TW202017923A - Novel compounds for the treatment, alleviation or prevention of disorders associated with tau aggregates - Google Patents

Abstract

The present invention relates to novel compounds that can be employed in the treatment, alleviation or prevention of a group of disorders and abnormalities associated with Tau (Tubulin associated unit) protein aggregates including, but not limited to, Neurofibrillary Tangles (NFTs), such as Alzheimer's disease (AD).

Description

Novel compounds for treating, ameliorating or preventing disorders associated with TAU aggregates

The present invention relates to novel compounds that can be used to treat, ameliorate, or prevent a group of disorders and abnormalities associated with Tau (tubulin-related units) protein aggregates, including but not limited to neurofibrillary tangles (NFT), such as Az Alzheimer's disease (AD).

Many aging diseases are based on or related to extracellular or intracellular deposition of amyloid or amyloid-like proteins, which contribute to the pathogenesis and disease progression. The best characterized amyloids forming extracellular aggregates are amyloid β (Aβ). Other examples of amyloid proteins that form extracellular aggregates are prion, ATTR (transthyretin) or Adan (ADanPP). Amyloid proteins that mainly form intracellular aggregates include (but are not limited to) Tau, α-synuclein, TAR DNA binding protein 43 (TDP-43), and huntingtin (htt). Diseases involving Tau aggregates are usually listed as tau protein lesions (eg AD).

Amyloid or amyloid deposits are caused by the misfolding of proteins and subsequent aggregation to produce β-pleated plate assemblies in which multiple peptides or proteins are held together by intermolecular hydrogen bonding. Although starch-like or amyloid-like proteins have different primary amino acid sequences, their deposits usually contain many shared molecular components, especially the presence of β-pleated quaternary structure. The link between starch-like deposits and disease is still largely unclear. It has been found that various protein aggregates (including both protein aggregates related to disease pathology and unrelated ones) are toxic, which indicates that the common molecular characteristics of starch-like are related to or cause disease onset (Bucciantini Et al., Nature, 2002, 416, 507-11). Various polymers of peptides or proteins aggregated by β-pleated plates are also related to the toxicity of different peptides or proteins ranging from dimers to soluble low molecular weight oligomers, fibrils or insoluble fibrous deposits.

Alzheimer's disease (AD) is a neurological disorder, which is mainly thought to be caused by amyloid plaques, that is, extracellular accumulation of abnormal deposits of (amyloid-β) Aβ aggregates in the brain. Other major neuropathological markers of AD are intracellular neurofibrillary tangles (NFT), which are derived from the aggregation of highly phosphorylated Tau protein, misfolded Tau or pathological Tau and its conformational isomers. AD shares its pathogenesis with many neurodegenerative tau protein lesions, especially with specified types of frontotemporal dementia (FTD). Tau protein is a soluble "natural unfolded" protein, which is tightly bound to the microtubule (MT) to promote its assembly and stability. MT is critical to the integrity of neuronal cell scaffolds, and thus to the proper formation and function of neuronal circuits, and therefore to learning and memory. The binding of Tau and MT is controlled by dynamic phosphorylation and dephosphorylation, as shown mainly in vitro and in non-neuronal cells. In the AD brain, Tau pathology (tau protein lesions) occurs later than amyloid pathology, but the industry is still discussing whether the Aβ protein is the causative agent of AD (this constitutes the essence of the so-called amyloid cascade hypothesis) Quite controversial (Hardy et al., Science 1992, 256, 184-185; Musiek et al., Nature Neurosciences 2015, 18(6), 800-806). The exact mechanism linking starch-like to Tau pathology is still largely unknown, but it is proposed to involve the activation of neuronal signaling pathways that act on GSK3 and cdk5 as the main "Tau-kinase" or By its role (Muyllaert et al., Rev. Neurol. (Paris), 2006, 162, 903-7; Muyllaert et al., Genes Brain and Behav. 2008, Supplement 1, 57-66). Even though the occurrence of tau protein lesions occurs later than starch-like, it is not only a harmless side effect, but the main pathological executor of AD. In the experimental mouse model, the cognitive defects caused by the amyloid pathology were almost completely improved due to the absence of Tau protein (Roberson et al., Science, 2007, 316(5825), 750-4), and the cognitive dysfunction and The severity of dementia is related to tau protein lesions, but not to amyloid pathology.

Diseases involving Tau aggregates are usually listed as tau protein lesions, and they include (but are not limited to) Alzheimer's disease (AD), familial AD, PART (primary age-related Tau protein lesions), library Creutzfeldt-Jacob disease, boxer-type dementia, Down's Syndrome, Gerstmann-Sträussler-Scheinker disease (GSS), inclusion bodies Myositis, Prion protein cerebral amyloid angiopathy, traumatic brain injury (TBI), amyotrophic lateral sclerosis (ALS), Guam Parkinsonism-dementia complex of Guam , Non-Guamanian motor neuron disease with neurofibrillary tangles, argyrophilic granulopathy, cortical basal nucleus degeneration (CBD), diffuse neurofibrillary tangles with calcification, and chromosome 17 Frontotemporal dementia with Parkinson's disease (FTDP-17), Hallervorden-Spatz disease (Hallervorden-Spatz disease), multiple system atrophy (MSA), C-type Niemann-Pick disease (Niemann- Pick disease type C), pale ball-pontine-melanotic degeneration, Pick's disease (Pick's disease, PiD), progressive subcortical glioma, progressive supranuclear palsy (PSP), subacute sclerotic pan-brain Inflammation, tangled dominant dementia, post-encephalitis Parkinson's disease, myotonic dystrophy, subacute sclerosing panencephalopathy, LRRK2 mutation, chronic traumatic brain disease (CTE), familial British dementia Disease (familial British dementia), familial Danish dementia, other frontotemporal degeneration, Guadeloupean Parkinsonism, neurodegeneration with iron accumulation in the brain, SLC9A6 correlation Mental retardation, white matter tau protein lesions with globular inclusions, epilepsy, Lewy body dementia (LBD), mild cognitive impairment (MCI), multiple sclerosis, Parkinson's disease ), HIV-related dementia, adult-onset diabetes, senile cardiac amyloidosis, glaucoma, ischemic stroke, psychosis in AD, and Huntington's disease. (Williams et al., Intern. Med. J., 2006, 36, 652-60; Kovacs et al., J Neuropathol Exp Neurol. 2008; 67(10): 963-975; Higuchi et al., Neuropsychopharmacology-5th Generation of Progress , 2002, Part 9, Chapter 94: 1339-1354; Hilton et al., Acta Neuropathol. 1995; 90(1): 101-6; Iqbal et al., Biochimica et Biophysica Acta 1739 (2005) 198-210; McQuaid et al. Human, Neuropathol Appl Neurobiol. April 1994; 20(2): 103-10; Vossel et al., Lancet Neurol 2017; 16: 311-22; Stephan et al., Molecular Psychiatry (2012) 17, 1056-1076; Anderson Et al. Brain (2008), 131, 1736-1748; Savica et al. JAMA Neurol. 2013; 70(7):859-866; Brown et al. Molecular Neurodegeneration 2014, 9:40; El Khoury et al. Front . Cell. Neurosci., 2014, Volume 8, Article No. 22: 1-18; Tanskann et al., Ann. Med. 2008; 40(3):232-9; Gupta et al., CAN J OPHTHALMOL, Volume 43 , No. 1, 2008: 53-60; Dickson et al., Int J Clin Exp Pathol 2010; 3(1): 1-23; Fernández-Nogales et al., Nature Medicine, 20, 881-885 (2014); Bi Et al., Nature Communications Volume 8, article number: 473 (2017); Murray et al., Biol Psychiatry. April 1, 2014; 75(7): 542-552).

Of all the agents used in the clinical trials for Alzheimer's disease in 2017, Tau-targeted agents are extremely scarce and account for only 8% of Phase II clinical trials (Cummings et al., Alzheimer's & Dementia: Translational Research & Clinical Interventions 3 (2017) 367-384). Current treatment methods targeting Tau protein mainly include antibody-based methods, of which the main limitation is to only target extracellular Tau. In methods using small molecules, several Tau kinase inhibitors have been developed, although they are extremely challenging in terms of toxicity and specificity. However, currently only one kinase inhibitor (ie, Nilotinib) is being tested in clinical trials. Finally, among Tau aggregation inhibitors, only one LMTX is currently in clinical trials (Cummings et al., 2017). Although Tau-based therapies have become the focus of increasing attention in recent years, there is still a great demand in the industry for other therapeutic agents that target pathological Tau conformers that are known or assumed to cause tau proteinopathy.

WO2011/128455 relates to specific compounds suitable for the treatment of disorders related to amyloid or amyloid.

WO2010/080253 relates to dipyridyl-pyrrole derivative compounds, which can be used to treat diseases suitable for the inhibition, regulation and/or regulation of protein kinase signal transduction.

The object of the present invention is to provide compounds that can be used to treat, ameliorate or prevent a group of disorders and abnormalities associated with Tau protein aggregates, such disorders and abnormalities including (but not limited to) NFT, such as Alzheimer's disease (AD ). In addition, there is a need in the industry for compounds that can be used as therapeutic agents to: (a) reduce Tau aggregates/NFT by recognizing aggregated Tau and depolymerizing Tau (eg, by changing the molecular configuration of Tau aggregates), And/or (b) prevent the formation of Tau aggregates, and/or (c) intracellularly interfere with Tau aggregates, and/or (d) reduce Tau misfolding and hyperphosphorylation in vivo and/or (f) reduce nerves Inflammation markers. The inventors have surprisingly found that these objectives can be achieved by compounds of formula (I) as set forth below.

The compounds of formula (I) (a) demonstrate the ability to reduce Tau aggregates by recognizing and deagglomerating Tau (eg by changing the molecular configuration of Tau aggregates), and/or (b) preventing The formation of Tau aggregates, and/or (c) intracellular interference with Tau aggregates, and/or (d) reducing Tau misfolding and hyperphosphorylation in vivo and/or (f) reducing neuroinflammation markers. Although not wishing to be bound by theory, it is assumed that the compound of formula (I) inhibits Tau aggregation or disaggregates pre-formed Tau aggregates, including when present in cells. Due to their unique design features, these compounds exhibit properties such as appropriate lipophilicity and molecular weight, brain uptake and pharmacokinetics, cell permeability, solubility, and metabolic stability to become useful for the treatment, improvement, or prevention of tau proteinopathy Of success.

Both histopathological analysis and in vivo Tau PET imaging have shown that the accumulation of Tau NFT lesions is fully correlated with cognitive deficits in AD. The compounds of the present invention can prevent the formation of Tau aggregates or depolymerize pre-existing Tau aggregates, and thus can be expected to prevent or reduce related cognitive defects in AD.

Ultrastructural analysis has shown that the Tau inclusion system consists of paired spiral filaments (PHF) or straight filaments (SF). High-resolution structural analysis has shown that the filaments are composed of the core region of amino acids 306-378 containing Tau, which adopts a crossed β/β-helical structure. The compounds of the present invention can recognize aggregated Tau and depolymerize Tau (eg, by changing the molecular configuration of Tau aggregates), and thus can be expected to promote Tau clearance.

In addition, it has been shown that Tau is able to spread from cell to cell and some forms of Tau (acting as a seed) can induce structural changes in natural Tau proteins in healthy cells to undergo misfolding and aggregation. It is believed that the gathered Tau is responsible for seeding and thus leads to the spread of Tau pathology. The compounds of the present invention can interfere with aggregated Tau intracellularly and thus can be expected to reduce Tau pathological spread and ultimately prevent or reduce related cognitive deficits in AD.

The Tau aggregation cascade starts from Tau misfolding and hyperphosphorylation. It is believed that these events precede the formation of intracellular Tau neuronal inclusion bodies, and therefore before the establishment and spread of Tau pathology. The compounds of the present invention can reduce Tau misfolding and hyperphosphorylation in vivo, and thus can be expected to be beneficial for treating, ameliorating, or preventing diseases related to Tau pathology.

Finally, the link between Tau pathology and neuroinflammation is now well established. Neuroinflammation is a key event in the early AD stage and is believed to be one of the reasons for triggering Tau aggregation in PHF. In addition, once Tau pathology is fully established in the brain, several mouse models of tau protein lesions show significant neuroinflammation, indicating that Tau pathology can also induce neuroinflammation. These two findings indicate that Tau pathology and neuroinflammation are related in a positive feedback loop. In Tau pathology, the compounds of the present invention reduce neuroinflammation markers.

The present invention discloses novel compounds of formula (I) having the following capabilities: (a) reducing Tau aggregates, recognizing aggregated Tau and depolymerizing Tau (eg, by changing the molecular configuration of Tau aggregates), and/or (b) Prevent the formation of Tau aggregates, and/or (c) intracellularly interfere with Tau aggregates, and/or (d) reduce Tau misfolding and hyperphosphorylation in vivo and/or (f) reduce neuroinflammation markers. The present invention provides a method of using the compound of formula (I) or a pharmaceutical composition thereof to treat conditions and abnormalities associated with Tau protein aggregates, such conditions including (but not limited to) NFT. The present invention further provides a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable carrier or excipient.

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。 9.     一種醫藥組合物，其包含如條目1至8中任一條目之化合物及視情況醫藥上可接受之載劑或賦形劑。 10.   如條目1至8中任一條目之化合物，其用作藥劑。 11.    一種式(I)化合物，

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及-NR³ R⁴ 組成之群；R^a 係選自由H及烷基組成之群；R^b 、R^c 、R^d 、R^e 、R^f 及R^g 獨立地選自由H及烷基組成之群，或R^b 、R^c 、R^d 、R^e 、R^f 及R^g 中之任兩者可連結以形成3員至8員環；R^j 獨立地選自由以下組成之群：-鹵素、-O-烷基、-CF₃ 、-CN、-NR³ R⁴ 、

及

，其中含C_1-2 碳原子之橋可存在於a碳原子與c或d碳原子之間或其中含C_1-2 碳原子之橋可存在於b碳原子與c或d碳原子之間；R¹ 係選自由H及烷基組成之群；R² 獨立地選自由烷基、F及=O組成之群，其中該烷基可視情況經鹵素、-OH或-O-烷基取代，且其中若兩個R² 係孿位的，則其可連結以形成3員至6員環；R³ 及R⁴ 獨立地選自由H及烷基組成之群，其中該烷基可視情況經鹵素、-OH或-O-烷基取代；R⁵ 係選自由H及烷基組成之群；n 係0、1、2、3或4；r 及s 獨立地係0、1、2或3；且t 及u 獨立地係1、2或3。 12.   如條目11所用之化合物，其中該式(I)化合物係如條目1至8中任一條目中所定義。 13.   如條目1至8或11中任一條目之化合物，其用於降低Tau聚集。 14.   如條目1至8或11中任一條目之化合物，其用於防止Tau聚集體之形成及/或用於抑制Tau聚集。 15.   如條目1至8或11中任一條目之化合物，其用於細胞內干擾Tau聚集體。 16.   如條目1至8或11中任一條目之化合物，其用於降低活體內Tau錯誤摺疊及過度磷酸化。 17.   如條目1至8或11中任一條目之化合物，其用於減少神經發炎標記物。 18.   一種治療、預防或改善與Tau蛋白聚集體有關的病症或異常之方法，該方法包含向有需要之個體投與有效量之如條目1至8或11中任一條目之化合物。 19.   一種降低Tau聚集之方法，該方法包含向有需要之個體投與有效量之如條目1至8或11中任一條目之化合物。 20.   一種防止Tau聚集體之形成及/或抑制Tau聚集之方法，該方法包含向有需要之個體投與有效量之如條目1至8或11中任一條目之化合物。 21.   一種細胞內干擾Tau聚集體之方法，該方法包含向有需要之個體投與有效量之如條目1至8或11中任一條目之化合物。 22.   一種降低活體內Tau錯誤摺疊及過度磷酸化之方法，該方法包含向有需要之個體投與有效量之如條目1至8或11中任一條目之化合物。 23.   一種減少神經發炎標記物之方法，該方法包含向有需要之個體投與有效量之如條目1至8或11中任一條目之化合物。 24.   一種如條目1至8或11中任一條目之化合物之用途，其用於製造用於治療、預防或改善與Tau蛋白聚集體有關的病症或異常之藥劑。 25.   一種如條目1至8或11中任一條目之化合物之用途，其用於製造用於降低Tau聚集之藥劑。 26.   一種如條目1至8或11中任一條目之化合物之用途，其用於製造用於防止Tau聚集體之形成及/或用於抑制Tau聚集之藥劑。 27.   一種如條目1至8或11中任一條目之化合物之用途，其用於製造用於細胞內干擾Tau聚集體之藥劑。 28.   一種如條目1至8或11中任一條目之化合物之用途，其用於製造用於降低活體內Tau錯誤摺疊及過度磷酸化之藥劑。 29.   一種如條目1至8或11中任一條目之化合物之用途，其用於製造用於減少神經發炎標記物之藥劑。 29.   一種混合物，其包含如條目1至8中任一條目之化合物及至少一種其他生物活性化合物，該至少一種其他生物活性化合物係選自與如條目1至8中任一條目之化合物不同之治療劑， 其中該混合物進一步包含醫藥上可接受之載劑、稀釋劑及賦形劑中之至少一者。 30.   如條目29之混合物，其中該其他生物活性化合物係用於治療類澱粉變性之化合物。 31.   如條目29或30之混合物，其中該化合物及/或該其他生物活性化合物係以治療有效量存在。 32.   如條目29至31中任一條目之混合物，其中該其他生物活性化合物係選自由以下組成之群：抗氧化壓力之化合物；抗細胞凋亡化合物；金屬螯合劑；DNA修復抑制劑，例如哌侖西平(pirenzepine)及代謝物；3-胺基-1-丙烷磺酸(3APS)；1,3-丙烷二磺酸鹽(1,3PDS)；α-分泌酶活化劑；β-分泌酶及γ-分泌酶抑制劑；肝醣合酶激酶3抑制劑；O連接之N-乙醯葡糖胺水解酶(O-glcnacase，OGA)抑制劑；神經傳遞質；β褶板碎裂劑；類澱粉β清除/消除細胞組分引誘劑；N末端截短之類澱粉β (包括焦麩胺酸化之類澱粉β 3-42 )之抑制劑，抗發炎分子，或膽鹼酯酶抑制劑(ChEI)，例如塔克寧(tacrine)、利凡斯的明(rivastigmine)、多奈派齊(donepezil)及/或加蘭他敏(galantamine)；M1激動劑；其他藥物，包括任何類澱粉或Tau改質藥物及營養補充劑；抗體，包括任何功能等效抗體或其功能部分；或疫苗。 33.   如條目32之混合物，其中該其他生物活性化合物係膽鹼酯酶抑制劑(ChEI)。 34.   如條目之混合物32，其中該其他生物活性化合物係選自由以下組成之群：塔克寧、利凡斯的明、多奈派齊、加蘭他敏、菸鹼酸及美金剛(memantine)。 35.   如條目32之混合物，其中該其他生物活性化合物係抗體、具體而言單株抗體，包括任何功能等效抗體或其功能部分。 36.   如條目29至35中任一條目之混合物，其中該化合物及/或該其他生物活性化合物係以治療有效量存在。 37.   如條目11所用之化合物、如條目18之方法或如條目24之用途，其中該病症係選自阿茲海默氏病(AD)、家族性AD、原發性年齡相關性Tau蛋白病變(PART)、庫賈氏病、拳擊手型失智症、唐氏症候群、傑茨曼-斯脫司勒-史茵克病(GSS)、包涵體肌炎、普里昂蛋白腦類澱粉血管病變、創傷性腦損傷(TBI)、肌肉萎縮性脊髓側索硬化症(ALS)、關島帕金森氏症-失智複合症、非關島運動神經元病伴神經纖維纏結、嗜銀顆粒病、皮質基底核退化症(CBD)、瀰漫性神經纖維纏結伴鈣化、與染色體17相關之額顳葉失智症伴帕金森氏症(FTDP-17)、哈勒沃登-施帕茨病、多系統萎縮(MSA)、C型尼曼匹克症、蒼白球-腦橋-黑質退化、匹克氏病(PiD)、進行性皮質下神經膠瘤病、進行性核上性麻痺(PSP)、亞急性硬化性泛腦炎、纏結主導型失智症、腦炎後帕金森氏症、肌強直性營養不良、亞急性硬化性泛腦病變、LRRK2突變、慢性創傷性腦病變(CTE)、家族性英國型失智症、家族性丹麥型失智症、其他額顳葉退化、瓜德羅普帕金森氏症、神經退化伴腦內鐵累積、SLC9A6相關性智力遲鈍、白質tau蛋白病變伴球狀膠質細胞包涵體、癲癇、路易氏體失智症(LBD)、輕度認知損害(MCI)、多發性硬化、帕金森氏病、HIV相關性失智症、成人發作型糖尿病、老年性心臟類澱粉變性、青光眼、缺血性中風、AD中之精神病及亨庭頓氏病，較佳地阿茲海默氏病(AD)、皮質基底核退化症(CBD)、匹克氏病(PiD)及進行性核上性麻痺(PSP)。 38.   一種如條目1至8或11中任一條目之化合物之用途，其用作分析參考或活體外篩選工具。The present invention is summarized in the following items: 1. A compound of formula (I),

And all its stereoisomers, racemic mixtures, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, and polymorphs; where A is selected from the group consisting of:

and

,among them

and

Can be connected to Q at any available location, where

and

Substituted by one or more substituents R ^j , and wherein

and

Optionally substituted with one or more substituents R ; B is selected from the group consisting of O and NR ^a ; E and V are independently selected from the group consisting of: N, NR ⁵ , O, and S; G is selected from the group consisting of Group consisting of: benzene ring, pyrimidine ring and pyridine ring; J is selected from the group consisting of O and NR ¹ ; Q is selected from the group consisting of N and CR ¹ ; Y is selected from the group consisting of CZ and N, conditional When Y is N and Y ¹ , Y ² and Y ³ are CZ, B is N-alkyl or O; Y ¹ is selected from the group consisting of CZ and N; Y ² is selected from the group consisting of CZ and N; Y ³ is selected from the group consisting of CZ and N; Z is independently selected from the group consisting of H, halogen, O-alkyl, alkyl and CN; R is independently selected from

And -NR ³ R ⁴ ; R ^a is selected from the group consisting of H and alkyl; R ^b , R ^c , R ^d , R ^e , R ^f and R ^{g are} independently selected from the group consisting of H and alkyl Group, or any two of R ^b , R ^c , R ^d , R ^e , R ^f and R ^g may be linked to form a ring of 3 to 8 members; R ^{j is} independently selected from the group consisting of-halogen, -O-alkyl, -CF ₃ , -CN, -NR ³ R ⁴ ,

and

, In which the bridge containing C _1-2 carbon atoms may exist between a carbon atom and c or d carbon atom or the bridge containing C _1-2 carbon atoms may exist between b carbon atom and c or d carbon atom ; R ¹ is selected from the group consisting of H and alkyl; R ^{2 is} independently selected from the group consisting of alkyl, F, and =O, where the alkyl is optionally substituted with halogen, -OH, or -O-alkyl, And if two R ² are twins, they can be connected to form a 3-member to 6-member ring; R ³ and R ^{4 are} independently selected from the group consisting of H and alkyl, where the alkyl is optionally halogenated , -OH or -O-alkyl substitution; R ⁵ is selected from the group consisting of H and alkyl; n is 0, 1, 2, 3 or 4. r and s are independently 0, 1, 2, or 3; and t and u are independently 1, 2, or 3. 2. The compound of item 1 is a compound of formula (Ia):

Wherein ^{A, B, R b, R} c, R d, R e, R f, R g, Y and Z are as defined in the system as an entry. 3. The compound of item 1 is a compound of formula (Ib):

Wherein ^{A, B, R b, R} c, R d, R e, R f, R g and Z entry system as defined in 1. 4. Compounds as in any of items 1 to 3, where A is

and

,among them

and

Can be connected to Q or N at any available location, where

Substituted by one or more substituents R ^j , and wherein

Optionally substituted with one or more substituents R. 5. Compounds as in any of items 1 to 3, where A is

,among them

Can be connected to Q or N at any available location, and where

Optionally substituted with one or more substituents R. 6. A compound of any of items 1 to 5 is a compound of formula (Ic):

Where E , R , V and Z are as defined in item 1. 7. The compound of any one of items 1 to 4 is a compound of formula (Id):

Where R ^a , R ^j and Z are as defined in item 1 and p is 1 or 2. 8. The compound of item 1, wherein the compound is selected from the group consisting of:

. 9. A pharmaceutical composition comprising a compound according to any one of items 1 to 8 and optionally a pharmaceutically acceptable carrier or excipient. 10. The compound as in any one of items 1 to 8 is used as a medicament. 11. A compound of formula (I),

And all its stereoisomers, racemic mixtures, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates and polymorphs; it is used to treat, improve or prevent aggregation with Tau protein Body-related diseases or abnormalities, where A is selected from the group consisting of:

and

,among them

and

Can be connected to Q at any available location, where

and

Substituted by one or more substituents R ^j , and wherein

and

Optionally substituted with one or more substituents R ; B is selected from the group consisting of O and NR ^a ; E and V are independently selected from the group consisting of: N, NR ⁵ , O, and S; G is selected from the group consisting of Group consisting of: benzene ring, pyrimidine ring and pyridine ring; J is selected from the group consisting of O and NR ¹ ; Q is selected from the group consisting of N and CR ¹ ; Y is selected from the group consisting of CZ and N; Y ¹ Is selected from the group consisting of CZ and N; Y ² is selected from the group consisting of CZ and N; Y ³ is selected from the group consisting of CZ and N; Z is independently selected from the group consisting of: H, halogen, O- Alkyl, alkyl and CN; R is independently selected from

And -NR ³ R ⁴ ; R ^a is selected from the group consisting of H and alkyl; R ^b , R ^c , R ^d , R ^e , R ^f and R ^{g are} independently selected from the group consisting of H and alkyl Group, or any two of R ^b , R ^c , R ^d , R ^e , R ^f and R ^g may be linked to form a ring of 3 to 8 members; R ^{j is} independently selected from the group consisting of-halogen, -O-alkyl, -CF ₃ , -CN, -NR ³ R ⁴ ,

and

, In which the bridge containing C _1-2 carbon atoms may exist between a carbon atom and c or d carbon atom or the bridge containing C _1-2 carbon atoms may exist between b carbon atom and c or d carbon atom ; R ¹ is selected from the group consisting of H and alkyl; R ^{2 is} independently selected from the group consisting of alkyl, F, and =O, where the alkyl is optionally substituted with halogen, -OH, or -O-alkyl, And if two R ² are twins, they can be connected to form a 3-member to 6-member ring; R ³ and R ^{4 are} independently selected from the group consisting of H and alkyl, where the alkyl is optionally halogenated , -OH or -O-alkyl substitution; R ⁵ is selected from the group consisting of H and alkyl; n is 0, 1, 2, 3 or 4; r and s are independently 0, 1, 2 or 3; And t and u are independently 1, 2, or 3. 12. The compound as used in item 11, wherein the compound of formula (I) is as defined in any one of items 1 to 8. 13. A compound as in any of items 1 to 8 or 11, which is used to reduce Tau aggregation. 14. A compound according to any one of items 1 to 8 or 11, which is used to prevent the formation of Tau aggregates and/or to inhibit Tau aggregation. 15. The compound of any one of items 1 to 8 or 11, which is used for intracellular interference with Tau aggregates. 16. A compound according to any one of items 1 to 8 or 11, which is used to reduce Tau misfolding and hyperphosphorylation in vivo. 17. The compound of any one of items 1 to 8 or 11, which is used as a marker for reducing neuroinflammation. 18. A method of treating, preventing or ameliorating a condition or abnormality associated with Tau protein aggregates, the method comprising administering an effective amount of a compound such as any one of items 1 to 8 or 11 to an individual in need. 19. A method for reducing Tau aggregation, which method comprises administering an effective amount of a compound such as any of items 1 to 8 or 11 to an individual in need. 20. A method for preventing the formation of Tau aggregates and/or inhibiting Tau aggregation, the method comprising administering an effective amount of a compound such as any one of items 1 to 8 or 11 to an individual in need. 21. A method of intracellular interference with Tau aggregates, the method comprising administering an effective amount of a compound such as any of items 1 to 8 or 11 to an individual in need. 22. A method of reducing Tau misfolding and hyperphosphorylation in vivo, which method comprises administering an effective amount of a compound such as any one of items 1 to 8 or 11 to an individual in need. 23. A method of reducing neuroinflammation markers, the method comprising administering an effective amount of a compound such as any of items 1 to 8 or 11 to an individual in need. 24. The use of a compound according to any one of items 1 to 8 or 11 for the manufacture of a medicament for the treatment, prevention or improvement of a disease or abnormality associated with Tau protein aggregates. 25. Use of a compound according to any one of items 1 to 8 or 11 for the manufacture of a medicament for reducing Tau aggregation. 26. The use of a compound according to any one of items 1 to 8 or 11 for the manufacture of a medicament for preventing the formation of Tau aggregates and/or for inhibiting Tau aggregation. 27. Use of a compound according to any one of items 1 to 8 or 11 for the manufacture of a medicament for intracellular interference with Tau aggregates. 28. The use of a compound as described in any of items 1 to 8 or 11 for the manufacture of a medicament for reducing Tau misfolding and hyperphosphorylation in vivo. 29. The use of a compound according to any one of items 1 to 8 or 11 for the manufacture of a medicament for reducing neuroinflammation markers. 29. A mixture comprising a compound according to any one of items 1 to 8 and at least one other biologically active compound selected from compounds different from any one of items 1 to 8 The therapeutic agent, wherein the mixture further comprises at least one of a pharmaceutically acceptable carrier, diluent and excipient. 30. The mixture of item 29, wherein the other biologically active compound is a compound used to treat amyloidosis. 31. The mixture of item 29 or 30, wherein the compound and/or the other biologically active compound is present in a therapeutically effective amount. 32. The mixture of any one of items 29 to 31, wherein the other biologically active compound is selected from the group consisting of: anti-oxidative stress compounds; anti-apoptotic compounds; metal chelating agents; DNA repair inhibitors, for example Pirenzepine and metabolites; 3-amino-1-propane sulfonic acid (3APS); 1,3-propane disulfonate (1,3PDS); α-secretase activator; β-secretase And γ-secretase inhibitors; glycogen synthase kinase 3 inhibitors; O-linked N-acetylglucosamine hydrolase (O-glcnacase, OGA) inhibitors; neurotransmitters; beta pleated plate fragmentation agent; Starch beta-eliminating/eliminating cell component attractants; inhibitors of N-terminal truncated starch beta (including pyroglutamated starch beta 3-42), anti-inflammatory molecules, or cholinesterase inhibitors ( ChEI), such as tacrine, rivastigmine, donepezil and/or galantamine; M1 agonists; other drugs, including any starch-like or Tau modified drugs and nutritional supplements; antibodies, including any functionally equivalent antibodies or functional parts thereof; or vaccines. 33. The mixture of item 32, wherein the other biologically active compound is a cholinesterase inhibitor (ChEI). 34. A mixture 32 as in the item, wherein the other biologically active compound is selected from the group consisting of: Tacrine, Levistamine, Donepezil, Galantamine, Nicotinic Acid, and Memantine ). 35. The mixture of item 32, wherein the other biologically active compound is an antibody, specifically a monoclonal antibody, including any functionally equivalent antibody or functional part thereof. 36. The mixture of any of items 29 to 35, wherein the compound and/or the other biologically active compound is present in a therapeutically effective amount. 37. The compound as used in item 11, the method as in item 18, or the use as in item 24, wherein the disorder is selected from Alzheimer's disease (AD), familial AD, primary age-related Tau proteinopathy (PART), Kuja's disease, boxer-type dementia, Down's syndrome, Jetzman-Strosler-Schenck disease (GSS), inclusion body myositis, prion protein cerebral amyloid angiopathy, Traumatic brain injury (TBI), amyotrophic lateral sclerosis (ALS), Guam Parkinson's disease-dementia complex, non-Guam motor neuron disease with neurofibrillary tangles, argyrophilic granulopathy, cortical base Nuclear degeneration (CBD), diffuse neurofibrillary tangles with calcification, frontotemporal dementia associated with chromosome 17 with Parkinson's disease (FTDP-17), Hallerwoden-Spartz disease, multiple system atrophy (MSA), Niemann-Pick disease type C, pale ball-pontine-melanosis, Pick's disease (PiD), progressive subcortical glioma, progressive supranuclear palsy (PSP), subacute sclerosis Pan-encephalitis, tangled dominant dementia, post-encephalitis Parkinson's disease, myotonic dystrophy, subacute sclerosing pan-encephalopathy, LRRK2 mutation, chronic traumatic brain disease (CTE), familial British type Dementia, familial Danish dementia, other frontotemporal lobe degeneration, Guadeloupe Parkinson's disease, neurodegeneration with iron accumulation in the brain, SLC9A6-related mental retardation, white matter tau protein lesions with glial cells Inclusion bodies, epilepsy, Lewy body dementia (LBD), mild cognitive impairment (MCI), multiple sclerosis, Parkinson's disease, HIV-related dementia, adult-onset diabetes, senile cardiac amyloidosis , Glaucoma, ischemic stroke, psychosis in AD and Huntington's disease, preferably Alzheimer's disease (AD), cortical basal nucleus degeneration (CBD), Pick's disease (PiD) and progressive Supranuclear palsy (PSP). 38. The use of a compound as described in any of items 1 to 8 or 11 as an analytical reference or in vitro screening tool.

