.

[0141] Intermediate Preparation Example 4: Preparation of 5-bromo-6-methoxy-3,3-dimethyl-2,3-dihydrothiophene[2,3-b]pyridine (can be used in Example 2)

[0142] Step 1: Preparation of 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropyl methanesulfonate

[0143] 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropan-1-ol (600.0 mg) was dissolved in 1,2-dichloroethane (10 mL). Triethylamine (848.4 mg), 4-dimethylaminopyridine (34.1 mg), and methanesulfonic anhydride (584.6 mg) were added sequentially and allowed to react at room temperature for 30 minutes. TLC indicated the reaction was complete. The reaction system was added to 30 mL of water and extracted three times with dichloromethane. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 820 mg of crude product.

[0144] MS (ESI) m / z (M+H) + =294.0.

[0145] Step 2: Preparation of 6-methoxy-3,3-dimethyl-2,3-dihydrothieno[2,3-b]pyridine

[0146] Dissolve crude 2-(2-chloro-6-methoxypyridin-3-yl)-2-methylpropyl methanesulfonate (850.0 mg) in N,N-dimethylformamide (10 mL), add sodium hydrosulfide (487.4 mg), and heat at 130°C for 3 hours. TLC indicates completion of the reaction. Add 30 mL of water to the reaction system, extract three times with ethyl acetate, combine the organic phases, backwash twice with purified water and once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 450.0 mg of the title compound.

[0147] MS (ESI) m / z (M+H) + =196.1.

[0148] Step 3: Preparation of 5-bromo-6-methoxy-3,3-dimethyl-2,3-dihydrothiophene[2,3-b]pyridine

[0149] Dissolve 6-methoxy-3,3-dimethyl-2,3-dihydrothieno[2,3-b]pyridine (450.0 mg) in N,N-dimethylformamide (5 mL), add N-bromosuccinimide (452.0 mg), and react at room temperature for 10 minutes. TLC indicates completion of the reaction. Add 30 mL of water to the reaction system, extract three times with ethyl acetate, combine the organic phases, backwash twice with purified water and once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 300.0 mg of the title compound.

[0150] MS (ESI) m / z (M+H) + =274.0,276.0.

[0151] Intermediate Preparation Example 5: Preparation of Methyl (R)-3-(7-methyl-1H-indazol-5-yl)-2-((phenoxycarbonyl)amino)propanoate (can be used in Example 2)

[0152] In an ice-water bath, dissolve (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)propionic acid methyl ester trifluoroacetate (990 mg) in dichloromethane (20 mL), and add N,N-diisopropylethylamine (1.16 g) dropwise. Phenyl chloroformate (469.71 mg) is then dissolved in dichloromethane (10 mL) and added dropwise to the reaction system. The reaction mixture is allowed to react for 1 hour. After completion of the reaction, the reaction solution is poured into water and extracted with ethyl acetate. The organic phases are combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by silica gel column chromatography to obtain 1 g of the title compound.

[0153] MS (ESI) m / z (M+H) + =354.1.

[0154] Intermediate Preparation Example 6: Preparation of 4-(2-chloro-6-methoxypyridin-3-yl)butan-1-ol (can be used in Intermediate Preparation Example 7)

[0155] Step 1: Preparation of 4-(2-chloro-6-methoxypyridin-3-yl)but-3-yn-1-ol

[0156] 3-Bromo-2-chloro-6-methoxypyridine (3.0 g) was dissolved in tetrahydrofuran (20 mL) and triethylamine (10 mL). 3-Butyn-1-ol (2.9 g) was diluted with tetrahydrofuran (5 mL) and placed in a constant pressure dropping funnel. After purging the system with nitrogen, bistriphenylphosphine palladium dichloride (476.0 mg) and cuprous iodide (64.0 mg) were added. After purging the system with nitrogen again, the temperature was raised to 50°C, and 3-butyn-1-ol was slowly added dropwise at this temperature. The addition was complete within two hours, and the reaction was continued at this temperature for 1 hour. TLC indicated that the reaction was complete. 30 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography to yield 1.8 g of the title compound.

[0157] MS (ESI) m / z (M+H) + =212.0.

[0158] Step 2: Preparation of 4-(2-chloro-6-methoxypyridin-3-yl)butan-1-ol

[0159] Dissolve 4-(2-chloro-6-methoxypyridin-3-yl)but-3-yn-1-ol (1.8 g) in methanol (100 mL), add palladium on carbon (10%, 180.0 mg), replace the air, and continue to flow hydrogen gas. Allow to react at room temperature for 1 hour. TLC indicates the reaction is complete. Filter, collect the filtrate, and concentrate. The crude product is purified by column chromatography to yield 1.5 g of the title compound.

[0160] MS (ESI) m / z (M+H) + =216.0.

[0161] Intermediate Preparation Example 7: Preparation of 7-bromo-8-methoxy-2,3,4,5-tetrahydrooxepin[2,3-b]pyridine (can be used in Example 3)

[0162] Step 1: Preparation of 8-methoxy-2,3,4,5-tetrahydrooxepin[2,3-b]pyridine

[0163] Dissolve 4-(2-chloro-6-methoxypyridin-3-yl)butan-1-ol (430.0 mg) in 1,4-dioxane (6 mL), add potassium tert-butoxide (448.0 mg), and heat to 85°C for 1 hour. TLC indicates the reaction is complete. Add dilute hydrochloric acid at 0°C and adjust the pH to approximately 7. Extract three times with ethyl acetate, combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate to yield 0.3 g of crude product.

[0164] MS (ESI) m / z (M+H) + =180.1.

[0165] Step 2: Preparation of 7-bromo-8-methoxy-2,3,4,5-tetrahydrooxepin[2,3-b]pyridine

[0166] Dissolve crude 8-methoxy-2,3,4,5-tetrahydrooxepino[2,3-b]pyridine (300.0 mg) in anhydrous N,N-dimethylformamide (4 mL), add N-bromosuccinimide (298.0 mg), and allow to react at room temperature for 30 minutes. TLC indicates completion of the reaction. Add 30 mL of water to the reaction system, extract three times with ethyl acetate, combine the organic phases, backwash twice with purified water and once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 195.0 mg of the title compound.

[0167] MS (ESI) m / z (M+H) +=258.0,260.0.

[0168] Intermediate Preparation Example 8: Preparation of 7-bromo-8-methoxy-2,3,4,5-tetrahydrothiop[2,3-b]pyridine (can be used in Example 4)

[0169] Step 1: Preparation of S-4-(2-chloro-6-methoxypyridin-3-yl)butyl)thioethyl ester

[0170] In an ice-water bath, 4-(2-chloro-6-methoxypyridin-3-yl)butan-1-ol (1.4 g) and triphenylphosphine (2.6 g) were dissolved in dry tetrahydrofuran (20 mL). Diisopropyl azodicarboxylate (2.0 g) was added and the mixture was allowed to react at room temperature for 10 minutes. Thioacetic acid (741.7 mg) was added to the mixture in an ice-water bath and the mixture was allowed to react at room temperature for 30 minutes. TLC indicated that the reaction was complete. 30 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 1.0 g of the title compound.

[0171] MS (ESI) m / z (M+H) + =274.1.

[0172] Step 2: Preparation of 4-(2-chloro-6-methoxypyridin-3-yl)butane-1-thiol hydrochloride

[0173] Dissolve S-4-(2-chloro-6-methoxypyridin-3-yl)butylthioethyl ester (1.0 g) in 1,4-dioxane-hydrochloric acid solution (4 M, 10 mL) and allow to react at 40°C for 1 hour. TLC indicated the reaction was complete, and the system was concentrated to yield 930.0 mg of crude product.

[0174] MS (ESI) m / z (M+H) + =232.0.

[0175] Step 3: Preparation of 8-methoxy-2,3,4,5-tetrahydrothiop[2,3-b]pyridine

[0176] Crude 4-(2-chloro-6-methoxypyridin-3-yl)butane-1-thiol hydrochloride (930.0 mg) was dissolved in tetrahydrofuran (20 mL) and slowly added dropwise to a suspension of sodium hydride (288.0 mg) in tetrahydrofuran (5 mL). The temperature was raised to 80°C and the reaction was allowed to proceed for 1 hour. TLC indicated the reaction was complete. The solution was then added dropwise to 30 mL of water in an ice-water bath and extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 830.0 mg of the crude product.

[0177] MS (ESI) m / z (M+H) + =196.1.

[0178] Step 4: Preparation of 7-bromo-8-methoxy-2,3,4,5-tetrahydrothiop[2,3-b]pyridine

[0179] Dissolve crude 8-methoxy-2,3,4,5-tetrahydrothiophene[2,3-b]pyridine (800.0 mg) in acetonitrile (9 mL), add N-bromosuccinimide (721.6 mg), and react at room temperature for 10 minutes. TLC indicates completion of the reaction. Add 30 mL of water to the reaction system, extract three times with ethyl acetate, combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 336.0 mg of the title compound.

[0180] MS (ESI) m / z (M+H) + =274.0,276.0.

[0181] Intermediate Preparation Example 9: Preparation of tert-Butyl 4-(2-Toluenesulfonylhydrazone)piperidine-1-carboxylate (can be used in Example 4)

[0182] Dissolve tert-butyl 4-oxopiperidine-1-carboxylate (4.5 g) in methanol (50 mL), add 4-methylbenzenesulfonylhydrazide (4.2 g), and heat at 70°C for 4 hours. Monitor the reaction by TLC until complete. Concentrate the reaction solution and purify the crude product by column chromatography to obtain 7.0 g of the title compound.

[0183] MS (ESI) m / z (M+H) + =368.1.

[0184] Intermediate Preparation Example 10: Preparation of 6-bromo-7-methoxy-3,4-dihydro-2H-pyrano[2,3-b]pyridine (can be used in Example 5)

[0185] The title compound was prepared using the corresponding common commercial reagents and the similar preparation methods as described in Intermediate Preparation Examples 6 and 7.

[0186] MS (ESI) m / z (M+H) + =244.0,246.0.

[0187] Intermediate Preparation Example 11: Preparation of 6-bromo-7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine (can be used in Example 6)

[0188] Step 1: Preparation of methyl 3-(2,6-dichloropyridin-3-yl)acrylate

[0189] In an ice-water bath under a nitrogen atmosphere, slowly add methyl 2-(dimethoxyphosphoryl)acetate (4.66 g) to anhydrous tetrahydrofuran (10 mL) containing sodium hydride (1.02 g) and allow to react for half an hour. Slowly add 2,6-dichloronicotinaldehyde (3.00 g) in anhydrous tetrahydrofuran (20 mL) dropwise to the system and allow to react at room temperature for 1 hour. LCMS indicates the reaction is complete. Add an appropriate amount of water and extract three times with ethyl acetate. The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 3.7 g of the title compound.

[0190] MS (ESI) m / z (M+H) + =231.9.

[0191] Step 2: Preparation of methyl 3-(2,6-dichloropyridin-3-yl)propionate

[0192] Dissolve methyl 3-(2,6-dichloropyridin-3-yl)acrylate (3.7 g) in ethanol (40 mL), add platinum dioxide (400 mg), replace the atmosphere with hydrogen three times, and allow to react at room temperature for 1 hour. LCMS indicates complete reaction. Filter the reaction mixture, collect the filtrate, and concentrate to yield 3.7 g of crude product.

[0193] MS (ESI) m / z (M+H) + =233.9.

[0194] Step 3: Preparation of 3-(2,6-dichloropyridin-3-yl)propan-1-ol

[0195] Under a nitrogen atmosphere at -20°C, methyl 3-(2,6-dichloropyridin-3-yl)propanoate (3.7 g) was dissolved in dichloromethane (20 mL). Diisobutylaluminum hydride (1.5 M, 21.09 mL) was slowly added and the mixture was allowed to react at room temperature for 1 hour. LCMS indicated complete reaction. The reaction mixture was quenched with saturated ammonium chloride solution (10 mL) and extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 2.2 g of the title compound.

[0196] MS (ESI) m / z (M+H) + =205.9.

[0197] Step 4: Preparation of S-(3-(2,6-dichloropyridin-3-yl)propyl)ethyl sulfate

[0198] Under a nitrogen atmosphere, 3-(2,6-dichloropyridin-3-yl)propan-1-ol (2.2 g) and triphenylphosphine (4.2 g) were dissolved in anhydrous tetrahydrofuran (20 mL). Diisopropyl azodicarboxylate (3.24 g) and thioacetic acid (1.22 g) were added sequentially. The mixture was allowed to react at room temperature for 1 hour. LCMS indicated the reaction was complete. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 2.0 g of the title compound.

[0199] MS (ESI) m / z (M+H) + =263.9.

[0200] Step 5: Preparation of 3-(2,6-dichloropyridin-3-yl)propane-1-thiol hydrochloride

[0201] Dissolve S-(3-(2,6-dichloropyridin-3-yl)propyl)ethyl sulfate (2.0 g) in 1,4-dioxane hydrochloride (4 M, 20 mL) and allow to react overnight at room temperature. LCMS indicated complete reaction. The reaction mixture was concentrated to yield 1.6 g of crude product.

[0202] MS (ESI) m / z (M+H) + =221.9.

[0203] Step 6: Preparation of 7-chloro-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine

[0204] Under a nitrogen atmosphere and an ice-water bath, slowly add a solution of 3-(2,6-dichloropyridin-3-yl)propane-1-thiol hydrochloride (1.6 g) in anhydrous N,N-dimethylformamide (10 mL) to a solution of sodium hydride (864.3 mg) in anhydrous N,N-dimethylformamide. Allow to react for 1 hour. LCMS indicates complete reaction of the starting material. The reaction mixture is quenched with saturated ammonium chloride solution and extracted three times with ethyl acetate. The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by column chromatography to yield 1.2 g of the title compound.

[0205] MS (ESI) m / z (M+H) + =186.1.

[0206] Step 7: Preparation of 7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine

[0207] Dissolve 7-chloro-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine (700 mg) and sodium methoxide (814.7 mg) in N-methylpyrrolidone (10 mL) and heat at 110°C for 15 hours. LCMS indicates complete reaction. Add an appropriate amount of water to the reaction solution and extract three times with ethyl acetate. The combined organic phases are dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 580 mg of the title compound.

[0208] MS (ESI) m / z (M+H) + =181.9.

[0209] Step 8: Preparation of 6-bromo-7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine

[0210] Dissolve 7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine (420 mg) in acetonitrile (4 mL), add N-bromosuccinimide (412.4 mg), and react at room temperature for 0.5 hours. LCMS indicates complete reaction. Add 30 mL of water to the reaction system, extract three times with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 580 mg of the title compound.

[0211] MS (ESI) m / z (M+H) + =260.1,262.1.

[0212] Intermediate Preparation Example 12: Preparation of 3-bromo-2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopenta[b]pyridine (can be used in Example 7)

[0213] Step 1: Preparation of 4-(6-methoxy-2-methylpyridin-3-yl)isoxazole

[0214] Under a nitrogen atmosphere, 3-bromo-6-methoxy-2-methylpyridine (10.0 g), potassium carbonate (10.25 g), and 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (2.54 g) were dissolved in 1,4-dioxane (70 mL) and water (20 mL). The temperature was raised to 90°C, and a solution of 4-isoxazoleboronic acid pinacol ester (9.65 g dissolved in 80 mL of 1,4-dioxane) was added dropwise over 3 hours. The reaction was continued for 1.5 hours. The reaction mixture was brought to room temperature, quenched with water, and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 7.0 g of the title compound.

[0215] MS (ESI) m / z (M+H) + =191.1.

[0216] Step 2: Preparation of 2-(6-methoxy-2-methylpyridin-3-yl)acetonitrile

[0217] Dissolve 4-(6-methoxy-2-methylpyridin-3-yl)isoxazole (7.0 g) in methanol (80 mL), add potassium fluoride (10.7 g), and heat in a sealed autoclave at 120°C for 4 hours. Cool the reaction to room temperature and concentrate to remove the solvent. Quench the reaction with water and extract three times with ethyl acetate. Combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The crude product is purified by column chromatography to obtain 5.1 g of the title compound.

[0218] MS (ESI) m / z (M+H) + =163.1.

[0219] Step 3: Preparation of 2-(6-methoxy-2-methylpyridin-3-yl)-2-methylpropionitrile

[0220] Under a nitrogen atmosphere, 2-(6-methoxy-2-methylpyridin-3-yl)acetonitrile (5.1 g) was dissolved in N,N-dimethylformamide (50 mL). Sodium hydride (3.78 g) was added at -20°C and the reaction was allowed to proceed for 10 minutes. Iodomethane (11.18 g) was added and the reaction was continued for 1.5 hours. The reaction was quenched by the addition of saturated ammonium chloride solution and extracted three times with ethyl acetate. The organic phases were combined, washed sequentially with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 5.8 g of the title compound.

[0221] MS (ESI) m / z (M+H) + =191.1.

[0222] Step 4: Preparation of 2-methoxy-5,5-dimethyl-5,7-dihydro-6H-cyclopenta[b]pyridin-6-one

[0223] Under a nitrogen atmosphere, 2-(6-methoxy-2-methylpyridin-3-yl)-2-methylpropionitrile (5.8 g) was dissolved in anhydrous tetrahydrofuran (60 mL). Potassium bis(trimethylsilyl)amide (1 M, 45 mL) was added dropwise at -40°C. The mixture was allowed to react at room temperature for 0.5 hours. The reaction was quenched by the addition of saturated ammonium chloride solution. The mixture was extracted three times with ethyl acetate. The organic phases were combined, washed sequentially with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 5.02 g of the title compound.

[0224] MS (ESI) m / z (M+H) + =192.1.

[0225] Step 5: Preparation of 2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopentadien[b]pyridin-6-ol

[0226] Dissolve 2-methoxy-5,5-dimethyl-5,7-dihydro-6H-cyclopenta[b]pyridin-6-one (4.3 g) in methanol (40 mL) and add sodium borohydride (1.08 g) in portions. Let the mixture react at room temperature for 1 hour. After completion of the reaction, monitor the reaction by LCMS and remove the solvent by concentration. Quench the reaction by adding saturated ammonium chloride solution and extract three times with ethyl acetate. The combined organic phases are washed sequentially with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by column chromatography to yield 4.1 g of the title compound.

[0227] MS (ESI) m / z (M+H) + =194.1.

[0228] Step 6: Preparation of 2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopentadienyl[b]pyridin-6-yl 4-methylbenzenesulfonate

[0229] Dissolve 2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopentadien[b]pyridin-6-ol (4.1 g) in 1,2-dichloroethane (30 mL). Add N-methylimidazole (3.48 g) and p-toluenesulfonyl chloride (5.26 g) sequentially. Heat at 70°C for 3 hours. Add water (80 mL) to stop the reaction. Extract three times with dichloromethane. Combine the organic phases, wash sequentially with water and saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The crude product is purified by column chromatography to yield 6.3 g of the title compound.

[0230] MS (ESI) m / z (M+H) + =348.1.

[0231] Step 7: Preparation of 2-methoxy-5,5-dimethyl-5H-cyclopentadienyl[b]pyridine

[0232] Dissolve 2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopentadienyl[b]pyridin-6-yl 4-methylbenzenesulfonate (6.3 g) in dimethyl sulfoxide (50 mL), add potassium tert-butoxide (4.07 g), and heat at 50°C for 2 hours. After the reaction, concentrate the system, quench with saturated ammonium chloride solution, and extract three times with ethyl acetate. The organic phases are combined, washed sequentially with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by column chromatography to yield 2.85 g of the title compound.

[0233] MS (ESI) m / z (M+H) + =176.1.

[0234] Step 8: Preparation of 2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopentadienyl[b]pyridine

[0235] Dissolve 2-methoxy-5,5-dimethyl-5H-cyclopentadien[b]pyridine (0.4 g) in methanol (10 mL), add palladium on carbon (10%, 100 mg), replace the air in the system, and heat at 50°C under a hydrogen atmosphere for 2 hours. After the reaction is complete, filter, collect the filtrate, and concentrate to obtain 400 mg of the crude product.

[0236] MS (ESI) m / z (M+H) + =178.1.

[0237] Step 9: Preparation of 3-bromo-2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopenta[b]pyridine

[0238] Dissolve 2-methoxy-5,5-dimethyl-6,7-dihydro-5H-cyclopentadien[b]pyridine (0.4 g) in N,N-dimethylformamide (8 mL), add N-bromosuccinimide (407 mg), and heat at 50°C for 1 hour. After the reaction, add an appropriate amount of water and extract three times with ethyl acetate. The organic phases are combined, washed sequentially with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by column chromatography to yield 520 mg of the title compound.

[0239] MS (ESI) m / z (M+H) + =256.1,258.1.

[0240] Intermediate Preparation Example 13: Preparation of 5-bromo-6-methoxy-2,3-dihydrothieno[2,3-b]pyridine (can be used in Example 8)

[0241] Step 1: Preparation of 2,6-dichloro-3-(2-methoxyvinyl)pyridine

[0242] In an ice-water bath and nitrogen atmosphere, dissolve (methoxymethyl)triphenylphosphonium chloride (20.0 g) in tetrahydrofuran (160 mL) and add potassium tert-butoxide (6.53 g in 58.3 mL of tetrahydrofuran). Allow to react for 30 minutes. Add 2,6-dichloronicotinaldehyde (6.0 g in 50 mL of tetrahydrofuran) dropwise and allow to react for 20 minutes. Bring the mixture to room temperature and continue the reaction for 1 hour. Quench the reaction with water, extract three times with ethyl acetate, combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The crude product is purified by column chromatography to yield 6.1 g of the title compound.

[0243] MS (ESI) m / z (M+H) + =204.1.

[0244] Step 2: Preparation of 2-(2,6-dichloropyridin-3-yl)acetaldehyde

[0245] Dissolve 2,6-dichloro-3-(2-methoxyvinyl)pyridine (6.1 g) in tetrahydrofuran (41 mL) and add aqueous hydrogen chloride (4.0 M, 24 mL). Heat the mixture under reflux for 3 hours. Cool the reaction mixture to room temperature and concentrate to remove the solvent. Dissolve the residue in ethyl acetate and wash with saturated sodium bicarbonate solution and saturated sodium chloride solution, sequentially. Dry over anhydrous sodium sulfate, filter, and concentrate to obtain 6.1 g of crude product.

[0246] MS (ESI) m / z (M+H) + =190.1.

[0247] Step 3: Preparation of 2-(2,6-dichloropyridin-3-yl)ethanol

[0248] In an ice-water bath, dissolve the crude 2-(2,6-dichloropyridin-3-yl)acetaldehyde (6.1 g) in methanol (51 mL). Add sodium borohydride (1.0 g) and allow to react for 30 minutes. Quench with saturated ammonium chloride solution, concentrate to remove methanol, and extract three times with ethyl acetate. The combined organic phases are dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is purified by column chromatography to yield 2.3 g of the title compound.

[0249] MS (ESI) m / z (M+H) + =192.1.

[0250] Step 4: Preparation of S-(2-(2,6-dichloropyridin-3-yl)ethyl)ethyl sulfate

[0251] In an ice-water bath, dissolve 2-(2,6-dichloropyridin-3-yl)ethanol (2.3 g) and triphenylphosphine (6.3 g) in anhydrous tetrahydrofuran (60 mL). Add diethyl azodicarboxylate (3.8 mL). After 10 minutes, add thioacetic acid (1.72 mL). Bring to room temperature and react for 2 hours. Dilute the reaction solution with n-hexane, filter to remove the precipitate, collect the filtrate, concentrate, and purify the crude product by column chromatography to obtain 2.5 g of the title compound.

[0252] MS (ESI) m / z (M+H) + =250.1.

[0253] Step 5: Preparation of 2-(2,6-dichloropyridin-3-yl)ethane-1-thiol

[0254] Dissolve S-(2-(2,6-dichloropyridin-3-yl)ethyl)ethyl sulfate (1.5 g) in methanol (40 mL) and add acetyl chloride (2.36 g). Allow to react at room temperature for 24 hours. After completion of the reaction, monitor by LCMS, and concentrate to remove the solvent to obtain 1.6 g of a crude product.

[0255] MS (ESI) m / z (M+H) + =208.1.

[0256] Step 6: Preparation of 6-chloro-2,3-dihydrothieno[2,3-b]pyridine

[0257] Under a nitrogen atmosphere and in an ice-water bath, crude 2-(2,6-dichloropyridin-3-yl)ethane-1-thiol (1.6 g) was dissolved in N,N-dimethylformamide (30 mL). Sodium hydride (720 mg) was added and allowed to react for 1.5 hours. Water (80 mL) was added to stop the reaction. The mixture was extracted three times with ethyl acetate. The organic phases were combined, washed sequentially with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 1.0 g of the title compound.

[0258] MS (ESI) m / z (M+H) + =172.1.