Definitions Within the meaning of this application, the following definitions apply: "Alkyl" means a saturated linear or branched organic part composed of carbon and hydrogen atoms. Examples of suitable alkyl groups have 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms, and include methyl, ethyl, propyl, isopropyl, n-butyl, tertiary butyl and isobutyl . The alkyl group is optionally substituted with halogen (preferably F), -OH or -O alkyl (preferably -OMe). "Hal" or "halogen" means F, Cl, Br and I. "3-member to 8-member ring" means a 3-member, 4-member, 5-member, 6-member, 7-member or 8-member ring, in which there are no carbon atoms in the ring, and one or more carbon atoms are already 1 or 2 (for 3 member ring), 1, 2 or 3 (for 4 member ring), 1, 2, 3 or 4 (for 5 member ring) or 1, 2, 3, 4 or 5 (for 6 member ring), 1, 2, 3, 4, 5 or 6 (for 7-membered rings) or 1, 2, 3, 4, 5, 6 or 7 (for 8-membered rings) the same or different heteroatoms are substituted, of which The heteroatom system is selected from O, N and S.

Compounds of the invention having one or more optically active carbons can exist as: racemates and racemic mixtures (including mixtures in all ratios), stereoisomers (including non-mirromeric mixtures and individual non-isomeric mixtures) Mirror isomers, mirror isomer mixtures and single mirror isomers, mixtures of configuration isomers and single configuration isomers), tautomers, atropisomers and rotamers. All isomeric forms are included in the present invention. The compounds described in the present invention containing olefinic double bonds include E and Z geometric isomers. The invention also includes all pharmaceutically acceptable salts, prodrugs, polymorphs, hydrates and solvates.

The term "polymorph" refers to various crystal structures of the compounds of the present invention. This may include, but is not limited to, crystal morphology (and amorphous materials) and all lattice forms. The salts of the present invention can be crystalline and can exist as more than one polymorph.

The invention also encompasses solvates, hydrates and anhydrous forms of salts. The solvent included in the solvate is not specifically limited and may be any pharmaceutically acceptable solvent. Examples include water and C _1-4 alcohols (such as methanol or ethanol).

"Pharmaceutically acceptable salt" is defined as a derivative of the disclosed compound, wherein the parent compound is modified by preparing its acidic or basic salt. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues (eg, amines), basic or organic salts of acidic residues (eg, carboxylic acids), and the like. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from, for example, non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and from organic Salts prepared by acids, such as (but not limited to) acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, Hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, Isethionic acid and the like. The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of the compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two. Organic solvents include, but are not limited to, non-aqueous media, such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts can be found in Remington’s Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, PA, 1990, p. 1445, the disclosure of which is incorporated herein by reference.

The compounds of the present invention can also be provided in the form of prodrugs, that is, compounds that are metabolized into active metabolites in vivo. As used in the following description of the invention and the scope of patent applications, the term "prodrug" means any covalently bonded compound that releases the active parent drug due to in vivo biotransformation. Goodman and Gilman's references to prodrugs (The Pharmacological Basis of Therapeutics, 8th Edition, McGraw-Hill, Int. Ed. 1992, "Biotransformation of Drugs", pages 13-15) are usually incorporated by reference In this article.

"Pharmaceutically acceptable" is defined as suitable for use in contact with human and animal tissues within reasonable medical judgment without excessive toxicity, irritation, allergic reactions or other problems or complications, and is commensurate with a reasonable benefit/risk ratio Their compounds, materials, compositions and/or dosage forms.

The patient or individual of the present invention is usually an animal, specifically a mammal, and more specifically a human.

"Tau" as used herein refers to a highly soluble microtubule binding protein mainly found in neurons, and includes mainly 6 isoforms, cleaved or truncated forms, and other modified forms, such as derived from phosphorylation, sugar Glycosylation, saccharification, prolamidoyl isomerization, nitration, acetylation, polyamination, ubiquitination, small ubiquitination (sumoylation) and oxidation.

"Aggregated Tau" refers to aggregated monomers of Tau peptides or proteins folded into an oligomeric or polymeric structure.

"Nerve fiber tangles" (NFT) as used herein refers to insoluble aggregates of highly phosphorylated Tau protein containing paired helical filaments (PHF) and straight filaments. Its presence is a hallmark of AD and other diseases called tauopathy.

As used herein, the term "antibody (antibody or antibodies)" is an industry-recognized term and should be understood to refer to molecules or active fragments of molecules that bind to known antigens, or specifically to immunoglobulin molecules and immunoglobulins The antigen-binding portion of a protein molecule. Specifically, the mixture of the present invention includes a compound of the present invention and an anti-Tau or anti-Abeta antibody.

The term "functionally equivalent antibody or functional part thereof" as used herein should be understood to refer to an equivalent molecule or active fragment of the molecule that binds to a known antigen, or specifically refers to immunoglobulin molecules and immunoglobulins The antigen-binding portion of a molecule has substantially the same (biological) activity as the antibody from which it is derived.

The "vaccines" or "vaccines" reported in the mixture of the present invention are specifically Tau or Abeta vaccines.

Unless otherwise stated, the definitions and preferred definitions given in the "Definitions" section apply to all embodiments set forth below.

The compounds of the present invention will be explained below. It should be understood that all possible combinations of the following definitions are also contemplated.

In one embodiment, the invention relates to compounds of formula (I):

And all its stereoisomers, racemic mixtures, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates and polymorphs.

A preferred embodiment of the compound of formula (I) is

Another preferred embodiment of the compound of formula (I) is

Another preferred embodiment of the compound of formula (I) is

In another preferred embodiment, the compound of formula (I) is

或

Even better, the compound of formula (I)

or

Even better,

The following definitions apply to the A of the formula (I) compounds of preferred embodiments.

及

，其中G 係選自苯環、嘧啶環及吡啶環。因此

涵蓋以下較佳實施例：

及

。 且因此，

涵蓋以下實施例：

及

。在較佳實施例中，

係選自

及

。 可設想以下各環：

及

。較佳實施例包括

及

。在該等結構式中，環G 未完全顯示，但僅由部分鍵

指示。 A is selected from the group consisting of:

and

, Where G is selected from benzene ring, pyrimidine ring and pyridine ring. therefore

Covers the following preferred embodiments:

and

. And therefore,

The following examples are covered:

and

. In a preferred embodiment,

Selected from

and

. The following rings can be imagined:

and

. Preferred embodiments include

and

. In these structural formulas, the ring G is not fully shown, but only part of the bond

Instructions.

及

。In a preferred embodiment, A is selected from

and

.

及

。在一個較佳實施例中，A 係

。In another preferred embodiment, A

and

. In a preferred embodiment, A

.

及

。在更佳實施例中，A 係選自

及

。In a more preferred embodiment, A is selected from

and

. In a more preferred embodiment, A is selected from

and

.

及

。In another preferred embodiment, A is selected from

and

.

。In an even better embodiment, the A series

.

及

可在任一可用位置處連接至Q。Defined above and A of the preferred embodiment,

and

Can be connected to Q at any available location.

及

經一或多個取代基R^j 取代。Defined above and A of the preferred embodiment,

and

Substitution by one or more substituents R ^j .

及

可視情況經一或多個取代基R 取代。Defined above and A of the preferred embodiment,

and

Optionally substituted with one or more substituents R.

If appropriate, the following definitions apply to formula (I) and its preferred embodiments.

B selected from the group consisting of O and NR ^a group consisting of. More preferably, B is NR ^a and NH is the best.

E and V are independently selected from the group consisting of: N, NR ⁵ , O, and S. Since the ring system containing E and V is unsaturated, at least one of E and V is N.

J is selected from the group consisting of O and NR ¹ , more preferably O.

Q is selected from the group consisting of N and CR ¹ . Preferably, Q is selected from the group consisting of N and CH, more preferably Q is N.

Y is selected from the group consisting of CZ and N. More preferably, Y is selected from the group consisting of CH and N. Even better, Y is CH. In one embodiment, if Y is N and Y ¹ , Y ² and Y ³ are CZ, then B is N-alkyl or O, preferably B is N-alkyl.

Y ¹ is selected from the group consisting of CZ and N. Preferably, Y ¹ is CZ.

Y ² is selected from the group consisting of CZ and N. Preferably, Y ² is CZ.

Y ³ is selected from the group consisting of CZ and N. Preferably, Y ³ is CZ.

Z is independently selected from the group consisting of H, halogen (preferably F), O-alkyl, alkyl and CN, preferably selected from H, halogen (preferably F) and O-alkyl. In a preferred embodiment, one Z is independently halogen (preferably F) or O-alkyl, and the other Z is H. In a more preferred embodiment, one Z is halogen (preferably F) and the other Z is H.

及-NR³ R⁴ 以下組成之群，較佳地R 係選自由

及-NR³ R⁴ 組成之群，更佳地R 係

，例如

。在該等實施例中，

較佳係

。 R is independently selected from

And -NR ³ R ⁴ or less, preferably R is selected from

And -NR ³ R ⁴ group, better R series

,E.g

. In these embodiments,

Better

.

R ^a group selected from the group consisting of H and alkyl groups, more preferably H and based Me, even more preferably H. Department

R ^b , R ^c , R ^d , R ^e , R ^f , R ^{g are} independently selected from the group consisting of H and alkyl, or any one of R ^b , R ^c , R ^d , R ^e , R ^f , R ^g Both (eg, they are connected to the same or adjacent ring atoms) can be joined to form a 3 to 8 member ring. More preferably, R ^b , R ^c , R ^d , R ^e , R ^f , R ^{g are} independently H or alkyl, and even more preferably H.

及

，其中含C_1-2 碳原子之橋可存在於a碳原子與c或d碳原子之間或其中含C_1-2 碳原子之橋可存在於b碳原子與c或d碳原子之間。更佳地，R^j 係選自由以下組成之群：-鹵素、-O-烷基、-NR³ R⁴ 及

，甚至更佳地R^j 係

。 R ^{j is} independently selected from the group consisting of: -halogen, -O-alkyl, -CF ₃ , -CN, -NR ³ R ⁴ ,

and

, In which the bridge containing C _1-2 carbon atoms can exist between a carbon atom and c or d carbon atom or the bridge containing C _1-2 carbon atoms can exist between b carbon atom and c or d carbon atom . More preferably, R ^j is selected from the group consisting of: -halogen, -O-alkyl, -NR ³ R ⁴ and

, Or even better, R ^j

.

R ¹ is selected from the group consisting of H and alkyl, preferably alkyl, and more preferably CH ₃ .

R ^{2 is} independently selected from the group consisting of alkyl, F, and =O, where the alkyl is optionally substituted with halogen, -OH, or -O-alkyl, and where two R ² are twins, then Can be connected to form a ring of 3 to 6 members. In one embodiment, R ² is optionally substituted alkyl, in another embodiment, R ² is F, and in another embodiment, R ² is =O.

R ³ and R ^{4 are} independently selected from the group consisting of H and alkyl, wherein the alkyl is optionally substituted with halogen, -OH or -O-alkyl. In one embodiment, R ³ or R ⁴ is optionally substituted alkyl and the other is H. In another embodiment, R ³ is alkyl and R ⁴ is optionally substituted alkyl. In another embodiment, R ³ and R ⁴ are H.

R ⁵ is selected from the group consisting of H and alkyl. In one embodiment, R ⁵ is H, and in another embodiment, R ⁵ is alkyl.

n is 0, 1, 2, 3 or 4, preferably n is 0 or 1, more preferably n is 0.

p is 1 or 2, more preferably 1.

r and s are independently 0, 1, 2 or 3.

t and u are independently 1, 2, or 3.

Preferred compounds of formula (I) are

Preferred compounds are also illustrated in the examples.

The present invention also contemplates any combination of the embodiments disclosed herein, preferred embodiments, and better embodiments.

Pharmaceutical composition Although the compound of the present invention can be administered alone, it is preferably formulated into a pharmaceutical composition according to standard pharmaceutical practice. Therefore, the present invention also provides a pharmaceutical composition comprising a mixture of a therapeutically effective amount of a compound of formula (I) as appropriate and a pharmaceutically acceptable carrier, diluent, adjuvant or excipient.

Pharmaceutically acceptable excipients are well known in pharmaceutical technology and are described in, for example, Remington's Pharmaceutical Sciences, 15th Edition, Mack Publishing Co., New Jersey (1975). Pharmaceutical excipients can be selected according to the intended route of administration and standard pharmaceutical practice. The excipient must be acceptable in the sense that it is not harmful to its recipient.

The pharmaceutically acceptable excipients that can be used to formulate the pharmaceutical composition of the present invention can include, for example, carriers; vehicles; diluents; solvents, such as monohydric alcohols (such as ethanol, isopropyl alcohol) and polyhydric alcohols (such as dihydric alcohols) Alcohol) and edible oils (eg soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil), oily esters (eg ethyl oleate, isopropyl myristate), binders; adjuvants; solubilizers; Thickener; stabilizer; disintegrant; glidant; lubricant; buffer; emulsifier; wetting agent; suspending agent; sweetener; colorant; flavoring agent; coating agent; preservative; antioxidant ; Treatment agents; drug delivery modifiers and enhancers, such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, right Spinose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low melting wax and ion exchange resin.

The administration (delivery) route of the compound of the present invention includes, but is not limited to, one or more of the following: oral (for example, as a lozenge, capsule, or as an ingestible solution), topical, and transmucosal (for example, as an inhalation solution) Nasal spray or aerosol), nasal, parenteral (e.g. by injectable form), gastrointestinal, intraspinal, intraperitoneal, intramuscular, intravenous, intrauterine, intraocular, intradermal, intracranial, tracheal Inner, intravaginal, intraventricular, intracerebral, subcutaneous, transocular (including intravitreal or anterior chamber), transdermal, rectal, transbuccal, epidural, and sublingual.

For example, the compound may be administered orally in the form of lozenges, capsules, vaginal suppositories (ovules), elixirs, solutions or suspensions containing flavoring or coloring agents for immediate, delayed, modified, sustained, pulsed or Controlled release application.

Lozenges may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine; disintegrants such as starch (preferably corn, potato or tapioca starch), hydroxyl Sodium starch acetate, croscarmellose sodium and certain composite silicates; and granulated binders, such as polyvinylpyrrolidone, hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose, gelatin and gum arabic. In addition, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc can be incorporated. Similar types of solid compositions can also be used as fillers in gelatin capsules. In this regard, preferred excipients include lactose, starch, cellulose, lactose or high molecular weight polyethylene glycol. For aqueous suspensions and/or elixirs, the agent can be combined with the following: various flavorings or flavoring agents, coloring substances or dyes, emulsifiers and/or suspending agents and diluents (eg water, ethanol, propylene glycol and glycerin) And combinations.

If the compound of the present invention is administered parenterally, examples of this administration include one or more of the following: intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, cranial These compounds are administered internally, intramuscularly or subcutaneously; and/or by using infusion techniques. For parenteral administration, it is best to use the compound in the form of a sterile aqueous solution, which may contain other substances, such as sufficient salt or glucose, to make the solution isotonic with blood. The aqueous solution should be appropriately buffered as needed (preferably to a pH of 3 to 9). It is easy to prepare suitable parenteral formulations under sterile conditions by standard pharmaceutical techniques well known to those skilled in the art.

As indicated, the compounds of the present invention can be administered intranasally or by inhalation, and use such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, hydrofluoroalkane (for example, 1, 1,1,2-tetrafluoroethane (HFA134AT) or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EA)), carbon dioxide or other suitable gas, etc. Pressurized containers, pumps, sprays or nebulizers are conveniently delivered in the form of dry powder inhaler or aerosol spray delivery. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve to deliver the meter. The pressurized container, pump, spray or atomizer may contain a solution or suspension of the active compound, for example using a mixture of ethanol and propellant as a solvent, which may additionally contain a lubricant, such as sorbitan trioleate. Capsules and cartridges (for example, made from gelatin) used in inhalers or insufflators can be formulated to contain a powder mixture of the compound and a suitable powder base (such as lactose or starch).

Alternatively, the compounds of the present invention can be administered in the form of suppositories or pessaries, or they can be applied topically in the form of gels, hydrogels, lotions, solutions, creams, ointments, or powders. The compounds of the present invention can also be administered transdermally or through the skin, for example by using skin patches.

It can also be administered via the lung or rectal route. It can also be administered by eye. For ocular use, the compound can be formulated as a micronized suspension in isotonic, pH-adjusted sterile saline, or preferably in an isotonic, pH-adjusted sterile saline, depending on the situation and the preservative (Eg benzalkonium chloride) combination. Alternatively, it can be formulated in ointment (eg paraffin).

For topical application to the skin, the compound of the present invention may be formulated as a suitable ointment containing the active compound suspended or dissolved in, for example, a mixture of one or more of: mineral oil, liquid paraffin, white petrolatum, propylene glycol, emulsifying wax And water. Alternatively, it can be formulated as a suitable lotion or cream, suspended or dissolved in, for example, a mixture of one or more of: mineral oil, sorbitan monostearate, polyethylene glycol, liquid paraffin, Polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

Generally, the physician will determine the actual dose that is most suitable for the individual. The specific dose and dose frequency for any particular individual may vary and will depend on a variety of factors, including the activity of the specific compound used, the metabolic stability of the compound and the length of action, age, weight, overall health status, gender, diet , The way and time of administration, the rate of excretion, the combination of drugs, the severity of a specific condition and the individual undergoing therapy.

The recommended dosage of the compound of the present invention for administration to humans (approximately 70 kg body weight) is 0.1 mg to 3 g, 0.1 mg to 2 g, 0.1 mg to 1 g, preferably 1 mg to 500 mg of active ingredient per unit dose. The unit dose can be administered, for example, 1 to 4 times per day. The dosage will depend on the route of administration. It should be understood that the dosage may need to be routinely changed depending on the age and weight of the patient and the severity of the condition to be treated. The precise dosage and route of administration will ultimately be determined by the attending physician or veterinarian.

The compounds of the present invention can also be used in combination with other therapeutic agents. When the compound of the present invention is used in combination with a second therapeutic agent having activity against the same disease, the dose of each compound may be different from that when the compound is used alone.

The combinations mentioned above can be conveniently presented and used in the form of pharmaceutical formulations. The individual components of these combinations can be administered sequentially or simultaneously by any convenient route, alone or in combination. When administered sequentially, the compound of the present invention or the second therapeutic agent may be administered first. When administered simultaneously, the combination can be administered with the same or different pharmaceutical compositions. When combined in the same formulation, it should be understood that the two compounds must be stable and compatible with each other and with other components of the formulation. When formulated separately, it can be provided in any convenient formulation, conveniently in a manner known in the art for such compounds.

The pharmaceutical composition of the present invention can be produced in a manner known to those skilled in the art as described in, for example, Remington's Pharmaceutical Sciences, 15th Edition, Mack Publishing Co., New Jersey (1975).

The diseases or conditions that can be treated, ameliorated, or prevented by the compounds of the present invention are disorders or abnormalities associated with Tau protein aggregates, such as neurodegenerative disorders. Examples of diseases and conditions that can be treated, ameliorated or prevented are caused by or related to the formation of nerve fiber lesions. This is the main brain pathology of tau protein lesions. These diseases and conditions include neurodegenerative diseases or conditions of heterogeneous populations, including those that show the coexistence of Tau and amyloid pathology.

Examples of diseases and conditions that can be treated, improved, or prevented include, but are not limited to, Alzheimer's disease (AD), familial AD, PART (primary age-related Tau proteinopathy), Kuja disease, Boxer-type dementia, Down's syndrome, Jetzman-Strodler-Schenck disease (GSS), inclusion body myositis, prion protein cerebral amyloid angiopathy, traumatic brain injury (TBI), Amyotrophic lateral sclerosis (ALS), Parkinson's disease-dementia complex of Guam, non-Guam motor neuron disease with neurofibrillary tangles, argyrophilic granulopathy, cortical basal nucleus degeneration (CBD), diffuse Neurofibrillary tangles with calcification, frontotemporal dementia associated with chromosome 17 with Parkinson's disease (FTDP-17), Hallerwalden-Spatz disease, multiple system atrophy (MSA), Niemann type C Pick's disease, globus pallidus-pontine-melanosis, Pick's disease (PiD), progressive subcortical glioma, progressive supranuclear palsy (PSP), subacute sclerosing panencephalitis, tangled dominant type Dementia, Parkinson's disease after encephalitis, myotonic dystrophy, subacute sclerosing panencephalopathy, LRRK2 mutation, chronic traumatic brain disease (CTE), familial British type dementia, familial Danish type Dementia, other frontotemporal lobar degeneration, Guadeloupe Parkinson's disease, neurodegeneration with iron accumulation in the brain, SLC9A6-related mental retardation, white matter tau protein lesions with glial cell inclusions, epilepsy, Lewy body Dementia (LBD), mild cognitive impairment (MCI), multiple sclerosis, Parkinson's disease, HIV-related dementia, adult-onset diabetes, senile cardiac amyloidosis, glaucoma, ischemic stroke, Psychosis and Huntington's disease in AD. Preferably, the diseases and conditions that can be treated, improved or prevented include Alzheimer's disease (AD), and other neurodegenerative tau protein lesions, such as Kuja's disease, boxer-type dementia, muscle atrophy Lateral sclerosis (ALS), argyrophilic granulopathy, cortical basal nucleus degeneration (CBD), frontotemporal dementia associated with chromosome 17 with Parkinson's disease (FTDP-17), Pick's disease (PiD ), progressive supranuclear palsy (PSP), tangled dominant dementia, Parkinson’s disease dementia syndrome in Guam, Hallerwalden-Spatts disease, chronic traumatic brain disease (CTE), trauma Brain injury (TBI) and other frontotemporal degeneration. More preferred are Alzheimer's disease (AD), cortical basal nucleus degeneration (CBD), Pick's disease (PiD) and progressive supranuclear palsy (PSP).

The compounds of the invention can also be used to reduce protein aggregation, specifically Tau aggregation. The ability of a compound to reduce Tau aggregation can be determined, for example, using ThT analysis (Hudson et al., FEBS J., 2009, 5960-72).

The compounds of the present invention can be used to treat a variety of disorders related to the misfolding and/or pathological aggregation of Tau protein in the process of neuroinflammation.

The compounds of the present invention can be used as analytical reference or in vitro screening tools for characterizing tissues with Tau pathology and for testing compounds targeting Tau pathology on this tissue.

The compounds of the present invention can be used as optical probes for cross-linking compounds with targets and in in vitro screening assays to identify and characterize the mode of action of compounds, including localization of binding sites. The following reports some examples of optical probes of the compounds of the present invention:

The compounds of the present invention may also be provided in the form of a mixture with at least one other biologically active compound and/or pharmaceutically acceptable carriers and/or diluents and/or excipients. These compounds and/or other biologically active compounds are preferably present in a therapeutically effective amount.

The nature of other biologically active compounds will depend on the intended use of the mixture. Other biologically active substances or compounds can exert their biological effects by the same or similar mechanisms as the compounds of the present invention or by an unrelated mechanism of action or by a variety of related and/or unrelated mechanisms of action.

Generally, other biologically active compounds may include neurotransmission enhancers, psychotherapeutics, acetylcholinesterase inhibitors, calcium channel blockers, biogenic amines, benzodiazepine sedatives, acetylcholine synthesis, Storage or release enhancers, acetylcholine post-synaptic receptor agonists, monoamine oxidase-A or monoamine oxidase-B inhibitors, N-methyl-D-aspartate glutamate receptor antagonists, non- Steroid anti-inflammatory drugs, antioxidants and serotonin receptor antagonists. In particular, other biologically active compounds can be selected from the group consisting of: compounds for the treatment of amyloidosis; compounds that resist oxidative stress; anti-apoptotic compounds; metal chelating agents; DNA repair inhibitors, such as pirenzepine And metabolites, 3-amino-1-propanesulfonic acid (3APS), 1,3-propanedisulfonate (1,3PDS); α-secretase activator; β-secretase and γ-secretase inhibition Agents; glycogen synthase kinase 3 inhibitors; O-linked N-acetylglucosamine hydrolase (OGA) inhibitors; neurotransmitters; beta pleated plate fragmenting agent; starch-like beta clearance/elimination of cellular components Agents; N-terminal truncated starch beta (including pyroglutaminated starch beta 3-42) inhibitors; anti-inflammatory molecules, or cholinesterase inhibitors (ChEI), such as Tacrine, Lifan Stemin, Donepezil and/or Galantamine; M1 agonists; other drugs, including any starch-like or Tau-modified drugs and nutritional supplements; antibodies, including any functionally equivalent antibodies or functional parts thereof; Or vaccine.

In another embodiment, the mixture of the invention may comprise nicotinic acid or memantine and the compound of the invention and optionally pharmaceutically acceptable carriers and/or diluents and/or excipients.

In another embodiment of the present invention, a mixture is provided, which contains, for example, for treating positive and negative psychotic symptoms, including hallucinations, delusions, and thinking disorders (embodied as obvious incoherence, derailment of thinking, speech bifurcation Tangentiality) and bizarre or deconstructive behaviors, as well as alopecia, flattened affect, indifference and social withdrawal, clozapine, ziprasidone, risperidone, and Aripiprazole or olanzapine as another biologically active compound "atypical antipsychotic", and the compounds of the present invention and optionally pharmaceutically acceptable carriers and/or diluents and/or excipient.

Other compounds that can be suitably used in the mixture in combination with the compounds of the present invention are described in, for example, WO 2004/058258 (see especially pages 16 and 17), which includes therapeutic drug targets (pages 36 to 39), Alkane sulfonic acid and alkanol sulfuric acid (pages 39 to 51), cholinesterase inhibitors (pages 51 to 56), NMDA receptor antagonists (pages 56 to 58), estrogen (pages 58 to 59) ), nonsteroidal anti-inflammatory drugs (pages 60 and 61), antioxidants (pages 61 and 62), peroxisome proliferator-activated receptor (PPAR) agonists (pages 63 to 67), cholesterol-lowering agents (pages 68-75), starch-like inhibitors (pages 75-77), starch-like formation inhibitors (pages 77-78), metal chelators (pages 78-79), antipsychotics and antidepressants (pages 78-79) Pages 80 to 82), nutritional supplements (pages 83 to 89) and compounds that increase the availability of bioactive substances in the brain (see pages 89 to 93) and prodrugs (pages 93 and 94), the document is Incorporated by reference.

The invention also includes all suitable isotopic variations of the compounds of the invention. The isotopic variations of the compounds of the present invention are defined as those in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from the atomic mass usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, and chlorine, such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ³⁵ S, ¹⁸ F and ³⁶ Cl. Certain isotopic variations of the invention (such as those in which radioisotopes (eg, ³ H or ¹⁴ C) are incorporated) can be used in drug and/or substrate tissue distribution studies. Tritium (ie ³ H) and carbon-14 (ie ¹⁴ C) isotopes are particularly preferred because of their ease of preparation and detectability. ¹⁸ F labeled compounds are particularly suitable for imaging applications such as PET. Further, substitution with isotopes such as deuterium (i.e., ² H) or the like having a substituent because of the greater metabolic stability may afford certain therapeutic advantages such that, for example, in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. The isotopic variations of the compounds of the present invention can generally be prepared by conventional procedures, for example by illustrative methods or by the preparation described in the examples and preparations below by preparing appropriate isotopic variations using suitable reagents.

The compound of the present invention can be synthesized by one of the general methods shown in the following scheme. These methods are given for illustrative purposes only and should not be interpreted as limiting.

General synthetic scheme for preparing the building unit of the present invention:

plan 1

Was heated in a suitable solvent commercially available phenylhydrazine derivatives under acidic conditions (Z = H, F, Cl , Me , or ^{OMe; R a = H, CH} 3) and commercially available 4-oxo-hexahydro-l -Tert-butyl formate (synthesized by Fischer-Indole), after purification, tricyclic 2,3,4,5-tetrahydro-1 H -pyrido[4,3- b ] Indole building block. In the case of using 2- or 3-substituted phenylhydrazine derivatives, regioisomers are separated by supercritical fluid chromatography (SFC). The tricyclic building block with the NH- moiety on the indole ring was further treated with Boc ₂ O to selectively protect the aliphatic secondary amine moiety and obtained after purification.

為避免因採用2-或3-取代之苯基肼衍生物所致之區域異構物形成，使用具有毗鄰肼部分之額外鹵素原子(Br、Cl)之相應苯基肼衍生物。因此，利用市售4-側氧基六氫吡啶-1-甲酸第三丁基酯於適宜溶劑中在酸性條件下之費舍爾吲哚合成在純化後僅得到具有額外鹵素原子之單一產物。脂肪族二級胺經Boc保護且在純化後獲得產物。然後藉由使用適宜鹼於適當溶劑中利用鈀觸媒進行氫化來去除額外鹵素原子，以在純化後得到期望三環2,3,4,5-四氫-1H -吡啶并[4,3-b ]吲哚構建單元。Scenario 2

To avoid the formation of regioisomers due to the use of 2- or 3-substituted phenylhydrazine derivatives, the corresponding phenylhydrazine derivatives with additional halogen atoms (Br, Cl) adjacent to the hydrazine moiety are used. Therefore, the Fischer-Indole synthesis using commercially available tert-butyl 4-oxohexahydropyridine-1-carboxylate in a suitable solvent under acidic conditions only yields a single product with additional halogen atoms after purification. The aliphatic secondary amine is protected by Boc and the product is obtained after purification. Then, by using a suitable base in a suitable solvent and hydrogenation with a palladium catalyst to remove additional halogen atoms, to obtain the desired tricyclic 2,3,4,5-tetrahydro-1 H -pyrido[4,3 -b ] Indole building block.

然後於適當溶劑中使用適宜鹼利用碘甲烷或甲苯磺醯氯處理吲哚部分之NH-部分，以在純化後得到N-甲基或N-甲苯磺醯基衍生物。於適當溶劑中藉由酸處理去除Boc-保護基團，以在純化後得到期望三環2,3,4,5-四氫-1H -吡啶并[4,3-b ]吲哚構建單元。在無鹼處理之情形下，獲得相應鹽。Scheme 3

The NH-moiety of the indole moiety is then treated with iodomethane or tosyl chloride in a suitable solvent using a suitable base to obtain N-methyl or N-toluenesulfonyl derivatives after purification. Remove the Boc-protecting group by acid treatment in an appropriate solvent to obtain the desired tricyclic 2,3,4,5-tetrahydro-1 H -pyrido[4,3- b ]indole building block after purification . In the absence of alkali treatment, the corresponding salt is obtained.

於適當溶劑中利用甲基肼處理在3位、4位或5位具有F原子且在2位具有額外鹵素原子(Br、Cl)之市售氟吡啶衍生物，以在純化後得到相應N-甲基肼衍生物。於適宜溶劑中在酸性條件下利用市售4-側氧基六氫吡啶-1-甲酸第三丁基酯之費舍爾吲哚合成在純化後得到期望三環9-甲基-6,7,8,9-四氫-5H -吡咯并[2,3-b :4,5-c ']二吡啶衍生物。Scheme 4

Treatment of commercially available fluoropyridine derivatives with an F atom at the 3, 4, or 5 position and an additional halogen atom (Br, Cl) at the 2 position with methylhydrazine in an appropriate solvent to obtain the corresponding N- after purification Methylhydrazine derivatives. Fischer indole synthesis using commercially available 4-butyloxyhexahydropyridine-1-carboxylic acid tertiary butyl ester in an appropriate solvent under acidic conditions yields the desired tricyclic 9-methyl-6,7 after purification ,8,9-tetrahydro-5 H -pyrrolo[2,3- b :4,5- c ']dipyridine derivative.

於適宜溶劑中在酸性條件下加熱市售苯基肼衍生物(Z = F)與市售3-側氧基-8-氮雜二環[3.2.1]辛烷-8-甲酸第三丁基酯(費舍爾吲哚合成)，在純化後得到三環5,6,7,8,9,10-六氫-7,10-環亞胺基環庚[b ]吲哚衍生物。在2-或3-取代之苯基肼衍生物之情形下，區域異構物必須藉由超臨界流體層析(SFC)分離。然後使脂肪族二級胺部分經Boc保護且在純化後獲得產物。然後使用適宜鹼於適當溶劑中利用甲苯磺醯氯處理吲哚部分之NH-部分，以在純化後得到N-甲苯磺醯基衍生物。於適當溶劑中藉由酸處理去除Boc-保護基團，以在純化後得到期望三環5,6,7,8,9,10-六氫-7,10-環亞胺基環庚[b ]吲哚構建單元。在無鹼處理之情形下，獲得相應鹽。Scheme 5

Heating a commercially available phenylhydrazine derivative ( Z = F) in a suitable solvent under acidic conditions with a commercially available 3-pentoxy-8-azabicyclo[3.2.1]octane-8-carboxylic acid third butyl Ester ester (Fisher indole synthesis), after purification to obtain tricyclic 5,6,7,8,9,10-hexahydro-7,10-cyclic iminocyclohepta [ b ] indole derivatives. In the case of 2- or 3-substituted phenylhydrazine derivatives, regioisomers must be separated by supercritical fluid chromatography (SFC). The aliphatic secondary amine moiety is then protected by Boc and the product is obtained after purification. The NH-moiety of the indole moiety is then treated with tosyl chloride in a suitable solvent with a suitable base to obtain N-toluenesulfonyl derivatives after purification. Remove the Boc-protecting group by acid treatment in an appropriate solvent to obtain the desired tricyclic 5,6,7,8,9,10-hexahydro-7,10-cycloiminocycloheptane after purification [ b ] Indole building blocks. In the absence of alkali treatment, the corresponding salt is obtained.

利用N-羥基酞醯亞胺及氯化銅(I)及吡啶於適當溶劑中處理市售苯基

酸酯衍生物(Z = F)，以在純化後得到含有酞醯亞胺保護基團之相應羥胺衍生物。利用水合肼裂解酞醯亞胺保護基團且獲得呈鹽形式之相應羥胺衍生物。於適宜溶劑中在酸性條件下加熱羥胺衍生物與市售1,4-二氧雜-8-氮雜螺[4.5]癸烷(費舍爾吲哚合成)，在純化後得到三環1,2,3,4-四氫苯并呋喃并[3,2-c ]吡啶衍生物。Scheme 6

Treatment of commercially available phenyl groups with N-hydroxyphthalimide, copper(I) chloride and pyridine in appropriate solvents

Ester derivatives (Z = F) to obtain the corresponding hydroxylamine derivatives containing phthalimide protecting groups after purification. The protective group of phthalimide is cleaved with hydrazine hydrate and the corresponding hydroxylamine derivative in salt form is obtained. Heat a hydroxylamine derivative and commercially available 1,4-dioxa-8-azaspiro[4.5]decane (Fisher indole synthesis) in a suitable solvent under acidic conditions to obtain tricyclic 1, after purification 2,3,4-Tetrahydrobenzofuro[3,2- c ]pyridine derivatives.

利用一級或二級胺於適當溶劑中且利用額外鹼處理含有兩個鹵素(Br、Cl)原子之市售苯并[d ]噻唑(G = Ph)或苯并[d ]噁唑(G = Ph)衍生物。脫離基X經由親核取代由一級或二級胺替代，以在純化後得到相應胺基取代之苯并[d ]噻唑或苯并[d ]噁唑衍生物。在反應性胺較少之情形下，藉由在微波條件下實施親核取代反應獲得期望苯并[d ]噻唑或苯并[d ]噁唑衍生物。利用嗎啉於適當溶劑中且利用額外鹼處理含有兩個鹵素(Br、Cl)原子之相應噻唑并[5,4-b ]吡啶(G = Py)及噻唑并[4,5-b ]吡啶(G = Py)衍生物，以在純化後得到相應嗎啉基-噻唑并[5,4-b ]吡啶(G = Py)及嗎啉基-噻唑并[4,5-b ]吡啶(G = Py)衍生物。利用嗎啉於適當溶劑中且利用額外鹼處理含有兩個鹵素(Br、Cl)原子之相應噻唑并[5,4-d ]嘧啶(G =嘧啶)衍生物，以在純化後得到相應嗎啉基-噻唑并[5,4-d ]嘧啶(G =嘧啶)衍生物。Scheme 7

Treatment of commercially available benzo[ d ]thiazole (G = Ph) or benzo[ d ]oxazole (G = Ph) derivatives. The leaving group X is replaced by a primary or secondary amine via nucleophilic substitution to obtain the corresponding amine substituted benzo[ d ]thiazole or benzo[ d ]oxazole derivative after purification. In the case of less reactive amines, the desired benzo[ d ]thiazole or benzo[ d ]oxazole derivatives are obtained by performing a nucleophilic substitution reaction under microwave conditions. Treatment of the corresponding thiazolo[5,4- b ]pyridine (G = Py) and thiazolo[4,5- b ]pyridine containing two halogen (Br, Cl) atoms using morpholine in a suitable solvent and additional base (G = Py) derivatives to give the corresponding morpholino-thiazolo[5,4- b ]pyridine (G = Py) and morpholino-thiazolo[4,5- b ]pyridine (G = Py) derivatives. Use morpholine in a suitable solvent and treat the corresponding thiazolo[5,4- d ]pyrimidine (G = pyrimidine) derivative containing two halogen (Br, Cl) atoms with an additional base to obtain the corresponding morpholine after purification -Thiazolo[5,4- d ]pyrimidine (G = pyrimidine) derivative.

利用嗎啉在微波條件下處理含有兩個鹵素(Br、Cl)原子之市售[1,2,4]三唑并[1,5-a ]吡啶衍生物，以在純化後得到親核置換產物。Scheme 8

Treatment of commercially available [1,2,4]triazolo[1,5- a ]pyridine derivatives containing two halogen (Br, Cl) atoms under morpholine under microwave conditions to obtain nucleophilic substitution after purification product.

利用嗎啉、3-氧雜-8-氮雜二環[3.2.1]辛烷或8-氧雜-3-氮雜二環[3.2.1]辛烷在微波條件下處理含有兩個溴原子之市售3,6-二溴嗒嗪及2,5-二溴吡嗪衍生物，以在純化後得到親核置換產物。Scheme 8a

Treatment with morpholine, 3-oxa-8-azabicyclo[3.2.1]octane or 8-oxa-3-azabicyclo[3.2.1]octane under microwave conditions contains two bromines Atomic's commercially available 3,6-dibromopyridazine and 2,5-dibromopyrazine derivatives to obtain nucleophilic replacement products after purification.

利用嗎啉在微波條件下處理含有兩個鹵素(Br、Cl)原子之市售2,7-二氯喹啉、2-溴-6-氯-1,7-萘啶、2-溴-6-氯-1,8-萘啶及2,6-二氯喹啉衍生物，以在純化後得到親核置換產物。Option 8b

Treatment of commercially available 2,7-dichloroquinoline, 2-bromo-6-chloro-1,7-naphthyridine, 2-bromo-6- using morpholine under microwave conditions with two halogen (Br, Cl) atoms Chloro-1,8-naphthyridine and 2,6-dichloroquinoline derivatives to obtain nucleophilic replacement products after purification.

General synthetic scheme for preparing compounds of the present invention:

將具有B = NCH₃ 或B = O之三環構建單元與胺基取代之苯并[d ]噻唑或苯并[d ]噁唑衍生物或經取代之吡啶衍生物經由鈀化學利用適宜鈀觸媒(氯-(2-二環己基膦基-2′,6′-二異丙氧基-1,1′-聯苯)[2-(2-胺基乙基)苯基]鈀(II)-甲基-第三丁基醚加成物；Pd(RuPhos) G1)、配體(2-二環己基膦基-2′,6′-二異丙氧基聯苯；RuPhos)及鹼(雙(三甲基矽基)胺基鋰；LiHMDS)於適宜溶劑中(四氫呋喃；THF)偶合，以在純化後得到期望之式(I)化合物。Scheme 9

The benzo[ d ]thiazole or benzo[ d ]oxazole derivatives or substituted pyridine derivatives substituted by tricyclic building blocks with B=NCH ₃ or B=O and amine groups are used by palladium chemistry for suitable palladium contact Medium (chloro-(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2-aminoethyl)phenyl]palladium(II )-Methyl-third butyl ether adduct; Pd(RuPhos) G1), ligand (2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl; RuPhos) and base (Lithium bis(trimethylsilyl)amide; LiHMDS) is coupled in a suitable solvent (tetrahydrofuran; THF) to obtain the desired compound of formula (I) after purification.

將在吲哚/氮雜吲哚部分處含有N-甲苯磺醯基之三環構建單元與胺基取代之苯并[d ]噻唑、苯并[d ]噁唑、噻唑并[5,4-b ]吡啶(G = Py)、噻唑并[4,5-b ]吡啶(G = Py)衍生物、噻唑并[5,4-d ]嘧啶(G =嘧啶)或胺基取代之[1,2,4]三唑并[1,5-a ]吡啶衍生物或胺基取代之吡啶(L = Py)、吡嗪(L =吡嗪)、嗒嗪(L =嗒嗪)、嘧啶(L =嘧啶)衍生物或胺基取代之異喹啉、萘啶、喹唑啉衍生物經由鈀化學利用適宜鈀觸媒(參(二亞苄基丙酮)二鈀(0)；Pd₂ (dba)₃ )、配體(2-二環己基膦基-2′,6′-二異丙氧基聯苯；RuPhos)及鹼(第三丁醇鈉；NaOtBu)於適宜溶劑中(1,4-二噁烷)偶合，以在純化後得到期望之式(I)化合物。或者，將在吲哚/氮雜吲哚部分處含有N-甲苯磺醯基之三環構建單元與胺基取代之苯并[d ]噻唑、苯并[d ]噁唑、噻唑并[5,4-b ]吡啶(G = Py)、噻唑并[4,5-b ]吡啶(G = Py)衍生物、噻唑并[5,4-d ]嘧啶(G =嘧啶)或胺基取代之[1,2,4]三唑并[1,5-a ]吡啶衍生物或胺基取代之吡啶(L = Py)、吡嗪(L =吡嗪)、嗒嗪(L =嗒嗪)、嘧啶(L =嘧啶)衍生物或胺基取代之異喹啉、萘啶、喹唑啉衍生物經由鈀化學利用適宜鈀觸媒(參(二亞苄基丙酮)二鈀(0)；Pd₂ (dba)₃ )、配體(2-二環己基膦基-2′,6′-二異丙氧基聯苯；RuPhos)及弱鹼(碳酸銫；Cs₂ CO₃ )於適宜溶劑(1,4-二噁烷)中偶合，以在純化後得到經N-甲苯磺醯基保護之化合物。然後使用適宜鹼(碳酸銫；Cs₂ CO₃ )於適宜溶劑中(2-甲基THF、甲醇)在升高溫度(回流)下去除甲苯磺醯基保護基團，以在純化後得到期望之式(I)化合物。Scheme 10

The tricyclic building block containing N-tosylate at the indole/azaindole part and benzo[ d ]thiazole, benzo[ d ]oxazole, thiazolo[5,4- b ] pyridine (G = Py), thiazolo [4,5- b ] pyridine (G = Py) derivative, thiazolo [5,4- d ] pyrimidine (G = pyrimidine) or amine substituted [1, 2,4]triazolo[1,5- a ]pyridine derivative or amine substituted pyridine (L = Py), pyrazine (L = pyrazine), pyrazine (L = pyrazine), pyrimidine (L = Pyrimidine) derivatives or amine substituted isoquinoline, naphthyridine, quinazoline derivatives via palladium chemistry using a suitable palladium catalyst (see (dibenzylideneacetone) dipalladium (0); Pd ₂ (dba) ₃ ), ligand (2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl; RuPhos) and base (sodium third butoxide; NaOtBu) in a suitable solvent (1,4- Dioxane) coupling to obtain the desired compound of formula (I) after purification. Alternatively, the tricyclic building block containing N-tosylate at the indole/azaindole moiety and amine substituted benzo[ d ]thiazole, benzo[ d ]oxazole, thiazolo[5, 4- b ]pyridine (G = Py), thiazolo[4,5- b ]pyridine (G = Py) derivative, thiazolo[5,4- d ]pyrimidine (G =pyrimidine) or amine substituted[ 1,2,4]triazolo[1,5- a ]pyridine derivative or amine substituted pyridine (L = Py), pyrazine (L = pyrazine), pyrazine (L = pyrazine), pyrimidine (L = pyrimidine) derivative or amine substituted isoquinoline, naphthyridine, quinazoline derivatives via palladium chemistry using a suitable palladium catalyst (see (dibenzylideneacetone) dipalladium (0); Pd ₂ ( dba) ₃ ), ligand (2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl; RuPhos) and weak base (cesium carbonate; Cs ₂ CO ₃ ) in a suitable solvent (1, 4-dioxane) to provide N-toluenesulfonyl-protected compounds after purification. Then use a suitable base (cesium carbonate; Cs ₂ CO ₃ ) in a suitable solvent (2-methylTHF, methanol) at elevated temperature (reflux) to remove the tosyl protecting group to obtain the desired after purification Compound of formula (I).

利用適宜鹼(碳酸鉀；K₂ CO₃ )於適宜溶劑(DMF)中使在吲哚部分處含有N-甲苯磺醯基之三環構建單元與二-鹵化苯并[d ]噻唑及二-鹵化苯并[d ]噁唑偶合，以得到親核置換產物。然後使產物經由鈀化學與適宜鈀觸媒(參(二亞苄基丙酮)二鈀(0)；Pd₂ (dba)₃ )、配體(2-二環己基膦基-2′,6′-二異丙氧基聯苯；RuPhos)及鹼(第三丁醇鈉；NaOtBu)於適宜溶劑中(1,4-二噁烷)中偶合，以在純化後得到期望之式(I)化合物。Scheme 11

Use a suitable base (potassium carbonate; K ₂ CO ₃ ) in a suitable solvent (DMF) to make the tricyclic building block containing N-tosylsulfonyl at the indole part and the di-halogenated benzo[ d ]thiazole and di- The halogenated benzo[ d ]oxazole is coupled to obtain a nucleophilic replacement product. The product is then subjected to palladium chemistry with a suitable palladium catalyst (see (dibenzylideneacetone) dipalladium (0); Pd ₂ (dba) ₃ ), ligand (2-dicyclohexylphosphino-2′,6′ -Diisopropoxybiphenyl; RuPhos) and base (sodium third butoxide; NaOtBu) are coupled in a suitable solvent (1,4-dioxane) to obtain the desired compound of formula (I) after purification .

General synthetic scheme for preparing tritium-labeled compounds of the present invention:

將觸媒添加至氚反應容器，之後添加本發明化合物於二氯甲烷中之溶液。將容器附接至氚管線並在-200℃下對氚氣加壓。將溶液在室溫下攪拌8小時，冷卻至-200℃且去除過量氣體。用甲醇沖洗反應燒瓶且使合併之有機相在真空下蒸發。藉由HPLC純化粗製材料，使移動相在真空下蒸發且將產物重新溶解於無水乙醇中。藉由質譜測定比活性。 可使用業內已知之任何適宜分析來量測Tau K18及全長(fl) Tau之解聚。闡述用於量測解聚能力之標準活體外分析。Scheme 12

The catalyst was added to the tritium reaction vessel, and then a solution of the compound of the invention in dichloromethane was added. Attach the vessel to the tritium line and pressurize the tritium gas at -200°C. The solution was stirred at room temperature for 8 hours, cooled to -200°C and excess gas was removed. The reaction flask was rinsed with methanol and the combined organic phase was evaporated under vacuum. The crude material was purified by HPLC, the mobile phase was evaporated under vacuum and the product was redissolved in absolute ethanol. The specific activity was determined by mass spectrometry. The disaggregation of Tau K18 and full-length (fl) Tau can be measured using any suitable analysis known in the industry. Explain the standard in vitro analysis used to measure the depolymerization ability.

Examples All reagents and solvents were obtained from commercial sources and used without further purification. ¹ H NMR spectra were recorded in deuterated solvents on Bruker AV 300 and 400 MHz spectrometers. The chemical shift (δ) is reported in parts per million and the coupling constant (J value) is reported in hertz. Spin multiplicity is indicated by the following symbols: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), bs (broad singlet). Mass spectra were obtained on an Agilent 1290 Infinity II spectrometer with 6130 Chemstation and an Agilent 1200 Infinity II spectrometer with 6130 Chemstation. An Agilent 7890B gas chromatograph and a 5977B mass spectrometer were used to collect GC-MS data. Infrared spectra were obtained on a PerkinElmer spectrometer. Chromatography was performed using silicone (Fluka: silicone 60, 0.063-0.2 mm) and suitable solvents as indicated in the specific examples. Rapid purification was performed using the solvent gradient indicated in Biotage Isolera (Biotage) with HP-Sil or KP-NH SNAP columns and specific examples. Thin layer chromatography (TLC) was performed on the silicone plate using UV detection.

步驟 A 在冰浴溫度下向4-氟苯基肼(1 g, 7.9 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(1.2 g, 8.3 mmol)於1,4-二噁烷(10 mL)中之溶液添加濃H₂ SO₄ (1 mL)。然後將反應混合物在110℃下加熱3 h。將該反應混合物冷卻至室溫，將沈澱物過濾出。將固體溶解於水中，經NaOH溶液鹼化並用DCM (二氯甲烷)萃取。將有機相分離且經Na₂ SO₄ 乾燥並去除溶劑，得到呈淡黃色固體之標題化合物(950 mg, 59%)。 MS: 191 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 10.91 (s, 1H), 7.23-7.24 (m, 1H), 7.09-7.09 (m, 1H), 6.80-6.81 (m, 1H), 3.91 (s, 2H), 3.11 (t,J = 5.56 Hz, 2H), 2.75 (d,J = 4.96 Hz, 2H)。步驟 B 向來自上文步驟A之標題化合物(0.95 g, 4.77 mmol)於THF (四氫呋喃)中之溶液添加二碳酸二-第三丁基酯(Boc₂ O) (1.5 g)，且將混合物攪拌過夜。如藉由TLC所證實，在反應完成後，將溶劑去除且使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到呈淡黃色黏性液體之標題化合物(1.1 g, 78%)。 MS: 291 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.00 (s, 1H), 7.26 (q,J = 4.52 Hz, 1H), 7.18 (t,J = 8.12 Hz, 1H), 6.83-6.83 (m, 1H), 4.49 (s, 2H), 3.69 (t,J = 5.64 Hz, 2H), 2.76 (s, 2H), 1.43 (s, 9H)。步驟 C 向來自上文步驟B之標題化合物(0.41 g, 1.41 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.15 g, 6.25 mmol)，之後添加對甲苯磺醯氯(TsCl) (0.29 g, 1.45 mmol)。將反應混合物攪拌10 min。將該混合物溶解於EtOAc (20 ml)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(20/80 =＞ 80/20)，在矽膠管柱上純化粗產物，得到標題化合物(0.45 g, 72%)。 MS: 445 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.03-8.04 (m, 1H), 7.77 (d,J = 8.20 Hz, 2H), 7.36-7.38 (m, 3H), 7.15-7.16 (m, 1H), 4.43 (s, 2H), 3.69 (t,J = 5.64 Hz, 2H), 3.08 (s, 2H), 2.32 (s, 3H), 1.43 (s, 9H)。步驟 D 向來自上文步驟C之標題化合物(0.42 g, 0.915 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2N HCl (5 mL)。將反應混合物攪拌過夜。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之標題化合物(0.2 g, 58%)。 MS: 345 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 9.61 (s, 1H), 8.01-8.02 (m, 1H), 7.82 (d,J = 8.40 Hz, 2H), 7.45-7.45 (m, 1H), 7.39 (d,J = 8.40 Hz, 2H), 7.20-7.21 (m, 1H), 4.25 (s, 2H), 3.49 (s, 2H), 3.35 (d, J = 3.60 Hz, 2H), 2.34 (s, 3H)。步驟 E 向來自上文步驟D之標題化合物(5.0 g, 13 mmol)於二氯甲烷(50 mL)中之溶液添加三乙胺(5 mL)並攪拌10 min。將反應混合物用二氯甲烷(20 mL)稀釋，用水(2 × 30 mL)及NaCl飽和溶液(30 mL)洗滌。使合併之有機層經硫酸鈉乾燥並在真空下濃縮，得到呈游離鹼之標題化合物(定量產率)。 Preparation Example 1

Step A: To 4-fluorophenylhydrazine (1 g, 7.9 mmol) and tert-butyl 4-oxohexahydropyridine-1-carboxylate (1.2 g, 8.3 mmol) at 1,4 at an ice bath temperature -A solution in dioxane (10 mL) was added concentrated H ₂ SO ₄ (1 mL). The reaction mixture was then heated at 110 °C for 3 h. The reaction mixture was cooled to room temperature, and the precipitate was filtered off. The solid was dissolved in water, basified with NaOH solution and extracted with DCM (dichloromethane). The organic phase was separated and dried over Na ₂ SO ₄ and the solvent was removed to give the title compound (950 mg, 59%) as a light yellow solid. MS: 191 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.91 (s, 1H), 7.23-7.24 (m, 1H), 7.09-7.09 (m, 1H), 6.80-6.81 (m, 1H), 3.91 (s, 2H), 3.11 (t, J = 5.56 Hz, 2H), 2.75 (d, J = 4.96 Hz, 2H). Step B To a solution of the title compound (0.95 g, 4.77 mmol) in THF (tetrahydrofuran) from Step A above, di-tert-butyl dicarbonate (Boc ₂ O) (1.5 g) was added, and the mixture was stirred overnight. As confirmed by TLC, after the reaction was completed, the solvent was removed and the crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/heptane gradient (10/80 => 80/20), The title compound (1.1 g, 78%) was obtained as a pale yellow viscous liquid. MS: 291 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.00 (s, 1H), 7.26 (q, J = 4.52 Hz, 1H), 7.18 (t, J = 8.12 Hz, 1H), 6.83-6.83 ( m, 1H), 4.49 (s, 2H), 3.69 (t, J = 5.64 Hz, 2H), 2.76 (s, 2H), 1.43 (s, 9H). Step C To the solution of the title compound (0.41 g, 1.41 mmol) in THF (5 mL) from Step B above was added sodium hydride (0.15 g, 6.25 mmol), followed by p-toluenesulfonyl chloride (TsCl) (0.29 g, 1.45 mmol). The reaction mixture was stirred for 10 min. The mixture was dissolved in EtOAc (20 ml) and washed with water and brine and dried over Na ₂ SO ₄ . Using the Biotage Isolera One purification system, using an EtOAc/heptane gradient (20/80 => 80/20), the crude product was purified on a silica gel column to give the title compound (0.45 g, 72%). MS: 445 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.03-8.04 (m, 1H), 7.77 (d, J = 8.20 Hz, 2H), 7.36-7.38 (m, 3H), 7.15-7.16 (m , 1H), 4.43 (s, 2H), 3.69 (t, J = 5.64 Hz, 2H), 3.08 (s, 2H), 2.32 (s, 3H), 1.43 (s, 9H). Step D To a solution of the title compound (0.42 g, 0.915 mmol) from dichloromethane from step C above was added 2N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred overnight. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (0.2 g, 58%) as an off-white solid. MS: 345 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.61 (s, 1H), 8.01-8.02 (m, 1H), 7.82 (d, J = 8.40 Hz, 2H), 7.45-7.45 (m, 1H ), 7.39 (d, J = 8.40 Hz, 2H), 7.20-7.21 (m, 1H), 4.25 (s, 2H), 3.49 (s, 2H), 3.35 (d, J = 3.60 Hz, 2H), 2.34 (s, 3H). Step E To the solution of the title compound (5.0 g, 13 mmol) in dichloromethane (50 mL) from step D above was added triethylamine (5 mL) and stirred for 10 min. The reaction mixture was diluted with dichloromethane (20 mL), washed with water (2×30 mL) and saturated NaCl solution (30 mL). The combined organic layer was dried over sodium sulfate and concentrated under vacuum to give the title compound as a free base (quantitative yield).

步驟 A 向來自製備實例1步驟B之標題化合物(0.41 g, 1.41 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.067 g, 2.82 mmol)，將懸浮液在室溫下攪拌20分鐘。將反應混合物再次冷卻至0℃且添加碘甲烷(0.24 g, 1.45 mmol)。將反應混合物攪拌3 h。使反應混合物溶解於EtOAc (15 mL)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(20/80 =＞ 80/20)，在矽膠管柱上純化粗產物，得到標題化合物(0.3 g, 70%)。 MS: 304 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.38-7.39 (m, 1H), 7.22 (dd,J = 2.40, 10.00 Hz, 1H), 6.90-6.91 (m, 1H), 4.49 (s, 2H), 3.71 (t,J = 6.00 Hz, 2H), 3.62 (s, 3H), 2.79 (t,J = 5.20 Hz, 2H), 1.40 (s, 9H)。 步驟 B 向來自上文步驟A之標題化合物(0.3 mg, 0.986 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2 N HCl (5 mL)。將反應混合物攪拌過夜。使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之標題化合物(0.12 g, 60%)。 MS: 205.08 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 9.45 (br, 1H), 7.45-7.46 (m, 1H), 7.30-7.31 (m, 1H), 6.98-7.00 (m, 1H), 4.26 (s, 2H), 3.66 (s, 3H), 3.49 (d,J = 4.80 Hz, 2H), 3.06 (t,J = 6.00 Hz, 2H), 2.50 (s, 9H)。 Preparation Example 2

Step A To a solution of the title compound (0.41 g, 1.41 mmol) in THF (5 mL) from Step B of Preparation Example 1 was added sodium hydride (0.067 g, 2.82 mmol), and the suspension was stirred at room temperature for 20 minutes. The reaction mixture was cooled to 0 °C again and methyl iodide (0.24 g, 1.45 mmol) was added. The reaction mixture was stirred for 3 h. The reaction mixture was dissolved in EtOAc (15 mL) and washed with water and brine and dried over Na ₂ SO ₄ . Using the Biotage Isolera One purification system, using an EtOAc/hexane gradient (20/80 => 80/20), the crude product was purified on a silica gel column to give the title compound (0.3 g, 70%). MS: 304 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.38-7.39 (m, 1H), 7.22 (dd, J = 2.40, 10.00 Hz, 1H), 6.90-6.91 (m, 1H), 4.49 (s , 2H), 3.71 (t, J = 6.00 Hz, 2H), 3.62 (s, 3H), 2.79 (t, J = 5.20 Hz, 2H), 1.40 (s, 9H) . Step B To a solution of the title compound (0.3 mg, 0.986 mmol) from dichloromethane from step A above was added 2 N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred overnight. The reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (0.12 g, 60%) as an off-white solid. MS: 205.08 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.45 (br, 1H), 7.45-7.46 (m, 1H), 7.30-7.31 (m, 1H), 6.98-7.00 (m, 1H), 4.26 (s, 2H), 3.66 (s, 3H), 3.49 (d, J = 4.80 Hz, 2H), 3.06 (t, J = 6.00 Hz, 2H), 2.50 (s, 9H).