[0259] Step 7: Preparation of 6-methoxy-2,3-dihydrothieno[2,3-b]pyridine

[0260] Dissolve 6-chloro-2,3-dihydrothieno[2,3-b]pyridine (1.0 g) and sodium methoxide (1.08 g) in methanol (15 mL) and heat at 115°C for 8 hours using microwave initiation. After the reaction, concentrate the system and purify the crude product by column chromatography to obtain 730 mg of the title compound.

[0261] MS (ESI) m / z (M+H) + =168.1.

[0262] Step 8: Preparation of 5-bromo-6-methoxy-2,3-dihydrothieno[2,3-b]pyridine

[0263] Dissolve 6-methoxy-2,3-dihydrothieno[2,3-b]pyridine (0.7 g) in N,N-dimethylformamide (10 mL) and add N-bromosuccinimide (0.8 g). Allow to react for 1 hour. After completion of the reaction, add water and extract three times with ethyl acetate. Wash with water and then with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The crude product is purified by column chromatography to yield 910 mg of the title compound.

[0264] MS (ESI) m / z (M+H) + =246.1,248.1.

[0265] Preparation Example 14: Preparation of (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one trifluoroacetate (can be used in Example 10)

[0266] Step 1: Preparation of (R)-2-((tert-Butoxycarbonyl)amino)-3-(7-methyl-1H-indazol-5-yl)propanoic acid

[0267] Methyl (R)-2-((tert-Butoxycarbonyl)amino)-3-(7-methyl-1H-indazol-5-yl)propanoate (100 mg) was dissolved in tetrahydrofuran (3 mL) and water (1 mL), and sodium hydroxide (120 mg) was added. The mixture was allowed to react at room temperature overnight. After completion of the reaction, the pH of the system was adjusted to 5-6 with an appropriate amount of hydrochloric acid (0.5 N). The mixture was extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 90.0 mg of a crude product.

[0268] MS (ESI) m / z (M+H) + =320.1.

[0269] Step 2: Preparation of tert-butyl (R)-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)carbamate

[0270] (R)-2-((tert-Butoxycarbonyl)amino)-3-(7-methyl-1H-indazol-5-yl)propanoic acid (50 mg), 1-(1-methylpiperidin-4-yl)piperazine (57.18 mg), 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (60.02 mg), and N,N-diisopropylethylamine (40.32 mg) were dissolved in acetonitrile (10 mL) and reacted at room temperature for 4 hours. After completion of the reaction, the crude product was concentrated and purified by column chromatography to obtain 30 mg of the title compound.

[0271] MS (ESI) m / z (M+H) + =485.2.

[0272] Step 3: Preparation of (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one trifluoroacetate

[0273] Dissolve tert-butyl (R)-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)carbamate (30 mg) in dichloromethane (1 mL) and trifluoroacetic acid (0.5 mL) and react at room temperature for 1 hour. After the reaction, concentrate to obtain the crude product, which is used in the next reaction without purification.

[0274] MS (ESI) m / z (M+H) + =385.2.

[0275] Preparation Example 15: Preparation of 3-(piperidin-4-yl)-5,7-dihydrothieno[3,4-b]pyridin-2(1H)-one hydrochloride (can be used in Example 11)

[0276] Step 1: Preparation of 6-bromo-2-fluoro-3-iodopyridine

[0277] Dissolve 6-bromo-2-fluoropyridine (10.0 g) in tetrahydrofuran (70 mL), replace the nitrogen atmosphere, and cool to -78°C. Add lithium diisopropylamide (34.3 mL, 68.6 mmol) dropwise at this temperature. After complete addition, react at -78°C for 1 hour. Add a solution of iodine (14.5 g) in tetrahydrofuran (50 mL) and complete the reaction at -78°C for 1.5 hours. Quench the reaction by adding 10 mL of water at -60°C, bring to room temperature, and add the mixture to 100 mL of water. Extract three times with ethyl acetate. Combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, and filter. Concentrate. The resulting crude product is purified by column chromatography to yield 8.2 g of the title compound.

[0278] MS (ESI) m / z (M+H) + =301.8.

[0279] Step 2: Preparation of 6-bromo-3-iodo-2-methoxypyridine

[0280] Dissolve 6-bromo-2-fluoro-3-iodopyridine (8.2 g) in methanol (100 mL), add sodium methoxide (4.4 g), and heat to 45°C for 1 hour. TLC indicates completion of the reaction. Bring the reaction system to room temperature, add 500 mL of water, and extract three times with ethyl acetate. Combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 8.4 g of the title compound.

[0281] MS (ESI) m / z (M+H) + =313.9.

[0282] Step 3: Preparation of tert-butyl 6-bromo-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate

[0283] Under a nitrogen atmosphere, 6-bromo-3-iodo-2-methoxypyridine (8.3 g), N-tert-butyloxycarbonyl-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (7.8 g), and potassium carbonate (7.3 g) were dissolved in 1,4-dioxane (100 mL) and water (10 mL). 1,1-Bis(diphenylphosphino)diphenylferric palladium chloride (1.9 g) was added and the mixture was allowed to react at 70°C overnight. TLC indicated the reaction was complete. 100 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 7.3 g of the title compound.

[0284] MS (ESI) m / z (M+H) + =369.1.

[0285] Step 4: Preparation of tert-butyl 6-(diisopropylcarbamoyl)-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate

[0286] 6-Bromo-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylic acid tert-butyl ester (1.45 g) was placed in a 100 mL autoclave, and toluene (15 mL), diisopropylamine (1.2 g), triethylamine (2.0 g), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (445.0 mg), and 1,1-bis(diphenylphosphino)diphenylferric palladium dichloride (322.0 mg) were added. The autoclave was sealed, the atmosphere replaced with nitrogen, and then carbon monoxide was introduced. The pressure was maintained at 13 atmospheres, and the temperature was raised to 90°C for 10 hours. TLC indicated that the reaction was complete. 50 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 1.5 g of the title compound.

[0287] MS (ESI) m / z (M+H) + =418.3.

[0288] Step 5: Preparation of tert-butyl 4-(6-(diisopropylcarbamoyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0289] Dissolve tert-butyl 6-(diisopropylcarbamoyl)-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate (6.0 g) in methanol (100 mL). Add palladium on carbon (0.6 g). After nitrogen replacement, continue bubbling with hydrogen and allow to react at room temperature for 1 hour. TLC indicates the reaction is complete. Filter and concentrate to obtain 6.0 g of crude product.

[0290] MS (ESI) m / z (M+H) + =420.3.

[0291] Step 6: Preparation of ethyl 5-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-(diisopropylcarbamoyl)-6-methoxynicotinate

[0292] Dissolve tert-butyl 4-(6-(diisopropylcarbamoyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (5.8 g) in anhydrous tetrahydrofuran (60 mL). After replacing the nitrogen atmosphere, cool the mixture to -78°C and add a 2.5 M hexane solution of n-butyllithium (6.6 mL) dropwise. Allow to react for 0.5 hour. Slowly add the above solution dropwise to a tetrahydrofuran solution of ethyl chloroformate (7.5 g) (40 mL) at -78°C and allow to react for 0.5 hour. TLC indicates completion of the reaction. The mixture is brought to room temperature and added to 50 mL of water. Extract the mixture three times with ethyl acetate. Combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 3.2 g of the title compound.

[0293] MS (ESI) m / z (M+H) + =492.3.

[0294] Step 7: Preparation of tert-butyl 4-(6-(diisopropylcarbamoyl)-5-(hydroxymethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0295] In an ice-water bath, ethyl 5-(1-(tert-butoxycarbonyl)piperidin-4-yl)-2-(diisopropylcarbamoyl)-6-methoxynicotinate (1.0 g) was dissolved in anhydrous tetrahydrofuran (5 mL) and slowly added dropwise to a suspension of lithium aluminum tetrahydride (77 mg) in anhydrous tetrahydrofuran (5 mL). The mixture was allowed to react for 20 minutes. TLC indicated the reaction was complete. In an ice-water bath, 77 μL of water, 77 μL of 15% aqueous NaOH, and 231 μL of water were added sequentially. The mixture was stirred at room temperature for 10 minutes. An appropriate amount of anhydrous sodium sulfate was added, and the mixture was stirred at room temperature for 10 minutes. Filtration and concentration afforded 0.9 g of the crude product.

[0296] MS (ESI) m / z (M+H) + =450.3.

[0297] Step 8: Preparation of tert-butyl 4-(2-methoxy-7-oxo-5,7-dihydrofuro[3,4-b]pyridin-3-yl)piperidine-1-carboxylate

[0298] Crude tert-butyl 4-(6-(diisopropylcarbamoyl)-5-(hydroxymethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (0.9 g) was dissolved in acetic acid (9 mL) and the temperature was raised to 90°C for 2 hours. TLC indicated the reaction was complete, and the system was concentrated. The crude product was purified by column chromatography to afford 502.0 mg of the title compound.

[0299] MS (ESI) m / z (M+H) + =349.2.

[0300] Step 9: Preparation of tert-butyl 4-(5,6-bis(hydroxymethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0301] Dissolve tert-butyl 4-(2-methoxy-7-oxo-5,7-dihydrofuro[3,4-b]pyridin-3-yl)piperidine-1-carboxylate (300.0 mg) in methanol (3 mL), add lithium borohydride (56.0 mg), and heat to 50°C for 2 hours. TLC indicates completion of the reaction. Adjust the pH of the reaction solution to neutral with 1M dilute hydrochloric acid in an ice-water bath. Concentrate the solution, and purify the crude product by column chromatography to obtain 210.0 mg of the title compound.

[0302] MS (ESI) m / z (M+H) + =353.2.

[0303] Step 10: Preparation of a mixture of tert-butyl 4-(5-(chloromethyl)-2-methoxy-6-((methylsulfonyl)oxymethyl)pyridin-3-yl)piperidine-1-carboxylate

[0304] Methanesulfonyl chloride (195.0 mg) was dissolved in dichloromethane (2 mL). A mixture of tert-butyl 4-(5,6-bis(hydroxymethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (150.0 mg) and N,N-diisopropylethylamine (219.0 mg) in dichloromethane (2 mL) was added under ice-water bath conditions. The mixture was stirred for 20 minutes and then allowed to react at room temperature for 1 hour. TLC indicated the reaction was complete. The system was concentrated and the crude product was purified by column chromatography to yield 180.0 mg of the mixture.

[0305] MS (ESI) m / z (M+H) + =449.1 and 389.1.

[0306] Step 11: Preparation of tert-butyl 4-(2-methoxy-5,7-dihydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxylate

[0307] Under a nitrogen atmosphere, a mixture of tert-butyl 4-(5-(chloromethyl)-2-methoxy-6-((methylsulfonyl)oxymethyl)pyridin-3-yl)piperidine-1-carboxylate (200.0 mg) was dissolved in N,N-dimethylformamide (5 mL). A suspension of sodium sulfide (37.0 mg) in N,N-dimethylformamide (2 mL) was slowly added dropwise. The mixture was allowed to react at room temperature for 1 hour. TLC indicated the reaction was complete. 30 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed twice with purified water and once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography to yield 67.0 mg of the title compound.

[0308] MS (ESI) m / z (M+H) + =351.2.

[0309] Step 12: Preparation of 3-(piperidin-4-yl)-5,7-dihydrothieno[3,4-b]pyridin-2(1H)-one hydrochloride

[0310] Dissolve tert-butyl 4-(2-methoxy-5,7-dihydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxylate (67.0 mg) in a 4M solution of hydrochloric acid in 1,4-dioxane (5 mL) and heat to 90°C for 3 hours. TLC indicated the reaction was complete, and the system was concentrated to yield 51.0 mg of the crude product.

[0311] MS (ESI) m / z (M+H) + =237.1.

[0312] Preparation Example 16: Preparation of 7-(piperidin-4-yl)-3,4-dihydro-2H-thiopyrano[3,2-b]pyridin-6(5H)-one hydrochloride (can be used in Example 12)

[0313] Step 1: Preparation of tert-butyl (E)-6-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-methoxy-3',6'-dihydro-[3,4'-bipyridine]-1'(2'H)-carboxylate

[0314] Under a nitrogen atmosphere, tert-butyl 6-bromo-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate (2.0 g), ethyl (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (1.23 g), potassium carbonate (1.11 g), and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (382 mg) were dissolved in 1,4-dioxane (15 mL) and water (1.5 mL) and reacted at 90°C for 3 hours. After the reaction, an appropriate amount of water was added to the system, and the mixture was extracted with ethyl acetate (200 mL). The organic phases were combined, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 2.0 g of the title compound.

[0315] MS (ESI) m / z (M+H) + =389.1.

[0316] Step 2: Preparation of tert-butyl 4-(6-(3-ethoxy-3-oxopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0317] Dissolve tert-butyl (E)-6-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-methoxy-3',6'-dihydro-[3,4'-bipyridine]-1'(2'H)-carboxylate (2.0 g) in methanol (30 mL), add 10% palladium on carbon (0.15 g), and heat at 55°C under a hydrogen atmosphere for 1 hour. After the reaction, remove the palladium on carbon by filtration, wash the solid with methanol (100 mL), combine the filtrates, and concentrate under reduced pressure to obtain 2.0 g of the crude product. MS (ESI) m / z (M+H) + =393.1.

[0318] Step 3: Preparation of tert-butyl 4-(5-bromo-6-(3-ethoxy-3-oxopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0319] Dissolve tert-butyl 4-(6-(3-ethoxy-3-oxopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (850 mg) in N,N-dimethylformamide (8 mL), add N-bromosuccinimide (386 mg), and allow to react at room temperature for 2 hours. After the reaction, add an appropriate amount of water and extract with ethyl acetate (100 mL). The organic phases are combined, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 1.1 g of crude product, which is used directly in the next step.

[0320] MS (ESI) m / z (M+H) + =471.1.

[0321] Step 4: Preparation of tert-butyl 4-(5-bromo-6-(3-hydroxypropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0322] Crude tert-butyl 4-(5-bromo-6-(3-ethoxy-3-oxopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (1.1 g) was dissolved in methanol (20 mL), and lithium borohydride (1.08 g) was added portionwise. The reaction was allowed to react at room temperature for 3 hours. After completion of the reaction, as monitored by LCMS, the solvent was removed by concentration, and an appropriate amount of saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 770 mg of the title compound.

[0323] MS (ESI) m / z (M+H) + =429.1.

[0324] Step 5: Preparation of tert-butyl 4-(6-(3-(acetylthio)propyl)-5-bromo-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0325] Under a nitrogen atmosphere and in an ice-water bath, tert-butyl 4-(5-bromo-6-(3-hydroxypropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (300 mg) and triphenylphosphine (368 mg) were dissolved in tetrahydrofuran (10 mL). Diethyl azodicarboxylate (284 mg) was added. After 10 minutes, thioacetic acid (106 mg) was added, and the mixture was allowed to react at room temperature for 1.5 hours. An appropriate amount of hexane was added to dilute the reaction, and the white precipitate was filtered off. The filtrate was collected, concentrated, and purified by column chromatography to obtain 330 mg of the title compound.

[0326] MS (ESI) m / z (M+H) + =487.1.

[0327] Step 6: Preparation of tert-butyl 4-(6-methoxy-3,4-dihydro-2H-thiopyrano[3,2-b]pyridin-7-yl)piperidine-1-carboxylate

[0328] Under a nitrogen atmosphere, tert-butyl 4-(6-(3-(acetylthio)propyl)-5-bromo-2-methoxypyridin-3-yl)piperidine-1-carboxylate (330 mg), cesium carbonate (443 mg), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (23 mg), and tris(dibenzylideneacetone)dipalladium (27 mg) were dissolved in 1,4-dioxane (3 mL) and heated at 100°C for 12 hours. After the reaction, an appropriate amount of water was added to the system, and extraction was performed with ethyl acetate (20 mL). The organic phases were combined, backwashed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 210 mg of the title compound.

[0329] MS (ESI) m / z (M+H) + =365.1.

[0330] Step 7: Preparation of 7-(piperidin-4-yl)-3,4-dihydro-2H-thiopyrano[3,2-b]pyridin-6(5H)-one hydrochloride

[0331] Dissolve tert-butyl 4-(6-methoxy-3,4-dihydro-2H-thiopyrano[3,2-b]pyridin-7-yl)piperidine-1-carboxylate (210 mg) in a 4.0 M hydrogen chloride / 1,4-dioxane solution (5 mL) and react at 90°C for 5 hours. Cool the reaction to room temperature and remove the solvent in vacuo to obtain 230 mg of a crude product.

[0332] MS (ESI) m / z (M+H) + =251.1.

[0333] Preparation Example 17: Preparation of 3-(piperidin-4-yl)-5,8-dihydro-1H-thiopyrano[3,4-b]pyridin-2(6H)-one hydrochloride (can be used in Example 13)

[0334] Step 1: Preparation of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate

[0335] To a suspension of sodium hydride (1.45 g) in anhydrous tetrahydrofuran (15 mL) under a nitrogen atmosphere and in an ice-water bath, slowly add a solution of dimethyl 3,3'-thiodipropionate (5.0 g) in anhydrous tetrahydrofuran (30 mL) dropwise. The mixture was allowed to react at room temperature overnight. LCMS indicated complete reaction. An appropriate amount of saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography to afford 3.65 g of the title compound.

[0336] MS (ESI) m / z (M+H) + =175.0.

[0337] Step 2: Preparation of methyl 4-(((trifluoromethyl)sulfonyl)oxy)-5,6-dihydro-2H-thiopyran-3-carboxylate

[0338] Under a nitrogen atmosphere and in an ice-water bath, slowly add a solution of methyl 4-oxotetrahydro-2H-thiopyran-3-carboxylate (3.65 g) in anhydrous tetrahydrofuran (30 mL) to a suspension of sodium hydride (1.09 g) in anhydrous tetrahydrofuran (10 mL). The mixture is allowed to react at room temperature for 1 hour. A solution of 1,1,1-trifluoro-N-phenyl-N-(trifluoromethyl)sulfonyl)methanesulfonamide (8.23 g) in anhydrous tetrahydrofuran (20 mL) is added and the mixture is allowed to react at room temperature overnight. LCMS indicates complete reaction of the starting material. The reaction is quenched by adding an appropriate amount of saturated ammonium chloride solution. The mixture is extracted three times with ethyl acetate, and the organic phases are combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 5.2 g of the title compound.

[0339] MS (ESI) m / z (M+H) + =306.9.

[0340] Step 3: Preparation of tert-butyl (E)-4-(2-(5-(methoxycarbonyl)-3,6-dihydro-2H-thiopyran-4-yl)vinyl)piperidine-1-carboxylate

[0341] Under a nitrogen atmosphere, tert-butyl 4-ethynylpiperidine-1-carboxylate (1.0 g) was dissolved in anhydrous tetrahydrofuran (4 mL). A solution of catecholborane (0.69 g) in anhydrous tetrahydrofuran (4 mL) was slowly added. The mixture was reacted at 70°C for 4 hours and then cooled to room temperature. Methyl 4-(((trifluoromethyl)sulfonyl)oxy)-5,6-dihydro-2H-thiopyran-3-carboxylate (1.5 g), 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) chloride (183 mg), and potassium carbonate (2.03 g) were dissolved in dioxane (10 mL) and water (1 mL) and slowly added dropwise to the reaction mixture. The temperature was raised to 70°C and the reaction was continued for 1 hour. LCMS indicated that the reaction was complete. The reaction mixture was filtered, the filtrate was collected, and concentrated. The crude product was purified by column chromatography to obtain 1.3 g of the title compound.

[0342] MS (ESI) m / z (M+H-100) + =267.9.

[0343] Step 4: Preparation of (E)-4-(2-(1-(tert-butoxycarbonyl)piperidin-4-yl)vinyl)-5,6-dihydro-2H-thiopyran-3-carboxylic acid

[0344] Dissolve tert-butyl (E)-4-(2-(5-(methoxycarbonyl)-3,6-dihydro-2H-thiopyran-4-yl)vinyl)piperidine-1-carboxylate (230 mg) in methanol (2 mL) and water (0.5 mL), add lithium hydroxide monohydrate (131.4 mg), and allow to react overnight. LCMS indicates the reaction is complete. The reaction solution is neutralized with 2M hydrochloric acid and extracted three times with ethyl acetate. The organic phases are combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 200 mg of the title compound.

[0345] MS (ESI) m / z (M+H-100) + =253.9.

[0346] Step 5: Preparation of tert-butyl 4-(2-oxo-2,5,6,8-tetrahydro-1H-thiopyrano[3,4-b]pyridin-3-yl)piperidine-1-carboxylate

[0347] Under a nitrogen atmosphere, (E)-4-(2-(1-(tert-Butoxycarbonyl)piperidin-4-yl)vinyl)-5,6-dihydro-2H-thiopyran-3-carboxylic acid (200 mg) and diphenylphosphoryl azide (155.7 mg) were dissolved in anhydrous toluene (3 mL). Triethylamine (85.9 mg) was added and the mixture was allowed to react at room temperature for 2 hours. The temperature was raised to 80°C and the reaction was continued for 1 hour. LCMS indicated that the starting material had reacted completely. The reaction solution was concentrated, and the crude product was purified by column chromatography to obtain 100 mg of the title compound.

[0348] MS (ESI) m / z (M+H-100) + =250.9.

[0349] Step 6: Preparation of 3-(piperidin-4-yl)-5,8-dihydro-1H-thiopyrano[3,4-b]pyridin-2(6H)-one hydrochloride

[0350] Dissolve tert-butyl 4-(2-oxo-2,5,6,8-tetrahydro-1H-thiopyrano[3,4-b]pyridin-3-yl)piperidine-1-carboxylate (100 mg) in dichloromethane (2 mL). Add a 4M hydrochloric acid / 1,4-dioxane solution (1 mL). Allow to react at room temperature for 1 hour. LCMS indicated complete reaction. The reaction mixture was concentrated to yield 80 mg of crude product, which was used directly in the next step.

[0351] MS (ESI) m / z (M+H) + =250.9.

[0352] Preparation Example 18: Preparation of 3-(piperidin-4-yl)-5,7-dihydrothieno[3,4-b]pyridin-2(1H)-one-7,7-d2 Hydrochloride (can be used in Example 14)

[0353] Step 1: Preparation of tert-butyl 4-(5-(hydroxymethyl)-6-(hydroxymethyl-d2)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0354] Under ice-water conditions, tert-butyl 4-(2-methoxy-7-oxo-5,7-dihydrofuro[3,4-b]pyridin-3-yl)piperidine-1-carboxylate (200.0 mg) was dissolved in anhydrous tetrahydrofuran (1 mL) and slowly added dropwise to a suspension of lithium aluminum deuterate (24.0 mg) in anhydrous tetrahydrofuran (1 mL). The mixture was allowed to react for 20 minutes. TLC indicated the reaction was complete. 24 μL of water, 24 μL of 15% aqueous NaOH, and 72 μL of water were added sequentially. The mixture was stirred at room temperature for 10 minutes. An appropriate amount of anhydrous sodium sulfate was added, and the mixture was stirred at room temperature for 10 minutes. The filtrate was filtered, collected, and concentrated to yield 166.0 mg of the crude product.

[0355] MS (ESI) m / z (M+H) + =355.2.

[0356] The title compound was prepared by using the corresponding common commercial reagents and subsequently using a preparation method similar to the above intermediate preparation example 15.

[0357] MS (ESI) m / z (M+H) + =239.1.

[0358] Preparation Example 19: Preparation of 3-(piperidin-4-yl)-1,5,7,8-tetrahydro-2H-thiopyrano[4,3-b]pyridin-2-one hydrochloride (can be used in Example 15)

[0359] Step 1: Preparation of tert-butyl 2-methoxy-6-methyl-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate

[0360] Under a nitrogen atmosphere, tert-butyl 6-bromo-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate (3 g) was dissolved in 1,4-dioxane (30 mL) and water (3 mL). Methylboric acid (1.5 g), potassium carbonate (2.3 g), and [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride (0.53 g) were added and reacted at 80°C for 2 hours. LCMS indicated the reaction was complete. The reaction mixture was brought to room temperature, a small amount of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 2.4 g of the title compound.

[0361] MS (ESI) m / z (M+H) + =305.1.

[0362] Step 2: Preparation of tert-butyl 4-(2-methoxy-6-methylpyridin-3-yl)piperidine-1-carboxylate

[0363] Dissolve tert-butyl 2-methoxy-6-methyl-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate (2.4 g) in methanol (30 mL), add palladium on carbon (100 mg), and purge the reaction system with hydrogen. Allow to react at room temperature for 2 hours. LCMS indicates the reaction is complete. Filter the reaction solution, collect the filtrate, and remove excess solvent by rotary evaporation. The resulting crude product is purified by column chromatography to yield 2.3 g of the title compound.

[0364] MS (ESI) m / z (M+H) + =307.1.