步驟 A 在冰浴溫度下向1-(4-氟苯基)-1-甲基肼(2 g, 14.0 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(2.84 g, 14.0 mmol)於1,4-二噁烷(10 mL)中之溶液添加濃H₂ SO₄ (1 mL)。然後將反應混合物在110℃下加熱過夜。將該反應混合物冷卻至室溫，將沈澱物過濾出。棄掉濾液。將固體溶解於水(10 mL)中，利用NaOH溶液將pH調整至14，並用二氯甲烷(150 mL)萃取混合物。將有機相用水及鹽水洗滌且經Na₂ SO₄ 乾燥。將溶劑去除，得到呈游離鹼之標題化合物(1.3 g, 46%)。 MS: 205.08 (M+H)⁺ 。 Preparation Example 3

Step A To 1-(4-fluorophenyl)-1-methylhydrazine (2 g, 14.0 mmol) and 4-pentoxyhexahydropyridine-1-carboxylic acid tert-butyl ester (2.84) at ice bath temperature g, 14.0 mmol) in 1,4-dioxane (10 mL) was added concentrated H ₂ SO ₄ (1 mL). The reaction mixture was then heated at 110°C overnight. The reaction mixture was cooled to room temperature, and the precipitate was filtered off. Discard the filtrate. The solid was dissolved in water (10 mL), the pH was adjusted to 14 with NaOH solution, and the mixture was extracted with dichloromethane (150 mL). The organic phase was washed with water and brine and dried over Na ₂ SO ₄ . The solvent was removed to give the title compound as a free base (1.3 g, 46%). MS: 205.08 (M+H) ⁺ .

步驟 A 在冰浴溫度下向(4-甲氧基苯基)肼(1 g, 5.6 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(1.13 g, 5.6 mmol)於1,4-二噁烷(10 mL)中之溶液添加濃H₂ SO₄ (1 mL)。然後將反應混合物在110℃下加熱3 h。使該反應混合物冷卻至室溫，將沈澱物過濾出。將固體溶解於水中，經NaOH溶液鹼化並用二氯甲烷萃取。將有機相分離且經Na₂ SO₄ 乾燥，過濾並將溶劑去除，得到呈淡黃色黏性液體之標題化合物(0.60 g, 53%)。粗產物原樣用於下一步驟。 MS: 203 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(0.60 mg, 2.9 mmol)於THF中之溶液添加二碳酸二-第三丁基酯(0.65 g, 2.9 mmol)，且將混合物攪拌過夜。將溶劑去除且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(80/20)，在矽膠管柱上純化粗產物，得到呈淡黃色固體之標題化合物(0.55 g, 62%)。 MS: 303 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 10.71 (bs, 1H), 7.17 (d,J = 8.40 Hz, 1H), 6.89 (s, 1H), 6.65-6.66 (m, 1H), 4.49 (s, 2H), 3.75 (s, 3H), 3.69 (t,J = 5.60 Hz, 2H), 2.74 (t,J = 5.60 Hz, 2H), 1.44 (s, 9H)。步驟 C 向來自上文步驟B之標題化合物(0.55 g, 1.8 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.087 g, 3.6 mmol)，之後添加對甲苯磺醯氯(0.342 g, 1.8 mmol)。將反應混合物攪拌45分鐘。在反應完成後，將該反應混合物溶解於EtOAc (200 mL)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(20/80)，在矽膠管柱上純化粗產物，得到呈灰白色固體之標題化合物(0.45 g ,45%)。 MS: 457 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.92 (d,J = 9.20 Hz, 1H), 7.71 (d,J = 8.40 Hz, 2H), 7.35 (d,J = 8.00 Hz, 2H), 7.01 (s, 1H), 6.91-6.92 (m, 1H), 4.42 (s, 2H), 3.77 (s, 3H), 3.68 (t,J = 5.60 Hz, 2H), 3.05 (bs, 2H), 2.31 (s, 3H), 1.43 (s, 9H)。步驟 D 向來自上文步驟C之標題化合物(0.450 g, 0.986 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2 N HCl (5 mL)。將反應混合物攪拌過夜。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之化合物(0.23 g, 65%)。 MS: 357 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 9.67 (bs, 1H), 7.89 (d,J = 9.20 Hz, 1H), 7.77 (d,J = 8.40 Hz, 2H), 7.36 (d,J = 8.00 Hz, 2H), 7.11 (d,J = 2.40 Hz, 1H), 6.93-6.94 (m, 1H), 4.24 (s, 2H), 3.77 (s, 3H), 3.48-3.49 (m, 2H), 3.34-3.35 (m, 2H), 2.33 (s, 3H)。 Preparation Example 4

Step A To (4-methoxyphenyl)hydrazine (1 g, 5.6 mmol) and 4-pentoxyhexahydropyridine-1-carboxylic acid tert-butyl ester (1.13 g, 5.6 mmol) at ice bath temperature To a solution in 1,4-dioxane (10 mL) was added concentrated H ₂ SO ₄ (1 mL). The reaction mixture was then heated at 110 °C for 3 h. The reaction mixture was allowed to cool to room temperature, and the precipitate was filtered off. The solid was dissolved in water, basified with NaOH solution and extracted with dichloromethane. The organic phase was separated and dried over Na ₂ SO ₄ , filtered and the solvent was removed to give the title compound (0.60 g, 53%) as a pale yellow viscous liquid. The crude product was used as is in the next step. MS: 203 (M+H) ⁺ . Step B To a solution of the title compound (0.60 mg, 2.9 mmol) in THF from Step A above was added di-tert-butyl dicarbonate (0.65 g, 2.9 mmol), and the mixture was stirred overnight. The solvent was removed and the crude product was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/hexane gradient (80/20) to give the title compound (0.55 g, 62%) as a light yellow solid. MS: 303 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.71 (bs, 1H), 7.17 (d, J = 8.40 Hz, 1H), 6.89 (s, 1H), 6.65-6.66 (m, 1H), 4.49 (s, 2H), 3.75 (s, 3H), 3.69 (t, J = 5.60 Hz, 2H), 2.74 (t, J = 5.60 Hz, 2H), 1.44 (s, 9H). Step C To the solution of the title compound (0.55 g, 1.8 mmol) in THF (5 mL) from Step B above was added sodium hydride (0.087 g, 3.6 mmol), followed by p-toluenesulfonyl chloride (0.342 g, 1.8 mmol). The reaction mixture was stirred for 45 minutes. After the reaction was completed, the reaction mixture was dissolved in EtOAc (200 mL), and washed with water and brine and dried over Na ₂ SO ₄ . The crude product was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/hexane gradient (20/80) to give the title compound (0.45 g, 45%) as an off-white solid. MS: 457 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.92 (d, J = 9.20 Hz, 1H), 7.71 (d, J = 8.40 Hz, 2H), 7.35 (d, J = 8.00 Hz, 2H) , 7.01 (s, 1H), 6.91-6.92 (m, 1H), 4.42 (s, 2H), 3.77 (s, 3H), 3.68 (t, J = 5.60 Hz, 2H), 3.05 (bs, 2H), 2.31 (s, 3H), 1.43 (s, 9H). Step D To the solution of the title compound (0.450 g, 0.986 mmol) from dichloromethane from Step C above was added 2 N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred overnight. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the compound as an off-white solid (0.23 g, 65%). MS: 357 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.67 (bs, 1H), 7.89 (d, J = 9.20 Hz, 1H), 7.77 (d, J = 8.40 Hz, 2H), 7.36 (d, J = 8.00 Hz, 2H), 7.11 (d, J = 2.40 Hz, 1H), 6.93-6.94 (m, 1H), 4.24 (s, 2H), 3.77 (s, 3H), 3.48-3.49 (m, 2H ), 3.34-3.35 (m, 2H), 2.33 (s, 3H).

步驟 A 在0℃下向來自製備實例4步驟B之標題化合物(0.55 g, 1.8 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.087 g, 3.6 mmol)。將懸浮液在室溫下攪拌20分鐘。將反應混合物再次冷卻至0℃且添加碘甲烷(0.255 g, 1.8 mmol)。將反應混合物攪拌45分鐘。在反應完成後，將該反應混合物溶解於EtOAc (20 ml)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥，過濾並濃縮。使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(80/20)，在矽膠管柱上純化粗製混合物，得到呈淡棕色固體之標題化合物(0.40 g, 45%)。 MS: 317 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.29 (d,J = 8.80 Hz, 2H), 6.93 (s, 1H), 6.72-6.73 (m, 1H), 4.50 (s, 2H), 3.76 (s, 3H), 3.70-3.72 (m, 2H), 3.59 (s, 3H), 2.78-2.79 (m, 2H), 1.44 (s, 9H)。步驟 B 向來自上文步驟A之標題化合物(0.4 g, 1.26 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2 N HCl (5 mL)。將反應混合物攪拌過夜。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈白色固體之標題化合物(0.2 g, 73%)。 MS: 217 (M+H)⁺ 。 Preparation Example 5

Step A To a solution of the title compound (0.55 g, 1.8 mmol) from Preparation Example 4, Step B in THF (5 mL) was added sodium hydride (0.087 g, 3.6 mmol) at 0°C. The suspension was stirred at room temperature for 20 minutes. The reaction mixture was cooled to 0 °C again and iodomethane (0.255 g, 1.8 mmol) was added. The reaction mixture was stirred for 45 minutes. After the reaction was completed, the reaction mixture was dissolved in EtOAc (20 ml), and washed with water and brine and dried over Na ₂ SO ₄ , filtered and concentrated. Using a Biotage Isolera One purification system, using an EtOAc/hexane gradient (80/20), the crude mixture was purified on a silica gel column to give the title compound (0.40 g, 45%) as a light brown solid. MS: 317 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.29 (d, J = 8.80 Hz, 2H), 6.93 (s, 1H), 6.72-6.73 (m, 1H), 4.50 (s, 2H), 3.76 (s, 3H), 3.70-3.72 (m, 2H), 3.59 (s, 3H), 2.78-2.79 (m, 2H), 1.44 (s, 9H). Step B To a solution of the title compound (0.4 g, 1.26 mmol) from dichloromethane from step A above was added 2 N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred overnight. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (0.2 g, 73%) as a white solid. MS: 217 (M+H) ⁺ .

步驟 A 於微波爐中將2-氯-5-氟吡啶(10 g, 76.34 mmol)及N- 甲基肼(6 mL)之混合物在180℃下輻照1 h。在反應完成後，使反應混合物冷卻並將反應混合物傾倒於冰冷水中且用二氯甲烷(2 × 50 mL)萃取。將有機相分離且經Na₂ SO₄ 乾燥並去除溶劑，得到呈淡棕色固體之標題化合物(10 g，粗製)。粗產物原樣用於下一步驟。 MS: 143 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(10 g, 70.84 mmol)於甲醇(100 mL)中之溶液添加4-側氧基六氫吡啶-1-甲酸第三丁基酯(16.9 g, 85 mmol)並攪拌過夜。將溶劑去除，使用Biotage Isolera One純化系統，採用甲醇/DCM梯度(1/99)，在矽膠管柱上純化粗製反應混合物，得到呈棕色黏性油狀物之標題化合物(5 g, 22%)。 MS: 323 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.82-7.84 (m, 1H), 7.13-7.14 (m, 1H), 6.72-6.73 (m, 1H), 4.51 (bs, 2H), 3.77 (s, 3H), 3.71-3.73 (m, 2H), 2.76-2.78 (m, 2H), 1.43 (s, 9H)。步驟 C 於微波爐中將來自上文步驟B之標題化合物(5 g, 15.50 mmol)及二乙二醇(5 mL)之混合物在180℃下輻照1 h。在反應完成後，使反應混合物冷卻並將反應混合物傾倒於冰冷水中並用二氯甲烷(2 × 50 mL)萃取。將有機相分離且經Na₂ SO₄ 乾燥並去除溶劑，藉由製備型HPLC (管柱：Phenomenex Gemini C18 (150*4.6)mm, 3.0 μm。移動相A：於Milli-Q水中之10 mM乙酸銨。移動相B：乙腈。流速：1.0 ml\min)純化粗製混合物，得到呈棕色固體之標題化合物(0.9 g, 29%)。 MS: 206 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.87-7.89 (m, 1H), 7.46-7.47 (m, 1H), 4.53 (bs, 2H), 3.77 (s, 3H), 3.71-3.73 (m, 2H), 2.76-2.78 (m, 2H)。 Preparation Example 6

Step A In a microwave oven, a mixture of 2-chloro-5-fluoropyridine (10 g, 76.34 mmol) and N -methylhydrazine (6 mL) was irradiated at 180°C for 1 h. After the reaction was completed, the reaction mixture was cooled and poured into ice cold water and extracted with dichloromethane (2×50 mL). The organic phase was separated and dried over Na ₂ SO ₄ and the solvent was removed to give the title compound (10 g, crude) as a light brown solid. The crude product was used as is in the next step. MS: 143 (M+H) ⁺ . Step B To a solution of the title compound from Step A above (10 g, 70.84 mmol) in methanol (100 mL) was added tert-butyl 4-oxohexahydropyridine-1-carboxylate (16.9 g, 85 mmol) and stirred overnight. The solvent was removed and the crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using a methanol/DCM gradient (1/99) to obtain the title compound (5 g, 22%) as a brown viscous oil . MS: 323 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.82-7.84 (m, 1H), 7.13-7.14 (m, 1H), 6.72-6.73 (m, 1H), 4.51 (bs, 2H), 3.77 (s, 3H), 3.71-3.73 (m, 2H), 2.76-2.78 (m, 2H), 1.43 (s, 9H). Step C The mixture of the title compound (5 g, 15.50 mmol) and diethylene glycol (5 mL) from Step B above was irradiated at 180°C for 1 h in a microwave oven. After the reaction was completed, the reaction mixture was cooled and poured into ice-cold water and extracted with dichloromethane (2×50 mL). The organic phase was separated and dried over Na ₂ SO ₄ and the solvent was removed by preparative HPLC (column: Phenomenex Gemini C18 (150*4.6) mm, 3.0 μm. Mobile phase A: 10 mM acetic acid in Milli-Q water Ammonium. Mobile phase B: acetonitrile. Flow rate: 1.0 ml\min) The crude mixture was purified to give the title compound (0.9 g, 29%) as a brown solid. MS: 206 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.87-7.89 (m, 1H), 7.46-7.47 (m, 1H), 4.53 (bs, 2H), 3.77 (s, 3H), 3.71-3.73 (m, 2H), 2.76-2.78 (m, 2H).

步驟 A 在0℃下向3-(氟苯基)肼(1 g, 6.1 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(1.2 g, 6.1 mmol)於1,4-二噁烷(10 mL)中之溶液添加濃H₂ SO₄ (1 mL)。然後將反應混合物升溫至25℃並在110℃下加熱3 h。將該反應混合物冷卻至室溫且將沈澱物過濾出。將固體溶解於水中，經NaOH溶液鹼化並用二氯甲烷萃取。將有機相分離且經Na₂ SO₄ 乾燥並去除溶劑，得到呈淡黃色固體之區域異構物混合物(0.65 g, 56%)。 MS: 191.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 10.87 (bs, 1H), 7.26-7.30 (m, 1H), 7.02-7.05 (m, 1H), 6.74-6.79 (m, 1H), 3.83 (bs, 2H), 2.99-3.02 (m, 2H), 2.65-2.66 (m, 2H)。步驟 B 向區域異構物混合物(0.65 g, 3.15 mmol)於THF中之溶液添加二碳酸二-第三丁基酯(0.757 g, 3.47 mmol)，且將混合物攪拌12 h。在反應完成後(藉由TLC監測)，將溶劑在減壓下濃縮以產生粗產物。藉由矽膠(60-120目)管柱層析使用己烷: EtOAc (70:30)純化該粗產物，得到呈黃色固體之區域異構物7-氟-1,3,4,5-四氫-2H -吡啶并[4,3-b ]吲哚-2-甲酸第三丁基酯及9-氟-1,3,4,5-四氫-2H -吡啶并[4,3-b ]吲哚-2-甲酸第三丁基酯之混合物(0.750 g, 61%)，其比率為分別約70:30。 MS: 291.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.01 (bs, 1H), 7.36-7.39 (m, 1H), 7.06-7.09 (m, 1H), 6.79-6.84 (m, 1H), 4.51 (bs, 2H), 3.68-3.71 (m, 2H), 2.74-2.76 (m, 2H), 1.38 (s, 9H)。步驟 C 藉由SFC手性管柱(Chiracel OJ-H；管柱：X-bridge C8 (50 × 4.6) mm, 3.5 μm，移動相A：於水中之0.1% TFA，移動相B：0.1% TFA乙腈)分離區域異構物混合物(0.750 mg, 70:30)，得到具有100%手性純度之呈淡黃色固體之第二溶析標題化合物7-氟-1,3,4,5-四氫-2H -吡啶并[4,3-b ]吲哚-2-甲酸第三丁基酯(0.4 mg, 53%)。第一溶析標題化合物9-氟-1,3,4,5-四氫-2H -吡啶并[4,3-b ]吲哚-2-甲酸第三丁基酯分離為具有100%手性純度之淡黃色固體(0.25 g, 33%)。 第二溶析標題化合物： MS: 291.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.01 (bs, 1H), 7.36-7.39 (m, 1H), 7.06-7.09 (m, 1H), 6.79-6.84 (m, 1H), 4.51 (bs, 2H), 3.68-3.71 (m, 2H), 2.74-2.77 (m, 2H), 1.44 (s, 9H)。 RT=2.08 min。 第一溶析標題化合物： MS: 291.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.22 (s, 1H), 7.13 (d,J = 8.08 Hz, 1H), 6.96-6.97 (m, 1H), 6.69-6.71 (m, 1H), 4.63 (s, 2H), 3.69-3.70 (m, 2H), 2.68-2.76 (m, 2H), 1.44 (s, 9H)。 RT=1.74 min。步驟 D 向來自上文步驟C之第二溶析標題化合物(0.4 g, 1.37 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.099 mg, 4.137 mmol)，之後添加對甲苯磺醯氯(0.288 g, 1.51 mmol)。將反應混合物攪拌30分鐘。將該混合物溶解於EtOAc (20 ml)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(20/80 =＞ 80/20)，在矽膠管柱上純化粗產物，得到標題化合物(0.3 g, 49%)。 MS: 445 (M+H)⁺ 。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.92-7.94 (m, 1H), 7.68-7.70 (m, 1H), 7.25-7.29 (m, 4H), 7.00-7.04 (m, 1H), 4.50 (bs, 2H), 3.76 (bs, 2H), 3.12 (bs, 2H), 2.38 (s, 3H), 1.51 (s, 9H)。步驟 E 向來自上文步驟D之標題化合物(0.3 g, 0.676 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2 N HCl (5 mL)。將反應混合物攪拌12 h。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之標題化合物(0.2 g 78%)。 MS: 345 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 9.50 (bs, 2H), 7.80-7.87 (m, 2H), 7.78 (d,J = 2.00 Hz, 1H), 7.57-7.61 (m, 1H), 7.40 (d,J = 8.16 Hz, 2H), 7.18-7.23 (m, 1H), 4.27 (bs, 2H), 3.56 (bs, 2H), 3.47 (bs, 2H), 2.34 (s, 3H)。 Preparation Example 7

Step A: To 3-(fluorophenyl)hydrazine (1 g, 6.1 mmol) and tert-butyl 4-oxohexahydropyridine-1-carboxylate (1.2 g, 6.1 mmol) at 1, To the solution in 4-dioxane (10 mL) was added concentrated H ₂ SO ₄ (1 mL). The reaction mixture was then warmed to 25 °C and heated at 110 °C for 3 h. The reaction mixture was cooled to room temperature and the precipitate was filtered off. The solid was dissolved in water, basified with NaOH solution and extracted with dichloromethane. The organic phase was separated and dried over Na ₂ SO ₄ and the solvent was removed to give a mixture of regioisomers as a light yellow solid (0.65 g, 56%). MS: 191.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.87 (bs, 1H), 7.26-7.30 (m, 1H), 7.02-7.05 (m, 1H), 6.74-6.79 (m, 1H), 3.83 (bs, 2H), 2.99-3.02 (m, 2H), 2.65-2.66 (m, 2H). Step B To a solution of the regioisomer mixture (0.65 g, 3.15 mmol) in THF was added di-tert-butyl dicarbonate (0.757 g, 3.47 mmol), and the mixture was stirred for 12 h. After the reaction was completed (monitored by TLC), the solvent was concentrated under reduced pressure to produce a crude product. The crude product was purified by silica gel (60-120 mesh) column chromatography using hexane:EtOAc (70:30) to obtain the regioisomer 7-fluoro-1,3,4,5-tetrazol as a yellow solid Hydrogen- 2H -pyrido[4,3- b ] indole-2-carboxylic acid tert-butyl ester and 9-fluoro-1,3,4,5-tetrahydro- 2H -pyrido[4,3 -b ] A mixture of tert-butyl indole-2-carboxylic acid (0.750 g, 61%), the ratio of which is about 70:30 respectively. MS: 291.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.01 (bs, 1H), 7.36-7.39 (m, 1H), 7.06-7.09 (m, 1H), 6.79-6.84 (m, 1H), 4.51 (bs, 2H), 3.68-3.71 (m, 2H), 2.74-2.76 (m, 2H), 1.38 (s, 9H). Step C: By SFC chiral column (Chiracel OJ-H; column: X-bridge C8 (50 × 4.6) mm, 3.5 μm, mobile phase A: 0.1% TFA in water, mobile phase B: 0.1% TFA Acetonitrile) to separate the regioisomer mixture (0.750 mg, 70:30) to obtain the second lysed title compound 7-fluoro-1,3,4,5-tetrahydro with a 100% chiral purity as a pale yellow solid -2 H - pyrido [4,3- b] indole-2-carboxylic acid tert-butyl ester (0.4 mg, 53%). The first elution of the title compound 9-fluoro-1,3,4,5-tetrahydro- 2H -pyrido[4,3- b ]indole-2-carboxylic acid tert-butyl ester was isolated with 100% Sexual purity of light yellow solid (0.25 g, 33%). Second elution of the title compound: MS: 291.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.01 (bs, 1H), 7.36-7.39 (m, 1H), 7.06-7.09 (m, 1H), 6.79-6.84 (m, 1H), 4.51 (bs, 2H), 3.68-3.71 (m, 2H), 2.74-2.77 (m, 2H), 1.44 (s, 9H). RT=2.08 min. First elute the title compound: MS: 291.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.22 (s, 1H), 7.13 (d, J = 8.08 Hz, 1H), 6.96-6.97 (m, 1H), 6.69-6.71 (m, 1H ), 4.63 (s, 2H), 3.69-3.70 (m, 2H), 2.68-2.76 (m, 2H), 1.44 (s, 9H). RT=1.74 min. Step D To the second solution of the title compound (0.4 g, 1.37 mmol) in THF (5 mL) from Step C above was added sodium hydride (0.099 mg, 4.137 mmol), followed by p-toluenesulfonyl chloride ( 0.288 g, 1.51 mmol). The reaction mixture was stirred for 30 minutes. The mixture was dissolved in EtOAc (20 ml) and washed with water and brine and dried over Na ₂ SO ₄ . The crude product was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/heptane gradient (20/80 => 80/20) to give the title compound (0.3 g, 49%). MS: 445 (M+H) ⁺ . ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.92-7.94 (m, 1H), 7.68-7.70 (m, 1H), 7.25-7.29 (m, 4H), 7.00-7.04 (m, 1H), 4.50 (bs, 2H), 3.76 (bs, 2H), 3.12 (bs, 2H), 2.38 (s, 3H), 1.51 (s, 9H). Step E To a solution of the title compound (0.3 g, 0.676 mmol) in dichloromethane from step D above was added 2 N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred for 12 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (0.2 g 78%) as an off-white solid. MS: 345 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.50 (bs, 2H), 7.80-7.87 (m, 2H), 7.78 (d, J = 2.00 Hz, 1H), 7.57-7.61 (m, 1H ), 7.40 (d, J = 8.16 Hz, 2H), 7.18-7.23 (m, 1H), 4.27 (bs, 2H), 3.56 (bs, 2H), 3.47 (bs, 2H), 2.34 (s, 3H) .

步驟 A 在0℃下向(2-氯-3-氟苯基)肼(10 g, 62.5 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(12 g, 62.5 mmol)於1,4-二噁烷(100 mL)中之溶液添加濃H₂ SO₄ (10 mL)。然後將反應混合物升溫至25℃並在110℃下加熱3 h。使該反應混合物冷卻至室溫且將沈澱物過濾出。將固體溶解於水中，經NaOH溶液鹼化並用二氯甲烷萃取。將有機相分離且經Na₂ SO₄ 乾燥並去除溶劑，得到呈淡黃色固體之標題化合物(10 g, 72%)。 MS: 225 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.23 (bs, 1H), 7.27-7.28 (m, 1H), 6.94-6.96 (m, 1H), 3.82 (s, 2H), 2.98-3.00 (m, 2H), 2.68 (d,J = 4.72 Hz, 2H)。步驟 B 向來自上文步驟A之標題化合物(10 g, 44.5 mmol)於THF (100 mL)中之溶液添加二碳酸二-第三丁基酯(10.5 g, 46.5 mmol)，且將混合物攪拌12 h。在反應完成後(藉由TLC監測)，將溶劑在減壓下濃縮以產生粗產物。藉由矽膠(60-120目)管柱層析純化該粗產物，得到呈黃色固體之標題化合物(12 g, 85%)。 MS: 325.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.43 (s, 1H), 7.36-7.38 (m, 1H), 6.97-7.00 (m, 1H), 4.51 (s, 2H), 3.68-3.69 (m, 2H), 2.76-2.78 (m, 2H), 1.43 (s, 9H)。步驟 C 向來自上文步驟B之標題化合物(5 g, 15.3 mmol)於無水甲醇(50 mL)中之溶液添加三乙胺(6.74 mL, 46.18 mmol)及10% Pd/C (0.2 mg, 20% wt)。在10巴壓力下進行氫化達16小時。經由矽藻土墊過濾反應混合物並在真空下濃縮，得到標題化合物(4 g, 90%)。 MS: 291.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.01 (bs, 1H), 7.36-7.39 (m, 1H), 7.06-7.09 (m, 1H), 6.79-6.84 (m, 1H), 4.51 (bs, 2H), 3.68-3.71 (m, 2H), 2.74-2.77 (m, 2H), 1.44 (s, 9H)。步驟 D 向來自上文步驟C之標題化合物(4 g, 13.7 mmol)於THF (40 mL)中之溶液添加氫化鈉(9.9 g, 41.23 mmol)，之後添加對甲苯磺醯氯(2.88 g, 15.1 mmol)。將反應混合物攪拌30 min。將該混合物溶解於EtOAc (200 ml)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(20/80 =＞ 80/20)，在矽膠管柱上純化粗產物，得到標題化合物(5 g, 82%)。 MS: 445 (M+H)⁺ 。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.92-7.94 (m, 1H), 7.68-7.70 (m, 1H), 7.25-7.29 (m, 4H), 7.00-7.04 (m, 1H), 4.50 (bs, 2H), 3.76 (bs, 2H), 3.12 (bs, 2H), 2.38 (s, 3H), 1.51 (s, 9H)。步驟 E 向來自上文步驟D之標題化合物(3 g, 6.76 mmol)於二氯甲烷(30 mL)中之溶液添加於1,4-二噁烷中之2 N HCl (15 mL)。將反應混合物攪拌12 h。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之標題化合物(2 g, 78%)。 MS: 345 (M+H)⁺ 。¹ H NMR (400 MHz, DMSO-d ₆ ) δ = 9.50 (bs, 2H), 7.80-7.87 (m, 2H), 7.78 (d,J = 2.00 Hz, 1H), 7.57-7.61 (m, 1H), 7.40 (d,J = 8.16 Hz, 2H), 7.18-7.23 (m, 1H), 4.27 (bs, 2H), 3.56 (bs, 2H), 3.47 (bs, 2H), 2.34 (s, 3H)。 Preparation Example 8

Step A To (2-chloro-3-fluorophenyl)hydrazine (10 g, 62.5 mmol) and tert-butyl 4-oxohexahydropyridine-1-carboxylate (12 g, 62.5 mmol) at 0°C ) A solution in 1,4-dioxane (100 mL) was added concentrated H ₂ SO ₄ (10 mL). The reaction mixture was then warmed to 25 °C and heated at 110 °C for 3 h. The reaction mixture was allowed to cool to room temperature and the precipitate was filtered off. The solid was dissolved in water, basified with NaOH solution and extracted with dichloromethane. The organic phase was separated and dried over Na ₂ SO ₄ and the solvent was removed to give the title compound (10 g, 72%) as a light yellow solid. MS: 225 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.23 (bs, 1H), 7.27-7.28 (m, 1H), 6.94-6.96 (m, 1H), 3.82 (s, 2H), 2.98-3.00 (m, 2H), 2.68 (d, J = 4.72 Hz, 2H). Step B To a solution of the title compound (10 g, 44.5 mmol) in THF (100 mL) from Step A above, di-tert-butyl dicarbonate (10.5 g, 46.5 mmol) was added, and the mixture was stirred for 12 h. After the reaction was completed (monitored by TLC), the solvent was concentrated under reduced pressure to produce a crude product. The crude product was purified by silica gel (60-120 mesh) column chromatography to obtain the title compound (12 g, 85%) as a yellow solid. MS: 325.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.43 (s, 1H), 7.36-7.38 (m, 1H), 6.97-7.00 (m, 1H), 4.51 (s, 2H), 3.68-3.69 (m, 2H), 2.76-2.78 (m, 2H), 1.43 (s, 9H). Step C To the solution of the title compound (5 g, 15.3 mmol) in anhydrous methanol (50 mL) from Step B above was added triethylamine (6.74 mL, 46.18 mmol) and 10% Pd/C (0.2 mg, 20 % wt). The hydrogenation is carried out at a pressure of 10 bar for 16 hours. The reaction mixture was filtered through a pad of celite and concentrated under vacuum to give the title compound (4 g, 90%). MS: 291.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.01 (bs, 1H), 7.36-7.39 (m, 1H), 7.06-7.09 (m, 1H), 6.79-6.84 (m, 1H), 4.51 (bs, 2H), 3.68-3.71 (m, 2H), 2.74-2.77 (m, 2H), 1.44 (s, 9H). Step D To the solution of the title compound (4 g, 13.7 mmol) in THF (40 mL) from Step C above was added sodium hydride (9.9 g, 41.23 mmol), followed by p-toluenesulfonyl chloride (2.88 g, 15.1 mmol). The reaction mixture was stirred for 30 min. The mixture was dissolved in EtOAc (200 ml) and washed with water and brine and dried over Na ₂ SO ₄ . Using the Biotage Isolera One purification system, using an EtOAc/heptane gradient (20/80 => 80/20), the crude product was purified on a silica gel column to give the title compound (5 g, 82%). MS: 445 (M+H) ⁺ . ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.92-7.94 (m, 1H), 7.68-7.70 (m, 1H), 7.25-7.29 (m, 4H), 7.00-7.04 (m, 1H), 4.50 (bs, 2H), 3.76 (bs, 2H), 3.12 (bs, 2H), 2.38 (s, 3H), 1.51 (s, 9H). Step E To a solution of the title compound (3 g, 6.76 mmol) in dichloromethane (30 mL) from step D above was added 2 N HCl (15 mL) in 1,4-dioxane. The reaction mixture was stirred for 12 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (2 g, 78%) as an off-white solid. MS: 345 (M+H) ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ = 9.50 (bs, 2H), 7.80-7.87 (m, 2H), 7.78 (d, J = 2.00 Hz, 1H), 7.57-7.61 (m, 1H) , 7.40 (d, J = 8.16 Hz, 2H), 7.18-7.23 (m, 1H), 4.27 (bs, 2H), 3.56 (bs, 2H), 3.47 (bs, 2H), 2.34 (s, 3H).

步驟 A 向來自製備實例8步驟C之標題化合物(0.4 mg, 1.37 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.099 g, 4.137 mmol)，之後添加碘甲烷(0.102 ml, 1.64 mmol)。將反應混合物攪拌2 h。將該混合物溶解於EtOAc (20 ml)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(20/80 =＞ 80/20)，在矽膠管柱上純化粗產物，得到標題化合物(0.3 mg, 73%)。 MS: 305.37 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(0.3 mg, 0.986 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2 N HCl (5 mL)。將反應混合物攪拌過夜。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之標題化合物(0.18 g 75%)。 MS: 205 (M+H)⁺ 。 Preparation Example 9

Step A To a solution of the title compound (0.4 mg, 1.37 mmol) in THF (5 mL) from Preparation Example 8, Step C was added sodium hydride (0.099 g, 4.137 mmol), followed by iodomethane (0.102 ml, 1.64 mmol) . The reaction mixture was stirred for 2 h. The mixture was dissolved in EtOAc (20 ml) and washed with water and brine and dried over Na ₂ SO ₄ . The crude product was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/heptane gradient (20/80 => 80/20) to give the title compound (0.3 mg, 73%). MS: 305.37 (M+H) ⁺ . Step B To a solution of the title compound (0.3 mg, 0.986 mmol) from dichloromethane from step A above was added 2 N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred overnight. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (0.18 g 75%) as an off-white solid. MS: 205 (M+H) ⁺ .

步驟 A 向來自製備實例7步驟C之第一溶析標題化合物(0.2 mg, 0.67 mmol)於THF (5 mL)中之溶液添加氫化鈉(0.048 g, 2.137 mmol)，之後添加對甲苯磺醯氯(0.144 g, 0.76 mmol)。將反應混合物攪拌30分鐘。將該混合物溶解於EtOAc (20 ml)中，並用水及鹽水洗滌且經Na₂ SO₄ 乾燥。使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(20/80 =＞ 80/20)，在矽膠管柱上純化粗產物，得到標題化合物(0.155 g, 50%)。 MS: 445 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.80-7.82 (m, 3H), 7.31-7.32 (m, 3H), 7.07-7.09 (m, 1H), 4.56 (s, 2H), 3.68-3.69 (m, 2H), 3.09 (bs, 2H), 2.33 (s, 3H), 1.43 (s, 9H)。步驟 B 向來自上文步驟A之標題化合物(0.15 g, 0.337 mmol)於二氯甲烷中之溶液添加於1,4-二噁烷中之2 N HCl (5 mL)。將反應混合物攪拌12 h。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈灰白色固體之標題化合物(0.1 g, 71%)。 MS: 345 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 9.49 (bs, 2H), 7.85-7.87 (m, 3H), 7.35-7.36 (m, 3H), 7.12-7.14 (m, 1H), 4.39 (s, 2H), 3.48 (bs, 2H), 3.17 (bs, 2H), 2.35 (s, 3H)。 Preparation Example 10

Step A To the first leaching solution of the title compound (0.2 mg, 0.67 mmol) in THF (5 mL) from Step C of Preparation Example 7 was added sodium hydride (0.048 g, 2.137 mmol), followed by p-toluenesulfonyl chloride (0.144 g, 0.76 mmol). The reaction mixture was stirred for 30 minutes. The mixture was dissolved in EtOAc (20 ml) and washed with water and brine and dried over Na ₂ SO ₄ . Using the Biotage Isolera One purification system, using an EtOAc/heptane gradient (20/80 => 80/20), the crude product was purified on a silica gel column to give the title compound (0.155 g, 50%). MS: 445 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.80-7.82 (m, 3H), 7.31-7.32 (m, 3H), 7.07-7.09 (m, 1H), 4.56 (s, 2H), 3.68 -3.69 (m, 2H), 3.09 (bs, 2H), 2.33 (s, 3H), 1.43 (s, 9H). Step B To a solution of the title compound from Step A above (0.15 g, 0.337 mmol) in dichloromethane was added 2 N HCl (5 mL) in 1,4-dioxane. The reaction mixture was stirred for 12 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (0.1 g, 71%) as an off-white solid. MS: 345 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.49 (bs, 2H), 7.85-7.87 (m, 3H), 7.35-7.36 (m, 3H), 7.12-7.14 (m, 1H), 4.39 (s, 2H), 3.48 (bs, 2H), 3.17 (bs, 2H), 2.35 (s, 3H).

步驟 A 向2-(4-氟苯基)-4,4,5,5-四甲基-1,3,2-二氧雜硼雜環戊烷(5g, 22.51 mmol)於二氯乙烷(250 mL)中之溶液添加N-羥基酞醯亞胺(7.34 g, 45.03 mmol)、氯化銅(I)(2.22 g, 22.51 mmol)、吡啶(2.75 mL, 33.75 mmol)、分子篩(5 g)，且將反應混合物加熱至70℃持續12 h。在反應完成後，經由矽藻土床過濾該反應混合物並用乙酸乙酯洗滌。將乙酸乙酯層濃縮且使用Biotage Isolera One純化系統，採用己烷/EtOAc梯度(90/10 =＞ 80/20)，在矽膠上純化粗產物，得到呈白色固體之標題化合物(1.4 g, 24%)。 MS: 258.22 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.92-7.93 (m, 4H), 7.33-7.34 (m, 2H), 7.20-7.22 (m, 2H)。步驟 B 向來自上文步驟A之標題化合物(1.4g, 5.44 mmol)於氯仿:甲醇(9:1, 100 mL)中之溶液添加水合肼(0.81 g, 16.32 mmol)，且將反應混合物在25℃下攪拌12 h。在反應完成後，經由矽藻土床過濾該反應混合物並用二氯甲烷洗滌。將二氯甲烷層濃縮，且在0℃下將粗產物添加至乙醚(5 mL)及於乙醚(2 mL)中之2 N HCl，然後將反應混合物在25℃下攪拌30 min。在反應完成後，將該反應混合物過濾，在真空下乾燥，得到灰白色固體(0.44 g, 50%)。 MS: 128.12 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.03-8.04 (m, 2H), 7.76-7.76 (m, 2H)。步驟 C 在0℃下向來自上文步驟B之標題化合物(0.44 g, 2.7 mmol)及1,4-二氧雜-8-氮雜螺[4.5]癸烷(0.44 g, 3.07 mmol)於1,4-二噁烷(5 mL)中之溶液添加濃H₂ SO₄ (0.5 mL)。將反應混合物升溫至25℃，且然後在微波條件下在150℃下進一步加熱1 h。使反應混合物冷卻至25℃，且將沈澱物過濾出。將固體溶解於水中，經NaOH溶液鹼化並用二氯甲烷萃取。將有機相分離且經Na₂ SO₄ 乾燥，並將溶劑濃縮，粗產物原樣用於下一步驟(0.270 g, 52%)。 MS: 192.21 (M+H)⁺ 。 Preparation Example 11

Step A To 2-(4-fluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5g, 22.51 mmol) in dichloroethane (250 mL) solution was added N-hydroxyphthalimide (7.34 g, 45.03 mmol), copper (I) chloride (2.22 g, 22.51 mmol), pyridine (2.75 mL, 33.75 mmol), molecular sieve (5 g ), and the reaction mixture was heated to 70° C. for 12 h. After the reaction was completed, the reaction mixture was filtered through a bed of celite and washed with ethyl acetate. The ethyl acetate layer was concentrated and the crude product was purified on silica gel using a Biotage Isolera One purification system using a hexane/EtOAc gradient (90/10 => 80/20) to give the title compound as a white solid (1.4 g, 24 %). MS: 258.22 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.92-7.93 (m, 4H), 7.33-7.34 (m, 2H), 7.20-7.22 (m, 2H). Step B To a solution of the title compound (1.4g, 5.44 mmol) from chloroform: methanol (9:1, 100 mL) from step A above was added hydrazine hydrate (0.81 g, 16.32 mmol), and the reaction mixture was added at 25 Stir for 12 h at ℃. After the reaction was completed, the reaction mixture was filtered through a bed of celite and washed with dichloromethane. The dichloromethane layer was concentrated, and the crude product was added to diethyl ether (5 mL) and 2 N HCl in diethyl ether (2 mL) at 0°C, and then the reaction mixture was stirred at 25°C for 30 min. After the reaction was completed, the reaction mixture was filtered and dried under vacuum to obtain an off-white solid (0.44 g, 50%). MS: 128.12 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.03-8.04 (m, 2H), 7.76-7.76 (m, 2H). Step C: At 0°C, add the title compound (0.44 g, 2.7 mmol) and 1,4-dioxa-8-azaspiro[4.5]decane (0.44 g, 3.07 mmol) from Step B above to 1 , 4-dioxane (5 mL) was added concentrated H ₂ SO ₄ (0.5 mL). The reaction mixture was warmed to 25°C, and then further heated at 150°C for 1 h under microwave conditions. The reaction mixture was cooled to 25°C, and the precipitate was filtered off. The solid was dissolved in water, basified with NaOH solution and extracted with dichloromethane. The organic phase was separated and dried over Na ₂ SO ₄ , and the solvent was concentrated, and the crude product was used as it is in the next step (0.270 g, 52%). MS: 192.21 (M+H) ⁺ .

步驟 A 在0℃下向4-(氟苯基)肼鹽酸鹽(10 g, 0.061 mol)及3-側氧基-8-氮雜二環[3.2.1]辛烷-8-甲酸第三丁基酯(13.9 g, 0.61 mol)於1,4-二噁烷(100 mL)中之溶液添加濃H₂ SO₄ (10 mL)，且然後將混合物加熱至100℃持續12 h。在反應完成後，將反應混合物冷卻至室溫，並藉由高真空將溶劑去除以得到粗製材料。使用30%氫氧化鈉溶液使粗製材料鹼化，且沈澱出固體。將沈澱固體過濾，用水(100 mL)及二乙醚(100 mL)洗滌，然後將固體在管線真空下乾燥16 h，得到呈淡棕色固體之標題化合物(11 g, 83%)。 MS: 216.10 (M+H)⁺ 。步驟 B 在0℃下向來自上文步驟A之標題化合物(11 g, 0.051 mol)於THF (50 mL)中之溶液添加三乙胺(11 mL, 0.076 mol)及二碳酸二-第三丁基酯(11.13 g, 0.051 mol)，且然後在室溫下攪拌12 h。如藉由TLC所證實，在反應完成後，將溶劑去除且藉由矽膠管柱使用於石油醚中之40%至50%乙酸乙酯純化粗製反應混合物，得到呈淡黃色固體之標題化合物(7.4 g, 46%)。 MS: 316.15 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 10.94 (bs, 1H), 7.23-7.24 (m, 2H), 6.80-6.81 (m, 1H), 5.09-5.11 (m, 1H), 4.46 (bs, 1H), 3.23-3.27 (m, 1H), 2.51-2.57 (m, 1H), 2.22-2.26 (m, 1H), 2.00-2.07 (m, 1H), 1.59-1.60 (m, 1H), 1.41-1.44 (m, 2H), 1.20-1.27 (m, 9H)。步驟 C 在0℃下向來自上文步驟B之標題化合物(3 g, 0.009 mol)於THF (30 mL)中之溶液逐份添加氫化鈉(60%於礦物油中；0.57 g, 0.014 mol)。添加完成後，將反應混合物在室溫下攪拌30分鐘，且然後將反應混合物再次冷卻至0℃。在0℃下向此混合物逐滴添加溶解於THF (20 mL)中之對甲苯磺醯氯(2.16 g, 0.11 mol)。添加完成後，將反應混合物在室溫下攪拌2 h。如藉由TLC所證實，在反應完成後，將反應混合物冷卻至0℃並用冰水淬滅，之後使用乙酸乙酯(100 mL)萃取。將乙酸乙酯層用水(30 mL)及鹽水溶液(30 mL)洗滌。使有機層經Na₂ SO₄ 乾燥，過濾並蒸發以得到粗產物，其藉由矽膠管柱使用於石油醚中之15%至25%乙酸乙酯進行純化，得到呈灰白色固體之標題化合物(2.9 g, 65%)。 MS: 470.16 (M+H)⁺ 。步驟 D 在0℃下向來自上文步驟C之標題化合物(2.9 g , 0.006 mol)於二氯甲烷(10 mL)中之溶液添加於1,4-二噁烷中之4 N HCl (20 mL)。將反應混合物在環境溫度下攪拌12 h。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚(10 mL)洗滌，得到呈灰白色固體之標題化合物(2.4 g, 98%)。 MS: 370.15 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.07-8.08 (m, 1H), 7.78-7.80 (m, 1H), 7.30-7.31 (m, 3H), 7.10-7.11 (m, 1H), 5.13 (d,J = 4.80 Hz, 1H), 4.52 (d,J = 4.80 Hz, 1H), 3.64-3.65 (m, 4H), 3.30-3.30 (m, 1H), 2.22-2.24 (m, 7H), 1.92 (bs, 1H)。 Preparation Example 12

Step A: To 4-(fluorophenyl)hydrazine hydrochloride (10 g, 0.061 mol) and 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylic acid A solution of tributyl ester (13.9 g, 0.61 mol) in 1,4-dioxane (100 mL) was added concentrated H ₂ SO ₄ (10 mL), and then the mixture was heated to 100° C. for 12 h. After the reaction was completed, the reaction mixture was cooled to room temperature, and the solvent was removed by high vacuum to obtain a crude material. A 30% sodium hydroxide solution was used to alkalize the crude material and a solid precipitated. The precipitated solid was filtered, washed with water (100 mL) and diethyl ether (100 mL), and then the solid was dried under line vacuum for 16 h to obtain the title compound (11 g, 83%) as a light brown solid. MS: 216.10 (M+H) ⁺ . Step B To a solution of the title compound (11 g, 0.051 mol) in THF (50 mL) from Step A above at 0°C was added triethylamine (11 mL, 0.076 mol) and di-tert-butyl dicarbonate Ester (11.13 g, 0.051 mol), and then stirred at room temperature for 12 h. As confirmed by TLC, after the reaction was completed, the solvent was removed and the crude reaction mixture was purified by silica gel column using 40% to 50% ethyl acetate in petroleum ether to obtain the title compound (7.4 g, 46%). MS: 316.15 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.94 (bs, 1H), 7.23-7.24 (m, 2H), 6.80-6.81 (m, 1H), 5.09-5.11 (m, 1H), 4.46 (bs, 1H), 3.23-3.27 (m, 1H), 2.51-2.57 (m, 1H), 2.22-2.26 (m, 1H), 2.00-2.07 (m, 1H), 1.59-1.60 (m, 1H) , 1.41-1.44 (m, 2H), 1.20-1.27 (m, 9H). Step C To a solution of the title compound (3 g, 0.009 mol) in THF (30 mL) from Step B above was added sodium hydride (60% in mineral oil; 0.57 g, 0.014 mol) portionwise at 0°C . After the addition was completed, the reaction mixture was stirred at room temperature for 30 minutes, and then the reaction mixture was cooled to 0°C again. To this mixture was added p-toluenesulfonyl chloride (2.16 g, 0.11 mol) dissolved in THF (20 mL) dropwise at 0°C. After the addition was complete, the reaction mixture was stirred at room temperature for 2 h. As confirmed by TLC, after the reaction was completed, the reaction mixture was cooled to 0°C and quenched with ice water, followed by extraction with ethyl acetate (100 mL). The ethyl acetate layer was washed with water (30 mL) and brine solution (30 mL). The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated to obtain the crude product, which was purified by silica gel column using 15% to 25% ethyl acetate in petroleum ether to give the title compound (2.9 g, 65%). MS: 470.16 (M+H) ⁺ . Step D To the solution of the title compound (2.9 g, 0.006 mol) in dichloromethane (10 mL) from Step C above was added 4 N HCl (20 mL in 1,4-dioxane) at 0°C ). The reaction mixture was stirred at ambient temperature for 12 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether (10 mL) to obtain the title compound (2.4 g, 98%) as an off-white solid. MS: 370.15 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.07-8.08 (m, 1H), 7.78-7.80 (m, 1H), 7.30-7.31 (m, 3H), 7.10-7.11 (m, 1H) , 5.13 (d, J = 4.80 Hz, 1H), 4.52 (d, J = 4.80 Hz, 1H), 3.64-3.65 (m, 4H), 3.30-3.30 (m, 1H), 2.22-2.24 (m, 7H ), 1.92 (bs, 1H).

步驟 A 向2,6-二氯苯并[d]噻唑(5 g, 24.5 mmol)於無水二氯甲烷(50 mL)中之溶液添加嗎啉(3.19 g, 36.6 mmol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(3.71 g, 36.7 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(5 g, 96%)。 MS: 213.4 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.80 (d,J = 8.00 Hz, 1H), 7.53(d,J = 2.00 Hz, 1H), 7.13-7.14 (m, 1H), 3.74-3.75 (m, 4H), 3.56-3.57 (m, 4H)。 Preparation Example 13

Step A To a solution of 2,6-dichlorobenzo[d]thiazole (5 g, 24.5 mmol) in anhydrous dichloromethane (50 mL) was added morpholine (3.19 g, 36.6 mmol), and the mixture was cooled to 0℃. To this cold reaction mixture, triethylamine (3.71 g, 36.7 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (5 g, 96%). MS: 213.4 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.80 (d, J = 8.00 Hz, 1H), 7.53 (d, J = 2.00 Hz, 1H), 7.13-7.14 (m, 1H), 3.74- 3.75 (m, 4H), 3.56-3.57 (m, 4H).

步驟 A 向2,5-二氯苯并[d]噻唑(5 g, 24.5 mmol)於無水二氯甲烷(50 mL)中之溶液添加嗎啉(3.19 g, 36.6 mmol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(3.71 g, 36.7 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(4.5 g, 86%)。 MS: 255.4 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.82 (d,J = 8.00 Hz, 1H), 7.50 (d,J = 2.00 Hz, 1H), 7.11-7.11 (m, 1H), 3.72-3.73 (m, 4H), 3.55-3.56 (m, 4H)。 Preparation Example 14

Step A To a solution of 2,5-dichlorobenzo[d]thiazole (5 g, 24.5 mmol) in anhydrous dichloromethane (50 mL) was added morpholine (3.19 g, 36.6 mmol), and the mixture was cooled to 0℃. To this cold reaction mixture, triethylamine (3.71 g, 36.7 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (4.5 g, 86%). MS: 255.4 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.82 (d, J = 8.00 Hz, 1H), 7.50 (d, J = 2.00 Hz, 1H), 7.11-7.11 (m, 1H), 3.72- 3.73 (m, 4H), 3.55-3.56 (m, 4H).

步驟 A 向2,6-二氯苯并[d]噁唑(5 g, 26.8 mmol)於無水二氯甲烷(50 mL)中之溶液添加嗎啉(3.50 g, 40.3 mmol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(4.0 g, 39.6 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(5 g, 78%)。 MS: 239.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.59 (d,J = 2.80 Hz, 1H), 7.30 (d,J = 11.20 Hz, 1H), 7.21 (dd,J = 2.80, 11.20 Hz, 1H), 3.71-3.74 (m, 4H), 3.57-3.60 (m, 4H)。 Preparation Example 15

Step A To a solution of 2,6-dichlorobenzo[d]oxazole (5 g, 26.8 mmol) in anhydrous dichloromethane (50 mL) was added morpholine (3.50 g, 40.3 mmol), and the mixture was cooled To 0°C. To this cold reaction mixture, triethylamine (4.0 g, 39.6 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (5 g, 78%). MS: 239.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.59 (d, J = 2.80 Hz, 1H), 7.30 (d, J = 11.20 Hz, 1H), 7.21 (dd, J = 2.80, 11.20 Hz, 1H), 3.71-3.74 (m, 4H), 3.57-3.60 (m, 4H).

步驟 A 向2,5-二氯苯并[d]噁唑(5 g, 26.8 mmol)於無水二氯甲烷(50 mL)中之溶液添加嗎啉(3.50 g, 40.3 mmol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(4.0 g, 39.6 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(5.2 g, 81%)。 MS: 239.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.44 (d,J = 8.40 Hz, 1H), 7.36 (d,J = 2.40 Hz, 1H), 7.06 (dd,J = 2.00, 8.40 Hz, 1H), 3.71-3.73 (m, 4H), 3.59-3.61 (m, 4H)。 Preparation Example 16

Step A To a solution of 2,5-dichlorobenzo[d]oxazole (5 g, 26.8 mmol) in anhydrous dichloromethane (50 mL) was added morpholine (3.50 g, 40.3 mmol), and the mixture was cooled To 0°C. To this cold reaction mixture, triethylamine (4.0 g, 39.6 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (5.2 g, 81%). MS: 239.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.44 (d, J = 8.40 Hz, 1H), 7.36 (d, J = 2.40 Hz, 1H), 7.06 (dd, J = 2.00, 8.40 Hz, 1H), 3.71-3.73 (m, 4H), 3.59-3.61 (m, 4H).

步驟 A 向2,6-二氯苯并[d]噻唑(5 g, 24.5 mmol)於無水二氯甲烷(50 mL)中之溶液添加於THF中之2 M二甲胺(18.37 mL, 36.65 mol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(6.8 mL, 49 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(4.8 g, 94%)。 MS: 213.4 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.88 (d,J = 2.10 Hz, 1H), 7.41 (d,J = 8.70 Hz, 1H), 7.25-7.26 (m, 1H), 3.14 (s, 6H)。 Preparation Example 17

Step A To a solution of 2,6-dichlorobenzo[d]thiazole (5 g, 24.5 mmol) in anhydrous dichloromethane (50 mL) was added 2 M dimethylamine (18.37 mL, 36.65 mol in THF) ), and the mixture was cooled to 0°C. To this cold reaction mixture, triethylamine (6.8 mL, 49 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (4.8 g, 94%). MS: 213.4 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.88 (d, J = 2.10 Hz, 1H), 7.41 (d, J = 8.70 Hz, 1H), 7.25-7.26 (m, 1H), 3.14 ( s, 6H).

步驟 A 向2,5-二氯苯并[d]噻唑(5 g, 24.5 mmol)於無水二氯甲烷(50 mL)中之溶液添加於THF中之2 M二甲胺(18.37 mL, 36.65 mol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(6.8 mL, 49 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(4.5 g, 88%)。 MS: 213.4 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.77 (d,J = 11.20 Hz, 1H), 7.46 (d,J = 2.40 Hz, 1H), 7.05-7.05 (m, 1H), 3.14 (s, 6H)。 Preparation Example 18

Step A To a solution of 2,5-dichlorobenzo[d]thiazole (5 g, 24.5 mmol) in anhydrous dichloromethane (50 mL) was added 2 M dimethylamine (18.37 mL, 36.65 mol in THF) ), and the mixture was cooled to 0°C. To this cold reaction mixture, triethylamine (6.8 mL, 49 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (4.5 g, 88%). MS: 213.4 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.77 (d, J = 11.20 Hz, 1H), 7.46 (d, J = 2.40 Hz, 1H), 7.05-7.05 (m, 1H), 3.14 ( s, 6H).

步驟 A 向6-溴-2-氯苯并[d]噻唑(0.45 g, 1.81 mmol)於乙醇(12 mL)中之溶液添加2 M二甲胺溶液(3 mL)，且使用Biotage微波爐將混合物在100℃下加熱60分鐘。將反應混合物冷卻至室溫。將溶劑去除，使用乙酸乙酯及庚烷梯度(40/60 =＞ 60/40)，在矽膠管柱上純化粗產物，得到呈固體之標題化合物(0.441 g, 95%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.70 (d,J = 1.9 Hz, 1H), 7.43 - 7.35 (m, 2H), 3.20 (s, 6H)。 Preparation Example 19

Step A To a solution of 6-bromo-2-chlorobenzo[d]thiazole (0.45 g, 1.81 mmol) in ethanol (12 mL) was added 2 M dimethylamine solution (3 mL), and the mixture was converted using a Biotage microwave oven Heat at 100°C for 60 minutes. The reaction mixture was cooled to room temperature. The solvent was removed, and the crude product was purified on a silica gel column using a gradient of ethyl acetate and heptane (40/60 => 60/40) to obtain the title compound (0.441 g, 95%) as a solid. ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.70 (d, J = 1.9 Hz, 1H), 7.43-7.35 (m, 2H), 3.20 (s, 6H).

步驟 A 向2,6-二氯苯并[d]噁唑(5 g, 26.6 mmol)於無水二氯甲烷(50 mL)中之溶液添加於THF中之2 M二甲胺(26.6 mL, 53.2 mmol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(5.6 mL, 39.9 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(5 g, 96%)。 MS: 197.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.56 (s, 1H), 7.23-7.24 (m, 1H), 7.16-7.16 (m, 1H), 3.13 (s, 6H)。 Preparation Example 20

Step A To a solution of 2,6-dichlorobenzo[d]oxazole (5 g, 26.6 mmol) in anhydrous dichloromethane (50 mL) was added 2 M dimethylamine (26.6 mL, 53.2) in THF mmol), and the mixture was cooled to 0°C. To this cold reaction mixture, triethylamine (5.6 mL, 39.9 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (5 g, 96%). MS: 197.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.56 (s, 1H), 7.23-7.24 (m, 1H), 7.16-7.16 (m, 1H), 3.13 (s, 6H).

步驟 A 向2,5-二氯苯并[d]噁唑(5 g, 26.6 mmol)於無水二氯甲烷(50 mL)中之溶液添加於THF中之2 M二甲胺(26.6 mL, 53.2 mmol)，且將混合物冷卻至0℃。向此冷的反應混合物逐滴添加三乙胺(5.6 mL, 39.9 mmol)，且將混合物在室溫下攪拌4 h。在反應完成後，利用H₂ O (2 × 20 mL)處理反應混合物並用二氯甲烷萃取。將有機層分離，經Na₂ SO₄ 乾燥，過濾並蒸發，得到白色固體，將其與二乙醚一起研磨，得到標題化合物(4.9 g, 94%)。 MS: 197.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.40 (d,J = 8.40 Hz, 1H), 7.30 (d,J = 2.00 Hz, 1H), 6.99-6.99 (m, 1H), 3.13 (s, 6H)。 Preparation Example 21

Step A To a solution of 2,5-dichlorobenzo[d]oxazole (5 g, 26.6 mmol) in anhydrous dichloromethane (50 mL) was added 2 M dimethylamine (26.6 mL, 53.2) in THF mmol), and the mixture was cooled to 0°C. To this cold reaction mixture, triethylamine (5.6 mL, 39.9 mmol) was added dropwise, and the mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was treated with H ₂ O (2×20 mL) and extracted with dichloromethane. The organic layer was separated, dried over Na ₂ SO ₄ , filtered and evaporated to give a white solid, which was triturated with diethyl ether to give the title compound (4.9 g, 94%). MS: 197.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.40 (d, J = 8.40 Hz, 1H), 7.30 (d, J = 2.00 Hz, 1H), 6.99-6.99 (m, 1H), 3.13 ( s, 6H).