[0365] Step 3: Preparation of tert-butyl 4-(5-bromo-2-methoxy-6-methylpyridin-3-yl)piperidine-1-carboxylate

[0366] Dissolve tert-butyl 4-(2-methoxy-6-methylpyridin-3-yl)piperidine-1-carboxylate (2.3 g) in N,N-dimethylformamide (25 mL), add N-bromosuccinimide (1.6 g), and react at room temperature for 1 hour. LCMS indicates the reaction is complete. Add a small amount of water and extract three times with ethyl acetate. The combined organic phases are backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 2.6 g of the title compound.

[0367] MS (ESI) m / z (M+H) + =385.0.

[0368] Step 4: Preparation of tert-butyl 4-(5-bromo-2-methoxy-6-(2-methoxy-2-oxoethyl)pyridin-3-yl)piperidine-1-carboxylate

[0369] Under a nitrogen atmosphere at -78°C, dissolve tert-butyl 4-(5-bromo-2-methoxy-6-methylpyridin-3-yl)piperidine-1-carboxylate (2.6 g) in tetrahydrofuran (30 mL). Slowly add lithium diisopropylamide (1 M, 10.2 mL) dropwise and stir for 20 minutes. Dissolve dimethyl carbonate (1.3 g) in tetrahydrofuran (10 mL) and slowly add dropwise to the reaction mixture. Stir for 20 minutes. TLC indicates completion of the reaction. Bring the reaction mixture to room temperature, add an appropriate amount of saturated aqueous ammonium chloride, and extract three times with ethyl acetate. Combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 90 mg of the title compound.

[0370] MS (ESI) m / z (M+H) + =443.1.

[0371] Step 5: Preparation of methyl 5-(1-(tert-butoxycarbonyl)piperidin-4-yl)-6-methoxy-2-(2-methoxy-2-oxoethyl)nicotinate

[0372] Dissolve tert-butyl 4-(5-bromo-2-methoxy-6-(2-methoxy-2-oxoethyl)pyridin-3-yl)piperidine-1-carboxylate (1 g) in toluene (10 mL) and methanol (1 mL). Add triethylamine (0.7 g), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (0.14 g), and [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride (0.16 g). The reaction system is saturated with carbon monoxide and allowed to react at 100°C for 10 hours. LCMS indicates completion of the reaction. The reaction mixture is brought to room temperature, a small amount of water is added, and the mixture is extracted three times with ethyl acetate. The organic phases are combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 750 mg of the title compound.

[0373] MS (ESI) m / z (M+H) + =423.1.

[0374] Step 6: Preparation of tert-butyl 4-(6-(2-hydroxyethyl)-5-(hydroxymethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0375] Methyl 5-(1-(tert-butoxycarbonyl)piperidin-4-yl)-6-methoxy-2-(2-methoxy-2-oxoethyl)nicotinate (750 mg) was dissolved in tetrahydrofuran (10 mL) under ice-water conditions. Lithium borohydride (112 mg) was added and the mixture was allowed to react at room temperature for 1 hour. LCMS indicated the reaction was complete. An appropriate amount of saturated aqueous ammonium chloride was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography to yield 560 mg of the title compound.

[0376] MS (ESI) m / z (M+H) + =367.1.

[0377] Step 7: Preparation of tert-butyl 4-(5-(chloromethyl)-2-methoxy-6-(2-((methylsulfonyl)oxy)ethyl)pyridin-3-yl)piperidine-1-carboxylate

[0378] Dissolve tert-butyl 4-(6-(2-hydroxyethyl)-5-(hydroxymethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (560 mg) in dichloromethane (8 mL) in an ice-water bath. Add triethylamine (455 mg) and methanesulfonyl chloride (431 mg). Allow to react at room temperature for 1 hour. LCMS indicates the reaction is complete. The reaction solution is rotary evaporated to remove excess solvent, and the resulting crude product is purified by column chromatography to yield 620 mg of the title compound.

[0379] MS (ESI) m / z (M+H) + =463.0.

[0380] Step 8: Preparation of tert-butyl 4-(2-methoxy-7,8-dihydro-5H-thiopyrano[4,3-b]pyridin-3-yl)piperidine-1-carboxylate

[0381] At room temperature, tert-butyl 4-(5-(chloromethyl)-2-methoxy-6-(2-((methylsulfonyl)oxy)ethyl)pyridin-3-yl)piperidine-1-carboxylate (180 mg) was dissolved in ethanol (20 mL), sodium sulfide (70 mg) was added, and the mixture was allowed to react at 80°C for 0.5 hours. LCMS indicated the reaction was complete. The reaction solution was rotary evaporated to remove excess solvent, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography to afford 110 mg of the title compound.

[0382] MS (ESI) m / z (M+H) + =365.1.

[0383] Step 9: Preparation of 3-(piperidin-4-yl)-1,5,7,8-tetrahydro-2H-thiopyrano[4,3-b]pyridin-2-one hydrochloride

[0384] At room temperature, tert-butyl 4-(2-methoxy-7,8-dihydro-5H-thiopyrano[4,3-b]pyridin-3-yl)piperidine-1-carboxylate (100.0 mg, 0.3 mmol) was dissolved in 4N dioxane hydrochloride (1 ml). The reaction was incubated at 90°C for 1 hour. LCMS analysis indicated a complete reaction. The reaction solution was directly spin-dried to obtain a pale yellow solid (110 mg, crude).

[0385] MS (ESI) m / z (M+H) + =251.0.

[0386] Preparation Example 20: Preparation of 6-(piperidin-4-yl)-2,3-dihydrothieno[3,2-b]pyridin-5(4H)-one hydrochloride (can be used in Example 17)

[0387] Step 1: Preparation of tert-butyl 4-(5-bromo-6-(2-hydroxyethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0388] Tert-butyl 4-(5-bromo-2-methoxy-6-(2-methoxy-2-oxoethyl)pyridin-3-yl)piperidine-1-carboxylate (120 mg) was dissolved in methanol (5 mL), and lithium borohydride (36 mg) was added portionwise. The mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, as monitored by LCMS, the solvent was removed in vacuo, and an appropriate amount of saturated ammonium chloride solution was added to quench the mixture. The mixture was extracted three times with ethyl acetate, and the organic phases were combined, backwashed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 100 mg of the title compound.

[0389] MS (ESI) m / z (M+H) + =415.1.

[0390] Step 2: Preparation of tert-butyl 4-(6-(2-(acetylthio)ethyl)-5-bromo-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0391] Under a nitrogen atmosphere and in an ice-water bath, tert-butyl 4-(5-bromo-6-(2-hydroxyethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (100 mg) and triphenylphosphine (131 mg) were dissolved in anhydrous tetrahydrofuran (5 mL). Diethyl azodicarboxylate (101 mg) was added. After 10 minutes, thioacetic acid (38 mg) was added and the mixture was allowed to react at room temperature for 1.5 hours. An appropriate amount of hexane was added to dilute the reaction, the white precipitate was filtered, and the filtrate was collected, concentrated, and purified by column chromatography to give 80 mg of the title compound.

[0392] MS (ESI) m / z (M+H) + =473.1.

[0393] Step 3: Preparation of tert-butyl 4-(5-methoxy-2,3-dihydrothieno[3,2-b]pyridin-6-yl)piperidine-1-carboxylate

[0394] Under a nitrogen atmosphere, tert-butyl 4-(6-(2-(acetylthio)ethyl)-5-bromo-2-methoxypyridin-3-yl)piperidine-1-carboxylate (80 mg), cesium carbonate (110 mg), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (23 mg), and tris(dibenzylideneacetone)dipalladium (27 mg) were dissolved in 1,4-dioxane (3 mL) and reacted at 100°C for 12 hours. After the reaction, an appropriate amount of water was added to the system, and extraction was performed with ethyl acetate (10 mL). The organic phases were combined, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 50 mg of the title compound.

[0395] MS (ESI) m / z (M+H) + =351.1.

[0396] Step 4: Preparation of 6-(piperidin-4-yl)-2,3-dihydrothieno[3,2-b]pyridin-5(4H)-one hydrochloride

[0397] Dissolve tert-butyl 4-(5-methoxy-2,3-dihydrothieno[3,2-b]pyridin-6-yl)piperidine-1-carboxylate (50 mg) in a 4.0 M hydrogen chloride / 1,4-dioxane solution (5 mL) and react at 90°C for 5 hours. Cool the reaction to room temperature and remove the solvent in vacuo to obtain 55 mg of the crude product, which was used directly in the next step.

[0398] MS (ESI) m / z (M+H) + =237.1.

[0399] Preparation Example 21: Preparation of 6-(piperazin-1-yl)-2,3,4,8-tetrahydro-7H-thiopyrano[2,3-b]pyridin-7-one hydrochloride (can be used in Example 18)

[0400] Step 1: Preparation of tert-butyl 4-(7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperazine-1-carboxylate

[0401] Under a nitrogen atmosphere, 6-bromo-7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine (50.0 mg), tert-butyl piperazine-1-carboxylate (43.0 mg), tris(dibenzylideneacetone)dipalladium (17.4 mg), 2-dicyclohexylphosphino-2',6'-diisopropylbiphenyl (17.7 mg), and lithium tert-butoxide (30.4 mg) were dissolved in dioxane (1 mL) and reacted at 100°C for 1 hour. LCMS showed complete reaction of the starting material. The reaction mixture was filtered, and the filtrate was collected and concentrated. The crude product was purified by column chromatography to obtain 60.0 mg of the title compound.

[0402] MS (ESI) m / z (M+H) + =365.9.

[0403] Step 2: Preparation of 6-(piperazin-1-yl)-2,3,4,8-tetrahydro-7H-thiopyrano[2,3-b]pyridin-7-one hydrochloride

[0404] Dissolve tert-butyl 4-(7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperazine-1-carboxylate (60 mg) in concentrated hydrochloric acid (2 mL) and react at 95°C for 2 hours. LCMS indicated the reaction was complete. The reaction solution was concentrated to yield 40 mg of crude product, which was used directly in the next step.

[0405] MS (ESI) m / z (M+H) + =251.9.

[0406] Preparation Example 22: Preparation of 4'-methyl-1,4'-bipiperidine hydrochloride (can be used in Example 22)

[0407] Step 1: Preparation of tert-butyl 4'-cyano-[1,4'-bipiperidinyl]-1'-carboxylate

[0408] Dissolve tert-butyl 4-oxopiperidine-1-carboxylate (2g), piperidine (1.7g), and trimethylsilyl cyanide (1.29g) in 1,2-dichloroethane (20mL) and react at 70°C overnight. After the reaction, add a small amount of water to the reaction solution and extract with ethyl acetate. The combined organic phases are washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product is separated and purified by silica gel column chromatography to obtain 1.1g of the title compound.

[0409] MS (ESI) m / z (M+H) + =294.2.

[0410] Step 2: Preparation of tert-butyl 4'-methyl-[1,4'-bipiperidinyl]-1'-carboxylate

[0411] Under a nitrogen atmosphere and in an ice-water bath, tert-butyl 4'-cyano-[1,4'-bipiperidinyl]-1'-carboxylate (700 mg) was dissolved in anhydrous tetrahydrofuran (5 mL). Methylmagnesium bromide solution (3N, 2.4 mL) was slowly added dropwise. The mixture was allowed to react for 1 hour. The reaction was then allowed to return to room temperature and continued for another hour. After completion of the reaction, a small amount of water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography to obtain 480 mg of the title compound.

[0412] MS (ESI) m / z (M+H) + =283.2.

[0413] Step 3: Preparation of 4'-methyl-1,4'-bipiperidine hydrochloride

[0414] Dissolve tert-butyl 4'-methyl-[1,4'-bipiperidinyl]-1'-carboxylate (100 mg) in a hydrochloric acid / 1,4-dioxane solution (4 N, 4 mL) and react at room temperature for 2 hours. After the reaction, concentrate to obtain 70 mg of a crude product.

[0415] MS (ESI) m / z (M+H) + =183.2.

[0416] Preparation Example 23: Preparation of 4-methoxy-3-((7-methyl-1H-indazol-5-yl)methyl)-4-oxobutanoic acid (can be used in Example 27)

[0417] Step 1: Preparation of 7-methyl-1H-indazole-5-carbaldehyde

[0418] Under a nitrogen atmosphere at -78°C, 5-bromo-7-methyl-1H-indazole (500 mg) was dissolved in tetrahydrofuran (5 mL). n-Butyl lithium (1.5 M, 2.4 mL) and sec-butyl lithium (1 M, 3.6 mL) were added and stirred for 1 hour. N,N-dimethylformamide (533 mg) was added dropwise and the reaction continued for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was brought to room temperature, and an appropriate amount of water was added. The mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 350 mg of the title compound.

[0419] MS (ESI) m / z (M+H) + =161.1.

[0420] Step 2: Preparation of 3-(methoxycarbonyl)-4-(7-methyl-1H-indazol-5-yl)but-3-enoic acid

[0421] Dissolve 7-methyl-1H-indazole-5-carbaldehyde (350 mg) in tert-butanol (5 mL), add sodium tert-butoxide (422.4 mg) and dimethyl succinate (481.8 mg), and react at 50°C for 5 hours. LCMS analysis indicates the reaction is complete. Bring the reaction mixture to room temperature, add an appropriate amount of water, and extract three times with ethyl acetate. Combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate to yield 800 mg of a crude product.

[0422] MS (ESI) m / z (M+H) + =275.1

[0423] Step 3: Preparation of 4-methoxy-3-((7-methyl-1H-indazol-5-yl)methyl)-4-oxobutanoic acid

[0424] 3-(Methoxycarbonyl)-4-(7-methyl-1H-indazol-5-yl)but-3-enoic acid (800 mg) was dissolved in methanol (10 mL) and ethyl acetate (10 mL). Palladium on carbon (100 mg) was added, and the reaction system was filled with hydrogen gas. The reaction was allowed to react at room temperature overnight. LCMS confirmed the reaction was complete. The reaction solution was filtered, and the filtrate was collected and rotary evaporated to remove excess solvent to yield 800 mg of crude product.

[0425] MS (ESI) m / z (M+H) + =277.1.

[0426] Preparation Example 24: Preparation of (S)-methyl 3-(7-methyl-1H-indazol-5-yl)-2-((phenoxycarbonyl)amino)propanoate (can be used in Example 28)

[0427] The title compound was prepared by using the corresponding common commercial reagents and subsequently using the preparation method similar to the above intermediate preparation examples 3 and 5.

[0428] MS (ESI) m / z (M+H) + =354.1.

[0429] Preparation Example 25: Preparation of 1-(1-(methyl-d3)piperidin-4-yl)piperazine hydrochloride (can be used in Example 29)

[0430] Step 1: Preparation of tert-butyl 4-(1-(methyl-d3)piperidin-4-yl)piperazine-1-carboxylate

[0431] Dissolve 1-Boc-4-(piperidin-4-yl)-piperazine (355 mg) and potassium carbonate (365.0 mg) in tetrahydrofuran (5 mL). Slowly add d3-toluenesulfonylmethyl ester (250.0 mg) dropwise in batches. Allow to react at room temperature for 10 hours. TLC indicates completion of the reaction. The system is filtered, the mother liquor is concentrated, and the crude product is purified by column chromatography to yield 210.0 mg of the title compound.

[0432] MS (ESI) m / z (M+H) + =287.2.

[0433] Step 2: Preparation of 1-(1-(methyl-d3)piperidin-4-yl)piperazine hydrochloride

[0434] Dissolve tert-butyl 4-(1-(methyl-d3)piperidin-4-yl)piperazine-1-carboxylate (210.0 mg) in a 4M solution of hydrochloric acid in 1,4-dioxane (5 mL) and allow to react at room temperature for 1 hour. TLC indicated the reaction was complete, and the system was concentrated to yield 180.0 mg of the crude product.

[0435] MS (ESI) m / z (M+H) + =187.2.

[0436] Preparation Example 26: Preparation of 5'-(piperidin-4-yl)-2'H-spiro[cyclopropane-1,3'-thieno[2,3-b]pyridine]-6'(7'H)-one hydrochloride (can be used in Example 33)

[0437] Step 1: Preparation of 3-bromo-6-chloro-2-methoxypyridine

[0438] Dissolve 3-bromo-6-chloro-2-fluoropyridine (10 g) in methanol (100 mL), add sodium methoxide (5.2 g), and react at 50°C for 2 hours. TLC shows the reaction is complete. The reaction solution is rotary evaporated to remove excess solvent, and an appropriate amount of water is added. The mixture is extracted three times with ethyl acetate. The organic phases are combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to obtain 10.1 g of the title compound.

[0439] Step 2: Preparation of tert-butyl 6-chloro-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate

[0440] Under a nitrogen atmosphere, 3-bromo-6-chloro-2-methoxypyridine (10.1 g) was dissolved in 1,4-dioxane (100 mL) and water (10 mL). Potassium carbonate (12.4 g), [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride (3 g), and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (15.4 g) were added. The mixture was allowed to react at 80°C for 3 hours. LCMS indicated the reaction was complete. The reaction mixture was brought to room temperature, and an appropriate amount of water was added. The mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain 11.0 g of the title compound.

[0441] MS (ESI) m / z (M+H) + =325.1.

[0442] Step 3: Preparation of tert-butyl 4-(6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0443] Dissolve tert-butyl 6-chloro-2-methoxy-3',6'-dihydro-[3,4'-bipyridyl]-1'(2'H)-carboxylate (9 g) in methanol (50 mL), add palladium on carbon (150 mg), and purge the reaction system with hydrogen. Allow to react at room temperature overnight. LCMS indicates the reaction is complete. Filter the reaction solution, collect the filtrate, and remove excess solvent by rotary evaporation. The resulting crude product is purified by column chromatography to yield 9.0 g of the title compound.

[0444] MS (ESI) m / z (M+H) + =327.1.

[0445] Step 4: Preparation of 3-bromo-2-chloro-6-methoxy-5-(piperidin-4-yl)pyridine

[0446] Under ice-water bath conditions, tert-butyl 4-(6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate (9 g) was dissolved in acetonitrile (60 mL). A solution of ferric chloride (4.5 g dissolved in 10 mL of acetonitrile) was slowly added dropwise. N-bromosuccinimide (4.8 g) was added and the mixture was allowed to react at room temperature for 16 hours. LCMS showed that the reaction was complete. An appropriate amount of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed three times with saturated sodium thiosulfate solution and sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to give 11 g of crude product.

[0447] MS (ESI) m / z (M+H) + =305.0.

[0448] Step 5: Preparation of tert-butyl 4-(5-bromo-6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0449] Dissolve 3-bromo-2-chloro-6-methoxy-5-(piperidin-4-yl)pyridine (11 g) in dichloromethane (200 mL), add triethylamine (9.1 g) and di-tert-butyl dicarbonate (15.7 g), and react at room temperature for 1 hour. LCMS indicates completion of the reaction. Remove excess solvent from the reaction solution by rotary evaporation, filter, and wash the solid residue several times with ethyl acetate. Collect the filtrate, wash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 4.8 g of the title compound.

[0450] MS (ESI) m / z (M+H) + =405.0.

[0451] Step 6: Preparation of tert-butyl 4-(6-chloro-5-(isoxazol-4-yl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0452] Under a nitrogen atmosphere, tert-butyl 4-(5-bromo-6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate (3.9 g) was dissolved in 1,4-dioxane (100 mL) and water (10 mL). Potassium carbonate (2.7 g), [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride (650 mg), and 4-isoxazoleboronic acid pinacol ester (550 mg) were added. The mixture was allowed to react at 80°C for 3 hours. LCMS indicated the reaction was complete. The reaction mixture was brought to room temperature, a small amount of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 2.2 g of the title compound.

[0453] MS (ESI) m / z (M+H) + =394.1.

[0454] Step 7: Preparation of tert-butyl 4-(6-chloro-5-(cyanomethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0455] Dissolve tert-butyl 4-(6-chloro-5-(isoxazol-4-yl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (2.2 g) in methanol (20 mL), add potassium fluoride (0.67 g), and react at 90°C for 10 hours. LCMS indicates the reaction is complete. Bring the reaction mixture to room temperature, remove excess solvent by rotary evaporation, add a small amount of water, and extract three times with ethyl acetate. The organic phases are combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product is purified by column chromatography to yield 2.0 g of the title compound.

[0456] MS (ESI) m / z (M+H) + =366.1.

[0457] Step 8: Preparation of tert-butyl 4-(6-chloro-5-(1-cyanocyclopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0458] Dissolve tert-butyl 4-(6-chloro-5-(cyanomethyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (2 g) in dimethyl sulfoxide (20 mL), add diphenyl(vinyl)sulfonium trifluoromethanesulfonate (2.4 g) and 1,8-diazabicyclo[5.4.0]undec-7-ene (2.5 g), and allow to react at room temperature for 1 hour. LCMS indicates completion of the reaction. Add a small amount of water to the reaction solution, extract three times with ethyl acetate, combine the organic phases, backwash once with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate. The resulting crude product is purified by column chromatography to yield 1.8 g of the title compound.

[0459] MS (ESI) m / z (M+H) + =392.2.

[0460] Step 9: Preparation of tert-butyl 4-(6-chloro-5-(1-formylcyclopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0461] Under a nitrogen atmosphere at -78°C, tert-butyl 4-(6-chloro-5-(1-cyanocyclopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (1.8 g) was dissolved in dichloromethane (20 mL). Diisobutylaluminum hydride (1 M, 5 mL) was slowly added and allowed to react for 1 hour. LCMS indicated the reaction was complete. The reaction mixture was brought to room temperature, a small amount of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 1.4 g of the title compound.

[0462] MS (ESI) m / z (M+H) + =395.2.

[0463] Step 10: Preparation of tert-butyl 4-(6-chloro-5-(1-(hydroxymethyl)cyclopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0464] Under ice-water conditions, tert-butyl 4-(6-chloro-5-(1-formylcyclopropyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (1.4 g) was dissolved in tetrahydrofuran (10 mL) and methanol (10 mL). Sodium borohydride (356.4 mg) was added and the mixture was allowed to react at room temperature for 1 hour. LCMS indicated the reaction was complete. A small amount of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting crude product was purified by column chromatography to yield 1.3 g of the title compound.

[0465] MS (ESI) m / z (M+H) + =397.2.

[0466] The title compound was prepared using the corresponding common commercial reagents and a similar preparation method to the above intermediate preparation example 19.

[0467] MS (ESI) m / z (M+H) + =263.1.

[0468] Preparation Example 27: Preparation of 3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octane hydrochloride (can be used in Example 34)

[0469] Step 1: Preparation of tert-butyl 3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate

[0470] Dissolve N-tert-Butyloxycarbonyl-nortropinone (600 mg) and hexahydropyridine (226 mg) in dichloromethane (10 mL), add sodium triacetoxyborohydride (848 mg), and react at room temperature for 3 hours. After the reaction, add an appropriate amount of water and extract with ethyl acetate (200 mL). The organic phases are combined, washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 430 mg of the title compound.

[0471] MS (ESI) m / z (M+H) + =295.1.

[0472] Step 2: Preparation of 3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octane hydrochloride

[0473] Dissolve tert-butyl 3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octane-8-carboxylate (200 mg) in a 4.0 M hydrogen chloride / 1-4-dioxane solution (5 mL) and allow to react at room temperature for 1 hour. After monitoring the reaction for completion, the solvent was removed by concentration to yield 130 mg of a crude product.

[0474] MS (ESI) m / z (M+H) + =195.1.

[0475] Preparation Example 28: Preparation of 5-(piperidin-1-yl)-2-azabicyclo[2.2.1]heptane hydrochloride (can be used in Example 36)

[0476] The title compound was prepared using the corresponding common commercial reagents and a similar preparation method to the above intermediate preparation example 27.

[0477] MS (ESI) m / z (M+H) + =181.1.

[0478] Preparation 29: Preparation of (R)-methyl 3-(7-methyl-1H-indazol-5-yl-3-d)-2-((phenoxycarbonyl)amino)propanoate (can be used in Example 37)

[0479] Step 1: Preparation of 5-bromo-3-iodo-7-methyl-1H-indazole

[0480] 5-Bromo-7-methyl-1H-indazole (250 mg) and potassium hydroxide (146.2 mg) were dissolved in anhydrous N,N-dimethylformamide (3 mL) in an ice-water bath. Elemental iodine (451 mg) was added and the mixture was allowed to react for 1 hour. LCMS indicated that the reaction was complete. The reaction solution was quenched with an appropriate amount of saturated sodium sulfite solution and extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to afford 390 mg of the title compound.

[0481] MS (ESI) m / z (M+H) + =336.9.