步驟 A 向6-溴-2-氯苯并[d]噻唑(0.8 g, 3.2 mmol)於乙醇(12 mL)中之溶液添加異丙胺溶液(1 mL)，且使用Biotage微波爐將混合物在100℃下加熱45分鐘。將反應混合物冷卻至室溫。去除溶劑，將粗產物自EtOAc及正庚烷混合物結晶，得到呈固體之標題化合物(0.663 g, 75.8%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.69 (t,J = 1.3 Hz, 1H), 7.38 (d,J = 1.3 Hz, 2H), 7.27 (s, 1H), 5.51 - 5.26 (m, 1H), 3.92 (h,J = 6.5 Hz, 1H), 1.33 (d,J = 6.4 Hz, 6H)。 Preparation Example 22

Step A To a solution of 6-bromo-2-chlorobenzo[d]thiazole (0.8 g, 3.2 mmol) in ethanol (12 mL) was added an isopropylamine solution (1 mL), and the mixture was placed at 100°C using a Biotage microwave oven Heat for 45 minutes. The reaction mixture was cooled to room temperature. The solvent was removed, and the crude product was crystallized from a mixture of EtOAc and n-heptane to give the title compound (0.663 g, 75.8%) as a solid. ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.69 (t, J = 1.3 Hz, 1H), 7.38 (d, J = 1.3 Hz, 2H), 7.27 (s, 1H), 5.51-5.26 (m , 1H), 3.92 (h, J = 6.5 Hz, 1H), 1.33 (d, J = 6.4 Hz, 6H).

步驟 A 向6-溴-2-氯苯并[d]噻唑(1 g, 4.0 mmol)於乙醇(6 mL)中之溶液添加異丙胺溶液(1.5 mL)，且使用Biotage微波爐將混合物在100℃下加熱90分鐘。將反應混合物冷卻至室溫。去除溶劑，將粗產物溶解於二氯甲烷(150 mL)中，且用1 M NaOH溶液、水及鹽水洗滌並經Na₂ SO₄ 乾燥。將溶劑在減壓下去除以得到粗產物，使用EtOAc及庚烷梯度(20/80 =＞ 50/50)，在矽膠管柱上純化該粗產物，得到呈固體之標題化合物(0.35 g, 36%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.72 (s, 1H), 7.41 (s, 1H), 7.29 (d,J = 3.3 Hz, 1H), 5.40 (s, 1H), 3.13 (s, 3H)。 Preparation Example 23

Step A To a solution of 6-bromo-2-chlorobenzo[d]thiazole (1 g, 4.0 mmol) in ethanol (6 mL) was added isopropylamine solution (1.5 mL), and the mixture was placed at 100°C using a Biotage microwave oven Heat for 90 minutes. The reaction mixture was cooled to room temperature. The solvent was removed and the crude product was dissolved in dichloromethane (150 mL) and washed with 1 M NaOH solution, water and brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure to obtain the crude product, which was purified on a silica gel column using EtOAc and heptane gradient (20/80 => 50/50) to obtain the title compound as a solid (0.35 g, 36 %). ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.72 (s, 1H), 7.41 (s, 1H), 7.29 (d, J = 3.3 Hz, 1H), 5.40 (s, 1H), 3.13 (s , 3H).

步驟 A 向5-溴-2-氯苯并[d]噻唑(0.9 g, 3.62 mmol)於乙醇(12 mL)中之溶液添加4 M甲胺溶液(1 mL)，且使用Biotage微波爐將混合物在100℃下加熱90分鐘。將反應混合物冷卻至室溫。去除溶劑，使用EtOAc及庚烷梯度(20/80 =＞ 50/50)，在矽膠管柱上純化粗產物，得到呈固體之標題化合物(0.57 g, 65%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.68 (t,J = 2.0 Hz, 1H), 7.46 (d,J = 8.3 Hz, 1H), 7.22 (dd,J = 8.4, 1.9 Hz, 1H), 5.90 (s, 1H), 3.13 (s, 3H)。 Preparation Example 24

Step A To a solution of 5-bromo-2-chlorobenzo[d]thiazole (0.9 g, 3.62 mmol) in ethanol (12 mL) was added 4 M methylamine solution (1 mL), and the mixture was removed using a Biotage microwave oven Heat at 100°C for 90 minutes. The reaction mixture was cooled to room temperature. The solvent was removed, and the crude product was purified on a silica gel column using EtOAc and heptane gradient (20/80 => 50/50) to obtain the title compound (0.57 g, 65%) as a solid. ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.68 (t, J = 2.0 Hz, 1H), 7.46 (d, J = 8.3 Hz, 1H), 7.22 (dd, J = 8.4, 1.9 Hz, 1H ), 5.90 (s, 1H), 3.13 (s, 3H).

步驟 A 向5-溴-2-氯苯并[d]噻唑(0.87 g, 3.5 mmol)於乙醇(12 mL)中之溶液添加異丙胺溶液(1.5 mL)，且使用Biotage微波爐將混合物在100℃下加熱60分鐘。將反應混合物冷卻至室溫。去除溶劑，將粗產物溶解於二氯甲烷(150 mL)中，且用1 M NaOH溶液、水及鹽水洗滌並經Na₂ SO₄ 乾燥。將溶劑在減壓下去除以得到粗產物，使用EtOAc及庚烷梯度(20/80 =＞ 50/50)，在矽膠管柱上純化該粗產物，得到呈固體之標題化合物(0.75 g, 79%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.67 (d,J = 1.9 Hz, 1H), 7.43 (d,J = 8.5 Hz, 1H), 7.19 (dd,J = 8.3, 1.9 Hz, 1H), 5.47 (s, 1H), 4.04 - 3.79 (m, 1H), 1.34 (d,J = 6.5 Hz, 6H)。 Preparation Example 25

Step A To a solution of 5-bromo-2-chlorobenzo[d]thiazole (0.87 g, 3.5 mmol) in ethanol (12 mL) was added an isopropylamine solution (1.5 mL), and the mixture was placed at 100°C using a Biotage microwave oven Heat for 60 minutes. The reaction mixture was cooled to room temperature. The solvent was removed and the crude product was dissolved in dichloromethane (150 mL) and washed with 1 M NaOH solution, water and brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure to obtain the crude product, which was purified on a silica gel column using EtOAc and heptane gradient (20/80 => 50/50) to give the title compound as a solid (0.75 g, 79 %). ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.67 (d, J = 1.9 Hz, 1H), 7.43 (d, J = 8.5 Hz, 1H), 7.19 (dd, J = 8.3, 1.9 Hz, 1H ), 5.47 (s, 1H), 4.04-3.79 (m, 1H), 1.34 (d, J = 6.5 Hz, 6H).

步驟 A 將5-溴-3-碘吡啶-2-胺(5 g, 16.72 mmol)及異硫氰酸苯甲醯基酯(3.29, 20.07 mmol)於丙酮(10 mL)中之溶液在60℃下攪拌12小時，藉由TLC監測反應。將溶劑蒸發且將固體過濾，用正己烷(200 mL)洗滌並乾燥，得到呈灰白色固體之標題化合物(4 g, 52%)。 MS: 461.5 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 12.35 (s, 1H), 11.86 (s, 1H), 8.64-8.65 (m, 2H), 7.99-7.99 (m, 2H), 7.67 (s, 1H), 7.56 (d,J = 9.40 Hz, 2H)。步驟 B 向來自上文步驟A之標題化合物(4 g, 12.1 mmol)於1,4-二噁烷(60 mL)中之溶液添加碳酸鉀(2.5 g, 18.15 mmol)、L-脯胺酸(0.28 g, 2.43 mmol)及碘化銅(I)(0.462 g, 2.43 mmol)。然後，將反應混合物在80℃下攪拌16小時，藉由TLC監測反應。將反應混合物傾倒至1.0 L水及1.0 L NH₄ Cl飽和水溶液中。將懸浮液在室溫下攪拌1小時。將固體過濾，用NH₄ Cl之飽和水溶液(2 × 300 mL)及水(2 × 300 mL)洗滌並乾燥，得到呈灰白色固體之標題化合物(2.5 g，粗製)。 MS: 334.51 (M+H)⁺ 。步驟 C 將來自上文步驟B之標題化合物(2 g, 5.98 mmol)於70% H₂ SO₄ (6, 3.0 vol)中之懸浮液在120℃下加熱2小時。將反應混合物冷卻至室溫，且將該反應混合物緩慢傾倒至100 mL冷水(0℃)中。然後，藉由添加50% NaOH水溶液將該反應混合物調整至鹼性pH。然後，用EtOAc (6 × 150 mL)萃取化合物。使合併之有機層經Na₂ SO₄ 乾燥並過濾，然後將溶液濃縮，得到呈淺黃色固體之標題化合物(0.3 g, 23%)。 MS: 230.4 (M)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.27-8.31 (m, 2H), 8.11 (s, 2H)。步驟 D 在0℃下利用注射器經10 min之時期向來自上文步驟C之標題化合物(0.3 mg, 1.3 mmol)於乙腈(5 mL)中之懸浮液添加亞硝酸第三丁基酯( 0.2 ml, 1.95 mmol)。然後，逐份添加氯化銅(II)(0.2 g, 1.56 mmol)。在0℃下30分鐘後，將反應混合物升溫至室溫持續1小時並加熱至65℃，然後攪拌4小時。藉由TLC監測反應之進展。在反應完成後，將溶劑蒸發且用水(20 mL)及5% MeOH/DCM (3 × 20 mL)稀釋產物。將合併之有機物用鹽水(10 mL)洗滌，經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。藉由矽膠(60-120)管柱層析，利用1%甲醇/二氯甲烷進行溶析來純化粗製化合物，得到呈灰白色固體之標題化合物(0.15 g, 46%)。 MS: 250.9 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.91 (d,J = 2.40 Hz, 1H), 8.82 (d,J = 1.60 Hz, 1H)。步驟 E 向來自上文步驟D之標題化合物(0.18 g, 0.72 mmol)於無水二氯甲烷(5 mL)中之溶液添加三乙胺(0.3 mL, 2.16 mmol)及嗎啉(0.074 g, 0.86 mmol)，且將混合物在室溫下攪拌6小時。將反應混合物在真空下濃縮。藉由矽膠(60-120)管柱層析，利用石油醚/乙酸乙酯進行溶析來純化粗製化合物，得到呈灰黃色固體之標題化合物(0.18 g, 83%)。 MS: 300.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.49 (d,J = 2.00 Hz, 1H), 8.38 (d,J = 1.60 Hz, 1H), 3.72-3.74 (m, 4H), 3.61-3.62 (m, 4H)。 Preparation Example 26

Step A: A solution of 5-bromo-3-iodopyridin-2-amine (5 g, 16.72 mmol) and benzyl isothiocyanate (3.29, 20.07 mmol) in acetone (10 mL) at 60°C Stir for 12 hours and monitor the reaction by TLC. The solvent was evaporated and the solid was filtered, washed with n-hexane (200 mL) and dried to give the title compound (4 g, 52%) as an off-white solid. MS: 461.5 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 12.35 (s, 1H), 11.86 (s, 1H), 8.64-8.65 (m, 2H), 7.99-7.99 (m, 2H), 7.67 (s , 1H), 7.56 (d, J = 9.40 Hz, 2H). Step B To the solution of the title compound (4 g, 12.1 mmol) in 1,4-dioxane (60 mL) from step A above was added potassium carbonate (2.5 g, 18.15 mmol), L-proline ( 0.28 g, 2.43 mmol) and copper(I) iodide (0.462 g, 2.43 mmol). Then, the reaction mixture was stirred at 80°C for 16 hours, and the reaction was monitored by TLC. The reaction mixture was poured into 1.0 L of water and 1.0 L of NH ₄ Cl saturated aqueous solution. The suspension was stirred at room temperature for 1 hour. The solid was filtered, washed with a saturated aqueous solution of NH ₄ Cl (2×300 mL) and water (2×300 mL) and dried to give the title compound (2.5 g, crude) as an off-white solid. MS: 334.51 (M+H) ⁺ . Step C A suspension of the title compound from Step B above (2 g, 5.98 mmol) in 70% H ₂ SO ₄ (6, 3.0 vol) was heated at 120°C for 2 hours. The reaction mixture was cooled to room temperature, and the reaction mixture was slowly poured into 100 mL of cold water (0°C). Then, the reaction mixture was adjusted to basic pH by adding 50% NaOH aqueous solution. Then, the compound was extracted with EtOAc (6×150 mL). The combined organic layer was dried over Na ₂ SO ₄ and filtered, and then the solution was concentrated to give the title compound (0.3 g, 23%) as a pale yellow solid. MS: 230.4 (M) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.27-8.31 (m, 2H), 8.11 (s, 2H). Step D To the suspension of the title compound (0.3 mg, 1.3 mmol) from step C above in acetonitrile (5 mL) was added tert-butyl nitrite ( 0.2 ml) at 0°C using a syringe over a period of 10 min , 1.95 mmol). Then, copper(II) chloride (0.2 g, 1.56 mmol) was added in portions. After 30 minutes at 0°C, the reaction mixture was warmed to room temperature for 1 hour and heated to 65°C, and then stirred for 4 hours. The progress of the reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated and the product was diluted with water (20 mL) and 5% MeOH/DCM (3×20 mL). The combined organics were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude compound was purified by silica gel (60-120) column chromatography using 1% methanol/dichloromethane for leaching to obtain the title compound (0.15 g, 46%) as an off-white solid. MS: 250.9 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.91 (d, J = 2.40 Hz, 1H), 8.82 (d, J = 1.60 Hz, 1H). Step E To a solution of the title compound (0.18 g, 0.72 mmol) from anhydrous dichloromethane (5 mL) from step D above was added triethylamine (0.3 mL, 2.16 mmol) and morpholine (0.074 g, 0.86 mmol ), and the mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated under vacuum. The crude compound was purified by silica gel (60-120) column chromatography using petroleum ether/ethyl acetate for leaching to obtain the title compound (0.18 g, 83%) as a pale yellow solid. MS: 300.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.49 (d, J = 2.00 Hz, 1H), 8.38 (d, J = 1.60 Hz, 1H), 3.72-3.74 (m, 4H), 3.61- 3.62 (m, 4H).

步驟 A 將2-溴-6-氯吡啶-3-胺(5 g, 24.1 mmol)及硫氰酸鉀(7 g, 72.3 mmol)於乙醇(50 mL)中之溶液及濃鹽酸(37%, 100 mL)在100℃下攪拌40-45 h。藉由TLC確認反應完成(PE/EA = 7.5/2.5)。將反應混合物冷卻至室溫並濃縮以提供棕色固體，將其在二氯甲烷(150 mL)及1 N NaOH水溶液(50 mL)中分配。將固體過濾並乾燥，得到呈淺黃色固體之標題化合物(3.5 g，79%產率)。 MS: 186.1 (M+H)⁺ 。步驟 B 在0℃下利用注射器經10 min之時期向來自上文步驟A之標題化合物(1.5 g, 8.08 mmol)於乙腈(25 mL)中之懸浮液添加亞硝酸第三丁基酯( 1.4 ml, 12.12 mmol)。然後，逐份添加溴化銅(II)(2.16 g, 9.69 mmol)。在0℃下30分鐘後，使反應混合物升溫至室溫持續2分鐘。藉由TLC監測反應之進展。在反應完成後，將溶劑蒸發，且用水(20 mL)及5% MeOH/DCM (3 × 20 mL)稀釋混合物。將合併之有機物用鹽水(10 mL)洗滌，經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。藉由矽膠(60-120)管柱層析且利用1%甲醇/二氯甲烷進行溶析來純化粗製化合物，得到呈淡黃色固體之標題化合物(0.65 g, 32%)。 MS: 248.5 (M+H)⁺ 。步驟 C 向來自上文步驟B之標題化合物(0.65 g, 2.61 mmol)於無水二氯甲烷(5 mL)中之溶液添加三乙胺(1.1 mL, 7.83 mmol)及嗎啉(0.34 g, 3.91 mmol)，且將混合物在室溫下攪拌6小時。將反應混合物在真空下濃縮。藉由矽膠(60-120)管柱層析，利用石油醚/乙酸乙酯進行溶析來純化粗製化合物，得到呈灰黃色固體之標題化合物(0.6 g, 90%)。 MS: 256.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.83 (d,J = 8.40 Hz, 1H), 7.41 (d,J = 8.44 Hz, 1H), 3.72-3.74 (m, 2H), 3.59-3.60 (m, 2H)。 Preparation Example 27

Step A: A solution of 2-bromo-6-chloropyridin-3-amine (5 g, 24.1 mmol) and potassium thiocyanate (7 g, 72.3 mmol) in ethanol (50 mL) and concentrated hydrochloric acid (37%, 100 mL) was stirred at 100°C for 40-45 h. Confirm the completion of the reaction by TLC (PE/EA = 7.5/2.5). The reaction mixture was cooled to room temperature and concentrated to provide a brown solid, which was partitioned between dichloromethane (150 mL) and 1 N aqueous NaOH (50 mL). The solid was filtered and dried to give the title compound (3.5 g, 79% yield) as a light yellow solid. MS: 186.1 (M+H) ⁺ . Step B: Add a third butyl nitrite ( 1.4 ml) to the suspension of the title compound (1.5 g, 8.08 mmol) in acetonitrile (25 mL) from Step A above using a syringe at 0°C over a period of 10 min. , 12.12 mmol). Then, copper (II) bromide (2.16 g, 9.69 mmol) was added portionwise. After 30 minutes at 0°C, the reaction mixture was allowed to warm to room temperature for 2 minutes. The progress of the reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated, and the mixture was diluted with water (20 mL) and 5% MeOH/DCM (3×20 mL). The combined organics were washed with brine (10 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude compound was purified by silica gel (60-120) column chromatography and leaching with 1% methanol/dichloromethane to obtain the title compound (0.65 g, 32%) as a pale yellow solid. MS: 248.5 (M+H) ⁺ . Step C To the solution of the title compound (0.65 g, 2.61 mmol) from anhydrous dichloromethane (5 mL) from step B above was added triethylamine (1.1 mL, 7.83 mmol) and morpholine (0.34 g, 3.91 mmol ), and the mixture was stirred at room temperature for 6 hours. The reaction mixture was concentrated under vacuum. The crude compound was purified by silica gel (60-120) column chromatography using petroleum ether/ethyl acetate for leaching to obtain the title compound (0.6 g, 90%) as a pale yellow solid. MS: 256.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.83 (d, J = 8.40 Hz, 1H), 7.41 (d, J = 8.44 Hz, 1H), 3.72-3.74 (m, 2H), 3.59- 3.60 (m, 2H).

步驟 A 使2,7-二溴-[1,2,4]三唑并[1,5-a]吡啶(0.4 g, 1.44 mmol)及嗎啉(4 mL)之混合物在N₂ 氣氛下回流4小時。將反應混合物濃縮至乾燥。藉由矽膠(60-120)管柱層析，利用石油醚/乙酸乙酯10%-100%作為溶析液進行溶析來純化粗製化合物，得到呈白色固體之標題化合物(0.27 g, 66%)。 MS: 285.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.64 (d,J = 7.08 Hz, 1H), 7.83 (s, 1H), 7.13-7.14 (m, 1H), 3.69-3.71 (m, 2H), 3.45-3.46 (m, 2H)。 Preparation Example 28

Step A: A mixture of 2,7-dibromo-[1,2,4]triazolo[1,5-a]pyridine (0.4 g, 1.44 mmol) and morpholine (4 mL) was refluxed under N ₂ atmosphere 4 hours. The reaction mixture was concentrated to dryness. The crude compound was purified by silica gel (60-120) column chromatography using petroleum ether/ethyl acetate 10%-100% as the eluent to obtain the title compound (0.27 g, 66%) as a white solid ). MS: 285.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.64 (d, J = 7.08 Hz, 1H), 7.83 (s, 1H), 7.13-7.14 (m, 1H), 3.69-3.71 (m, 2H ), 3.45-3.46 (m, 2H).

將3,6-二溴嗒嗪(0.2 g, 0.841 mmol)溶解於乙腈(2.5 mL)中。然後添加嗎啉(0.110 mL, 1.261 mmol)及三乙胺(0.176 mL, 1.261 mmol)，且將懸浮液於微波中在160℃下輻照1 h及20分鐘。用二氯甲烷及水稀釋反應混合物。分離有機層，且將水層用二氯甲烷萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(100/0 -＞ 0/100)，在矽膠管柱上純化粗產物，得到呈白色固體之標題化合物(0.168 g, 82%)。 MS: 245.2 (M+H)⁺ 。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.35 (d,J = 9.5 Hz, 1H), 6.79 (d,J = 9.5 Hz, 1H), 3.88 - 3.79 (m, 4H), 3.66 - 3.55 (m, 4H)。 Preparation Example 29

3,6-Dibromopyridazine (0.2 g, 0.841 mmol) was dissolved in acetonitrile (2.5 mL). Then morpholine (0.110 mL, 1.261 mmol) and triethylamine (0.176 mL, 1.261 mmol) were added, and the suspension was irradiated in the microwave at 160° C. for 1 h and 20 minutes. The reaction mixture was diluted with dichloromethane and water. The organic layer was separated, and the aqueous layer was extracted twice with dichloromethane. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Using the Biotage Isolera One purification system with n-heptane/ethyl acetate gradient (100/0 -> 0/100), the crude product was purified on a silica gel column to give the title compound as a white solid (0.168 g, 82%) . MS: 245.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.35 (d, J = 9.5 Hz, 1H), 6.79 (d, J = 9.5 Hz, 1H), 3.88-3.79 (m, 4H), 3.66-3.55 (m, 4H).

在氮下在0℃下向氫化鈉(0.0404 g, 1.682 mmol)於無水THF (3 mL)中之懸浮液添加嗎啉(0.146 mL, 1.682 mmol)。將反應混合物在0℃下攪拌15分鐘，然後添加溶解於無水THF (3 mL)中之2,5-二溴吡嗪(0.400 g, 1.682 mmol)。將反應混合物在回流條件下攪拌過夜。用水使其淬滅且將產物用EtOAc萃取三次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(100/0 -＞ 0/100)，在矽膠管柱上純化粗產物，得到呈白色固體之標題化合物(0.245 g, 60%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 8.15 (d,J = 1.5 Hz, 1H), 7.86 (d,J = 1.5 Hz, 1H), 3.89 - 3.76 (m, 4H), 3.60 - 3.45 (m, 4H)。 Preparation Example 30

To a suspension of sodium hydride (0.0404 g, 1.682 mmol) in anhydrous THF (3 mL) was added morpholine (0.146 mL, 1.682 mmol) under nitrogen at 0 °C. The reaction mixture was stirred at 0°C for 15 minutes, and then 2,5-dibromopyrazine (0.400 g, 1.682 mmol) dissolved in anhydrous THF (3 mL) was added. The reaction mixture was stirred at reflux overnight. It was quenched with water and the product was extracted three times with EtOAc. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Using the Biotage Isolera One purification system with n-heptane/ethyl acetate gradient (100/0 -> 0/100), the crude product was purified on a silica gel column to give the title compound as a white solid (0.245 g, 60%) . ¹ H-NMR (400 MHz, chloroform- d ) δ = 8.15 (d, J = 1.5 Hz, 1H), 7.86 (d, J = 1.5 Hz, 1H), 3.89-3.76 (m, 4H), 3.60-3.45 (m, 4H).

步驟 A 在120℃下向2-氯-5-硝基-4-氰硫基嘧啶(10 g, 46.2 mmol)於乙酸(264 mL, 4617 mmol)中之熱溶液添加鐵(15.47 g, 277 mmol)。將混合物在相同溫度下攪拌1小時。使該混合物冷卻至室溫且藉由過濾去除不溶性材料並用乙酸洗滌。將濾液在減壓下濃縮並溶解於THF (200 mL)及EtOAc (400 mL)中，且用飽和NH₄ Cl水溶液(100 mL)、飽和NaHCO₃ 溶液(100 mL)洗滌，經Na₂ SO₄ 乾燥且在減壓下去除溶劑，得到標題化合物(6.82 g, 79%)。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 9.42 (s, 1H)。步驟 B 在0℃下經30分鐘向來自上文步驟A之標題化合物(8.2 g, 43 mmol)於乙腈(20 mL)中之懸浮液添加亞硝酸第三丁基酯(6.8 g, 65.9 mmol)。然後，分多次添加溴化銅(II)(11.78 g, 52.8 mmol)。在0℃下30分鐘後，使反應混合物升溫至室溫並攪拌12小時。添加水(50 mL)及EtOAc (100 mL)且分離各相。將水層用EtOAc萃取兩次且將有機層合併，經Na₂ SO₄ 乾燥且將溶劑在減壓下蒸發。使用Biotage Isolera One純化系統，採用EtOAc/正庚烷(20/80)在HP-Sil SNAP柱上純化殘餘物，得到標題化合物(4.94 g, 45%)。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.54 (s, 1H), 8.30 (s, 2H)。步驟 C 將來自上文步驟B之標題化合物(0.157 g, 0.627 mmol)溶解於嗎啉(3 mL, 0.627 mmol)中，且在室溫下攪拌2 h。用二氯甲烷及水稀釋反應混合物。分離有機層，且將水層用二氯甲烷萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(100/0 -＞ 0/100)，在矽膠管柱上純化粗產物，得到呈白色固體之標題化合物(0.048 g, 30%)。 MS: 258.6 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.71 (s, 1H), 3.74 (dd,J = 6.0, 3.6 Hz, 4H), 3.70 - 3.58 (m, 4H)。 Preparation Example 31

Step A Add iron (15.47 g, 277 mmol) to a hot solution of 2-chloro-5-nitro-4-cyanothiopyrimidine (10 g, 46.2 mmol) in acetic acid (264 mL, 4617 mmol) at 120°C ). The mixture was stirred at the same temperature for 1 hour. The mixture was allowed to cool to room temperature and insoluble materials were removed by filtration and washed with acetic acid. The filtrate was concentrated under reduced pressure and dissolved in THF (200 mL) and EtOAc (400 mL), and washed with saturated aqueous NH ₄ Cl solution (100 mL), saturated NaHCO ₃ solution (100 mL), and washed with Na ₂ SO ₄ Dry and remove the solvent under reduced pressure to give the title compound (6.82 g, 79%). ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.42 (s, 1H). Step B To a suspension of the title compound (8.2 g, 43 mmol) in acetonitrile (20 mL) from Step A above was added tert-butyl nitrite (6.8 g, 65.9 mmol) at 0°C for 30 minutes . Then, copper (II) bromide (11.78 g, 52.8 mmol) was added in portions. After 30 minutes at 0°C, the reaction mixture was warmed to room temperature and stirred for 12 hours. Water (50 mL) and EtOAc (100 mL) were added and the phases were separated. The aqueous layer was extracted twice with EtOAc and the organic layers were combined, dried over Na ₂ SO ₄ and the solvent was evaporated under reduced pressure. Using the Biotage Isolera One purification system, the residue was purified on an HP-Sil SNAP column using EtOAc/n-heptane (20/80) to give the title compound (4.94 g, 45%). ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.54 (s, 1H), 8.30 (s, 2H). Step C The title compound (0.157 g, 0.627 mmol) from Step B above was dissolved in morpholine (3 mL, 0.627 mmol) and stirred at room temperature for 2 h. The reaction mixture was diluted with dichloromethane and water. The organic layer was separated, and the aqueous layer was extracted twice with dichloromethane. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Using the Biotage Isolera One purification system with n-heptane/ethyl acetate gradient (100/0 -> 0/100), the crude product was purified on a silica gel column to give the title compound (0.048 g, 30%) as a white solid . MS: 258.6 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.71 (s, 1H), 3.74 (dd, J = 6.0, 3.6 Hz, 4H), 3.70-3.58 (m, 4H).

在氮下在0℃下向氫化鈉(0.077 g, 3.21 mmol)於無水THF (5 mL)中之懸浮液添加(1R ,5S )-3-氧雜-8-氮雜二環[3.2.1]辛烷鹽酸鹽(0.457 g, 3.06 mmol)。將反應混合物攪拌15分鐘，然後添加溶解於無水THF (5 mL)中之2,5-二溴吡嗪(0.800 g, 3.36 mmol)。將反應混合物在回流條件下攪拌過夜。用水使反應混合物淬滅且將產物用乙酸乙酯萃取三次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(100/0 -＞ 0/100)，在矽膠管柱上純化粗產物，得到呈白色固體之標題化合物(0.038 mg, 5%)。 MS: 271.8 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.24 (d,J = 1.4 Hz, 1H), 8.11 (d,J = 1.4 Hz, 1H), 4.49 (s, 2H), 3.62 (d,J = 10.9 Hz, 2H), 3.52 (d,J = 11.0 Hz, 2H), 2.02 - 1.95 (m, 2H), 1.91 (m, 2H)。 Preparation Example 32

To a suspension of sodium hydride (0.077 g, 3.21 mmol) in anhydrous THF (5 mL) was added (1 R ,5 S )-3-oxa-8-azabicyclo[3.2 .1] Octane hydrochloride (0.457 g, 3.06 mmol). The reaction mixture was stirred for 15 minutes, and then 2,5-dibromopyrazine (0.800 g, 3.36 mmol) dissolved in anhydrous THF (5 mL) was added. The reaction mixture was stirred at reflux overnight. The reaction mixture was quenched with water and the product was extracted three times with ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Using the Biotage Isolera One purification system with n-heptane/ethyl acetate gradient (100/0 -> 0/100), the crude product was purified on a silica gel column to give the title compound (0.038 mg, 5%) as a white solid . MS: 271.8 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.24 (d, J = 1.4 Hz, 1H), 8.11 (d, J = 1.4 Hz, 1H), 4.49 (s, 2H), 3.62 (d, J = 10.9 Hz, 2H), 3.52 (d, J = 11.0 Hz, 2H), 2.02-1.95 (m, 2H), 1.91 (m, 2H).

步驟 A 將乙酸鈀(II)(0.00651 g, 0.029 mmol)及2-二環己基膦基-2′,4′,6′-三異丙基聯苯(XPhos) (0.0415 g, 0.087 mmol)置於反應小瓶中，且添加經脫氣之1,4-二噁烷(2 mL)。將所得溶液短暫脫氣。將懸浮液在100℃下(在預加熱之加熱塊上)加熱少於1分鐘，直至溶液之色彩自橙色變為深粉色為止。然後，將小瓶自加熱塊移除，且添加來自製備實例1之標題化合物(0.110 g, 0.290 mmol)及來自製備實例29之標題化合物(0.078 g, 0.319 mmol)及碳酸銫(0.331 g, 1.015 mmol)。將反應小瓶填充氬，之後將其關閉。將反應混合物在100℃下加熱12 h。用乙酸乙酯及水稀釋該反應混合物。分離有機層，且將水層用乙酸乙酯再萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(90/10 -＞ 0/100)，在矽膠管柱上純化粗產物，得到呈黃色固體之標題化合物(0.072 g, 49%)。 MS: 508.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 8.03 (dd,J = 9.1, 4.5 Hz, 1H), 7.70 (d,J = 8.4 Hz, 2H), 7.42 (d,J = 9.9 Hz, 1H), 7.38 (dd,J = 8.9, 2.6 Hz, 1H), 7.32 (dd,J = 9.0, 6.1 Hz, 2H), 7.17 (td,J = 9.2, 2.6 Hz, 1H), 6.93 (s, 1H), 4.55 (s, 2H), 3.88 (t,J = 5.6 Hz, 2H), 3.78 - 3.65 (m, 4H), 3.41 - 3.34 (m, 4H), 3.21 - 3.14 (m, 2H), 2.30 (s, 3H)。 Preparation Example 33

Step A: Place palladium(II) acetate (0.00651 g, 0.029 mmol) and 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (XPhos) (0.0415 g, 0.087 mmol) In a reaction vial, and degassed 1,4-dioxane (2 mL) was added. The resulting solution was briefly degassed. The suspension was heated at 100°C (on the preheated heating block) for less than 1 minute until the color of the solution changed from orange to dark pink. Then, the vial was removed from the heating block, and the title compound from Preparation Example 1 (0.110 g, 0.290 mmol) and the title compound from Preparation Example 29 (0.078 g, 0.319 mmol) and cesium carbonate (0.331 g, 1.015 mmol) were added ). The reaction vial was filled with argon and then closed. The reaction mixture was heated at 100 °C for 12 h. The reaction mixture was diluted with ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice more with ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. Using the Biotage Isolera One purification system with n-heptane/ethyl acetate gradient (90/10 -> 0/100), the crude product was purified on a silica gel column to give the title compound as a yellow solid (0.072 g, 49%) . MS: 508.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.03 (dd, J = 9.1, 4.5 Hz, 1H), 7.70 (d, J = 8.4 Hz, 2H), 7.42 (d, J = 9.9 Hz, 1H), 7.38 (dd, J = 8.9, 2.6 Hz, 1H), 7.32 (dd, J = 9.0, 6.1 Hz, 2H), 7.17 (td, J = 9.2, 2.6 Hz, 1H), 6.93 (s, 1H ), 4.55 (s, 2H), 3.88 (t, J = 5.6 Hz, 2H), 3.78-3.65 (m, 4H), 3.41-3.34 (m, 4H), 3.21-3.14 (m, 2H), 2.30 ( s, 3H).

Preparation Examples 34 to 41f In addition to using the tricyclic amino derivatives and bromine/chlorine derivatives indicated in the following table, following the palladium coupling procedure as explained in Preparation Example 33, the following compounds were prepared: Table 1

步驟 A 將乙酸鈀(II)(0.0045 g, 0.020 mmol)及4,5-雙(二苯基膦基)-9,9-二甲基𠮿

(XANTPHOS) (0.035 g, 0.061 mmol)置於反應小瓶中，且添加經脫氣之1,4-二噁烷(4 mL)。將所得溶液短暫脫氣。將懸浮液在100℃下(在預加熱之加熱塊上)加熱少於1分鐘，直至溶液之色彩自橙色變為深粉色為止。然後，將小瓶自加熱塊移除，且添加來自製備實例1步驟E之標題化合物(70 mg, 0.203 mmol)及來自製備實例26之標題化合物(0.067 g, 0.224 mmol)及碳酸銫(0.232 g, 0.771 mmol)。將反應小瓶填充氬，之後將其關閉。將反應混合物在100℃下加熱18 h。用乙酸乙酯及水稀釋該反應混合物。分離有機層，且將水層用乙酸乙酯再萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發。在矽膠管柱上使用Biotage Isolera One純化系統，採用二氯甲烷/甲醇(100/00 -＞ 95/05)純化粗產物，得到呈黃色固體之標題化合物(0.052 g, 45%)。¹ H NMR (400 MHz，氯仿-d ) δ = 8.24 (d,J = 2.7 Hz, 1H), 8.16 - 8.03 (m, 1H), 7.64 - 7.59 (m, 2H), 7.55 (d,J = 2.7 Hz, 1H), 7.18 (d,J = 8.2 Hz, 2H), 7.02 (dd,J = 8.6, 1.9 Hz, 2H), 4.23 (d,J = 2.0 Hz, 2H), 3.84 - 3.78 (m, 4H), 3.69 - 3.63 (m, 4H), 3.59 (t,J = 5.6 Hz, 2H), 3.48 (s, 1H), 3.25 (dq,J = 5.7, 3.5, 2.8 Hz, 2H), 2.33 (s, 3H)。 Preparation Example 42

Step A: Palladium(II) acetate (0.0045 g, 0.020 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethyl 𠮿

(XANTPHOS) (0.035 g, 0.061 mmol) was placed in the reaction vial, and degassed 1,4-dioxane (4 mL) was added. The resulting solution was briefly degassed. The suspension was heated at 100°C (on the preheated heating block) for less than 1 minute until the color of the solution changed from orange to dark pink. Then, the vial was removed from the heating block, and the title compound from Preparation Example 1, Step E (70 mg, 0.203 mmol) and the title compound from Preparation Example 26 (0.067 g, 0.224 mmol) and cesium carbonate (0.232 g, 0.771 mmol). The reaction vial was filled with argon and then closed. The reaction mixture was heated at 100 °C for 18 h. The reaction mixture was diluted with ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice more with ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. Using a Biotage Isolera One purification system on a silica gel column, the crude product was purified using dichloromethane/methanol (100/00 -> 95/05) to obtain the title compound (0.052 g, 45%) as a yellow solid. ¹ H NMR (400 MHz, chloroform- d ) δ = 8.24 (d, J = 2.7 Hz, 1H), 8.16-8.03 (m, 1H), 7.64-7.59 (m, 2H), 7.55 (d, J = 2.7 Hz, 1H), 7.18 (d, J = 8.2 Hz, 2H), 7.02 (dd, J = 8.6, 1.9 Hz, 2H), 4.23 (d, J = 2.0 Hz, 2H), 3.84-3.78 (m, 4H ), 3.69-3.63 (m, 4H), 3.59 (t, J = 5.6 Hz, 2H), 3.48 (s, 1H), 3.25 (dq, J = 5.7, 3.5, 2.8 Hz, 2H), 2.33 (s, 3H).

步驟 A 將乙酸鈀(II)(0.0045 g, 0.020 mmol)及4,5-雙(二苯基膦基)-9,9-二甲基𠮿

(XANTPHOS) (0.035 g, 0.061 mmol)置於反應小瓶中，且添加經脫氣之1,4-二噁烷(4 mL)。將所得溶液短暫脫氣。將懸浮液在100℃下(在預加熱之加熱塊上)加熱少於1分鐘，直至溶液之色彩自橙色變為深粉色為止。然後，將小瓶自加熱塊移除，且添加來自製備實例1步驟E之標題化合物(70 mg, 0.203 mmol)及來自製備實例27之標題化合物 (0.067 g, 0.224 mmol)及碳酸銫(0.232 g, 0.771 mmol)。將反應小瓶填充氬，之後將其關閉。將反應混合物在100℃下加熱18 h。用乙酸乙酯及水稀釋該反應混合物。分離有機層，且將水層用乙酸乙酯再萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發。使用Biotage Isolera One純化系統，採用二氯甲烷/甲醇(100/00 -＞ 95/05)，在矽膠管柱上純化粗產物，得到呈黃色固體之標題化合物(0.048 g, 42%)。¹ H NMR (400 MHz，氯仿-d ) δ = 8.10 (dd,J = 9.0, 4.4 Hz, 1H), 7.65 (d,J = 8.9 Hz, 1H), 7.63 - 7.57 (m, 2H), 7.15 (d,J = 8.2 Hz, 2H), 7.09 - 6.97 (m, 2H), 6.75 (d,J = 8.9 Hz, 1H), 4.51 (d,J = 2.0 Hz, 2H), 3.94 (t,J = 5.6 Hz, 2H), 3.82 (q,J = 5.2 Hz, 4H), 3.57 (dd,J = 5.8, 4.0 Hz, 4H), 3.25 (td,J = 5.5, 2.7 Hz, 2H), 2.31 (s, 3H)。 Preparation Example 43

Step A: Palladium(II) acetate (0.0045 g, 0.020 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethyl 𠮿

(XANTPHOS) (0.035 g, 0.061 mmol) was placed in the reaction vial, and degassed 1,4-dioxane (4 mL) was added. The resulting solution was briefly degassed. The suspension was heated at 100°C (on the preheated heating block) for less than 1 minute until the color of the solution changed from orange to dark pink. Then, the vial was removed from the heating block, and the title compound (70 mg, 0.203 mmol) from Preparation Example 1, Step E and the title compound (0.067 g, 0.224 mmol) from Preparation Example 27 and cesium carbonate (0.232 g, 0.771 mmol). The reaction vial was filled with argon and then closed. The reaction mixture was heated at 100 °C for 18 h. The reaction mixture was diluted with ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice more with ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. The crude product was purified on a silica gel column using a Biotage Isolera One purification system using dichloromethane/methanol (100/00 -> 95/05) to obtain the title compound (0.048 g, 42%) as a yellow solid. ¹ H NMR (400 MHz, chloroform- d ) δ = 8.10 (dd, J = 9.0, 4.4 Hz, 1H), 7.65 (d, J = 8.9 Hz, 1H), 7.63-7.57 (m, 2H), 7.15 ( d, J = 8.2 Hz, 2H), 7.09-6.97 (m, 2H), 6.75 (d, J = 8.9 Hz, 1H), 4.51 (d, J = 2.0 Hz, 2H), 3.94 (t, J = 5.6 Hz, 2H), 3.82 (q, J = 5.2 Hz, 4H), 3.57 (dd, J = 5.8, 4.0 Hz, 4H), 3.25 (td, J = 5.5, 2.7 Hz, 2H), 2.31 (s, 3H ).

步驟 A 將3,6-二溴嗒嗪(0.2 g, 0.841 mmol)溶解於乙腈(2.5 mL)中。然後添加8-氧雜-3-氮雜二環[3.2.1]辛烷(0.105 g, 0.925 mmol)及三乙胺(0.176 mL, 1.261 mmol)，且將懸浮液於微波中輻照至160℃持續1 h及20分鐘。用二氯甲烷及水稀釋反應混合物。分離有機層，且將水層用二氯甲烷萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(100/0 -＞ 0/100)，在矽膠管柱上純化粗產物，得到呈米色固體之標題化合物(0.152 g, 67%)。 MS: 271.5 (M+H)⁺ 。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.32 (d,J = 9.5 Hz, 1H), 6.70 (d,J = 9.5 Hz, 1H), 4.60 - 4.43 (m, 2H), 3.83 (d,J = 12.8 Hz, 2H), 3.23 (dd,J = 12.4, 2.7 Hz, 2H), 2.08 - 1.94 (m, 2H), 1.90 - 1.76 (m, 2H)。 Preparation Example 44

Step A Dissolve 3,6-dibromopyridazine (0.2 g, 0.841 mmol) in acetonitrile (2.5 mL). Then 8-oxa-3-azabicyclo[3.2.1]octane (0.105 g, 0.925 mmol) and triethylamine (0.176 mL, 1.261 mmol) were added, and the suspension was irradiated in the microwave to 160 ℃ for 1 h and 20 minutes. The reaction mixture was diluted with dichloromethane and water. The organic layer was separated, and the aqueous layer was extracted twice with dichloromethane. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. Using the Biotage Isolera One purification system with n-heptane/ethyl acetate gradient (100/0 -> 0/100), the crude product was purified on a silica gel column to give the title compound as a beige solid (0.152 g, 67%) . MS: 271.5 (M+H) ⁺ . ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.32 (d, J = 9.5 Hz, 1H), 6.70 (d, J = 9.5 Hz, 1H), 4.60-4.43 (m, 2H), 3.83 (d , J = 12.8 Hz, 2H), 3.23 (dd, J = 12.4, 2.7 Hz, 2H), 2.08-1.94 (m, 2H), 1.90-1.76 (m, 2H).

步驟 A 向2,6-二氯苯并[d ]噁唑之溶液(1.2 g, 6.3 mmol)添加於無水乙醇(8 mL)中之33 wt.%甲胺溶液，且使用Biotage微波爐將混合物在100℃下加熱30分鐘。使反應混合物冷卻至室溫，且溶解於二氯甲烷(150 mL)中。將有機相用水、1 N NaOH溶液及鹽水洗滌，且然後經Na₂ SO₄ 乾燥。將溶劑去除，且獲得標題化合物(1.08 g, 94%)。¹ H NMR (400 MHz，氯仿-d ) δ 7.30 - 7.24 (m, 2H), 7.16 (dd,J = 8.4, 1.9 Hz, 1H), 5.35 (s, 1H), 3.14 (s, 3H)。步驟 B 在0℃下向來自上文步驟A之標題化合物(39 mg, 1.643 mmol)於無水THF (1.5 mL)中之攪拌懸浮液緩慢添加6-氯-N-甲基苯并[d]噁唑-2-胺(100 mg, 0.548 mmol)於無水THF (2 mL)中之溶液，且在相同溫度下攪拌30 min。然後在0℃下逐滴添加4-甲苯-1-磺醯氯(107 mg, 0.561 mmol)於無水THF (1.5 mL)中之溶液，且將反應混合物在0℃下攪拌2 h。利用冰水(4 mL)將反應混合物在0℃下淬滅。然後將產物用乙酸乙酯萃取三次。將合併之有機萃取物用水、鹽水洗滌，經Na₂ SO₄ 乾燥，過濾並在減壓下蒸發，得到呈米色固體之標題化合物(154 mg, 83%)。 MS: 337.14 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 7.93 - 7.89 (m, 2H), 7.88 (d,J = 2.0 Hz, 1H), 7.58 (d,J = 8.5 Hz, 1H), 7.49 - 7.44 (m, 2H), 7.36 (dd,J = 8.5, 2.0 Hz, 1H), 3.49 (s, 3H), 2.39 (s, 3H)。 Preparation Example 45

Step A To a solution of 2,6-dichlorobenzo[ d ]oxazole (1.2 g, 6.3 mmol) was added a 33 wt.% methylamine solution in absolute ethanol (8 mL), and the mixture was placed in a Biotage microwave oven. Heat at 100°C for 30 minutes. The reaction mixture was cooled to room temperature and dissolved in dichloromethane (150 mL). The organic phase was washed with water, 1 N NaOH solution and brine, and then dried over Na ₂ SO ₄ . The solvent was removed, and the title compound (1.08 g, 94%) was obtained. ¹ H NMR (400 MHz, chloroform- d ) δ 7.30-7.24 (m, 2H), 7.16 (dd, J = 8.4, 1.9 Hz, 1H), 5.35 (s, 1H), 3.14 (s, 3H). Step B To a stirred suspension of the title compound (39 mg, 1.643 mmol) in anhydrous THF (1.5 mL) from Step A above was slowly added 6-chloro-N-methylbenzo[d]oxo at 0°C. A solution of oxazol-2-amine (100 mg, 0.548 mmol) in anhydrous THF (2 mL) and stirred at the same temperature for 30 min. Then a solution of 4-toluene-1-sulfonyl chloride (107 mg, 0.561 mmol) in anhydrous THF (1.5 mL) was added dropwise at 0°C, and the reaction mixture was stirred at 0°C for 2 h. The reaction mixture was quenched with ice water (4 mL) at 0°C. The product was then extracted three times with ethyl acetate. The combined organic extracts were washed with water, brine, dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give the title compound (154 mg, 83%) as a beige solid. MS: 337.14 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.93-7.89 (m, 2H), 7.88 (d, J = 2.0 Hz, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.49- 7.44 (m, 2H), 7.36 (dd, J = 8.5, 2.0 Hz, 1H), 3.49 (s, 3H), 2.39 (s, 3H).

步驟 A 在0℃下向NaH (0.47 g, 19.55 mmol)於THF (20 mL)中之懸浮液逐滴添加市售1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(2.0 g, 7.35 mmol) (溶解於THF中)。然後將混合物在室溫下攪拌1 h。之後，在0℃下添加甲苯磺醯氯(1.5 g, 7.82 mmol) (溶解於THF中)，且然後將混合物在室溫下攪拌2 h。如藉由TLC所檢查在反應完成後，利用冰水將反應混合物淬滅，之後使用乙酸乙酯進行萃取。將有機層濃縮，得到標題化合物(1.8 g, 65%)。產物原樣用於下一步驟。 MS: 427.1 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(1.8 g, 4.22 mmol)於DCM (20 mL)中之溶液添加於二噁烷中之4 M HCl (5 mL)。將反應混合物攪拌過夜。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌殘餘物，得到呈灰白色固體之標題化合物(1.0 g, 66%)。 MS: 361.3 (M+H)⁺ 。 Preparation Example 46

Step A To a suspension of NaH (0.47 g, 19.55 mmol) in THF (20 mL) was added commercially available 1,3,4,5-tetrahydro-2H-pyrido[4,3- b] Tert-butyl indole-2-carboxylic acid (2.0 g, 7.35 mmol) (dissolved in THF). The mixture was then stirred at room temperature for 1 h. After that, tosyl chloride (1.5 g, 7.82 mmol) (dissolved in THF) was added at 0°C, and then the mixture was stirred at room temperature for 2 h. After completion of the reaction as checked by TLC, the reaction mixture was quenched with ice water, and then extracted with ethyl acetate. The organic layer was concentrated to give the title compound (1.8 g, 65%). The product was used as is in the next step. MS: 427.1 (M+H) ⁺ . Step B To a solution of the title compound (1.8 g, 4.22 mmol) in DCM (20 mL) from Step A above was added 4 M HCl (5 mL) in dioxane. The reaction mixture was stirred overnight. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and the residue was washed with diethyl ether to obtain the title compound (1.0 g, 66%) as an off-white solid. MS: 361.3 (M+H) ⁺ .

步驟 A 在0℃下向市售(2-氟苯基)肼鹽酸鹽(2 g, 12.34 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(2.45 g, 12.34 mmol)於二噁烷(20 mL)中之溶液添加濃H₂ SO₄ (2 mL)。然後將反應混合物在100℃下加熱4 h。如藉由TLC所證實，在反應完成後，將反應混合物冷卻至25℃且然後濃縮。藉由10% NaOH溶液使粗製混合物鹼化，且將沈澱物過濾出。將固體用水洗滌並在真空下乾燥，得到標題化合物(1.7 g, 75%)。 MS: 191.0 (M+H)⁺ 。步驟 B 在室溫下向來自上文步驟A之標題化合物(1.7g，粗製)於THF (20 mL)中之攪拌溶液添加TEA (3.76 mL, 26.82 mmol)及二碳酸二-第三丁基酯(2.34 mL, 10.73 mmol)。將混合物攪拌12 h。如藉由TLC所證實，在反應完成後，將溶劑去除且使用Biotage Isolera One純化系統，採用EtOAc/庚烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到呈淡黃色固體之標題化合物(700 mg, 27%)。 MS: 291.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 11.38 (bs, 1H), 7.22 (d,J = 7.60 Hz, 1H), 6.96-6.85 (m, 2H), 4.53 (s, 2H), 3.70-3.71 (m, 2H), 2.78 (bs, 2H), 1.44 (s, 9H)。步驟 C 在0℃下向NaH (144 mg, 3.61 mmol)於THF (15 mL)中之懸浮液逐滴添加來自上文步驟B之標題化合物(700 mg, 2.41 mmol) (溶解於THF中)。然後將混合物在室溫下攪拌1 h。之後，在0℃下添加甲苯磺醯氯(549 mg, 2.89 mmol) (溶解於THF中)，且然後將混合物在室溫下攪拌2 h。如藉由TLC所證實，在反應完成後，利用冰水將反應混合物淬滅，之後使用乙酸乙酯進行萃取。將有機層濃縮，且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到標題化合物(900 mg, 84%)。 MS: 345.1 (M-Boc)。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.79 (d,J = 8.04 Hz, 2H), 7.41 (d,J = 8.24 Hz, 2H), 7.34 (d,J = 7.68 Hz, 2H), 7.22-7.23 (m, 1H), 7.07-7.09 (m, 1H), 4.50 (s, 2H), 3.70-3.72 (m, 2H), 3.17 (bs, 2H), 2.36 (s, 3H), 1.40 (s, 9H)。步驟 D 向來自上文步驟C之標題化合物(900 mg, 2.02 mmol)於DCM (10 mL)中之溶液添加於二噁烷中之2 N HCl (5 mL)。將反應混合物在室溫下攪拌2 h。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌殘餘物，得到呈淡棕色固體之標題化合物(450 mg, 65%)。 MS: 345.1 (M+H)⁺ 。 Preparation Example 47

Step A To commercially available (2-fluorophenyl)hydrazine hydrochloride (2 g, 12.34 mmol) and tert-butyl 4-oxohexahydropyridine-1-carboxylate (2.45 g, 12.34) at 0°C A solution of mmol) in dioxane (20 mL) was added concentrated H ₂ SO ₄ (2 mL). The reaction mixture was then heated at 100 °C for 4 h. As confirmed by TLC, after the reaction was completed, the reaction mixture was cooled to 25°C and then concentrated. The crude mixture was basified by 10% NaOH solution and the precipitate was filtered off. The solid was washed with water and dried under vacuum to give the title compound (1.7 g, 75%). MS: 191.0 (M+H) ⁺ . Step B To a stirred solution of the title compound (1.7 g, crude) in THF (20 mL) from Step A above was added TEA (3.76 mL, 26.82 mmol) and di-tert-butyl dicarbonate at room temperature. (2.34 mL, 10.73 mmol). The mixture was stirred for 12 h. As confirmed by TLC, after the reaction was completed, the solvent was removed and the crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/heptane gradient (10/80 => 80/20), The title compound was obtained as a light yellow solid (700 mg, 27%). MS: 291.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.38 (bs, 1H), 7.22 (d, J = 7.60 Hz, 1H), 6.96-6.85 (m, 2H), 4.53 (s, 2H), 3.70-3.71 (m, 2H), 2.78 (bs, 2H), 1.44 (s, 9H). Step C To the suspension of NaH (144 mg, 3.61 mmol) in THF (15 mL) was added dropwise the title compound (700 mg, 2.41 mmol) from Step B above (dissolved in THF) at 0°C. The mixture was then stirred at room temperature for 1 h. After that, tosyl chloride (549 mg, 2.89 mmol) (dissolved in THF) was added at 0°C, and then the mixture was stirred at room temperature for 2 h. As confirmed by TLC, after the reaction was completed, the reaction mixture was quenched with ice water, and then extracted with ethyl acetate. The organic layer was concentrated, and the crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/hexane gradient (10/80 => 80/20) to obtain the title compound (900 mg, 84%) . MS: 345.1 (M-Boc). ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.79 (d, J = 8.04 Hz, 2H), 7.41 (d, J = 8.24 Hz, 2H), 7.34 (d, J = 7.68 Hz, 2H) , 7.22-7.23 (m, 1H), 7.07-7.09 (m, 1H), 4.50 (s, 2H), 3.70-3.72 (m, 2H), 3.17 (bs, 2H), 2.36 (s, 3H), 1.40 (s, 9H). Step D To the solution of the title compound (900 mg, 2.02 mmol) in DCM (10 mL) from Step C above was added 2 N HCl (5 mL) in dioxane. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and the residue was washed with diethyl ether to obtain the title compound (450 mg, 65%) as a light brown solid. MS: 345.1 (M+H) ⁺ .

步驟 A 在0℃下向NaH (300 mg, 12.39 mmol)於THF (15 mL)中之懸浮液逐滴添加來自製備實例47步驟B之標題化合物(1.2 g, 4.13 mmol) (溶解於THF中)。然後將反應混合物在室溫下攪拌1 h。之後，在0℃下添加碘甲烷(0.5 mL, 8.26 mmol)，且然後將混合物在室溫下攪拌2 h。完成之後，利用冰水將反應混合物淬滅，之後使用乙酸乙酯進行萃取。將有機層濃縮，且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到標題化合物(900 mg, 72%)。 MS: 305.3 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.23 (d,J = 9.60 Hz, 1H), 6.88-6.91 (m, 2H), 4.51 (s, 2H), 3.71-3.73 (m, 5H), 2.73-2.77 (m, 2H), 1.50 (s, 9H)。步驟 B 向來自上文步驟A之標題化合物(900 mg, 2.96 mmol)於DCM (10 mL)中之溶液添加於二噁烷中之4 M HCl (5 mL)。將反應混合物在室溫下攪拌2 h。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌殘餘物，得到呈淡棕色固體之標題化合物(500 mg, 83%)。 MS: 205.1 (M+H)⁺ 。 Preparation Example 48

Step A To a suspension of NaH (300 mg, 12.39 mmol) in THF (15 mL) was added dropwise the title compound (1.2 g, 4.13 mmol) from Preparation Example 47 Step B (dissolved in THF) at 0°C . The reaction mixture was then stirred at room temperature for 1 h. After that, iodomethane (0.5 mL, 8.26 mmol) was added at 0°C, and then the mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was quenched with ice water, and then extracted with ethyl acetate. The organic layer was concentrated, and the crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/hexane gradient (10/80 => 80/20) to obtain the title compound (900 mg, 72%) . MS: 305.3 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.23 (d, J = 9.60 Hz, 1H), 6.88-6.91 (m, 2H), 4.51 (s, 2H), 3.71-3.73 (m, 5H ), 2.73-2.77 (m, 2H), 1.50 (s, 9H). Step B To a solution of the title compound (900 mg, 2.96 mmol) in DCM (10 mL) from step A above was added 4 M HCl (5 mL) in dioxane. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and the residue was washed with diethyl ether to obtain the title compound (500 mg, 83%) as a light brown solid. MS: 205.1 (M+H) ⁺ .

步驟 A 向市售2-溴-6-氯-1,8-萘啶(0.2 g, 0.821 mmol)於無水乙腈(5 mL)中之溶液添加碳酸鉀(0.335 mg, 2.46 mmol)及嗎啉(0.11 g, 1.23 mmol)。將反應混合物加熱至100℃持續3小時。將反應混合物在真空下濃縮，且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到呈淡黃色固體之標題化合物(170 mg, 83%)。 MS: 250.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.71 (s, 1H), 8.07-8.08 (m, 1H), 7.82-7.95 (m, 1H), 7.55-7.56 (m, 1H), 3.72 (s, 8H)。 Preparation Example 49

Step A: To a solution of commercially available 2-bromo-6-chloro-1,8-naphthyridine (0.2 g, 0.821 mmol) in anhydrous acetonitrile (5 mL), add potassium carbonate (0.335 mg, 2.46 mmol) and morpholine ( 0.11 g, 1.23 mmol). The reaction mixture was heated to 100°C for 3 hours. The reaction mixture was concentrated under vacuum and the Biotage Isolera One purification system was used to purify the crude reaction mixture on a silica gel column using an EtOAc/hexane gradient (10/80 => 80/20) to give the title as a light yellow solid Compound (170 mg, 83%). MS: 250.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.71 (s, 1H), 8.07-8.08 (m, 1H), 7.82-7.95 (m, 1H), 7.55-7.56 (m, 1H), 3.72 (s, 8H).

步驟 A 向市售2-溴-6-氯-1,7-萘啶(0.5 g, 2.05 mmol)於無水乙腈(5 mL)中之溶液添加碳酸鉀(0.837 mg, 6.16 mmol)及嗎啉(0.27 g, 3.08 mmol)。將反應混合物加熱至100℃持續3小時。將反應混合物在真空下濃縮，且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到呈淡黃色固體之標題化合物(270 mg, 53%)。 MS: 250.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 9.01 (s, 1H), 7.91-7.93 (m, 2H), 6.92-6.93 (m, 1H), 3.71 (s, 8H)。 Preparation Example 50

Step A: To a solution of commercially available 2-bromo-6-chloro-1,7-naphthyridine (0.5 g, 2.05 mmol) in anhydrous acetonitrile (5 mL), add potassium carbonate (0.837 mg, 6.16 mmol) and morpholine ( 0.27 g, 3.08 mmol). The reaction mixture was heated to 100°C for 3 hours. The reaction mixture was concentrated under vacuum and the crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/hexane gradient (10/80 => 80/20) to give the title as a light yellow solid Compound (270 mg, 53%). MS: 250.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 9.01 (s, 1H), 7.91-7.93 (m, 2H), 6.92-6.93 (m, 1H), 3.71 (s, 8H).

步驟 A 向市售2,7-二氯喹啉(0.5 g, 2.52 mmol)於無水DMF (5 mL)中之溶液添加碳酸鉀(1 g, 7.52 mmol)及嗎啉(0.32 g, 3.78 mmol)。將反應混合物加熱至100℃持續3小時。將反應混合物在真空下濃縮，得到呈淡黃色固體之標題化合物(500 mg, 80%)。 MS: 249.1 (M+H)⁺ 。 Preparation Example 51

Step A To a solution of commercially available 2,7-dichloroquinoline (0.5 g, 2.52 mmol) in anhydrous DMF (5 mL) was added potassium carbonate (1 g, 7.52 mmol) and morpholine (0.32 g, 3.78 mmol). The reaction mixture was heated to 100°C for 3 hours. The reaction mixture was concentrated under vacuum to give the title compound (500 mg, 80%) as a light yellow solid. MS: 249.1 (M+H) ⁺ .

步驟 A 向市售2,6-二氯喹啉(0.5 g, 2.52 mmol)於無水DMF (5 mL)中之溶液添加碳酸鉀(1 g, 7.52 mmol)及嗎啉(0.32 g, 3.78 mmol)。將反應混合物加熱至100℃持續3小時。將反應混合物在真空下濃縮，且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(10/80 =＞ 80/20)，在矽膠管柱上純化粗製反應混合物，得到呈淡黃色固體之標題化合物(500 mg, 80%)。 MS: 249.1 (M+H)^{+ 1} H-NMR (400 MHz, DMSO-d₆ ) δ = 8.06 (d,J = 12.40 Hz, 1H), 7.84 (d,J = 2.40 Hz, 1H), 7.51-7.52 (m, 2H), 7.31 (d,J = 12.40 Hz, 1H), 3.66-3.67 (m, 8H)。 Preparation Example 52

Step A To a solution of commercially available 2,6-dichloroquinoline (0.5 g, 2.52 mmol) in anhydrous DMF (5 mL) was added potassium carbonate (1 g, 7.52 mmol) and morpholine (0.32 g, 3.78 mmol). The reaction mixture was heated to 100°C for 3 hours. The reaction mixture was concentrated under vacuum and the Biotage Isolera One purification system was used to purify the crude reaction mixture on a silica gel column using an EtOAc/hexane gradient (10/80 => 80/20) to give the title as a light yellow solid Compound (500 mg, 80%). MS: 249.1 (M+H) ^{+ 1} H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.06 (d, J = 12.40 Hz, 1H), 7.84 (d, J = 2.40 Hz, 1H), 7.51- 7.52 (m, 2H), 7.31 (d, J = 12.40 Hz, 1H), 3.66-3.67 (m, 8H).