[0482] Step 2: Preparation of 5-bromo-7-methyl-1H-indazole-3d

[0483] In an ice-water bath and nitrogen atmosphere, 5-bromo-3-iodo-7-methyl-1H-indazole (0.39 g) was dissolved in anhydrous tetrahydrofuran (4 mL). Isopropylmagnesium chloride-lithium chloride solution (1.96 mL, 1.3 M in THF) was slowly added dropwise. The reaction was allowed to react for 0.5 hour. Deuterated methanol (1 mL) was added, and the mixture was brought to room temperature and the reaction continued for 0.5 hour. LCMS indicated complete reaction of the starting material. The reaction solution was concentrated, and the crude product was purified by column chromatography to yield 0.2 g of the title compound.

[0484] MS (ESI) m / z (M+H) + =211.9.

[0485] The title compound was prepared by using the corresponding common commercial reagents and subsequently using the preparation method similar to the above intermediate preparation examples 3 and 5.

[0486] MS (ESI) m / z (M+H) + =354.9.

[0487] Preparation Example 30: Preparation of 2-hydroxy-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one (can be used in Example 41)

[0488] Step 1: Preparation of (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)propionic acid hydrochloride

[0489] Dissolve methyl (R)-2-((tert-Butoxycarbonyl)amino)-3-(7-methyl-1H-indazol-5-yl)propanoate (500.0 mg) in concentrated hydrochloric acid (3 mL) and react at 90°C for 2 hours. LCMS indicated complete reaction. The reaction mixture was concentrated to yield 300 mg of crude product, which was used directly in the next step.

[0490] MS (ESI) m / z (M+H) + =220.1.

[0491] Step 2: Preparation of (R)-2-hydroxy-3-(7-methyl-1H-indazol-5-yl)propanoic acid

[0492] Under ice-water conditions, (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)propionic acid hydrochloride (300 mg) was dissolved in water (3 mL) and concentrated sulfuric acid (0.45 mL) was slowly added dropwise. Sodium nitrite (567 mg) was added in batches and the mixture was allowed to react at 40°C for 16 hours. LCMS indicated that the reaction was complete. The reaction solution was adjusted to pH 4 with 2N dilute hydrochloric acid and extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 200 mg of crude product, which was used directly in the next step.

[0493] MS (ESI) m / z (M+H) + =220.9.

[0494] Step 3: Preparation of 2-hydroxy-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one

[0495] Under ice-water conditions, (R)-2-hydroxy-3-(7-methyl-1H-indazol-5-yl)propanoic acid (200 mg), 1-(1-methylpiperidin-4-yl)piperazine (332.9 mg), and N,N-diisopropylethylamine (234.8 mg) were dissolved in N,N-dimethylformamide (2.0 mL). 1H-Benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (519.9 mg) was slowly added and allowed to react for 0.5 hour. LCMS indicated the reaction was complete. The reaction solution was concentrated, and the crude product was purified by preparative liquid chromatography to obtain 100 mg of the title compound.

[0496] MS (ESI) m / z (M+H) + =385.9.

[0497] Preparation Example 31: Preparation of tert-butyl (E)-4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)vinyl)piperidine-1-carboxylate

[0498] Dissolve 1-Boc-4-ethynylpiperidine (5.0 g) and bis(cyclopentadienyl)zirconium chloride hydride (307.32 mg) in toluene (50 mL), add 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.67 g) dropwise, and heat at 60°C to react overnight. After the reaction, concentrate the reaction solution, and purify the crude product by column chromatography to remove the other configuration, yielding 5.3 g of the title compound.

[0499] Preparation Example 32: Preparation of 3-(piperidin-4-yl)-5,6,7,8-tetrahydro-5,8-methanoquinolin-2(1H)-one hydrochloride (can be used in Example 45)

[0500] Step 1: Preparation of methyl 3-oxobicyclo[2.2.1]heptane-2-carboxylate

[0501] Under a nitrogen atmosphere, sodium hydride (726 mg) was dissolved in dimethyl carbonate (9 mL), and then bicyclo[2.2.1]heptan-2-one (1 g) was dissolved in dimethyl carbonate (6 mL). The mixture was slowly added dropwise to the sodium hydride system and allowed to react at 90°C for 2 hours. After completion of the reaction, the reaction solution was quenched with water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was separated and purified by column chromatography to obtain 1.10 g of the title compound.

[0502] MS (ESI) m / z (M+H) + =169.1.

[0503] Step 2: Preparation of methyl 3-((trifluoromethyl)sulfonyl)oxy)bicyclo[2.2.1]hept-2-ene-2-carboxylate

[0504] Under ice-water conditions, sodium hydride (385 mg) was dissolved in anhydrous tetrahydrofuran (10 mL). Methyl 3-oxobicyclo[2.2.1]heptane-2-carboxylate (1.08 g) was then dissolved in anhydrous tetrahydrofuran (5 mL) and slowly added dropwise to the sodium hydride system. The mixture was stirred for 30 minutes. N-phenylbis(trifluoromethanesulfonyl)imide (2.98 g) was then dissolved in anhydrous tetrahydrofuran (10 mL) and slowly added dropwise to the system. The mixture was allowed to react overnight at room temperature. After completion of the reaction, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was separated and purified by column chromatography to obtain 1.70 g of the title compound.

[0505] MS (ESI) m / z (M+H) + =301.1.

[0506] Step 3: Preparation of tert-butyl (E)-4-(2-(3-(methoxycarbonyl)bicyclo[2.2.1]hept-2-en-2-yl)vinyl)piperidine-1-carboxylate

[0507] Under a nitrogen atmosphere, methyl 3-((trifluoromethyl)sulfonyl)oxy)bicyclo[2.2.1]hept-2-ene-2-carboxylate (600 mg), tert-butyl (E)-4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)vinyl)piperidine-1-carboxylate (742 mg), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (163 mg), and potassium carbonate (690 mg) were dissolved in 1,4-dioxane (10 mL) and water (2 mL) and reacted at 80°C for 2 hours. After completion of the reaction, water was added to quench the reaction, the mixture was filtered, and the filtrate was collected and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product, which was separated and purified by column chromatography to obtain 660.00 mg of the title compound.

[0508] MS (ESI) m / z (M+H) + =362.2.

[0509] Step 4: Preparation of (E)-3-(2-(1-(tert-butoxycarbonyl)piperidin-4-yl)vinyl)bicyclo[2.2.1]hept-2-ene-2-carboxylic acid

[0510] Tert-butyl (E)-4-(2-(3-(methoxycarbonyl)bicyclo[2.2.1]hept-2-en-2-yl)vinyl)piperidine-1-carboxylate (200 mg) was dissolved in methanol (10 mL) and water (1 mL). Sodium hydroxide (155 mg) was added and the mixture was allowed to react at 50°C overnight. After completion of the reaction, the pH of the reaction system was adjusted to 5-6 with 1N hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 180 mg of a crude product.

[0511] MS (ESI) m / z (M+H) + =348.2.

[0512] Step 5: Preparation of tert-butyl 4-(2-oxo-1,2,5,6,7,8-hexahydro-5,8-methanoquinolin-3-yl)piperidine-1-carboxylate

[0513] Under a nitrogen atmosphere, (E)-3-(2-(1-(tert-Butoxycarbonyl)piperidin-4-yl)vinyl)bicyclo[2.2.1]hept-2-ene-2-carboxylic acid (180 mg) was dissolved in toluene (2 mL), and diphenylphosphoryl azide (154 mg) and triethylamine (78 mg) were added. The mixture was allowed to react at room temperature for 3 hours. After the reaction was complete, the system was pre-purified by preparative thin-layer chromatography, dissolved in toluene (2 mL), and the reaction was continued at 90°C for 3 hours. After the reaction was completed, the mixture was concentrated, and the crude product was separated and purified by preparative thin-layer chromatography to obtain 80.00 mg of the title compound.

[0514] MS (ESI) m / z (M+H) + =345.2.

[0515] Step 6: Preparation of 3-(piperidin-4-yl)-5,6,7,8-tetrahydro-5,8-methanoquinolin-2(1H)-one hydrochloride

[0516] Dissolve tert-butyl 4-(2-oxo-1,2,5,6,7,8-hexahydro-5,8-quinolin-3-yl)piperidine-1-carboxylate (80 mg) in a 4M hydrogen chloride / 1,4-dioxane solution (1 mL) and allow to react at room temperature for 1 hour. After the reaction, concentrate to obtain 78 mg of the crude product.

[0517] MS (ESI) m / z (M+H) + =245.1

[0518] Preparation Example 33: Preparation of 6'-(piperidin-4-yl)-4',8'-dihydro-2'H,7'H-spiro[cyclopropane-1,3'-thiopyrano[2,3-b]pyridine]-7'-one hydrochloride (can be used in Example 46)

[0519] Step 1: Preparation of methyl 2-(((1-(2-methoxy-2-oxoethyl)cyclopropyl)methyl)thio)acetate

[0520] Under ice-water conditions, methyl 2-(1-(mercaptomethyl)cyclopropyl)acetate (600 mg) was dissolved in methanol (12 mL). Sodium methoxide (750 mg) was added and stirred for 10 minutes. Ethyl bromoacetate (750 mg) was then added and allowed to react for 2 hours. LCMS indicated the reaction was complete. Hydrochloric acid (2N) was added to adjust the pH to 7-8. The mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 650 mg of the title compound.

[0521] MS (ESI) m / z (M+H) + = 233.1.

[0522] Step 2: Preparation of methyl 7-oxo-5-thiaspiro[2.5]octane-6-carboxylate

[0523] Methyl 2-(((1-(2-methoxy-2-oxoethyl)cyclopropyl)methyl)thio)acetate (630 mg) was dissolved in tetrahydrofuran (10 mL) in an ice-water bath. Potassium tert-butoxide (608 mg) was added portionwise and allowed to react for 1 hour. LCMS indicated the reaction was complete. Hydrochloric acid (2N) was added to adjust the pH to 5-6. The mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 380 mg of the title compound.

[0524] MS (ESI) m / z (M+H) + = 201.1.

[0525] The title compound was prepared by using the corresponding common commercial reagents and subsequently using a preparation method similar to the above intermediate preparation example 32.

[0526] MS (ESI) m / z (M+H) + =277.1.

[0527] Preparation Example 34: Preparation of 6-(piperidin-4-yl)-1a,2,4,7b-tetrahydrocyclopropane[4,5]thiopyrano[2,3-b]pyridin-5(1H)-one hydrochloride (can be used in Example 47)

[0528] Step 1: Preparation of 2-(methoxycarbonyl)cyclopropane-1-carboxylic acid

[0529] Dissolve triethylamine (901.8 mg) in methanol (15 mL) and add 3-oxabicyclo[3.1.0]hexane-2,4-dione (1 g) in portions. Allow to react at room temperature for 1 hour. After the reaction is complete, quench with water, remove excess methanol, adjust the pH to 3-4 with 1N hydrochloric acid, extract with ethyl acetate, combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to obtain 1.15 g of crude product.

[0530] Step 2: Preparation of methyl 2-(hydroxymethyl)cyclopropane-1-carboxylate.

[0531] Dissolve 2-(Methoxycarbonyl)cyclopropane-1-carboxylic acid (1.15 g) in anhydrous tetrahydrofuran (2 mL) in an ice-water bath. Add 1 M borane in tetrahydrofuran (16 mL) dropwise. Allow to react at room temperature for 3 hours. After completion, quench the reaction with an appropriate amount of methanol. Concentrate to obtain the crude product, which is then purified by column chromatography to afford 750 mg of the title compound.

[0532] MS (ESI) m / z (M+H) + =131.1.

[0533] Step 3: Preparation of methyl 2-(((methylsulfonyl)oxy)methyl)cyclopropane-1-carboxylate

[0534] At -10°C, methyl 2-(hydroxymethyl)cyclopropane-1-carboxylate (750 mg) was dissolved in dichloromethane (10 mL) and N,N-diisopropylethylamine (1.49 g) was added. Methanesulfonyl chloride (856 mg) was diluted with dichloromethane (3 mL) and added dropwise to the reaction system. The mixture was allowed to return to room temperature and allowed to react for 30 minutes. After completion of the reaction, water was added to quench the mixture, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain 1.1 g of the title compound.

[0535] MS (ESI) m / z (M+H) + =209.0.

[0536] Step 4: Preparation of methyl 2-(((2-ethoxy-2-oxoethyl)thio)methyl)cyclopropane-1-carboxylate

[0537] In an ice-water bath and nitrogen atmosphere, methyl 2-(((methylsulfonyl)oxy)methyl)cyclopropane-1-carboxylate (1.1 g) and ethyl 2-mercaptoacetate (936 mg) were dissolved in dry N,N-dimethylformamide (12 mL). Sodium methoxide (421 mg) was added and the mixture was allowed to warm to room temperature for 2 hours. After completion of the reaction, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain 1.10 g of the title compound.

[0538] MS (ESI) m / z (M+H) + =233.0.

[0539] Step 5: Preparation of ethyl 5-oxo-3-thiabicyclo[4.1.0]heptane-4-carboxylate

[0540] At -10°C under a nitrogen atmosphere, titanium tetrachloride (1.51 g) was dissolved in dry dichloromethane (20 mL). Isopropyl alcohol (518 mg) was added and stirred for 30 minutes. Methyl 2-(((2-ethoxy-2-oxoethyl)thio)methyl)cyclopropane-1-carboxylate (1.54 g) was dissolved in dry dichloromethane (5 mL) and slowly added dropwise to the reaction system, maintaining the internal temperature at -10°C and stirring for 30 minutes. Triethylamine (2.35 g) was added and the reaction continued for 1.5 hours. After completion of the reaction, an appropriate amount of 1N hydrochloric acid was added to quench the reaction. The mixture was extracted with dichloromethane, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain 740 mg of the title compound.

[0541] MS (ESI) m / z (M+H) + =201.0.

[0542] Step 6: Preparation of ethyl 5-(((trifluoromethyl)sulfonyl)oxy)-3-thiabicyclo[4.1.0]hept-4-ene-4-carboxylate

[0543] At -78°C, ethyl 5-oxo-3-thiabicyclo[4.1.0]heptane-4-carboxylate (570 mg) was dissolved in dry dichloromethane (15 mL), and N,N-diisopropylethylamine (1.1 g) was added. Trifluoromethanesulfonic anhydride (1.36 g) was diluted with dry dichloromethane (5 mL) and added dropwise to the above system. The reaction was continued for 30 minutes. After completion of the reaction, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to obtain 700 mg of the title compound.

[0544] MS (ESI) m / z (M+H) + =333.0.

[0545] The title compound was prepared by using the corresponding common commercial reagents and subsequently using a preparation method similar to the above intermediate preparation example 32.

[0546] MS (ESI) m / z (M+H) + =263.1.

[0547] Preparation Example 35: Preparation of 6-(piperidin-4-yl)-2,8-dihydro-7H-thiopyrano[2,3-b]pyridin-7-one hydrochloride (can be used in Example 48)

[0548] Step 1: Preparation of tert-butyl (E)-4-(6-chloro-5-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0549] Under a nitrogen atmosphere, tert-butyl 4-(5-bromo-6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate (1.4 g), potassium carbonate (1.19 g), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (281 mg) were dissolved in 1,4-dioxane (30 mL) and water (5 mL). The system was heated at 90°C, and a solution of ethyl (E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (1.17 g dissolved in 8 mL of 1,4-dioxane) was slowly added dropwise. The reaction mixture was allowed to react for 2 hours. The reaction mixture was brought to room temperature, quenched with a small amount of water, and filtered. The filtrate was collected and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography to obtain 1 g of the title compound.

[0550] MS (ESI) m / z (M+H) + =425.1.

[0551] Step 2: Preparation of tert-butyl (E)-4-(6-chloro-5-(3-hydroxyprop-1-enyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0552] At -40°C, tert-butyl (E)-4-(6-chloro-5-(3-ethoxy-3-oxoprop-1-en-1-yl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (980 mg) was dissolved in dry tetrahydrofuran (20 mL). Diisobutylaluminum hydride (1.5 M, 7.7 mL) was slowly added dropwise. The mixture was allowed to react at room temperature for 1.5 hours. The reaction mixture was quenched with a small amount of water and then treated with aqueous sodium hydroxide (2.5 N, 24 mL). The mixture was extracted with ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography to yield 380 mg of the title compound.

[0553] MS (ESI) m / z (M+H) + =383.1.

[0554] Step 3: Preparation of (E)-tert-butyl 4-(6-chloro-2-methoxy-5-(3-((methylsulfonyl)oxy)prop-1-en-1-yl)pyridin-3-yl)piperidine-1-carboxylate

[0555] Under ice-water conditions, tert-butyl (E)-4-(6-chloro-5-(3-hydroxyprop-1-enyl)-2-methoxypyridin-3-yl)piperidine-1-carboxylate (370 mg) was dissolved in dichloromethane (10 mL), and N,N-diisopropylethylamine (499 mg) was added. Methanesulfonyl chloride (172 mg) was dissolved in dichloromethane (2 mL) and slowly added dropwise to the reaction system. The reaction was allowed to react for 0.5 hours. The reaction solution was quenched with a small amount of water and extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to yield 442 mg of the crude product.

[0556] MS (ESI) m / z (M+H) + =461.1.

[0557] Step 4: Preparation of tert-butyl (E)-4-(5-(3-(acetylthio)prop-1-en-1-yl)-6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate

[0558] Under a nitrogen atmosphere, tert-butyl (E)-4-(6-chloro-2-methoxy-5-(3-((methylsulfonyl)oxy)prop-1-en-1-yl)pyridin-3-yl)piperidine-1-carboxylate (445 mg) was dissolved in N,N-dimethylformamide (10 mL), and potassium carbonate (398 mg) and potassium thioacetate (329 mg) were added. The mixture was allowed to react at room temperature for 2 hours. After completion of the reaction, a small amount of water was added to quench the reaction system, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by silica gel column chromatography to obtain 390 mg of the title compound. MS (ESI) m / z (M+H) + =441.1.

[0559] Step 5: Preparation of tert-butyl 4-(7-methoxy-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxylate

[0560] Under a nitrogen atmosphere, tert-butyl (E)-4-(5-(3-(acetylthio)prop-1-en-1-yl)-6-chloro-2-methoxypyridin-3-yl)piperidine-1-carboxylate (150 mg), tris(dibenzylideneacetone)dipalladium (155 mg), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (196 mg), and potassium carbonate (276 mg) were dissolved in 1,4-dioxane (10 mL). The reaction was initiated by microwave at 115°C for 5 hours. After completion of the reaction, the reaction solution was filtered through celite, and the filtrate was collected and concentrated. The crude product was purified by preparative high-performance liquid chromatography to obtain 11 mg of the title compound. MS (ESI) m / z (M+H) + =363.1.

[0561] Step 6: Preparation of 6-(piperidin-4-yl)-2,8-dihydro-7H-thiopyrano[2,3-b]pyridin-7-one hydrochloride

[0562] Dissolve tert-butyl 4-(7-methoxy-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxylate (11 mg) in a hydrochloric acid / 1,4-dioxane solution (4N, 3 mL) and react at 90°C for 4 hours. After the reaction, concentrate the reaction mixture to obtain 9 mg of crude product.

[0563] MS (ESI) m / z (M+H) + =249.1.

[0564] Preparation Example 36: Preparation of 3-(piperidin-4-yl)-1,5,6,7-tetrahydro-2H-cyclopenta[b]pyridin-2-one hydrochloride (can be used in Example 51)

[0565] The title compound was prepared by using the corresponding common commercial reagents and subsequently using a preparation method similar to the above intermediate preparation example 32.

[0566] MS (ESI) m / z (M+H) + =219.1.

[0567] Preparation Example 37: Preparation of 3-(piperidin-4-yl)-5,6,7,8-tetrahydroquinolin-2(1H)-one hydrochloride (can be used in Example 52)

[0568] The title compound was prepared by using the corresponding common commercial reagents and subsequently using a preparation method similar to the above intermediate preparation example 32.

[0569] MS (ESI) m / z (M+H) + =233.2.

[0570] The following illustrative methods for synthesizing the compounds of Formula I of the present invention are provided. The raw materials and reagents required for synthesizing the compounds of Formula I of the present invention in the following examples can be obtained through common commercial channels, or according to the synthesis methods of the intermediate compounds provided herein, or by combining conventional chemical reaction synthesis methods with methods reported in prior art literature.

[0571] Example 1: Preparation of (R)-4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuran[2,3-b]pyridin-5-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxyylidenepropyl-2-yl)piperidine-1-carboxamide

[0572] Step 1: Preparation of tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate

[0573] Under a nitrogen atmosphere, 5-bromo-6-methoxy-3,3-dimethyl-2,3-dihydrofuran[2,3-b]pyridine (610.0 mg), N-tert-butyloxycarbonyl-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (880.0 mg), and potassium carbonate (981.0 mg) were dissolved in 1,4-dioxane (7 mL) and water (0.7 mL). 1,1-Bis(diphenylphosphino)diphenylferric palladium chloride (173.0 mg) was added and the mixture was reacted at 90°C for 2 hours. TLC indicated the reaction was complete. 30 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 620.0 mg of the title compound.

[0574] MS (ESI) m / z (M+H) + =361.2.

[0575] Step 2: Preparation of tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-5-yl)piperidine-1-carboxylate

[0576] Dissolve tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (610.0 mg) in methanol (30 mL). Add palladium on carbon (10%, 61.0 mg). Replace the air with hydrogen and continue to flow through the mixture. Allow to react at room temperature for 1 hour. TLC indicates the reaction is complete. Filter the mixture, collect the filtrate, and concentrate. The crude product is purified by column chromatography to yield 550.0 mg of the title compound.

[0577] MS (ESI) m / z (M+H) + =363.2.

[0578] Step 3: Preparation of 3,3-dimethyl-5-(piperidin-4-yl)-3,7-dihydrofuran[2,3-b]pyridin-6(2H)-one hydrochloride

[0579] Dissolve tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrofuro[2,3-b]pyridin-5-yl)piperidine-1-carboxylate (200.0 mg) in 1,4-dioxane hydrochloride (4 M, 3 mL) and react at 90°C for 3 hours. TLC indicated the reaction was complete, and the system was concentrated to yield 230.0 mg of the crude product.

[0580] MS (ESI) m / z (M+H) + =249.2.

[0581] Step 4: Preparation of (R)-methyl 2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuro[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoate

[0582] Methyl (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)propanoate trifluoroacetate (187.9 mg) and N,N'-disuccinimidyl carbonate (206.4 mg) were dissolved in N,N-dimethylformamide (3 mL). Triethylamine (244.3 mg) was added and the mixture was allowed to react at room temperature for 30 minutes. 3,3-Dimethyl-5-(piperidin-4-yl)-3,7-dihydrofuro[2,3-b]pyridin-6(2H)-one hydrochloride (200.0 mg) was added to the above mixture and the reaction was allowed to react at room temperature for 1 hour. TLC indicated that the reaction was complete. The mixture was filtered, and the filtrate was collected and subjected to reverse phase purification to obtain 90.0 mg of the title compound.

[0583] MS (ESI) m / z (M+H) + =508.2.

[0584] Step 5: Preparation of (R)-2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuro[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoic acid

[0585] Methyl (R)-2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuran[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoate (90.0 mg) was dissolved in tetrahydrofuran (5 mL) and methanol (1 mL). A 1 mL solution of lithium hydroxide (8.2 mg) was added and allowed to react at room temperature for 1 hour. TLC indicated the reaction was complete. The pH of the system was adjusted to approximately 7 with 1 M dilute hydrochloric acid, and the mixture was concentrated to yield 100.0 mg of the crude product.

[0586] MS (ESI) m / z (M+H)+ =494.2.

[0587] Step 6: Preparation of (R)-4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuro[2,3-b]pyridin-5-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxyylidenepropyl-2-yl)piperidine-1-carboxamide

[0588] (R)-2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuran[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoic acid (100.0 mg) and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (78 mg) were dissolved in N,N-dimethylformamide (2 mL). N,N-diisopropylethylamine (77.4 mg) was added and the mixture was allowed to react at room temperature for 10 minutes. 1-(1-methylpiperidin-4-yl)piperazine (43.9 mg) was added to the reaction mixture and allowed to react at room temperature for 20 minutes. TLC indicated the reaction was complete. The mixture was filtered, and the filtrate was subjected to reverse phase purification and freeze-dried to obtain 36.0 mg of the title compound.

[0589] MS (ESI) m / z (M+H) + =659.4.

[0590] 1 H NMR (400MHz, DMSO-d6) δ13.02(s,1H),10.62(brs,1H),7.96(s,1H),7.36(s,1H),7.29(s,1H),7.00(s,1H),6.63(d,J=8 .0Hz,1H),4.78(q,J=7.7Hz,1H),4.16(s,2H),4.11(d,J=13.0Hz,2H),3.62–3.55(m,1H),3.22(t,J=9.9Hz,2H),3.12–3. 06(m,1H),3.00–2.86(m,2H),2.80–2.63(m,5H),2.47(s,3H),2.35(d,J=7.7Hz,1H),2.27–2.20(m,1H),2.09(s,3H),1. 98–1.85(m,2H),1.75–1.62(m,4H),1.50(t,J=9.1Hz,1H),1.43–1.34(m,4H),,1.25(d,J=2.2Hz,6H),1.21–1.09(m,2H).

[0591] Example 2: Preparation of (R)-4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxyylidenepropyl-2-yl)piperidine-1-carboxamide

[0592] Step 1: Preparation of tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrothieno[2,3-b]pyridin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate

[0593] Under a nitrogen atmosphere, 5-bromo-6-methoxy-3,3-dimethyl-2,3-dihydrothiophene[2,3-b]pyridine (300.0 mg), N-tert-butyloxycarbonyl-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (441.4 mg), and potassium carbonate (454.9 mg) were dissolved in 1,4-dioxane (4 mL) and water (0.4 mL). 1,1-Bis(diphenylphosphino)diphenylferric palladium chloride (80.4 mg) was added, and the temperature was raised to 90°C for 2 hours. TLC indicated the reaction was complete. 30 mL of water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 300.0 mg of the title compound.

[0594] MS (ESI) m / z (M+H) + =377.2.

[0595] Step 2: Preparation of tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxylate

[0596] Dissolve tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrothieno[2,3-b]pyridin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (300.0 mg) in methanol (20 mL). Add palladium on carbon (10%, 30.0 mg). Replace the air with hydrogen and continue to flow through the mixture. Allow to react at room temperature for 1 hour. TLC indicates the reaction is complete. Filter the mixture, collect the filtrate, and concentrate. The crude product is purified by column chromatography to yield 550.0 mg of the title compound.

[0597] MS (ESI) m / z (M+H) + =379.2.

[0598] Step 3: Preparation of 3,3-dimethyl-5-(piperidin-4-yl)-3,7-dihydrothieno[2,3-b]pyridin-6(2H)-one hydrochloride

[0599] Dissolve tert-butyl 4-(6-methoxy-3,3-dimethyl-2,3-dihydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxylate (300.0 mg) in 1,4-dioxane hydrochloride solution (10 mL) and react at 90°C for 3 hours. TLC indicated the reaction was complete, and the system was concentrated to yield 340.0 mg of crude product.

[0600] MS (ESI) m / z (M+H) + =265.1.

[0601] Step 4: Preparation of (R)-methyl 2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoate

[0602] Dissolve 3,3-dimethyl-5-(piperidin-4-yl)-3,7-dihydrothieno[2,3-b]pyridin-6(2H)-one hydrochloride (62.9 mg) and methyl (R)-3-(7-methyl-1H-indazol-5-yl)-2-((phenoxycarbonyl)amino)propanoate (67.2 mg) in acetonitrile (4 mL). Add triethylamine (121.2 mg) and heat to 60°C for 7 hours. TLC indicates completion of the reaction, and the system is concentrated. The crude product is purified by column chromatography to yield 90.0 mg of the title compound.

[0603] MS (ESI) m / z (M+H) + =524.2.

[0604] Step 5: Preparation of (R)-2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoic acid

[0605] Methyl (R)-2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoate (90.0 mg) was dissolved in tetrahydrofuran (3 mL) and methanol (0.6 mL). Lithium hydroxide (22.0 mg) in aqueous solution (0.6 mL) was added and allowed to react at room temperature for 1 hour. TLC indicated the reaction was complete. The pH of the system was adjusted to approximately 7 with 1 M dilute hydrochloric acid and the mixture was concentrated to yield 110.0 mg of the crude product.

[0606] MS (ESI) m / z (M+H) + =510.2.

[0607] Step 6: Preparation of (R)-4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxyylidenepropyl-2-yl)piperidine-1-carboxamide

[0608] (R)-2-(4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamido)-3-(7-methyl-1H-indazol-5-yl)propanoic acid (80.0 mg) and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (61 mg) were dissolved in N,N-dimethylformamide (2 mL). N,N-diisopropylethylamine (60.8 mg) was added and the mixture was allowed to react at room temperature for 10 minutes. 1-(1-methylpiperidin-4-yl)piperazine (34.5 mg) was added to the above mixture and the mixture was allowed to react at room temperature for 20 minutes. TLC indicated the reaction was complete. The mixture was filtered, and the filtrate was subjected to reverse phase purification and freeze-dried to obtain 20.0 mg of the title compound.

[0609] MS (ESI) m / z (M+H) + =675.4.

[0610] 1H NMR (400MHz, DMSO-d6) δ12.94(s,1H),11.37(s,1H),7.89(s,1H),7.29(s,1H),7.00(s,1H),6.93(s,1H),6.56(d,J=8.1Hz, 1H),4.70(q,J=7.7Hz,1H),4.04(d,J=12.9Hz,2H),3.51(d,J=13.1Hz,1H),3.13(s,3H),3.04–2.99(m,1H),2.92–2.87(m,1 H),2.84–2.79(m,1H),2.70–2.58(m,5H),2.40(s,3H),2.34–2.20(m,2H),2.17–2.14(m,1H),2.03(s,3H),1.86–1.78(m,2H ),1.66(t,J=11.6Hz,2H),1.57(d,J=12.3Hz,2H),1.43(t,J=9.4Hz,1H),1.34–1.28(m,4H),1.18(s,6H),1.14–1.07(m,2H).

[0611] Example 3: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxyprop-2-yl)-4-(8-oxo-2,3,4,5,8,9-hexahydrooxepin[2,3-b]pyridin-7-yl)piperidine-1-carboxamide

[0612] The title compound was prepared using a similar preparation method to that of Example 2 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0613] MS (ESI) m / z (M+H) + =659.4.

[0614] 1H NMR (400MHz, DMSO-d6) δ12.95(brs,1H),11.27(brs,1H),7.96(s,1H),7.36(s,1H),7.08(s,1H),7.00(d,J=1.3Hz,1H),6.64(d,J=8.0Hz ,1H),4.78(q,J=7.7Hz,1H),4.13–4.06(m,2H),4.02(dd,J=6.0,4.0Hz,2H),3.59–3.54(m,1H),3.25–3.19(m,1H),3.13–3.10(m,1H),2. 99–2.94(m,1H),2.89–2.86(m,1H),2.74–2.66(m,4H),2.66–2.61(m,1H),2.56–2.53(m,2H),2.47(s,3H),2.36–2.30(m,1H),2.25–2.20 (m,1H),2.10(s,3H),1.97–1.83(m,4H),1.76–1.70(m,2H),1.66–1.60(m,4H),1.55–1.48(m,1H),1.40–1.33(m,4H),1.28–1.09(m,3H).

[0615] Example 4: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxypropane-2-yl)-4-(8-oxo-2,3,4,5,8,9-hexahydrothiazole[2,3-b]pyridin-7-yl)piperidine-1-carboxamide

[0616] Step 1: Preparation of (8-methoxy-2,3,4,5-tetrahydrothiopino[2,3-b]pyridin-7-yl)boronic acid

[0617] Under a nitrogen atmosphere, 7-bromo-8-methoxy-2,3,4,5-tetrahydrothiop[2,3-b]pyridine (230.0 mg) was dissolved in tetrahydrofuran (3 mL). At -78°C, n-butyllithium (2.5 M, 0.4 mL) was added dropwise and allowed to react for 30 minutes. Trimethyl borate (113.9 mg) was added and the reaction continued for 40 minutes. TLC indicated the reaction was complete. The reaction system was brought to room temperature, 10 mL of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 90.0 mg of the title compound.

[0618] MS (ESI) m / z (M+H) + =240.1.

[0619] Step 2: Preparation of tert-butyl 4-(8-methoxy-2,3,4,5-tetrahydrothiazole[2,3-b]pyridin-7-yl)piperidine-1-carboxylate

[0620] Dissolve (8-methoxy-2,3,4,5-tetrahydrothiopino[2,3-b]pyridin-7-yl)boronic acid (90.0 mg), tert-butyl 4-(2-tosylhydrazone)piperidine-1-carboxylate (165.0 mg), and potassium carbonate (156.0 mg) in 1,4-dioxane (5 mL) and heat at 120°C for 8 hours. TLC indicated completion of the reaction. The reaction system was brought to room temperature, 10 mL of water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, backwashed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography to yield 78.0 mg of the title compound.

[0621] MS (ESI) m / z (M+H) + =379.2.

[0622] The subsequent steps used the corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and embodiments as raw materials, and used a preparation method similar to that of Example 2 to prepare the title compound.

[0623] MS (ESI) m / z (M+H) + =675.4.

[0624] 1 H NMR(400MHz,DMSO-d6)δ12.94(s,1H),11.45(s,1H),7.89(s,1H),7.29(s,1H),7.00(s,1H),6.93(s,1H),6.58(d,J =8.1Hz,1H),4.70(q,J=7.7Hz,1H),4.03(d,J=12.4Hz,2H),3.54–3.45(m,1H),3.16(dd,J=11.8,8.1Hz,1H),3.05– 3.00(m,1H),2.92–2.79(m,2H),2.75–2.71(m,2H),2.67–2.55(m,7H),2.40(s,3H),2.28–2.24(m,1H),2.19–2.12( m,1H),2.03(s,3H),1.91–1.80(m,4H),1.69–1.56(m,4H),1.53–1.42(m,3H),1.32–1.29(m,2H),1.26–1.04(m,5H).

[0625] Example 5: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxypropane-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-pyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0626] The title compound was prepared using a similar preparation method to that of Example 2 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0627] MS (ESI) m / z (M+H) + =645.4.

[0628] 1 H NMR (400MHz, DMSO-d6) δ12.99(s,1H),11.15(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H),6.96(s,1H),6.64(d,J=8.0 Hz,1H),4.87-4.69(m,1H),4.24-4.13(m,2H),4.09(d,J=12.0,2H),3.60-3.52(m,1H),3.30-3.19(m,2H),3.18-3.08(m, 1H),3.03-2.93(m,1H),2.92-2.82(m,1H),2.79-2.57(m,6H),2.47(s,4H),2.37-2.30(m,1H),2.26-2.18(m,1H),2.11(s ,3H),1.99-1.82(m,4H),1.80-1.70(m,2H),1.68-1.59(m,2H),1.58-1.48(m,1H),1.43-1.34(m,2H),1.31-1.14(m,4H).

[0629] Example 6: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxypropane-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0630] The title compound was prepared using a similar preparation method to that of Example 4 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0631] MS (ESI) m / z (M+H) + =660.8.

[0632] 1 H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.31(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H),6.86(s,1H),6.64(d,J=8. 0,1H),4.80-4.72(m,1H),4.14-4.02(m,2H),3.57-3.53(m,1H),3.27-3.18(m,2H),3.14-3.08(m,1H),3.04-3.00(m,2H ),2.98-2.93(m,1H),2.92-2.86(m,1H),2.74-2.60(m,6H),2.56-2.54(m,1H),2.47(s,3H),2.37-2.31(m,1H),2.25-2. 19(m,1H),2.09(s,3H),2.01-1.89(m,4H),1.74-1.61(m,4H),1.55-1.49(m,1H),1.40-1.34(m,2H),1.30-1.13(m,4H).

[0633] Example 7: Preparation of (R)-4-(5,5-dimethyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopentadienyl[b]pyridin-3-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropyl-2-yl)piperidine-1-carboxamide

[0634] The title compound was prepared using a similar preparation method to that of Example 2 using corresponding common commercial reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0635] MS (ESI) m / z (M+H) + =657.1.

[0636] 1H NMR (400MHz, DMSO-d6) δ13.02(s,1H),11.59(s,1H),7.96(s,1H),7.36(s,1H),7.07(s,1H),7.00(s,1H),6.61(d,J=8.0Hz ,1H),4.77(q,J=7.7Hz,1H),4.10(d,J=12.8Hz,2H),3.58(d,J=13.0Hz,1H),3.21(s,1H),3.12–3.05(m,1H),2.97–2.86(m ,2H),2.79–2.63(m,8H),2.47(s,3H),2.33(s,2H),2.22(s,1H),2.11(s,3H),1.97–1.87(m,2H),1.82(t,J=7.2Hz,2H),1. 76(s,1H),1.66(d,J=12.4Hz,2H),1.49(t,J=9.1Hz,1H),1.38(d,J=12.7Hz,2H),1.32–1.20(m,4H),1.14(d,J=1.9Hz,6H).

[0637] Example 8: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0638] The title compound was prepared using a similar preparation method to that of Example 4 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0639] MS (ESI) m / z (M+H) + =647.1.

[0640] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.38(s,1H),7.96(s,1H),7.36(s,1H),7.09(s,1H),7.00(s,1H),6.65(d,J=8.0Hz,1H),4.77(q,J= 7.7Hz,1H),4.09(d,J=12.5Hz,2H),3.55(d,J=13.6Hz,1H),3.40(s,2H),3.24(s,1H),3.11(s,1H),3.06(t,J=8.1Hz,2H),2.96(dd,J=13.1, 7.8Hz,1H),2.88(dd,J=13.1,7.1Hz,1H),2.74–2.67(m,4H),2.63(d,J=11.1Hz,1H),2.47(s,3H),2.37–2.31(m,1H),2.23(d,J=10.1Hz,1H ),2.11(s,3H),1.99–1.90(m,2H),1.74(t,J=11.5Hz,2H),1.67–1.59(m,2H),1.57–1.51(m,1H),1.38(d,J=11.9Hz,2H),1.27–1.18(m,5H).

[0641] Example 9: Preparation of 4-(3,3-dimethyl-1-oxido-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)-N-((R)-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropane-2-yl)piperidine-1-carboxamide

[0642] Step 1: Preparation of 3,3-dimethyl-5-(piperidin-4-yl)-3,7-dihydrothieno[2,3-b]pyridine-6(2H)-1-oxide hydrochloride

[0643] Dissolve 3,3-dimethyl-5-(piperidin-4-yl)-3,7-dihydrothieno[2,3-b]pyridin-6(2H)-one hydrochloride (140.0 mg) in dichloromethane (10 mL). Add dilute hydrochloric acid (2M, 0.01 mL) and N-bromosuccinimide (94.0 mg). Allow to react at room temperature for 30 minutes. TLC indicates the reaction is complete, and the system is concentrated to yield 210.0 mg of crude product.

[0644] MS (ESI) m / z (M+H) + =281.1.

[0645] The subsequent steps used the corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and embodiments as raw materials, and used a preparation method similar to that of Example 2 to prepare the title compound.

[0646] MS (ESI) m / z (M+H) + =691.4.

[0647] 1 H NMR(400MHz,DMSO-d6)δ12.95(s,1H),7.90(s,1H),7.47(s,1H),7.30(s,1H),6.93(s,1 H),6.60(d,J=8.0Hz,1H),4.71(q,J=7.8Hz,1H),4.09(d,J=12.9Hz,2H),3.54–3.51(m, 2H),3.23–3.16(m,3H),3.01(d,J=9.3Hz,1H),2.97–2.90(m,2H),2.82–2 .80(m,2H),2.68–2.63(m,4H),2.40(s,3H),2.30–2.24(m,1H),2.18–2.11 (m,1H),2.03(s,3H),1.88–1.78(m,2H),1.69–1.61(m,4H),1.42–1.40(m, 2H),1.38(s,3H),1.29(d,J=12.2Hz,2H),1.24(s,3H),1.16–1.06(m,2H).

[0648] Example 10: Preparation of (R)-4-(3,3-dimethyl-6-oxo-2,3,6,7-tetrahydrofuran[2,3-b]pyridin-5-yl)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropyl-2-yl)piperidine-1-thiocarboxamide

[0649] (R)-2-Amino-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one (23.0 mg) was dissolved in acetonitrile (1 mL). N,N-diisopropylethylamine (54.0 mg) and N,N'-thiocarbonyldiimidazole (11.0 mg) were added and allowed to react at room temperature for 1 hour. 3,3-Dimethyl-5-(piperidin-4-yl)-3,7-dihydrofuro[2,3-b]pyridin-6(2H)-one hydrochloride (15.0 mg) was added and the temperature was raised to 40°C for 1 hour. TLC indicated the reaction was complete. The mixture was filtered, and the filtrate was collected, purified by reverse phase preparative chromatography, and freeze-dried to obtain 2.0 mg of the title compound.

[0650] MS (ESI) m / z (M+H) + =675.4.

[0651] 1 H NMR (400MHz, DMSO-d6) δ12.97(s,1H),10.65(brs,1H),7.91(s,1H),7.58(d,J=7.5Hz,1H),7.32(s,1H),7.23(s,1H),6.95 (s,1H),5.47(q,J=7.5Hz,1H),4.76(t,J=13.3Hz,2H),4.10(s,2H),3.46–3.41(m,1H),3.22–3.20(m,2H),3.14–3.10(m,1H ),3.05–2.92(m,3H),2.93–2.78(m,3H),2.66–2.63(m,2H),2.41(s,3H),2.28(d,J=6.7Hz,1H),2.23–2.17(m,1H),2.03(s, 3H),2.00–1.93(m,1H),1.89–1.81(m,1H),1.69–1.63(m,4H),1.44–1.32(m,5H),1.18(d,J=2.9Hz,6H),1.14–1.09(m,1H).

[0652] Example 11: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxamide

[0653] Step 1: Preparation of (R)-methyl 3-(7-methyl-1H-indazol-5-yl)-2-(4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxamido)propanoate

[0654] 3-(Piperidin-4-yl)-5,7-dihydrothieno[3,4-b]pyridin-2(1H)-one hydrochloride (51.0 mg) and (R)-3-(7-methyl-1H-indazol-5-yl)-2-((phenoxycarbonyl)amino)propanoic acid methyl ester (68.0 mg) were dissolved in acetonitrile (3 mL). Triethylamine (64.0 mg) was added and the temperature was raised to 60°C for 8 hours. TLC indicated the reaction was complete. The system was concentrated and the crude product was purified by column chromatography to obtain 65.0 mg of the title compound.

[0655] MS (ESI) m / z (M+H) + =496.2.

[0656] Step 2: Preparation of (R)-3-(7-methyl-1H-indazol-5-yl)-2-(4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxamido)propanoic acid

[0657] Methyl (R)-3-(7-methyl-1H-indazol-5-yl)-2-(4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxamido)propanoate (30.0 mg) was dissolved in tetrahydrofuran (2 mL) and methanol (0.5 mL). Aqueous lithium hydroxide (7.6 mg) and 0.5 mL were added and allowed to react at room temperature for 1 hour. TLC indicated the reaction was complete. The pH of the system was adjusted to neutral with 1 M dilute hydrochloric acid, and the mother liquor was concentrated to yield 40.0 mg of the crude product.

[0658] MS (ESI) m / z (M+H) + =482.2.

[0659] Step 3: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxamide

[0660] (R)-3-(7-methyl-1H-indazol-5-yl)-2-(4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl)piperidine-1-carboxamido)propanoic acid (30.0 mg) and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (22.0 mg) were dissolved in N,N-dimethylformamide (2 mL). N,N-diisopropylethylamine (24.0 mg) was added and the mixture was allowed to react at room temperature for 10 minutes. 1-(1-methylpiperidin-4-yl)piperazine (17.0 mg) was then added and the mixture was allowed to react at room temperature for 20 minutes. TLC indicated the reaction was complete. The mixture was filtered, and the mother liquor was purified by reverse phase preparative chromatography and freeze-dried to yield 11.3 mg of the title compound.

[0661] MS (ESI) m / z (M+H) + =647.3.

[0662] 1H NMR(400MHz,DMSO-d6)δ13.01(s,1H),11.76(brs,1H),7.96(s,1H),7.36(s,1H),7.06(s,1H),7.00(s,1H),6.66( d,J=8.1Hz,1H),4.77(q,J=7.7Hz,1H),4.09(d,J=12.9Hz,2H),4.01–3.93(m,4H),3.58–3.52(m,1H),,3.27–3.22 (m,1H),3.15–3.09(m,1H),2.98–2.85(m,2H),2.78–2.62(m,5H),2.47(s,3H),2.36–2.29(m,1H),2.27–2.20(m,1 H),2.10(s,3H),1.99–1.86(m,2H),1.76–1.63(m,4H),1.57–1.53(m,1H),1.40–1.37(m,2H),,1.26–1.15(m,5H).

[0663] Example 12: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-3,4,5,6-tetrahydro-2H-thiopyrano[3,2-b]pyridin-7-yl)piperidine-1-carboxamide

[0664] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0665] MS (ESI) m / z (M+H) + =661.1.

[0666] 1H NMR (400MHz, DMSO-d6) δ12.94(s,1H),11.40(s,1H),7.89(s,1H),7.28(s,1H),6.93(s,1H),6.71(s,1H),6.57(d,J=8.0Hz,1H),4.70 (q,J=7.7Hz,1H),4.02(s,2H),3.49(d,J=13.5Hz,1H),3.15(t,J=9.6Hz,1H),3.03(t,J=9.4Hz,1H),2.92–2.79(m,4H),2.69–2.55(m, 5H),2.48(t,J=6.3Hz,2H),2.41(s,3H),2.26(s,1H),2.13(d,J=8.8Hz,1H),2.02(s,3H),1. 96–1.80(m,4H),1.69–1.57(m,4H),1.42(s,1H),1.30(d,J=12.0Hz,2H),1.21–1.07(m,5H).

[0667] Example 13: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-2,5,6,8-tetrahydro-1H-thiopyrano[3,4-b]pyridin-3-yl)piperidine-1-carboxamide

[0668] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0669] MS (ESI) m / z (M+H) + =660.8.

[0670] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.36(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H),6.89(s,1H),6.64(d,J=8.0 Hz,1H),4.82-4.72(m,1H),4.10(d,J=12.0Hz,2H),3.60-3.54(m,1H),3.50(s,2H),3.25-3.18(m,1H),3.15-3.08(m,1H ),2.98-2.92(m,1H),2.90-2.85(m,1H),2.81-2.78(m,3H),2.75-2.62(m,6H),2.47(s,3H),2.37-2.31(m,1H),2.25-2. 19(m,1H),2.09(s,3H),2.01-1.85(m,2H),1.75-1.64(m,4H),1.58-1.46(m,1H),1.43-1.32(m,2H),1.29-1.11(m,5H).

[0671] Example 14: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-1,2,5,7-tetrahydrothieno[3,4-b]pyridin-3-yl-7,7-d2)piperidine-1-carboxamide

[0672] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0673] MS (ESI) m / z (M+H) + =649.4.

[0674] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.75(brs,1H),7.96(s,1H),7.36(s,1H),7.06(s,1H),7.00(s,1H),6.65(d,J= 8.0Hz,1H),4.77(q,J=7.7Hz,1H),4.09(d,J=12.9Hz,2H),3.95(d,J=2.2Hz,2H),3.58–3.51(m,1H),3.26–3.21(m,1H) ,3.17–3.09(m,1H),2.99–2.93(m,1H),2.90–2.85(m,1H),2.79–2.62(m,5H),2.47(s,3H),2.36–2.31(m,1H),2.24–2. 20(m,1H),2.10(s,3H),1.99–1.87(m,2H),1.75–1.62(m,4H),1.58–1.53(m,1H),1.46–1.37(m,2H),1.27–1.12(m,5H).

[0675] Example 15: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-1,5,7,8-tetrahydro-2H-thiopyrano[4,3-b]pyridin-3-yl)piperidine-1-carboxamide

[0676] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0677] MS (ESI) m / z (M+H) + =661.3.

[0678] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.35(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H),6.90(s,1H),6.65(d,J=8.0Hz,1H) ,4.77(q,J=7.7Hz,1H),4.09(d,J=12.8Hz,2H),3.52(d,J=17.4Hz,3H),3.23(s,1H),3.15–3.06(m,1H),3.01–2.83(m,2H),2.82 (t,J=5.9Hz,2H),2.69(dt,J=20.1,8.4Hz,6H),2.47(s,3H),2.33(s,1H),2.23(d,J=10.4Hz,1H),2.10(s,3H),1.95(dd,J=25.7 ,15.2,9.4Hz,3H),1.75(d,J=11.4Hz,1H),1.71–1.61(m,2H),1.54(t,J=8.9Hz,1H),1.38(d,J=12.4Hz,2H),1.30–1.10(m,6H).

[0679] Example 16: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-thiocarboamide

[0680] The title compound was prepared using a similar preparation method to that of Example 10 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and embodiments as raw materials.

[0681] MS (ESI) m / z (M+H) + =663.3.

[0682] 1H NMR (400MHz, DMSO-d6) δ13.04(s,1H),11.49(brs,1H),7.98(s,1H),7.67(d,J=7.5Hz,1H),7.40(s,1H),7.10(s,1 H),7.02(s,1H),5.54(q,J=7.5Hz,1H),4.80(t,J=12.5Hz,2H),3.50–3.45(m,1H),3.42–3.38(m,2H),3.25–3.18(m ,2H),3.09–2.96(m,4H),2.99–2.83(m,3H),2.72(d,J=10.9Hz,2H),2.48(s,3H),2.35–2.23(m,2H),2.10(s,3H), 2.08–1.99(m,1H),1.97–1.89(m,1H),1.77–1.67(m,4H),1.65–1.59(m,1H),1.44–1.31(m,4H),1.29–1.10(m,3H).

[0683] Example 17: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(5-oxo-2,3,4,5-tetrahydrothieno[3,2-b]pyridin-6-yl)piperidine-1-carboxamide

[0684] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0685] MS (ESI) m / z (M+H) + =647.1.

[0686] 1H NMR (400MHz, DMSO-d6) δ12.94(s,1H),11.67(s,1H),7.89(s,1H),7.28(s,1H),7.07(s,1H),6.92(s,1H),6.58(d,J=8 .0Hz,1H),4.70(q,J=7.7Hz,1H),4.03(t,J=11.1Hz,2H),3.54–3.44(m,1H),3.24(s,2H),3.18–3.11(m,1H),3.00(t,J =8.1Hz,3H),2.90(dd,J=13.1,8.1Hz,1H),2.81(dd,J=13.1,6.8Hz,1H),2.72–2.54(m,5H),2.40(s,3H),2.29–2.22(m ,1H),2.17–2.10(m,1H),2.02(s,3H),1.91–1.77(m,2H),1.69–1.55(m,4H),1.41(t,J=9.1Hz,1H),1.32–1.05(m,7H).

[0687] Example 18: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperazine-1-carboxamide

[0688] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0689] MS (ESI) m / z (M+H) + =661.8.

[0690] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.41(s,1H),7.96(s,1H),7.36(s,1H),6.99(s,1H),6.78(d,J=8.0Hz,1H), 6.45(s,1H),4.82-4.72(m,1H),3.60-3.53(m,1H),3.40-3.35(m,3H),3.27-3.18(m,1H),3.15-3.05(m,1H),3.05- 2.92(m,3H),2.90-2.80(m,5H),2.75-2.62(m,2H),2.55-2.51(m,2H),2.47(s,3H),2.35-2.25(m,1H),2.23-2.18( m,1H),2.09(s,3H),2.01-1.85(m,4H),1.76-1.64(m,2H),1.55-1.46(m,1H),1.42-1.32(m,2H),1.29-1.09(m,4H).

[0691] Example 19: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0692] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0693] MS (ESI) m / z (M+H) + =647.1.

[0694] 1H NMR (400MHz, DMSO-d6) δ12.93(s,1H),11.40(s,1H),7.89(d,J=3.2Hz,1H),7.30(d,J=14.3Hz,1H),6.94(d,J=5.6Hz,2H),6.59(dd,J= 12.8,8.1Hz,1H),4.74(dq,J=15.2,7.6Hz,1H),4.26(d,J=12.7Hz,1H),4.03(s,2H),3.85(t,J=16.0Hz,1H),3.33(t,J=7.9Hz,2H),2.9 9(t,J=8.0Hz,2H),2.94–2.86(m,1H),2.81(dd,J=13.1,6.4Hz,1H),2.73–2.55(m,4H),2.40(d,J=4.1Hz,3H),2.34(s,1H),2.30–2.07( m,5H),2.04(d,J=5.1Hz,3H),1.95–1.81(m,3H),1.57(d,J=14.5Hz,3H),1.37(d,J=12.2Hz,1H),1.27–1.10(m,4H),0.82–0.42(m,1H).

[0695] Example 20: Preparation of (R)-N-(1-([1,4'-bipiperidinyl]-1'-yl)-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0696] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0697] MS (ESI) m / z (M+H) + =632.1.

[0698] 1H NMR (400MHz, DMSO-d6) δ12.99(s,1H),11.45(s,1H),7.96(d,J=3.1Hz,1H),7.37(d,J=14.4Hz,1H),7.01(d,J=5.0Hz,2H) ,6.65(dd,J=14.3,8.1Hz,1H),4.80(dq,J=24.1,7.9Hz,1H),4.34(d,J=12.1Hz,1H),4.10(s,2H),3.94(t,J=16.8Hz,1H) ,3.40(t,J=7.9Hz,2H),3.06(t,J=7.9Hz,2H),3.00–2.92(m,1H),2.88(dd,J=13.3,6.3Hz,1H),2.78–2.61(m,4H),2.47( d,J=4.7Hz,3H),2.38(s,2H),2.30–2.19(m,1H),2.02–1.79(m,3H),1.62(s,3H),1.45–1.20(m,10H),0.74–0.55(m,1H).

[0699] Example 21: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-1-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-4-carboxamide

[0700] Step 1: Preparation of 1-(7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-4-carboxylic acid

[0701] Under a nitrogen atmosphere, 6-bromo-7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridine (50.0 mg), ethyl piperidine-4-carboxylate (36.3 mg), tris(dibenzylideneacetone)dipalladium (17.4 mg), 2-dicyclohexylphosphino-2',6'-diisopropylbiphenyl (17.7 mg), and lithium tert-butoxide (30.4 mg) were dissolved in 1,4-dioxane (1 mL) and reacted at 100°C for 2 hours. LCMS showed that some starting material remained, and the reaction was terminated. The reaction solution was filtered, and the filtrate was collected and concentrated. The resulting crude product was purified by column chromatography to obtain 40.0 mg of the title compound.

[0702] MS (ESI) m / z (M+H) + =308.9.

[0703] Step 2: Preparation of 1-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-4-carboxylic acid hydrochloride

[0704] Dissolve 1-(7-methoxy-3,4-dihydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-4-carboxylic acid (40 mg) in concentrated hydrochloric acid (2 mL) and react at 95°C for 2 hours. LCMS indicated the reaction was complete. The reaction solution was concentrated to yield 38 mg of crude product, which was used directly in the next step.

[0705] MS (ESI) m / z (M+H) + =294.9.

[0706] Step 3: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-1-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-4-carboxamide

[0707] 1-(7-Oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-4-carboxylic acid hydrochloride (38 mg), N,N-diisopropylethylamine (50.0 mg), and (R)-2-amino-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one (54.6 mg) were dissolved in N,N-dimethylformamide (1.0 mL). 2-(1H-benzotriazol-L-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (62.2 mg) was added and allowed to react for 1 hour. LCMS indicated the reaction was complete. The reaction solution was concentrated, and the crude product was purified by preparative liquid chromatography and freeze-dried to obtain 17.3 mg of the title compound.

[0708] MS (ESI) m / z (M+H) + =660.9.

[0709] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.31(s,1H),8.21(d,J=8.0Hz,1H),7.96(s,1H),7.34(s,1H),6.99(s, 1H),6.45(s,1H),4.94-4.89(m,1H),3.60-3.45(m,3H),3.28-3.23(m,2H),3.20-3.08(m,1H),3.08-2.92(m,3 H),2.88-2.80(m,1H),2.75-2.65(m,2H),2.47(s,3H),2.37-2.30(m,3H),2.28-2.19(m,2H),2.09(s,3H),2.0 5-1.96(m,1H),1.95-1.86(m,3H),1.79-1.68(m,2H),1.65-1.48(m,5H),1.45-1.35(m,2H),1.33-1.08(m,4H).

[0710] Example 22: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4'-methyl-[1,4'-bipiperidinyl]-1'-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0711] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0712] MS (ESI) m / z (M+H) + =646.3.

[0713] 1H NMR (400MHz, DMSO-d6) δ13.02(s,1H),7.96(d,J=5.7Hz,1H),7.37(d,J=6.5Hz,1H),7.09 (d,J=13.4Hz,1H),6.99(d,J=15.0Hz,1H),6.61(t,J=6.9Hz,1H),4.78(s,1H),4.09(d,J= 13.0Hz,2H),3.40(s,1H),3.06(t,J=8.0Hz,2H),2.93(dd,J=19.0,9.0Hz,2H),2.74(d,J= 14.2Hz,1H),2.66(d,J=12.7Hz,2H),2.47(d,J=2.5Hz,3H),2.36–2.18(m,5H),2.00(q,J= 7.1Hz,1H),1.62(s,3H),1.52–1.39(m,4H),1.38–1.30(m,4H),1.30–1. 17(m,5H),0.85(t,J=6.5Hz,1H),0.82–0.68(m,2H),0.59–0.38(m,2H).

[0714] Example 23: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-thiocarboxamide

[0715] The title compound was prepared using a similar preparation method to that of Example 10 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0716] MS (ESI) m / z (M+H) + =677.3

[0717] 1H NMR (400MHz, DMSO-d6) δ13.03(s,1H),11.36(s,1H),7.97(s,1H),7.66(d,J=7.6Hz,1H),7.39(s,1H),7.02(s,1H),6.87(s,1H), 5.54(q,J=7.5Hz,1H),4.80(t,J=11.3Hz,2H),3.53–3.43(m,1H),3.32-3.25(m,2H),3.24–3.16(m,1H),3.07–2.90(m,6H),2.88 -2.76(m,1H),2.71(d,J=11.0Hz,2H),2.53(t,J=6.1Hz,2H),2.48(s,3H),2.36 -2.30(m,1H),2.29–2.20(m,1H),2.09(s,3H),2.08–2.00(m,1H),1.98-1.88(m, 3H),1.76–1.66(m,4H),1.64-1.58(m,1H),1.45–1.31(m,4H),1.29–1.10(m,2H).

[0718] Example 24: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4'-methyl-[1,4'-bipiperidinyl]-1'-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0719] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0720] MS (ESI) m / z (M+H) + =660.3.

[0721] 1H NMR(400MHz,DMSO-d6)δ13.01(s,1H),11.33(s,1H),7.96(s,0.5H),7.94(s,0.5H),7.36(s,0 .5H),7.35(s,0.5H),7.01(s,0.5H),6.97(s,0.5H),6.87(s,0.5H),6.84(s,0.5H),6.62-6.58 (m,1H),4.82-4.73(m,1H),4.09(d,J=13.0Hz,2H),3.80-3.75(m,1H),3.63-3.58(m,0.5H),3.39-3.3 5(m,0.5H),3.20(t,J=11.6Hz,0.5H),3.13–3.05(m,0.5H),3.05–2.83(m,5H),2.74–2.59(m,3H),2.5 5-2.52(m,1.5H),2.47(s,1.5H),2.46(s,1.5H),2.34-2.18(m,4H),1.98-1.91(m,2H),1.64-1.56(m, 3H),1.50–1.10(m,10.5H),0.77(s,1.5H),0.76-0.70(m,0.5H),0.47(s,1.5H),0.17-0.11(m,0.5H).

[0722] Example 25: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0723] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0724] MS (ESI) m / z (M+H) + =661.4.

[0725] 1H NMR (400MHz, DMSO-d6) δ12.99(s,1H),11.32(s,1H),7.96(d,J=3.1Hz,1H),7.36(d,J=13.7Hz,1H),7.00(d,J=3.8Hz,1H),6.90(s, 1H),6.65(dd,J=13.9,8.1Hz,1H),4.80(dq,J=25.6,7.7Hz,1H),4.34(d,J=12.5Hz,1H),4.10(s,2H),3.93(t,J=15.5Hz,1H),3.05 –2.99(m,2H),2.99–2.77(m,2H),2.76–2.55(m,4H),2.53(d,J=6.0Hz,4H),2.47(d,J=4.4Hz,3H),2.41–2.30(m,2H),2.23(dd,J=2 3.8,11.9Hz,1H),1.91(ddt,J=33.0,10.8,5.6Hz,4H),1.65(d,J=12.6Hz,3H),1.43(s,2H),1.37–0.81(m,8H),0.86–0.56(m,1H).

[0726] Example 26: Preparation of (R)-N-(1-([1,4'-bipiperidinyl]-1'-yl)-3-(7-methyl-1H-indazol-5-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0727] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0728] MS (ESI) m / z (M+H) + = 646.3.

[0729] 1H NMR (400MHz, DMSO-d6) δ13.00(s,1H),11.33(s,1H),7.96(d,J=3.3Hz,1H),7.36(d,J=13.7Hz,1H),7.01(d,J=4.2Hz,1H),6.85(d,J=38.7Hz,1 H),6.70–6.60(m,1H),4.80(dq,J=24.2,7.8Hz,1H),4.32(d,J=12.6Hz,1H),4.10(s,2H),3.91(t,J=14.3Hz,1H),3.09–2.92(m,3H),2.87(dt, J=13.1,6.2Hz,1H),2.69(td,J=25.3,22.8,11.9Hz,4H),2.54(t,J=6.2Hz,2H),2.47(d,J=3.7Hz,3H),2.43–2.31(m,2H),2.23(t,J=28.8Hz,4 H),2.10(d,J=4.6Hz,3H),1.94(td,J=10.8,5.7Hz,5H),1.65(d,J=12.9Hz,2H),1.44(d,J=12.3Hz,1H),1.36–1.15(m,4H),0.87–0.49(m,1H).

[0730] Example 27: Preparation of 2-((7-methyl-1H-indazol-5-yl)methyl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-4-(4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidin-1-yl)butane-1,4-dione

[0731] Step 1: Preparation of methyl 2-((7-methyl-1H-indazol-5-yl)methyl)-4-oxo-4-(4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidin-1-yl)butanoate

[0732] 4-Methoxy-3-((7-methyl-1H-indazol-5-yl)methyl)-4-oxobutanoic acid (80 mg), 6-(piperidin-4-yl)-2,3,4,8-tetrahydro-7H-thiopyrano[2,3-b]pyridin-7-one hydrochloride (72 mg), O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (111 mg), and N,N-diisopropylethylamine (112 mg) were dissolved in N,N-dimethylformamide (2.0 mL) and reacted at room temperature for 2 hours. LCMS showed the reaction was complete. The reaction solution was quenched with saturated ammonium chloride solution (5 mL) and extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative liquid chromatography to yield 45 mg of the title compound.

[0733] MS (ESI) m / z (M+H) + = 509.2.

[0734] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0735] MS (ESI) m / z (M+H) + = 660.4.

[0736] 1 H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.32(s,1H),7.96(s,1H),7.29(s,1H),6.94(d,J=7.8Hz,2H),4.44(d,J=12.6 Hz,1H),3.99–3.89(m,1H),3.67(d,J=12.9Hz,1H),3.43(s,1H),3.32(s,1H),3.11–2.97(m,4H),2.93–2.77(m,3H),2. 71(dd,J=19.5,9.4Hz,4H),2.54(d,J=6.1Hz,2H),2.48(s,3H),2.42–2.27(m,2H),2.16(d,J=11.1Hz,1H),2.08(s,3H) ,1.94(p,J=6.1Hz,2H),1.82–1.62(m,6H),1.43–1.34(m,1H),1.31–1.19(m,4H),1.11(ddt,J=20.4,11.9,5.8Hz,3H).

[0737] Example 28: Preparation of (S)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0738] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0739] MS (ESI) m / z (M+H) + =661.3.

[0740] 1 H NMR(400MHz,DMSO-d6)δ13.01(s,1H),11.27(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H ),6.86(s,1H),6.64(d,J=8.1Hz,1H),4.77(q,J=7.7Hz,1H),4.09(d,J=12.9Hz,2H),3.62 –3.51(m,1H),3.26–3.17(m,1H),3.15–3.06(m,1H),3.04–2.85(m,4H) ,2.76–2.59(m,5H),2.54(t,J=6.1Hz,2H),2.47(s,3H),2.37–2.29(m, 1H),2.26–2.18(m,1H),2.09(s,3H),2.00–1.85(m,4H),1.76–1.62(m, 4H),1.52(t,J=9.3Hz,1H),1.37(d,J=12.2Hz,2H),1.32-1.14(m,5H).

[0741] Example 29: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-(methyl-d3)piperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0742] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0743] MS (ESI) m / z (M+H)+ =650.4.

[0744] 1 H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.45(brs,1H),7.96(s,1H),7.36(s,1H),7.09(s,1H),7.00(s,1H),6.65(d,J=8.0 Hz,1H),4.77(q,J=7.7Hz,1H),4.14–4.05(m,2H),3.58–3.52(m,1H),3.42–3.38(m,2H),3.27–3.21(m,1H),3.16–3.10(m,1 H),3.10–3.02(m,3H),2.98–2.86(m,2H),2.75–2.70(m,3H),2.69–2.60(m,2H),2.47(s,3H),2.36–2.31(m,1H),2.26–2.19 (m,1H),1.98–1.89(m,2H),1.80–1.75(m,2H),1.68–1.60(m,2H),1.57–1.52(m,1H),1.41–1.38(m,1H),1.31–1.16(m,5H).

[0745] Example 30: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-(methyl-d3)piperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0746] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0747] MS (ESI) m / z (M+H) + =664.4.

[0748] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.32(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H),6.86(s,1H),6.64(d,J=8.1Hz,1H), 4.77(q,J=7.7Hz,1H),4.09(d,J=12.6Hz,2H),3.57–3.54(m,1H),3.25–3.20(m,1H),3.13–3.07(m,1H),3.04–3.01(m,2H),2.98 –2.86(m,2H),2.72–2.67(m,4H),2.64–2.61(m,1H),2.58–2.54(m,2H),2.47(s,3H),2.35–2.32(m,1H),2.25–2.19(m,1H),1.98 –1.93(m,3H),1.92–1.84(m,1H),1.75–1.70(m,2H),1.68–1.63(m,2H),1.55–1.51(m,1H),1.39–1.36(m,2H),1.29–1.14(m,5H).

[0749] Example 31: Preparation of 2-((7-methyl-1H-indazol-5-yl)methyl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-4-(4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidin-1-yl)butane-1,4-dione

[0750] The title compound was prepared using a similar preparation method to Example 27 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0751] MS (ESI) m / z (M+H) + =646.5.

[0752] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.46(s,1H),7.96(s,1H),7.29(s,1H),7.18(d,J=3.3Hz,1H),6.95(s,1H), 4.44(d,J=12.8Hz,1H),3.95(d,J=13.3Hz,1H),3.67(d,J=12.4Hz,1H),3.39(t,J=7.9Hz,3H),3.29(s,1H),3.04(q ,J=6.7,5.7Hz,3H),2.90–2.78(m,3H),2.71(dd,J=19.5,9.2Hz,4H),2.48(s,3H),2.40(dd,J=15.8,3.6Hz,1H),2. 31(d,J=12.4Hz,2H),2.08(s,4H),1.86–1.64(m,7H),1.38(d,J=12.3Hz,1H),1.32–1.21(m,5H),1.20-1.05(m,1H).

[0753] Example 32: Preparation of N-((R)-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(1-oxido-7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0754] The title compound was prepared using a similar preparation method to that of Example 9 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0755] MS (ESI) m / z (M+H) + = 677.4.

[0756] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),12.03(s,1H),7.96(d,J=1.6Hz,1H),7.36(s,1H),7.23(s,1H),7.00(s,1H),6.68(d,J=8.0 Hz,1H),4.77(q,J=7.6Hz,1H),4.12(d,J=13.2Hz,2H),3.56(d,J=14.0Hz,1H),3.22(d,J=10.4Hz,2H),3.12(dt,J=19.2,9.7Hz,2 H),3.04–2.93(m,2H),2.91–2.81(m,2H),2.77–2.63(m,5H),2.47(s,3H),2.33(dd,J=11.2,6.4Hz,1H),2.21(d,J=10.4Hz,2H),2 .10(s,3H),2.02–1.85(m,3H),1.79–1.60(m,4H),1.52(t,J=9.4Hz,1H),1.43–1.28(m,4H),1.19(ddd,J=23.6,12.4,3.2Hz,3H).

[0757] Example 33: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6'-oxo-6',7'-dihydro-2'H-spiro[cyclopropane-1,3'-thieno[2,3-b]pyridine]-5'-yl)piperidine-1-carboxamide

[0758] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0759] MS (ESI) m / z (M+H) + =673.4.

[0760] 1H NMR(400MHz,DMSO-d6)δ13.01(s,1H),11.31(s,1H),7.96(s,1H),7.35(s, 1H),6.99(s,1H),6.68(s,1H),6.61(d,J=8.1Hz,1H),4.76(q,J=7.8Hz,1H ),4.10(d,J=12.9Hz,2H),3.57(d,J=12.8Hz,1H),3.38(s,2H),3.20(t,J= 9.8Hz,1H),3.07(t,J=10.0Hz,1H),2.96(dd,J=13.1,8.1Hz,1H),2.87(dd ,J=13.1,6.8Hz,1H),2.74–2.59(m,5H),2.47(s,3H),2.38–2.29(m,1H),2 .21(d,J=11.4Hz,1H),2.09(s,3H),2.03–1.83(m,2H),1.72(tt,J=11.8,2 .9Hz,2H),1.60(d,J=12.6Hz,2H),1.47(t,J=9.1Hz,1H),1.34(tt,J=12.0 ,5.3Hz,4H),1.24(d,J=3.6Hz,1H),1.21–1.08(m,2H),0.97–0.89(m,4H).

[0761] Example 34: Preparation of N-((2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octan-8-yl)propan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0762] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0763] MS (ESI) m / z (M+H) + =658.1.

[0764] 1H NMR (400MHz, DMSO-d6) δ12.99(s,1H),11.38(s,1H),7.95(t,J=2.3Hz,1H),7.37(d,J=7.2Hz,1H),7. 14–6.98(m,2H),6.69(q,J=8.4Hz,1H),4.66–4.57(m,1H),4.53–4.38(m,1H),4.33–3.95(m,3H),3.4 0(t,J=7.9Hz,2H),3.05(t,J=7.9Hz,2H),2.90(d,J=7.1Hz,1H),2.87–2.62(m,4H),2.46(d,J=2.8Hz ,3H),2.34(d,J=20.2Hz,3H),2.11–1.96(m,1H),1.86–1.75(m,2H),1.61(s,4H),1.54–1.19(m,13H).

[0765] Example 35: Preparation of N-((R)-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(1-oxido-6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0766] The title compound was prepared using a similar preparation method to that of Example 9 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0767] MS (ESI) m / z (M+H) + =663.3.

[0768] 1H NMR (400MHz, DMSO-d6) δ13.03(s,1H),12.01(s,1H),7.97(d,J=2.7Hz,1H),7.45(d,J=2.8Hz,1H),7.38(s,1H),7.02(s,1H),6.71(d,J= 8.0Hz,1H),4.78(q,J=7.8Hz,1H),4.12(d,J=12.9Hz,2H),3.64–3.37(m,5H),3.26–3.02(m,6H),3.01–2.81(m,3H),2.78–2.53(m,5H), 2.48(s,3H),2.44–2.09(m,3H),1.99(q,J=13.2,7.3Hz,2H),1.79–1.53(m,3H),1.46(s,1H),1.40–1.14(m,5H).

[0769] Example 36: Preparation of N-((2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octan-8-yl)propan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0770] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0771] MS (ESI) m / z (M+H) + =644.3.

[0772] 1H NMR (400MHz, DMSO-d6) δ12.99(s,1H),11.46(s,1H),7.95(d,J=2.2Hz,1H),7.52–7.31(m,1H),7.12–6.97(m,2H ),6.70–6.39(m,1H),4.74–4.43(m,1H),4.29(d,J=23.3Hz,1H),4.08(t,J=12.2Hz,3H),3.47–3.37(m,5H),3.1 9–2.99(m,3H),2.96–2.81(m,2H),2.78–2.58(m,2H),2.47(d,J=4.4Hz,3H),2.31(d,J=64.9Hz,3H),2.04–1.88 (m,1H),1.74(t,J=11.0Hz,1H),1.60(d,J=12.5Hz,2H),1.45(d,J=10.8Hz,2H),1.40–1.28(m,6H),1.24(d,3H).

[0773] Example 37: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl-3-d)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0774] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0775] MS (ESI) m / z (M+H) + =661.8.

[0776] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.31(s,1H),7.36(s,1H),7.00(s,1H),6.86(s,1H),6.64(d,J=8.0Hz,1H),4 .80-4.72(m,1H),4.14-4.02(m,2H),3.57-3.53(m,1H),3.27-3.18(m,2H),3.14-3.08(m,1H),3.04-3.00(m,2H),2.9 8-2.93(m,1H),2.92-2.86(m,1H),2.74-2.60(m,6H),2.56-2.54(m,1H),2.47(s,3H),2.37-2.31(m,1H),2.25-2.19 (m,1H),2.09(s,3H),2.01-1.89(m,4H),1.74-1.61(m,4H),1.55-1.49(m,1H),1.40-1.34(m,2H),1.30-1.13(m,4H).

[0777] Example 38: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl-3-d)-1-(4-(1-(methyl-d3)piperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0778] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0779] MS (ESI) m / z (M+H) + =665.8.

[0780] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.31(s,1H),7.36(s,1H),7.00(s,1H),6.86(s,1H),6.64(d,J=8.0Hz,1H ),4.80-4.72(m,1H),4.14-4.02(m,2H),3.57-3.53(m,1H),3.27-3.18(m,2H),3.14-3.08(m,1H),3.04-3.00(m,2 H),2.98-2.93(m,1H),2.92-2.86(m,1H),2.74-2.60(m,6H),2.56-2.54(m,1H),2.47(s,3H),2.37-2.31(m,1H),2 .25-2.19(m,1H),2.01-1.89(m,4H),1.74-1.61(m,4H),1.55-1.49(m,1H),1.40-1.34(m,2H),1.30-1.13(m,4H).

[0781] Example 39: Preparation of N-((2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(3-(piperidin-1-yl)-8-azabicyclo[3.2.1]octan-8-yl)propan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0782] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0783] MS (ESI) m / z (M+H) + = 672.4.

[0784] 1H NMR (400MHz, DMSO-d6) δ12.99(s,1H),10.97(s,1H),8.05–7.84(m,1H),7.37(d,J=8.7Hz,1H),7.07–6.97(m,1H),6.91–6.80(m,1 H),6.68(dt,J=14.7,7.6Hz,1H),4.59(dd,J=17.1,8.3Hz,1H),4.40(dd,J=40.4,32.5Hz,2H),4.05(d,J=44.4Hz,2H),3.08–2.99 (m,2H),2.91(tt,J=23.6,6.7Hz,2H),2.75–2.61(m,3H),2.57–2.51(m,2H),2.46(d,J=2.6Hz,3H),2.43–2.21(m,3H),2.07(s,1H ),1.99–1.93(m,2H),1.87–1.73(m,2H),1.63(t,J=10.4Hz,4H),1.48–1.36(m,4H),1.25(d,J=11.2Hz,6H),0.94(d,J=6.4Hz,3H).

[0785] Example 40: Preparation of N-((2R)-3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(5-(piperidin-1-yl)-2-azabicyclo[2.2.1]hept-2-yl)propan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0786] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0787] MS (ESI) m / z (M+H) + = 658.3.

[0788] 1H NMR (400MHz, DMSO-d6) δ13.01(d,J=15.6Hz,1H),11.38(s,1H),7.95(d,J=2.4Hz,1H),7.46–7.35(m,1H),7.02(dd,J=13.2,10.4Hz,1 H),6.83(d,J=14.0Hz,1H),6.67–6.52(m,1H),4.58(ddt,J=30.2,22.6,7.6Hz,1H),4.43–4.24(m,1H),4.08(t,J=13.6Hz,2H),3.67–3 .37(m,1H),3.04–3.00(m,2H),2.94–2.84(m,2H),2.67(t,J=13.2Hz,3H),2.47(d,J=4.4Hz,3H),2.35–2.11(m,4H),1.95(s,3H),1.74 (t,J=10.0Hz,1H),1.62(d,J=12.6Hz,2H),1.45(d,J=11.3Hz,4H),1.32(d,J=17.6Hz,4H),1.23(s,5H),0.90(dd,J=34.6,6.4Hz,2H).

[0789] Example 41: Preparation of 3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl 4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxylate

[0790] Step 1: Preparation of tert-butyl 7-methyl-5-(3-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-2-(((4-nitrophenoxy)carbonyl)oxy)-3-oxopropyl)-1H-indazole-1-carboxylate

[0791] Under ice-water conditions, 2-hydroxy-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)propan-1-one (40.0 mg) and 4-dimethylaminopyridine (1.27 mg) were dissolved in anhydrous dichloromethane (2.0 mL). Di-tert-butyl dicarbonate (24.9 mg) was slowly added dropwise and allowed to react for 1 hour. p-Nitrophenyl chloroformate (31.4 mg) was added portionwise and the mixture was allowed to react at room temperature for 15 hours. LCMS indicated that the reaction was complete. Half of the reaction solution was withdrawn and used directly in the next step.

[0792] MS (ESI) m / z (M+H) + =651.9.

[0793] Step 2: Preparation of 3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxylate

[0794] N,N-Diisopropylethylamine (7.94 mg) and anhydrous dichloromethane (1.0 mL) were added to the above reaction solution, followed by 6-(piperidin-4-yl)-2,3,4,8-tetrahydro-7H-thiopyrano[2,3-b]pyridin-7-one hydrochloride (9.23 mg). The reaction was allowed to react for 1 hour. LCMS indicated the reaction was complete. The reaction solution was filtered, the filtrate collected, and concentrated. The crude product was purified by preparative liquid chromatography to yield 4.0 mg of the title compound.

[0795] MS (ESI) m / z (M+H) + =661.8.

[0796] 1 H NMR(400MHz,DMSO-d6)δ12.99(s,1H),7.98(s,1H),7.41(s,1H),7.04(s,1H), 7.05-6.75(m,1H),5.42-5.35(m,1H),4.25-3.89(m,3H),3.65-3.50(m,2H),3. 23-2.95(m,6H),2.94-2.66(m,5H),2.48(s,3H),2.43-2.24(m,2H),2.10(s,3H) ),2.06-1.88(m,4H),1.81-1.60(m,5H),1.50-1.35(m,2H),1.32-1.10(m,5H).

[0797] Example 42: Preparation of 3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxylate

[0798] The title compound was prepared using a similar preparation method to that of Example 41 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0799] MS (ESI) m / z (M+H) + =647.8.

[0800] 1 H NMR(400MHz,DMSO-d6)δ13.06(s,1H),11.44(s,1H),7.98(s,1H),7.42(s,1H),7.04(s, 1H),7.05-6.75(m,1H),5.42-5.31(m,1H),4.25-3.89(m,2H),3.75-3.50(m,1H),3.45- 23.35(m,3H),3.20-2.95(m,5H),2.92-2.66(m,4H),2.48(s,3H),2.43-2.24(m,2H),2. 10(s,3H),2.06-1.88(m,2H),1.81-1.60(m,4H),1.50-1.35(m,2H),1.35-1.10(m,7H).

[0801] Example 43: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl-3-d)-1-(4-(1-(methyl-d3)piperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0802] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0803] MS (ESI) m / z (M+H) + =650.8.

[0804] 1H NMR (400MHz, DMSO-d6) δ13.00(s,1H),11.43(s,1H),7.36(s,1H),7.09(s,1H),7.00(s,1H),6.65(d,J=8 .0Hz,1H),4.82-4.72(m,1H),4.09(d,J=16.0,2H),3.65-3.50(m,1H),3.45-3.35(m,2H),3.29-3.21(m, 1H),3.17-3.03(m,2H),3.00-2.83(m,2H),2.80-2.60(m,4H),2.47(s,3H),2.37-2.31(m,1H),2.25-2.1 9(m,1H),2.05-1.85(m,3H),1.80-1.69(m,2H),1.67-1.51(m,3H),1.44-1.34(m,2H),1.32-1.13(m,6H).

[0805] Example 44: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl-3-d)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0806] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0807] MS (ESI) m / z (M+H) + =647.8.

[0808] 1H NMR (400MHz, DMSO-d6) δ13.00(s,1H),11.44(s,1H),7.36(s,1H),7.09(s,1H),7.00(s,1H),6.65(d,J=8.0H z,1H),4.82-4.72(m,1H),4.09(d,J=12.0Hz,2H),3.65-3.50(m,1H),3.45-3.35(m,2H),3.29-3.21(m,1H),3 .13-3.02(m,2H),3.00-2.83(m,2H),2.79-2.61(m,4H),2.47(s,3H),2.37-2.31(m,1H),2.25-2.19(m,1H), 2.10(s,3H),2.05-1.85(m,2H),1.80-1.69(m,2H),1.67-1.51(m,3H),1.44-1.34(m,2H),1.32-1.13(m,7H).

[0809] Example 45: Preparation of N-((R)-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-1,2,5,6,7,8-hexahydro-5,8-methanoquinolin-3-yl)piperidine-1-carboxamide

[0810] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0811] MS (ESI) m / z (M+H) + =655.4

[0812] 1H NMR(400MHz,DMSO-d6)δ13.01(s,1H),11.72(s,1H),7.96(s,1H),7.36(s,1H),7.06(s,1H),7.00(s,1H),6.6 3(d,J=8.0Hz,1H),4.77(q,J=7.7Hz,1H),4.08(d,J=12.7Hz,2H),3.55(d,J=13.6Hz,1H),3.38-3.33(m,1H), 3.26-3.20(m,2H),3.18–3.05(m,2H),3.00–2.84(m,2H),2.76-2.60(m,5H),2.47(s,3H),2.37–2.29(m,1H), 2.27–2.18(m,1H),2.09(s,3H),1.99–1.52(m,10H),1.43–1.33(m,3H),1.29-1.21(m,4H),1.06-0.96(m,2H).

[0813] Example 46: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7'-oxo-7',8'-dihydro-2'H,4'H-spiro[cyclopropane-1,3'-thiopyrano[2,3-b]pyridine]-6'-yl)piperidine-1-carboxamide

[0814] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0815] MS (ESI) m / z (M+H) + = 687.4.

[0816] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.36(s,1H),7.95(s,1H),7.35(s,1H),7.00(s,1H),6.78(s,1H),6.64(d,J=8.0Hz,1H),4.76(q, J=7.6Hz,1H),4.08(d,J=12.8Hz,2H),3.55(d,J=13.2Hz,1H),3.23(s,1H),3.10(t,J=9.6Hz,1H),2.95(dd,J=13.2,7.6Hz,1H),2.88(d,J =16.0Hz,3H),2.67(dt,J=25.6,11.2Hz,5H),2.47(s,3H),2.41(s,2H),2.33(s,1H),2.22(s,1H),2.09(s,3H),1.92(dt,J=23.6,11.2Hz, 2H),1.72(t,J=11.6Hz,2H),1.64(d,J=12.6Hz,2H),1.54(t,J=9.2Hz,1H),1.38(d,J=12.0Hz,2H),1.32–1.09(m,5H),0.68–0.50(m,4H).

[0817] Example 47: Preparation of N-((R)-3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(5-oxo-1,1a,2,4,5,7b-hexahydrocyclopropano[4,5]thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0818] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0819] MS (ESI) m / z (M+H) + =673.3

[0820] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.31(s,1H),7.96(s,1H),7.36(s,1H),7.16(d,J=7.5Hz,1H),7.00(s,1H),6.65(d,J= 7.9Hz,1H),4.77(q,J=7.7Hz,1H),4.11(d,J=12.9Hz,2H),3.56(d,J=13.3Hz,1H),3.40-3.36(m,1H),3.24-3.19(m,1H),3.12- 3.06(m,3H),2.99-2.93(m,1H),2.90-2.84(m,1H),2.77–2.60(m,5H),2.47(s,3H),2.36-2.29(m,1H),2.26–2.17(m,1H),2.09 (s,3H),1.97–1.78(m,4H),1.76–1.61(m,4H),1.52(t,J=9.8Hz,1H),1.42–1.27(m,5H),1.24–1.10(m,2H),0.85-0.77(m,1H).

[0821] Example 48: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(7-oxo-7,8-dihydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0822] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0823] MS (ESI) m / z (M+H) + =659.3.

[0824] 1H NMR(400MHz,DMSO-d6)δ13.01(s,1H),11.57(s,1H),7.96(s,1H),7.36(s,1H),7.02(s,1H),6.99(s,1H),6.65(d ,J=7.9Hz,1H),6.33(d,J=9.9Hz,1H),5.62-5.57(m,1H),4.79-4.73(m,1H),4.10(d,J=12.5Hz,2H),3.63–3.50(m ,3H),3.30-3.18(m,1H),3.14–3.05(m,1H),3.05–2.84(m,3H),2.75–2.62(m,5H),2.47(s,3H),2.35-2.28(m,1H) ,2.24-2.16(m,1H),2.10(s,3H),2.00–1.86(m,2H),1.77–1.61(m,4H),1.51(t,J=8.7Hz,1H),1.39–1.11(m,6H).

[0825] Example 49: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(4-(piperidin-4-yl)piperazin-1-yl)propan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0826] Step 1: Preparation of (R)-tert-butyl 4-(4-(3-(7-methyl-1H-indazol-5-yl)-2-(4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamido)propanoyl)piperazin-1-yl)piperidine-1-carboxylate

[0827] (R)-3-(7-methyl-1H-indazol-5-yl)-2-(4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamido)propanoic acid (66 mg), tert-butyl 4-(piperazin-1-yl)piperidine-1-carboxylate (43 mg), and N,N-diisopropylethylamine (43 mg) were dissolved in N,N-dimethylformamide (2 mL). 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (43 mg) was added and reacted at room temperature for 4 hours. After completion of the reaction, the reaction solution was separated and purified by preparative HPLC to obtain 30 mg of the title compound.

[0828] MS (ESI) m / z (M+H) + =747.3.

[0829] Step 2: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(4-(piperidin-4-yl)piperazin-1-yl)propan-2-yl)-4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamide

[0830] Tert-butyl (R)-4-(4-(3-(7-methyl-1H-indazol-5-yl)-2-(4-(7-oxo-3,4,7,8-tetrahydro-2H-thiopyrano[2,3-b]pyridin-6-yl)piperidine-1-carboxamido)propionyl)piperazin-1-yl)piperidine-1-carboxylate (30 mg) was dissolved in acetic acid and water (2 mL, v / v = 1 / 5) and reacted at 100°C for 2 hours. After completion of the reaction, the mixture was concentrated, and the crude product was dissolved in an appropriate amount of methanol. The pH of the system was adjusted to 8-9 with saturated potassium carbonate solution, filtered, and the filtrate was separated and purified by preparative HPLC and freeze-dried to obtain 10.00 mg of the title compound.

[0831] MS (ESI) m / z (M+H) + =647.3.

[0832] 1H NMR (400MHz, DMSO-d6) δ13.02(s,1H),7.96(s,1H),7.36(s,1H),7.00(s,1H),6.86(s,1H),6.63(d,J=8.0Hz,1H),4.77(q,J=7.7Hz,1H),4 .08(d,J=12.6Hz,2H),3.60-3.47(m,1H),3.30-3.20(m,1H),3.15-3.06(m,2H),3.05–2.84(m,7H),2.75–2.59(m,3H),2.56-2.51(m,2H), 2.47(s,3H),2.41–2.18(m,4H),2.06-1.91(m,4H),1.70–1.52(m,3H),1.41–1.17(m,4H),1.09–0.97(m,2H).

[0833] Example 50: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-oxo-1-(4-(piperidin-4-yl)piperazin-1-yl)propan-2-yl)-4-(6-oxo-2,3,6,7-tetrahydrothieno[2,3-b]pyridin-5-yl)piperidine-1-carboxamide

[0834] The title compound was prepared using a similar preparation method to Example 49 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0835] MS (ESI) m / z (M+H) + =633.3.

[0836] 1 H NMR(400MHz,DMSO-d6)δ13.02(s,1H),7.96(s,1H),7.36(s,1H),7.08(s,1H),7.00(s,1H), 6.64(d,J=8.1Hz,1H),4.77(q,J=7.7Hz,1H),4.15–4.06(m,2H),3.63-3.48(m,1H),3.16–3 .10(m,1H),3.06(t,J=8.1Hz,2H),2.99-2.84(m,5H),2.75–2.60(m,4H),2.47(s,3H),2.38 –2.20(m,4H),2.05–1.95(m,2H),1.69–1.54(m,3H),1.39–1.20(m,7H),1.11-0.97(m,2H).

[0837] Example 51: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[b]pyridin-3-yl)piperidine-1-carboxamide

[0838] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0839] MS (ESI) m / z (M+H) + =629.3.

[0840] 1H NMR (400MHz, DMSO-d6) δ13.01(s,1H),11.66(s,1H),7.96(s,1H),7.36(s,1H),7.03(s,1H),7.00(d,J=1.2Hz,1H),6.6 4(d,J=8.1Hz,1H),4.77(q,J=7.7Hz,1H),4.16–4.03(m,2H),3.59–3.49(m,1H),3.38-3.36(m,1H),3.28–3.19(m,1H),3 .15-3.07(m,1H),2.99-2.92(m,1H),2.91-2.84(m,1H),2.79–2.56(m,9H),2.47(s,3H),2.37–2.28(m,1H),2.26–2.17( m,1H),2.09(s,3H),2.04–1.84(m,4H),1.78–1.60(m,4H),1.54(t,J=8.8Hz,1H),1.41-1.33(m,2H),1.29–1.09(m,4H).

[0841] Example 52: Preparation of (R)-N-(3-(7-methyl-1H-indazol-5-yl)-1-(4-(1-methylpiperidin-4-yl)piperazin-1-yl)-1-oxopropan-2-yl)-4-(2-oxo-1,2,5,6,7,8-hexahydroquinolin-3-yl)piperidine-1-carboxamide

[0842] The title compound was prepared using a similar preparation method to that of Example 11 using corresponding common commercially available reagents and the products of the aforementioned intermediate preparation examples and examples as raw materials.

[0843] MS (ESI) m / z (M+H) + = 643.4.

[0844] 1H NMR (400MHz, DMSO-d6) δ13.02(s,1H),11.23(s,1H),7.96(s,1H),7.37(s,1H),7.00(t,J=1.2Hz,1H),6.85(s,1H),6.64(d,J=8. 1Hz,1H),4.77(q,J=7.7Hz,1H),4.10(d,J=12.9Hz,2H),3.56(d,J=13.4Hz,1H),3.24(t,J=10.2Hz,1H),3.12(t,J=9.9Hz,1H),2 .97(dd,J=13.1,7.9Hz,1H),2.88(dd,J=13.1,7.0Hz,1H),2.82–2.54(m,5H),2.48(s,3H),2.39(d,J=19.6Hz,5H),2.23(d,J=11 .0Hz,1H),2.10(s,3H),2.03–1.80(m,2H),1.78–1.60(m,7H),1.55(t,J=9.2Hz,1H),1.38(d,J=12.3Hz,2H),1.32–1.08(m,6H).

[0845] Biological test data

[0846] Unless otherwise specified, the experimental materials, reagents, operations, and methods used in the following activity test examples can be obtained from commercial channels or can be easily known or prepared based on existing technologies.

[0847] Test Example 1: Cell Functional Antagonism Assay - cAMP Assay

[0848] 1. Experimental Principle

[0849] The CGRP receptor complex is coupled to Gs in the G protein. The binding of CGRP to the CGRP receptor complex leads to the activation of Gs and the production of cAMP (3',5'-cyclic adenosine monophosphate).

[0850] 2. Experimental Purpose

[0851] The compounds of the invention were determined for their ability to inhibit CGRP-stimulated cAMP formation in SK-N-MC cells.

[0852] 3. Experimental Materials

[0853] 3.1 Experimental cell lines:

[0854] SK-N-MC (neuroepithelioma cells), source: National Biomedical Experimental Cell Resource Bank.

[0855] 3.2 Reagents and consumables

[0856] 4. Experimental Procedure

[0857] 4.1 Cell preparation:

[0858] Resuscitate and culture SK-N-MC cells in advance. Before use, the cell confluence should be 70%-80%.

[0859] 4.2 Prepare sample diluent, test compound and α-CGRP (human).

[0860] 4.3 Cell plating and drug effects:

[0861] SK-N-MC cells were digested, centrifuged, resuspended in sample diluent, and plated in a 384-well plate. α-CGRP (human) and the test compound were added to each well and mixed thoroughly by pipetting. The plate was sealed with film and incubated in an ELISA incubator at 25°C.

[0862] 4.4 cAMP Assay: Follow the instructions for the LANCE Ultra cAMP Kit. Prepare the Eu-cAMP tracer and ULight anti-cAMP working solutions in a dark environment. Add each solution to the plate and pipette to mix thoroughly. Seal the plate with film and incubate in a microplate incubator at 25°C.

[0863] 4.5 Use the chemiluminescence module of the fully automatic microplate reader to read the luminescence value.

[0864] 4.6 Data Analysis

[0865] The IC50 value was calculated using the nonlinear formula with the LOG value of the compound concentration as the horizontal axis and the Inhibition as the vertical axis. The results are shown in the table below.

[0866] IC of compounds for inhibiting CGRP-stimulated cAMP production in SK-N-MC cells 50 value

[0867] 5. Conclusion

[0868] The novel compounds provided by the present invention can significantly inhibit the production of cAMP stimulated by CGRP and have a low IC 50 In particular, the compounds represented by Examples 1, 2, 3, 4, 5, 6, 8, and 9 have exceptionally significant inhibitory activity, with IC50 values ​​reaching pM levels.

[0869] Test Example 2: Liver microsome stability test

[0870] 1. Purpose of the experiment

[0871] The stability of the compounds of the present invention in rat, monkey and human liver microsomes was determined. The compounds of the present invention tested were the compounds of Examples 6 and 8, and the control drug was Zavegepant.

[0872] 2. Reagents and consumables

[0873] 3. Experimental Procedure

[0874] 3.1 Experimental incubation system

[0875] 3.2 Pipette an appropriate amount of liver microsome solution into a 1 mL 96-deep-well plate, add the test solution (or probe substrate solution), and pre-incubate in a 37°C thermomixer for 5 minutes. Remove two mixed solutions from the system, add 1×PBS instead of NADPH, and remove the mixed solutions at 0 and 60 minutes respectively, and add methanol (containing internal standard) to terminate the reaction. Add NADPH to each well of the remaining mixed system to start the reaction. Remove the mixed solution from the system at 0, 5, 15, 30, 45, and 60 minutes for the test group and 0, 30, and 60 minutes for the control group, and add methanol (containing internal standard) to terminate the reaction. Mix all the samples that have terminated the reaction, centrifuge at 3800 rpm for 15 minutes, and take the supernatant for LC-MS / MS analysis.

[0876] 3.3 Data Analysis

[0877] Peak areas were determined from the extracted ion chromatograms. The slope value k was determined by linear regression of the residual percentage of the parent drug against the natural logarithm of the incubation time curve. The in vitro half-life (t 1 / 2 ) and calculated the in vitro intrinsic clearance (CL int , expressed in μL / min / mg protein). The calculation formula is as follows:

[0878] t 1 / 2 =ln2 / k=0.693 / k; CL int =0.693 / t 1 / 2 / liver microsomal protein concentration

[0879] The experimental results are shown in the following table:

[0880] Stability data of the compounds of the present invention in rat, monkey and human liver microsomes

[0881] Test Example 3: Plasma protein binding test

[0882] 1. Purpose of the experiment

[0883] The protein binding rate of the compounds of the present invention in rat, monkey and human plasma was determined by equilibrium dialysis. The compounds of the present invention tested were the compounds of Examples 6 and 8.

[0884] 2. Test Matrix

[0885] 3. Experimental Procedure

[0886] 3.1 Rinse the dried dialysis membrane 2-3 times with ultrapure water, then soak it in phosphate buffer for 1 hour. Soak the polytetrafluoroethylene module in 20% ethanol for 30 minutes, blot the surface moisture with a clean paper, and air dry. Assemble the pretreated dialysis membrane into the dialysis plate according to the product instructions, and add 100 μL of receiving solution (100 mM phosphate buffer plus 0.002% Tween 80) to one side of the membrane (receiving chamber) in each dialysis well.

[0887] 3.2 Place plasma in a centrifuge tube and add the working solution of the substance to be tested to a final concentration of 1 μM. Mix by inverting the tube (perform this step on an ice bath). Transfer 20 μL of drug-containing plasma to two replicates in a 96-well sample plate as T0 samples and store in a -20°C refrigerator.

[0888] 3.3 Transfer 100 μL of the drug-containing plasma to the other side of the membrane (sample chamber) of the dialysis device. In duplicate, incubate at 37°C with constant shaking for 6 hours. After 6 hours of incubation, remove 20 μL from each of the equilibrated receiving and sample chambers to obtain samples B and A. Add the corresponding volume of blank plasma or phosphate buffer (containing 0.002% Tween 80) to sample B and sample A, respectively, so that the plasma to buffer volume ratio in each sample well is 1:1.

[0889] 3.4 Add 250 μL of methanol solution containing the internal standard to all sample wells, mix thoroughly, and centrifuge at 3800 rpm for 10 minutes. Take 20 μL of the supernatant and add 180 μL of methanol. Vortex mix thoroughly, and inject into the sample for LC-MS / MS analysis.

[0890] 3.5 Data Analysis

[0891] The plasma protein binding rate and recovery rate of the compound in plasma were calculated using the following formula: Free percentage (%) = C B / C A ; Plasma protein binding rate (f b %) = 1-free percentage (%); Recovery rate (%) = (C B +C A ) / C T0

[0892] Among them C B is the concentration of the compound in the receiving fluid after equilibrium dialysis; CA is the concentration of the compound in the plasma after equilibrium dialysis; C T0 is the initial concentration of the compound in plasma.

[0893] The experimental results are shown in the following table:

[0894] Test Example 4: In vivo pharmacokinetic study of the test compound administered intravenously and nasally to SD rats

[0895] 1. Experimental Animals

[0896] Species: Male SD rats, SPF grade. Source: Purchased from Chengdu Dashuo Laboratory Animal Co., Ltd., Laboratory Animal Production License No.: SCXK(Chuan)2020-030. Quantity: Two rats were administered intravenously, and four rats were administered intranasally.

[0897] 2. Preparation of test samples

[0898] 2.1 Accurately weigh an appropriate amount of drug and add 50 mM succinate buffer (diluted with D5W, pH = 5-6). Ultrasonicate and vortex to mix and dissolve thoroughly to obtain a 0.2 mg / mL dosing solution for intravenous (IV) administration.

[0899] 2.2 Accurately weigh an appropriate amount of drug, add 50 mM succinate buffer (diluted with purified water, pH = 5-6), and thoroughly mix and dissolve by ultrasonication and vortexing to obtain a 10 mg / mL dosing solution for intranasal (IN) administration.

[0900] 3. Experimental Design

[0901] The test drugs were Examples 6 and 8, and the control drug was Zavegepant.

[0902] 4. Blood collection time

[0903] 5min, 10min, 15min, 0.5h, 1h, 2h, 4h, 8h, and 24h after administration.

[0904] 5. Sample Collection and Disposal

[0905] Blood samples were collected via the jugular vein, approximately 0.2 mL per sample. After anticoagulation with EDTA-K2, the blood samples were placed on ice and centrifuged within 2 hours to separate the plasma (centrifugation conditions: 6000g, 5 min, 2-8°C). The collected plasma samples were stored at -80°C until analysis. After analysis, the remaining plasma samples were temporarily stored at -80°C.

[0906] 6. Bioanalysis and Data Processing

[0907] The plasma concentration of the designated compound was determined by LC-MS / MS, and the main pharmacokinetic parameters were calculated using the Winnolin 8.3 non-compartmental model. max The previous BLQ (including "No peak") is calculated as 0; C max BLQ (including "No peak") that appears later will not be included in the calculation.

[0908] In vivo pharmacokinetic study data of the test compound administered intravenously and intranasally to SD rats

[0909] Test Example 5: Study on the effect of different drug compositions with or without alkyl glycosides on PK characteristics in SD rats

[0910] The experimental animals in Test Example 5 were also SD rats, and the same experimental design, collection and biological analysis data processing methods and processes as those in Test Example 4 were adopted.

[0911] When preparing the test sample of Test Example 5, a dosing solution group containing 0.2% dodecyl-β-D-maltoside (DDM) was also prepared in parallel to compare and observe the changes in rat PK characteristics with the dosing solution group without dodecyl-β-D-maltoside (DDM).

[0912] The results of the effects of different drug compositions on rat PK in Test Example 5 are shown in the following table: Note: After adding 0.2% DDM to the solution group of Example 6 compound, T max The time taken for the compound of Example 8 and Zavegepant to be administered was shortened from 1 h to 0.167. When DDM was not added (0% DDM), the plasma concentration of the compound of Example 8 and Zavegepant reached the peak relatively quickly (T max 0.209h and 0.25h respectively), so there is limited room for further shortening the time to peak blood concentration after adding 0.2% DDM (T max 0.167h and 0.209h respectively).

[0913] It can be seen from the above table:

[0914] 1. When the compounds of Formula I represented by Examples 6 and 8 were administered in combination with an alkyl glycoside, several key PK parameters were surprisingly and significantly improved. For example, after administration of the compound of Example 6 in combination with an alkyl glycoside: max The time from 1 hour to 0.167 hours was shortened by 83.3%. The ultra-short peak time laid a good foundation for the rapid onset of the drug. maxThe results show that the drug content of the compound in Example 8 is significantly increased by 10 times, and the AUC is increased by about 5 times, which ensures a rapid onset of action while reducing the drug dosage and increasing safety. max and AUC 0-inf They increased by 27.6 times and 7.5 times respectively. max It was also significantly shortened by 20.1%, remaining at a shorter time level of 0.167h.

[0915] For the indications of migraine and neuralgic headache, rapid relief, treatment and prevention of acute attacks of the compound of formula I are particularly important. The combination of the compound of formula I represented by Examples 6 and 8 and the alkyl glycoside brings about the T max Shorten and C max The surprising improvement in PK properties indicates the great clinical application potential of this composition.

[0916] 2. Gempanic CGRP antagonists, such as those represented by Examples 6 and 8 and Zavigepan, also showed some improvement in multiple PK parameters when added to alkyl glycosides during nasal administration. This demonstrates the clinical potential of combining CGRP antagonists with alkyl glycosides in the formulation of nasal formulations.

[0917] Test Example 6: Study on the Effect of Different Drug Compositions Combined with or Without Alkyl Glycoside on PK Properties in Monkeys

[0918] 1. Experimental Animals

[0919] Species: Cynomolgus macaques, male, 2-5 kg. Source: Ankai Yibo (Zhanjiang) Biotechnology Co., Ltd., Laboratory Animal Production License No.: SCXK(Guangdong)2019-0046. Quantity: 3 animals administered intravenously and 3 animals administered nasal spray.

[0920] 2. Preparation of test samples

[0921] 2.1 Accurately weigh an appropriate amount of drug, add 50 mM succinate buffer (diluted with D5W, pH = 5-6), and thoroughly mix and dissolve by sonication and vortexing to obtain a 0.5 mg / mL dosing solution for intravenous administration.

[0922] 2.2 Accurately weigh an appropriate amount of drug and add a solvent (1:50 mM succinate buffer (diluted with purified water, pH = 5-6)). Ultrasonicate and vortex to mix and dissolve thoroughly to obtain a 20 mg / mL dosing solution. Place the solution in a nasal spray device for nasal spray administration at 100 μL / spray.

[0923] 2.3 Accurately weigh an appropriate amount of drug and add solvent 2: 50 mM succinate buffer (pH = 5-6) containing 0.2% dodecyl-β-D-maltoside. Sonicate and vortex to thoroughly mix and dissolve to obtain a 20 mg / mL dosing solution. Place the solution in a nasal spray device for nasal spray administration at 100 μL / spray.

[0924] 3. Experimental Design

[0925] 4. Blood collection time

[0926] 5min, 10min, 15min, 0.5, 1, 2, 4, 6, 8, 10, 12, and 24h after administration.

[0927] 5. Sample Collection and Disposal

[0928] Blood samples were collected from the saphenous or cephalic veins, approximately 0.6 mL per sample. After anticoagulation with EDTA-K2, the blood samples were placed on ice and centrifuged within 2 hours to separate the plasma (centrifugation conditions: 2000g, 10 min, 2-8°C). The collected plasma samples were stored at -80°C until analysis. After analysis, the remaining plasma samples were temporarily stored at -80°C.

[0929] 6. Bioanalysis and Data Processing

[0930] The plasma concentration of the designated compound was determined by LC-MS / MS, and the main pharmacokinetic parameters were calculated using the Winnolin 8.3 non-compartmental model. max The previous BLQ (including "No peak") is calculated as 0; C max BLQ (including "No peak") that appears later will not be included in the calculation.

[0931] In vivo pharmacokinetic study data of the test compound administered intravenously and nasally in cynomolgus monkeys

[0932] In vivo pharmacokinetics of different drug compositions administered intranasally in monkeys

[0933] Result analysis:

[0934] Similar to the rat PK characteristics study in Test Example 5, the monkey PK characteristics study showed that when the compounds of Formula I represented by Examples 6 and 8 were administered in combination with an alkyl glycoside, multiple key PK parameters were also surprisingly and significantly improved. This was manifested in that after the compounds of Examples 6 and 8 were administered in combination with an alkyl glycoside, C maxGreatly improved to 52.2, 30 times, while AUC 0-inf Increased to 17.1, 8.6 times, T max The onset of action was shortened from 1 hour to approximately 0.17 hours, a significant 83.3% reduction. This indicates that, compared to rats, the Formula I compound combined with an alkyl glycoside further demonstrated superior PK improvement in monkeys, a species more closely related to humans, demonstrating promising clinical application prospects.

[0935] Test Example 7: Study on the Effect of Different Alkyl Glycoside Concentrations in Pharmaceutical Compositions on PK Characteristics in SD Rats

[0936] The experimental animals in Test Example 7 were also SD rats, and the same experimental design, collection and biological analysis data processing methods and processes as those in Test Example 4 were adopted.

[0937] When preparing the test sample of Test Example 7, parallel groups of dosing solutions with 0.05% and 5% dodecyl-β-D-maltoside (DDM) were prepared to compare and observe the changes in the PK characteristics of rats in the dosing solution groups with the two concentrations of dodecyl-β-D-maltoside (DDM).

[0938] The results of the effects of different drug compositions on rat PK in Test Example 7 are shown in the following table:

[0939] It can be seen from the above table:

[0940] When the compounds of Formula I represented by Examples 6 and 8 were administered in combination with different concentrations of alkyl glycoside, multiple key PK parameters were surprisingly and significantly improved. For example, when the compound of Example 6 was administered in combination with 0.05% and 5% alkyl glycoside: max The time to peak was shortened from 1 h to 0.25 h and 0.083 h, respectively, which was 75% and 91.7% shorter. This indicates that the higher the concentration of alkyl glycoside, the shorter the time to peak. max The results showed that the drug content of the compound of Example 8 was increased by 6.51 times and 13.3 times, and the AUC was increased by 3.8 times and 3.6 times, which ensured a rapid onset of action while reducing the drug dosage and increasing safety. max The time to peak was shortened from 0.2h to 0.167h and 0.083h, respectively, which was 20% and 60% shorter, indicating that the higher the concentration of alkyl glycoside, the shorter the time to peak. max The results showed that the concentration of alkyl glycoside increased by 16.2 times and 73.0 times, and the AUC increased by 6.23 times and 13.5 times, indicating that the higher the concentration of alkyl glycoside, the stronger its penetration-enhancing effect.

[0941] The present invention is not limited to the above optional embodiments. Anyone can derive various other forms of products based on the teachings of the present invention. The above specific embodiments should not be construed as limiting the scope of protection of the present invention. The scope of protection of the present invention shall be based on the scope defined in the claims, and the description can be used to interpret the claims.

[0942] Cited Literature:

[0943] [1]Ashina M, Terwindt GM, Al-Karagholi MA, et al. Migraine: disease characterization, biomarkers, and precision medicine. Lancet.2021;397(10283):1496-1504.doi:10.1016 / S0140-6736(20)32162-0

[0944] [2] Chinese Medical Doctor Association Neurology Branch, Chinese Society of Research Hospitals Headache and Sensory Disorders Professional Committee. Chinese Guidelines for the Diagnosis and Treatment of Migraine (2022 Edition)[J]. Chinese Journal of Pain Medicine, 2022, 28(12):881-898.

[0945] [3]Ashina M,Katsarava Z,Do TP,et al.Migraine:epidemiology and systems of care.Lancet.2021;397(10283):1485-1495.doi:10.1016 / S0140-6736(20)32160-7

[0946] [4]Russo AF.Calcitonin gene-related peptide(CGRP): a new target for migraine.Annu Rev Pharmacol Toxicol.2015;55:533-552.doi:10.1146 / annurev-pharmtox-010814-124701

[0947] [5]Iyengar S,Ossipov MH,Johnson KW.The role of calcitonin gene-related peptide in peripheral and central pain mechanisms including migraine.Pain.2017;158(4):543-559.doi:10.1097 / j.pain.0000000000000831

[0948] [6]Hargreaves R,Olesen J.Calcitonin Gene-Related Peptide Modulators-The History and Renaissance of a New Migraine Drug Class.Headache.2019;59(6):951-970.doi:10.1111 / head.13510

[0949] [7]Russell FA,King R,Smillie SJ,Kodji X,Brain SD.Calcitonin gene-related peptide:physiology and pathophysiology.Physiol Rev.2014;94(4):1099-1142.doi:10.1152 / physrev.00034.2013

[0950] [8]Clemow DB,Johnson KW,Hochstetler HM,Ossipov MH,Hake AM,Blumenfeld AM.Lasmiditan mechanism of action-review of a selective 5-HT1F agonist.J Headache Pain.2020;21(1):71.Published 2020Jun 10.doi:10.1186 / s10194-020-01132-3.

Claims

1. A pharmaceutical composition containing: a. active ingredient and b. alkyl glycoside; wherein the active ingredient is a compound represented by the following formula I, or an isomer, pharmaceutically acceptable salt or solvate thereof, Where X 1 and X 2 , respectively, independently represent CH or N; X 3 and X 4 , respectively, independently represent C, CH or N; X 5 represents C, CH, CH2, NH or N; Y 1 represents CH, O, or NH; Y 2 represents O or S; R 2 in each instance, independently represents -H, -(C1-C6)alkyl, or -(C3-C8)cycloalkyl, and n is 0, 1, 2, 3, 4, 5, or 6, or two R's 2form a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 2 , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R is attached 2 , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S, or -(R 2 ) n forms a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 2 ) n , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 2 )n, and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S; R3 in each instance, independently represents -H, -(C1-C6)alkyl, or -(C3-C8)cycloalkyl, and o is 0, 1, 2, 3, 4, 5, or 6, or two R's 3 form a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 3 , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R is attached 3 , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S, or -(R 3 ) o forms a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 3 ) o, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 3 ) o , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S; A has a structure represented by the following formula II the G-chain group forms a 5- to 7-membered aliphatic ring or an aliphatic heterocyclic ring together with the two carbon atoms to which the G-chain group is attached, and the ring-forming heteroatoms in the aliphatic heterocyclic ring are -O-, -S-, or R 1 in each instance, independently represents -H, -(C1-C6)alkyl, or -(C3-C8)cycloalkyl, and m is 0, 1, 2, 3, 4, 5, or 6, or two R's 1form a 3- to 8-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 1 , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R is attached 1 , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S, or -(R 1 ) m forms a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 1 ) m wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 1 ) m , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S.

2. A pharmaceutical composition according to paragraph 1, characterized in that: Alkyl glycoside is mainly produced by covalently linking a hydrophobic alkyl carbon chain and a hydrophilic oligosaccharide, the covalently bonded linker group contains a glycosidic bond, a thioglycosidic bond, or an amide bond, wherein the hydrophobic alkyl carbon chain is selected from C5-C 16 alkyl chain, C 10 -WITH 16 alkyl chain, C 11 -WITH 14 alkyl chain, C 11 alkyl chain, C 12 alkyl chain, O 13 alkyl chain or C 14 alkyl chain, and wherein the hydrophilic oligosaccharide contains a monosaccharide or a disaccharide, wherein the monosaccharide contains glucose and the disaccharide contains maltose, trehalose or sucrose.

3. The pharmaceutical composition according to paragraph 1, characterized in that the alkyl glycoside is undecyl maltoside, dodecyl maltoside, tridecyl maltoside, or tetradecyl maltoside, and the weight percentage of alkyl glycoside in the pharmaceutical composition is 0.05-20%, 0.05-10%, 0.05-5%, 0.1-5%, 0.1-0.5%, 0.1-0.3%, 0.05%, 0.2% or 5%.

4. The pharmaceutical composition according to paragraph 1, characterized in that the pharmaceutical composition is prepared in a form for nasal administration, a form for oral administration, a form for transdermal administration or a form for transmucosal administration, wherein the form for nasal administration includes nasal drops, nasal spray, nasal aerosol, nasal gel or nasal cream, wherein the form for nasal administration is a single-dose form, a two-dose form or a multi-dose form, the delivery volume of the nasal spray is 50-200 µl / spray, and wherein the pharmaceutical composition is presented in the form of a solution and additionally in the form of an aqueous solution.

5. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition further comprises one or a combination of two or more of a buffer, an osmotic pressure regulator, a wetting agent, a thickener, a pH regulator, a metal chelating agent, and a preservative.

6. The pharmaceutical composition according to claim 1, characterized in that, compared to the pharmaceutical composition without the addition of b. alkyl glycoside, said pharmaceutical composition provides a 10-fold or more, 30-fold or more, 10-52.2-fold or 30-52.2-fold C value max active ingredient, and / or compared to the pharmaceutical composition without the addition of b. alkyl glycoside, said pharmaceutical composition provides 5.1-fold or more, 8.6-fold or more, 5.1-17.1-fold or 8.6-17.1-fold AUC value 0-infactive ingredient, and / or compared to the pharmaceutical composition without the addition of b. alkyl glycoside, the said pharmaceutical composition provides a T value max , which is reduced to 0.17 hours or less.

7. The pharmaceutical composition according to claim 1, characterized in that the weight percentage of the active ingredient in the pharmaceutical composition is from 0.05% to 20%, and / or the content of the active ingredient in the pharmaceutical composition is from 0.001 mg to 1000 mg.

8. The pharmaceutical composition according to claim 1, characterized in that the compound represented by formula I has a chiral structure represented by formula I-2 or formula I-3: where X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 , Y 2 , A, R 2 , R 3 , n and o have the meanings defined above.

9. A pharmaceutical composition according to paragraph 1 or 8, characterized in that in A: in case the G-chain group forms a 5- or 6-membered aliphatic ring together with the two carbon atoms to which the G-chain group is attached, the G-chain group may have 0 or 1 double bond, and all other bonds are single bonds; in the case where the G-chain group forms a 7-membered aliphatic ring together with the two carbon atoms to which the G-chain group is attached, the G-chain group may have 0, 1, or 2 double bonds, and all other bonds are single bonds; in the case where the G-chain group forms a 5-membered aliphatic heterocyclic ring together with the two carbon atoms to which the G-chain group is attached, all bonds in the G-chain group are single bonds; and in case the G-chain group forms a 6- or 7-membered aliphatic heterocyclic ring together with the two carbon atoms to which the G-chain group is attached, the G-chain group optionally has 0 or 1 double bond and all the remaining bonds are single bonds.

10. The pharmaceutical composition according to paragraph 9, characterized in that in A the G-chain group is selected from:

11. A pharmaceutical composition according to paragraph 1 or 8, characterized in that: in A R 1 in each instance, independently represents -H, -(C1-C3)alkyl, or -(C3-C6)cycloalkyl, and m is 0, 1, 2, or 3, or two R's 1 form a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 1, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R is attached 1 , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S, or -(R 1 ) m forms a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 1 ) m , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 1 ) m , and the heteroatoms in the aliphatic heterocyclic ring are O, N, or S.

12. The pharmaceutical composition according to paragraph 11, characterized in that in A R 1in each instance independently represents -H, methyl, ethyl, or cyclopropyl, and m is 0, 1, or 2, or two R1 form cyclopropane, cyclobutane, or cyclopentane together with the atoms to which R1 is attached, and the cyclopropane, cyclobutane, or cyclopentane form a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R1 is attached, or -(R 1 ) m forms cyclopropane, cyclobutane, or cyclopentane together with the atoms to which -(R) is attached 1 ) m , and cyclopropane, cyclobutane, or cyclopentane form a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 1 ) m .

13. The pharmaceutical composition according to paragraphs 1, 8, 11 or 12, characterized in that A has a structure represented by the following formula II-1, II-2 or II-3: in formulas II-1, II-2 and II-3 X 6 represents -O-, -S- or In formula II-2, two "---" refer to single bonds, or any one of the two "---" represents a double bond and the other represents a single bond; and In formula II-3, all three "-"s represent single bonds, or any one of the three "---"s represents a double bond and the other two represent single bonds; or A has a structure represented by the following formula II-4 or II-5: in formula II-4 or II-5 X 6 represents -O-, -S- or In formula II-4, "---" refers to a single bond or a double bond; and In formula II-5, two "---"s refer to single bonds, or any one of the two "---"s represents a double bond and the other represents a single bond.

14. A pharmaceutical composition according to paragraph 1 or 8, characterized in that: And chosen from preferably A is selected from more preferably, A is selected from: additionally preferably A is selected from:

15. A pharmaceutical composition according to paragraph 1 or 8, characterized in that: R 2 in each instance, independently represents -H, -(C1-C3)alkyl, or -(C3-C6)cycloalkyl, and n is 0, 1, 2, or 3, or two R's 2 form a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 2, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R is attached 2 , and both ring atoms provided by two R 2 in an aliphatic ring or an aliphatic heterocyclic ring, are C atoms; or -(R 2 )n forms a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 2 ) n , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 2 ) n , and all ring atoms provided by the -(R 2 )n in an aliphatic ring or an aliphatic heterocyclic ring are C atoms; R 3in each instance, independently represents -H, -(C1-C3)alkyl, or -(C3-C6)cycloalkyl, and n is 0, 1, 2, or 3, or two R's 3 form a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 3 , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which R is attached 3 , and both ring atoms provided by two R 3 in an aliphatic ring or an aliphatic heterocyclic ring, are C atoms; or -(R 3 ) o forms a 3-6-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 3 ) o, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure, a fused ring structure, or a bridged ring structure with the ring to which -(R 3 ) o , and all ring atoms provided by the -(R 3 ) o in an aliphatic ring or aliphatic heterocyclic ring, are C atoms.

16. The pharmaceutical composition according to paragraph 15, characterized in that R 2 in each instance independently represents -H, methyl, ethyl, cyclopropyl or cyclobutyl, and n is 0, 1 or 2, or two R's 2 form a 3-5-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached 2 , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure with the ring to which R is attached 2, and both ring atoms provided by two R 2 in an aliphatic ring or an aliphatic heterocyclic ring, are C atoms; or -(R 2 ) n forms a 3-5-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 2 ) n , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure with the ring to which -(R 2 ) n , and all ring atoms provided by the -(R 2 ) n in an aliphatic ring or aliphatic heterocyclic ring, are C atoms; R 3 in each instance, independently represents -H, -(C1-C3)alkyl, or -(C3-C6)cycloalkyl, and n is 0, 1, 2, or 3, or two R's 3 form a 3-5-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which R is attached3 , wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure with the ring to which R is attached 3 , and both ring atoms provided by two R 3 in an aliphatic ring or an aliphatic heterocyclic ring, are C atoms; or -(R 3 ) o forms a 3-5-membered aliphatic ring or an aliphatic heterocyclic ring together with the atoms to which -(R) is attached 3 )o, wherein the aliphatic ring or aliphatic heterocyclic ring optionally forms a spiro structure with the ring to which -(R 3 ) o , and all ring atoms provided by the -(R 3 ) o in an aliphatic ring or aliphatic heterocyclic ring, are C atoms.

17. A pharmaceutical composition according to claim 1 or 8, characterized in that: X 3 and X 4do not represent N simultaneously.

18. The pharmaceutical composition according to claim 1, characterized in that the active ingredient is a compound represented by the following formula I-1, or an isomer, pharmaceutically acceptable salt or solvate thereof where X 1 , X 2 , X 3 , X 4 , X 5 , Y 1 and Y 2 are as defined in paragraph 1, and A is as defined in paragraph 13 or 14; and R 2 and R 3 in each case independently represent -H, -CH3, or are absent.

19. A pharmaceutical composition according to claims 1, 8 or 18, characterized in that one or more H in the compound are replaced by D.

20. The pharmaceutical composition according to claim 19, characterized in that one or more H in the G-chain group or R 1 replaced by D.

21. The pharmaceutical composition according to claim 1, characterized in that the compound represented by formula I is selected from:

22. A pharmaceutical composition according to claim 1, characterized in that said pharmaceutical composition is presented in sterile or non-sterile form.

23. A preparation for nasal administration containing a CGRP antagonist and an alkyl glycoside, wherein the CGRP antagonist is selected from zavegepant, rimegepant, ubrogepant, atogepant, telcagepant, olcegepant or an active ingredient in a pharmaceutical composition according to any one of claims 1 and 8-21.

24. A preparation for nasal administration according to paragraph 23, characterized in that Alkyl glycoside is mainly produced by covalently linking a hydrophobic alkyl carbon chain and a hydrophilic oligosaccharide, the covalently bonded linker group contains a glycosidic bond, a thioglycosidic bond, or an amide bond, wherein the hydrophobic alkyl carbon chain is selected from C5-C 16 alkyl chain, C 10 -WITH 16 alkyl chain, C 11 -WITH 14 alkyl chain, C 11 alkyl chain, C 12 alkyl chain, C 13 alkyl chain or C 14 alkyl chain, and wherein the hydrophilic oligosaccharide contains a monosaccharide or a disaccharide, wherein the monosaccharide contains glucose and the disaccharide contains maltose, trehalose or sucrose.

25. A preparation for nasal administration according to paragraph 23, characterized in that the alkyl glycoside is undecyl maltoside, dodecyl maltoside, tridecyl maltoside, or tetradecyl maltoside, and the weight percentage of alkyl glycoside in the pharmaceutical composition is 0.05-20%, 0.05-10%, 0.05-5%, 0.1-5%, 0.1-0.5%, 0.1-0.3%, 0.05%, 0.2% or 5%.

26. Use of a pharmaceutical composition according to any one of claims 1-22 or a preparation for nasal administration according to any one of claims 23-25 ​​for the preparation of a medicament for the prevention, treatment or alleviation of a disease mediated / regulated by CGRP.

27. The use according to paragraph 26, characterized in that the said disease includes migraine and neuropathic pain.

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