步驟 A 將市售4,6-二氯吡啶-3-胺(8.0 g, 49.07 mmol)及異硫氰酸苯甲醯基酯(7.3 mL, 53.98 mmol)於丙酮(120 mL)中之溶液在60℃下攪拌3小時。藉由TLC監測反應。將溶劑蒸發且將固體過濾，用正己烷(100 mL)洗滌並乾燥，得到呈灰白色固體之期望產物(14.0 g, 87%)。 MS: 328.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ =12.39 (s, 1H), 12.02 (s, 1H), 8.74 (s, 1H), 7.98-7.99 (m, 3H), 7.67-7.68 (m, 1H), 7.56 (t,J = 7.60 Hz, 2H)。步驟 B 在0℃向來自上文步驟A之標題化合物(14.0 g, 42.94 mmol)於N-甲基-2-吡咯啶酮(NMP) (70 mL)中之溶液添加甲醇鈉(NaOMe) (4.6 g, 85.88 mmol)。然後將混合物加熱至120℃且繼續攪拌4小時。藉由TLC監測反應。將反應混合物傾倒至冷水(300 mL)中且獲得白色沈澱物。將固體過濾，用水(300 mL)及正己烷(200 mL)洗。將化合物在真空下乾燥6 h，得到呈白色固體之期望產物(14.0 g，粗製)。產物原樣用於下一步驟。 MS: 290.0 (M+H)⁺ 。步驟 C 將來自上文步驟B之標題化合物(14.0 g, 48.4 mmol)於70% H₂ SO₄ (50.0 mL)中之懸浮液在110℃加熱4小時。將反應混合物冷卻至室溫，且將反應混合物緩慢傾倒至200 mL冷水(0℃)中。然後，藉由添加50% NaOH水溶液將反應混合物調整至鹼性pH。然後，用EtOAc (6 × 100 mL)萃取化合物。使合併之有機層經Na₂ SO₄ 乾燥並過濾，然後將溶劑濃縮，得到呈淺黃色固體之期望產物(6 g, 67%)。 MS: 186.1 (M+H)^{+ 1} H-NMR (400 MHz, DMSO-d₆ ) δ = 8.30 (s, 1H), 7.86 (s, 1H)。步驟 D 在0℃利用注射器經10 min之時期向來自上文步驟C之標題化合物(5.0 g, 27.02 mmol)於乙腈(120 mL)中之懸浮液添加亞硝酸第三丁基酯( 4.8 mL, 40.54 mmol)。然後，逐份添加溴化銅(II)(9.0 g, 40.54 mmol)。在0℃下30分鐘後，將反應混合物升溫至室溫2.5小時，藉由TLC監測反應之進展。在反應完成後，將溶劑蒸發且用水(200 mL)及5% MeOH/ DCM (3 × 200 mL)稀釋。將合併之有機物用鹽水(50 mL)洗，經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮，得到呈白色固體之標題化合物(6.5 g)。產物原樣用於下一步驟。 MS: 250.9 (M+H)⁺ 步驟 E 向來自上文步驟D之標題化合物(6.5 g, 26.09 mmol)於無水DCM (100 mL)中之溶液添加三乙胺(11.2 mL, 81.5 mmol)及嗎啉(2.8 mL, 28.13 mmol)。將反應混合物在室溫攪拌4小時。將反應混合物在真空下濃縮。在矽膠管柱上使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(10/80 =＞ 80/20)純化粗製反應混合物，得到呈淡黃色固體之標題化合物(4.7g, 71%)。 MS: 256.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.48 (s, 1H), 8.05 (s, 1H), 3.73-3.74 (m, 4H), 3.60-3.61 (m, 4H)。 Preparation Example 53

Step A: A solution of commercially available 4,6-dichloropyridin-3-amine (8.0 g, 49.07 mmol) and benzyl isothiocyanate (7.3 mL, 53.98 mmol) in acetone (120 mL) Stir at 60°C for 3 hours. The reaction was monitored by TLC. The solvent was evaporated and the solid was filtered, washed with n-hexane (100 mL) and dried to give the desired product (14.0 g, 87%) as an off-white solid. MS: 328.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 12.39 (s, 1H), 12.02 (s, 1H), 8.74 (s, 1H), 7.98-7.99 (m, 3H), 7.67-7.68 (m , 1H), 7.56 (t, J = 7.60 Hz, 2H). Step B To a solution of the title compound (14.0 g, 42.94 mmol) in N-methyl-2-pyrrolidone (NMP) (70 mL) from Step A above was added sodium methoxide (NaOMe) (4.6 g, 85.88 mmol). The mixture was then heated to 120°C and stirring continued for 4 hours. The reaction was monitored by TLC. The reaction mixture was poured into cold water (300 mL) and a white precipitate was obtained. The solid was filtered and washed with water (300 mL) and n-hexane (200 mL). The compound was dried under vacuum for 6 h to give the desired product (14.0 g, crude) as a white solid. The product was used as is in the next step. MS: 290.0 (M+H) ⁺ . Step C A suspension of the title compound (14.0 g, 48.4 mmol) from 70% H ₂ SO ₄ (50.0 mL) from Step B above was heated at 110° C. for 4 hours. The reaction mixture was cooled to room temperature, and the reaction mixture was slowly poured into 200 mL of cold water (0°C). Then, the reaction mixture was adjusted to basic pH by adding 50% NaOH aqueous solution. Then, the compound was extracted with EtOAc (6×100 mL). The combined organic layer was dried over Na ₂ SO ₄ and filtered, and then the solvent was concentrated to give the desired product (6 g, 67%) as a pale yellow solid. MS: 186.1 (M+H) ^{+ 1} H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.30 (s, 1H), 7.86 (s, 1H). Step D To the suspension of the title compound (5.0 g, 27.02 mmol) in acetonitrile (120 mL) from Step C above was added tert-butyl nitrite ( 4.8 mL, using a syringe at 0°C over a period of 10 min. 40.54 mmol). Then, copper (II) bromide (9.0 g, 40.54 mmol) was added portionwise. After 30 minutes at 0°C, the reaction mixture was warmed to room temperature for 2.5 hours, and the progress of the reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated and diluted with water (200 mL) and 5% MeOH/DCM (3×200 mL). The combined organics were washed with brine (50 mL), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give the title compound (6.5 g) as a white solid. The product was used as is in the next step. MS: 250.9 (M+H) ⁺ Step E To the solution of the title compound (6.5 g, 26.09 mmol) in anhydrous DCM (100 mL) from Step D above was added triethylamine (11.2 mL, 81.5 mmol) and Porphyrin (2.8 mL, 28.13 mmol). The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated under vacuum. Using a Biotage Isolera One purification system on a silica gel column, the crude reaction mixture was purified using an EtOAc/hexane gradient (10/80 => 80/20) to obtain the title compound (4.7 g, 71%) as a light yellow solid. MS: 256.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.48 (s, 1H), 8.05 (s, 1H), 3.73-3.74 (m, 4H), 3.60-3.61 (m, 4H).

向2,5-二氯-1,3-苯并噁唑(0.25 g, 0.0013 mol)於乙腈(15 mL)中之攪拌溶液添加K₂ CO₃ (0.538 g, 0.0039 mol)及5-氧雜-8-氮雜螺[3.5]壬烷(0.178 g, 0.0014 mol)。之後，將反應混合物加熱至70℃持續12 h。根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，得到呈白色固體之標題化合物(0.30 g, 80.21%)。 MS: 279.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ =7.44 (d,J = 8.40 Hz, 1H), 7.36-7.37 (m, 1H), 7.06 (dd,J = 2.00, 8.40 Hz, 1H), 3.64-3.65 (m, 2H), 3.60 (s, 2H), 3.54-3.55 (m, 2H), 1.96-1.97 (m, 4H), 1.71-1.74 (m, 2H)。 Preparation Example 54

To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.25 g, 0.0013 mol) in acetonitrile (15 mL), add K ₂ CO ₃ (0.538 g, 0.0039 mol) and 5-oxa -8-azaspiro[3.5]nonane (0.178 g, 0.0014 mol). After that, the reaction mixture was heated to 70 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to give the title compound (0.30 g, 80.21%) as a white solid. MS: 279.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ =7.44 (d, J = 8.40 Hz, 1H), 7.36-7.37 (m, 1H), 7.06 (dd, J = 2.00, 8.40 Hz, 1H), 3.64-3.65 (m, 2H), 3.60 (s, 2H), 3.54-3.55 (m, 2H), 1.96-1.97 (m, 4H), 1.71-1.74 (m, 2H).

向2,5-二氯-1,3-苯并噁唑(0.60 g, 0.0032 mol)於乙腈(15 mL)中之攪拌溶液添加K₂ CO₃ (1.32 g, 0.0096 mol)及2-甲氧基乙烷-1-胺(0.266 g, 0.0035 mol)。之後，將反應混合物加熱至70℃持續12 h。根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，得到呈棕色固體之標題化合物(0.41 g, 56.24%)。 MS: 226.9 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.26 (s, 1H), 7.35 (d,J = 11.20 Hz, 1H), 7.28 (d,J = 2.80 Hz, 1H), 6.99 (dd,J = 2.80, 11.20 Hz, 1H), 3.48 (d,J = 7.20 Hz, 4H), 3.27 (s, 3H)。 Preparation Example 55

To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.60 g, 0.0032 mol) in acetonitrile (15 mL), add K ₂ CO ₃ (1.32 g, 0.0096 mol) and 2-methoxy Ethyl-1-amine (0.266 g, 0.0035 mol). After that, the reaction mixture was heated to 70 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to give the title compound (0.41 g, 56.24%) as a brown solid. MS: 226.9 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.26 (s, 1H), 7.35 (d, J = 11.20 Hz, 1H), 7.28 (d, J = 2.80 Hz, 1H), 6.99 (dd, J = 2.80, 11.20 Hz, 1H), 3.48 (d, J = 7.20 Hz, 4H), 3.27 (s, 3H).

向2,5-二氯-1,3-苯并噁唑(0.719 g, 0.0038 mol)於DCM (15 mL)中之攪拌溶液添加TEA (2.67 ml, 0.0191 mol)及(2R)-2-甲基嗎啉(0.5 g, 0.00494 mol)，且在25℃下攪拌12 h。根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，得到呈白色固體之標題化合物(0.6 g, 45.4%)。 MS: 253.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.43 (d,J = 11.20 Hz, 1H), 7.35 (d,J = 2.40 Hz, 1H), 7.05 (dd,J = 2.80, 11.20 Hz, 1H), 3.89-3.90 (m, 3H), 3.56-3.59 (m, 2H), 3.17-3.18 (m, 1H), 2.86-2.89 (m, 1H), 1.15 (d,J = 8.40 Hz, 3H)。 Preparation Example 56

To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.719 g, 0.0038 mol) in DCM (15 mL) was added TEA (2.67 ml, 0.0191 mol) and (2R)-2-methyl Morpholine (0.5 g, 0.00494 mol), and stirred at 25 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to give the title compound (0.6 g, 45.4%) as a white solid. MS: 253.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.43 (d, J = 11.20 Hz, 1H), 7.35 (d, J = 2.40 Hz, 1H), 7.05 (dd, J = 2.80, 11.20 Hz, 1H), 3.89-3.90 (m, 3H), 3.56-3.59 (m, 2H), 3.17-3.18 (m, 1H), 2.86-2.89 (m, 1H), 1.15 (d, J = 8.40 Hz, 3H) .

向2,5-二氯-1,3-苯并噁唑(0.360 g, 0.0019 mol)於DCM (15 mL)中之攪拌溶液添加TEA (1.33 ml, 0.0095 mol)及(2S)-2-甲基嗎啉(0.25 g, 0.00247 mol)，且在25℃下攪拌12 h。根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮。使用Biotage Isolera One純化系統，採用於石油醚中之20%-25%乙酸乙酯，在矽膠管柱上純化粗製反應混合物，得到呈白色固體之標題化合物(0.3 g, 45.5%)。 MS: 253.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.43 (d,J = 8.48 Hz, 1H), 7.34-7.35 (m, 1H), 7.05 (dd,J = 2.08, 8.46 Hz, 1H), 3.90-3.90 (m, 3H), 3.61-3.62 (m, 2H), 3.24-3.25 (m, 1H), 2.87-2.89 (m, 1H), 1.15 (d,J = 6.20 Hz, 3H)。 Preparation Example 57

To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.360 g, 0.0019 mol) in DCM (15 mL) was added TEA (1.33 ml, 0.0095 mol) and (2S)-2-methyl Morpholine (0.25 g, 0.00247 mol), and stirred at 25 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated. Using the Biotage Isolera One purification system, using 20%-25% ethyl acetate in petroleum ether, the crude reaction mixture was purified on a silica gel column to give the title compound (0.3 g, 45.5%) as a white solid. MS: 253.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.43 (d, J = 8.48 Hz, 1H), 7.34-7.35 (m, 1H), 7.05 (dd, J = 2.08, 8.46 Hz, 1H), 3.90-3.90 (m, 3H), 3.61-3.62 (m, 2H), 3.24-3.25 (m, 1H), 2.87-2.89 (m, 1H), 1.15 (d, J = 6.20 Hz, 3H).

在0℃下向2,5-二氯-1,3-苯并噁唑(1.20 g, 6.38 mmol)於無水DCM (50 mL)中之溶液添加(3R)-3-甲基嗎啉(0.775 g, 7.65 mmol)及Et₃ N (1.94 g, 19.10 mmol)，且在25℃下攪拌4 h。在反應完成後(藉由TLC監測)，用水(20 mL)稀釋反應混合物並用DCM (20 mL × 2)萃取。使合併之有機萃取物經Na₂ SO₄ 乾燥，過濾並在減壓下蒸發。 使用Biotage Isolera One純化系統，採用於石油醚中之10%-20%乙酸乙酯，在矽膠管柱上純化粗製反應混合物，得到標題化合物(1.0 g, 59.9%)。 MS: 252.9 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.43 (d,J = 11.20 Hz, 1H), 7.35 (d,J = 2.80 Hz, 1H), 7.04 (dd,J = 2.80, 11.40 Hz, 1H), 4.17-4.19 (m, 1H), 3.69-3.76 (m, 3H), 3.42-3.43 (m, 5H), 1.30 (d,J = 9.20 Hz, 3H)。 Preparation Example 58

To a solution of 2,5-dichloro-1,3-benzoxazole (1.20 g, 6.38 mmol) in anhydrous DCM (50 mL) was added (3R)-3-methylmorpholine (0.775 g, 7.65 mmol) and Et ₃ N (1.94 g, 19.10 mmol), and stirred at 25° C. for 4 h. After the reaction was completed (monitored by TLC), the reaction mixture was diluted with water (20 mL) and extracted with DCM (20 mL×2). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure. The crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using 10%-20% ethyl acetate in petroleum ether to obtain the title compound (1.0 g, 59.9%). MS: 252.9 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.43 (d, J = 11.20 Hz, 1H), 7.35 (d, J = 2.80 Hz, 1H), 7.04 (dd, J = 2.80, 11.40 Hz, 1H), 4.17-4.19 (m, 1H), 3.69-3.76 (m, 3H), 3.42-3.43 (m, 5H), 1.30 (d, J = 9.20 Hz, 3H).

在0℃下向2,5-二氯-1,3-苯并噁唑(1.20 g, 6.38 mmol)於無水DCM (50 mL)中之溶液添加(3S)-3-甲基嗎啉(0.775 g, 7.65 mmol)及Et₃ N (1.94 g, 19.10 mmol)，且在25℃下攪拌4 h。在反應完成後(藉由TLC監測)，用H₂ O (20 mL)稀釋反應混合物並用DCM (20 mL × 2)萃取。使合併之有機萃取物經Na₂ SO₄ 乾燥，過濾並在減壓下蒸發。 使用Biotage Isolera One純化系統，採用於石油醚中之10%-20%乙酸乙酯，在矽膠管柱上純化粗製反應混合物，得到標題化合物(0.8 g,49.6%)。 MS: 252.9 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.43 (d,J = 11.20 Hz, 1H), 7.35 (d,J = 2.80 Hz, 1H), 7.04 (dd,J = 2.80, 11.20 Hz, 1H), 4.17-4.19 (m, 1H), 3.73-3.75 (m, 4H), 3.46-3.47 (m, 2H), 1.30 (d,J = 9.20 Hz, 3H), 。 Preparation Example 59

To a solution of 2,5-dichloro-1,3-benzoxazole (1.20 g, 6.38 mmol) in anhydrous DCM (50 mL) was added (3S)-3-methylmorpholine (0.775 g, 7.65 mmol) and Et ₃ N (1.94 g, 19.10 mmol), and stirred at 25° C. for 4 h. After the reaction was completed (monitored by TLC), the reaction mixture was diluted with H ₂ O (20 mL) and extracted with DCM (20 mL×2). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure. The crude reaction mixture was purified on a silica gel column using a Biotage Isolera One purification system using 10%-20% ethyl acetate in petroleum ether to obtain the title compound (0.8 g, 49.6%). MS: 252.9 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.43 (d, J = 11.20 Hz, 1H), 7.35 (d, J = 2.80 Hz, 1H), 7.04 (dd, J = 2.80, 11.20 Hz, 1H), 4.17-4.19 (m, 1H), 3.73-3.75 (m, 4H), 3.46-3.47 (m, 2H), 1.30 (d, J = 9.20 Hz, 3H),.

向2,5-二氯-1,3-苯并噁唑(0.5 g, 0.00265 mol)於乙腈(10 mL)中之攪拌溶液添加K₂ CO₃ (1.1 g, 0.00797 mol)及2-甲氧基-N-甲基乙烷-1-胺(0.284 g, 0.00319 mol)。之後，將反應混合物加熱至70℃持續12 h。根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，得到呈棕色液體之標題化合物(0.61 g, 95%)。 MS: 241.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.40 (d,J = 11.20 Hz, 1H), 7.30 (d,J = 2.80 Hz, 1H), 7.00 (dd,J = 2.80, 11.20 Hz, 1H), 3.68 (t,J = 6.40 Hz, 2H), 3.58 (t,J = 7.20 Hz, 2H), 3.27 (s, 3H), 3.16 (s, 3H)。 Preparation Example 60

To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.5 g, 0.00265 mol) in acetonitrile (10 mL), add K ₂ CO ₃ (1.1 g, 0.00797 mol) and 2-methoxy -N-methylethane-1-amine (0.284 g, 0.00319 mol). After that, the reaction mixture was heated to 70 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to give the title compound (0.61 g, 95%) as a brown liquid. MS: 241.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.40 (d, J = 11.20 Hz, 1H), 7.30 (d, J = 2.80 Hz, 1H), 7.00 (dd, J = 2.80, 11.20 Hz, 1H), 3.68 (t, J = 6.40 Hz, 2H), 3.58 (t, J = 7.20 Hz, 2H), 3.27 (s, 3H), 3.16 (s, 3H).

步驟 A 向2-胺基-3-氯苯酚(4 g, 0.0280 mol)於乙醇(80 mL)中之攪拌溶液添加O-乙基二硫代碳酸酯鉀鹽(4.49 g, 0.0280 mol)，然後在N₂ 下加熱至85℃持續12 h。根據LCMS在反應完成後，將反應混合物濃縮，藉由使用乙酸(pH = 5)使粗產物酸化且將固體過濾，用水洗滌，在真空下乾燥6 h，得到呈淡棕色固體之4-氯苯并[d]噁唑-2-硫醇(4.6 g, 89%)。 MS: 186.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 14.44 (bs, 1H), 7.46 (d,J = 10.80 Hz, 1H), 7.35 (d,J = 11.20 Hz, 1H), 7.23 (t,J = 10.80 Hz, 1H)。步驟 B 在0℃下向來自上文步驟A之標題化合物(4.6 g, 0.0244 mol)於DCM (90 mL)中之溶液添加草醯氯(3.20 mL, 0.0374 mol)，之後添加DMF (1 mL)，然後在25℃下攪拌1 h。然後在0℃下添加TEA (10 mL, 0.0734 mol)及嗎啉(2.5 mL, 0.0293 mol)且在氮下在25℃下攪拌2 h。在藉由TLC跟蹤反應完成後，添加水(40 mL)並用DCM (2×50 mL)萃取。將有機層濃縮且使用Biotage Isolera One純化系統，採用於石油醚中之10%-15%乙酸乙酯，在矽膠管柱上純化粗製物，得到呈淡黃色固體之標題化合物(3.4 g, 58%)。 MS: 239.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.40 (d,J = 10.80 Hz, 1H), 7.23 (d,J = 10.40 Hz, 1H), 7.03 (t,J = 10.80 Hz, 1H), 3.71-3.73 (m, 4H), 3.60-3.62 (m, 4H)。 Preparation Example 61

Step A To a stirred solution of 2-amino-3-chlorophenol (4 g, 0.0280 mol) in ethanol (80 mL) was added O-ethyldithiocarbonate potassium salt (4.49 g, 0.0280 mol), then Heat to 85 °C under N ₂ for 12 h. After completion of the reaction according to LCMS, the reaction mixture was concentrated, the crude product was acidified by using acetic acid (pH=5) and the solid was filtered, washed with water, and dried under vacuum for 6 h to give 4-chlorobenzene as a light brown solid And [d] oxazole-2-thiol (4.6 g, 89%). MS: 186.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 14.44 (bs, 1H), 7.46 (d, J = 10.80 Hz, 1H), 7.35 (d, J = 11.20 Hz, 1H), 7.23 (t, J = 10.80 Hz, 1H). Step B To a solution of the title compound (4.6 g, 0.0244 mol) in DCM (90 mL) from Step A above was added oxalyl chloride (3.20 mL, 0.0374 mol) at 0°C, followed by DMF (1 mL) And then stirred at 25 °C for 1 h. Then TEA (10 mL, 0.0734 mol) and morpholine (2.5 mL, 0.0293 mol) were added at 0°C and stirred at 25°C for 2 h under nitrogen. After tracking the completion of the reaction by TLC, water (40 mL) was added and extracted with DCM (2×50 mL). The organic layer was concentrated and the crude product was purified on a silica gel column using a Biotage Isolera One purification system using 10%-15% ethyl acetate in petroleum ether to obtain the title compound (3.4 g, 58%) as a light yellow solid ). MS: 239.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.40 (d, J = 10.80 Hz, 1H), 7.23 (d, J = 10.40 Hz, 1H), 7.03 (t, J = 10.80 Hz, 1H) , 3.71-3.73 (m, 4H), 3.60-3.62 (m, 4H).

步驟 A 在0℃下向NaH (0.47 g,19.8 mmol)於THF (10 mL)中之懸浮液逐滴添加1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(1.8 g, 6.61 mmol) (溶解於THF中)，且然後在室溫下攪拌1 h。之後，在0℃下添加碘甲烷(0.7 mL, 11.76 mmol)，且然後在室溫下攪拌2 h。在反應完成後(藉由TLC監測)，利用冰水將反應混合物淬滅，之後使用乙酸乙酯進行萃取。將有機層濃縮以得到粗製反應混合物5-甲基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(1.5 g，粗製)。產物原樣用於下一步驟。 MS: 287.2 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(1.5 g, 5.24 mmol)於DCM (20 mL)中之溶液添加於二噁烷中之4.0 M HCl (5 mL)。將反應混合物攪拌過夜。在反應完成後，使該反應混合物蒸發以去除溶劑並用二乙醚洗滌，得到呈棕色固體之標題化合物(800 mg，粗製)。 MS: 187.1 (M+H)⁺ 。 Preparation Example 62

Step A To a suspension of NaH (0.47 g, 19.8 mmol) in THF (10 mL) was added dropwise 1,3,4,5-tetrahydro-2H-pyrido[4,3-b] at 0°C Indole-2-carboxylic acid tert-butyl ester (1.8 g, 6.61 mmol) (dissolved in THF), and then stirred at room temperature for 1 h. After that, iodomethane (0.7 mL, 11.76 mmol) was added at 0°C, and then stirred at room temperature for 2 h. After the reaction was completed (monitored by TLC), the reaction mixture was quenched with ice water, and then extracted with ethyl acetate. The organic layer was concentrated to obtain a crude reaction mixture 5-methyl-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indole-2-carboxylic acid tert-butyl ester (1.5 g , Crude). The product was used as is in the next step. MS: 287.2 (M+H) ⁺ . Step B To a solution of the title compound (1.5 g, 5.24 mmol) in DCM (20 mL) from Step A above was added 4.0 M HCl (5 mL) in dioxane. The reaction mixture was stirred overnight. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and washed with diethyl ether to obtain the title compound (800 mg, crude) as a brown solid. MS: 187.1 (M+H) ⁺ .

步驟 A 向(3-甲氧基苯基)肼鹽酸鹽(10 g, 57.4 mmol)及4-側氧基六氫吡啶-1-甲酸第三丁基酯(11.3 g, 57.4 mmol)於乙醇(100 mL)中之溶液添加酒石酸。添加二甲基脲(30:70, 10 g)，且在70℃下加熱16 h。將反應混合物冷卻至25℃並在真空下濃縮。將殘餘物溶解於水中且利用NaOH溶液(30%)鹼化至pH = 14並用DCM萃取。將有機相分離且經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發，得到呈淡黃色膠狀物之7-甲氧基-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(2.5 g, 21%)。粗產物原樣用於下一步驟。 MS: 203.0 (M+H)⁺ 。步驟 B 在25℃下向來自上文步驟A之標題化合物(600 mg, 2.9 mmol)於THF (6 mL)中之溶液添加三乙胺(0.8 mL, 5.8 mmol)及Boc酸酐(650 mg, 3 mmol)，且攪拌12 h。在反應完成後(藉由TLC監測)，將反應混合物在減壓下濃縮且藉由急速管柱層析使用己烷: EtOAc (80:20)純化粗產物，得到呈淡黃色固體之7-甲氧基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(610 mg, 68%)。 MS: 303.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 10.69 (bs, 1H), 7.25 (d,J = 11.60 Hz, 1H), 6.80 (d,J = 2.80 Hz, 1H), 6.61 (dd,J = 2.80, 11.60 Hz, 1H), 4.48 (s, 2H), 3.81 (s, 3H), 3.67-3.68 (m, 2H), 2.73 (bs, 2H), 1.44 (s, 9H)。 MS: 187.1 (M+H)⁺ 。步驟 C 在0℃下向來自上文步驟B之標題化合物(550 mg, 1.8 mmol)於THF (5 mL)中之溶液添加NaH (60%礦物油，0.14 g, 3.6 mmol)，且攪拌30 min。然後添加對甲苯磺醯氯(342 mg, 1.8 mmol)並攪拌45 min。在反應完成後(藉由TLC監測)，利用水將反應混合物淬滅並用EtOAc (20 mL × 3)萃取。將合併之有機萃取物用水、鹽水洗滌，且經Na₂ SO₄ 乾燥。過濾且使溶劑在減壓下蒸發，產生呈灰白色固體之7-甲氧基-5-甲苯磺醯基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(550 mg, 66%)。產物原樣用於下一步驟。 MS: 357.0 (M+H)⁺ 。步驟 D 在0℃下向來自上文步驟C之標題化合物(550 mg, 1.204 mmol)於無水DCM (5 mL)中之溶液緩慢添加於二噁烷中之HCl (g) (2 M, 5 mL)，且在25℃下攪拌12 h。在反應完成後(藉由TLC監測)，使反應混合物在減壓下蒸發以產生粗產物。用二乙醚洗滌粗產物，得到呈灰白色固體之7-甲氧基-5-甲苯磺醯基-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(350 mg, 81%)。 MS: 357.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.73 (d,J = 8.40 Hz, 1H), 7.56 (s, 1H), 7.37 (d,J = 8.00 Hz, 2H), 7.27 (d,J = 8.40 Hz, 1H), 6.85-6.86 (m, 1H), 3.82 (s, 3H), 3.72 (s, 2H), 2.90-2.97 (m, 4H), 2.32 (s, 3H)。 Preparation Example 63

Step A To (3-methoxyphenyl)hydrazine hydrochloride (10 g, 57.4 mmol) and tert-butyl 4-oxohexahydropyridine-1-carboxylate (11.3 g, 57.4 mmol) in ethanol (100 mL) was added tartaric acid. Dimethylurea (30:70, 10 g) was added and heated at 70°C for 16 h. The reaction mixture was cooled to 25°C and concentrated under vacuum. The residue was dissolved in water and basified with NaOH solution (30%) to pH = 14 and extracted with DCM. The organic phase was separated and dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure to give 7-methoxy-2,3,4,5-tetrahydro-1H-pyridine as a pale yellow gum And [4,3-b]indole (2.5 g, 21%). The crude product was used as is in the next step. MS: 203.0 (M+H) ⁺ . Step B To a solution of the title compound (600 mg, 2.9 mmol) in THF (6 mL) from Step A above was added triethylamine (0.8 mL, 5.8 mmol) and Boc anhydride (650 mg, 3 mmol), and stirred for 12 h. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure and the crude product was purified by flash column chromatography using hexane: EtOAc (80:20) to give 7-A as a light yellow solid Oxy-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indole-2-carboxylic acid tert-butyl ester (610 mg, 68%). MS: 303.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.69 (bs, 1H), 7.25 (d, J = 11.60 Hz, 1H), 6.80 (d, J = 2.80 Hz, 1H), 6.61 (dd, J = 2.80, 11.60 Hz, 1H), 4.48 (s, 2H), 3.81 (s, 3H), 3.67-3.68 (m, 2H), 2.73 (bs, 2H), 1.44 (s, 9H). MS: 187.1 (M+H) ⁺ . Step C To the solution of the title compound (550 mg, 1.8 mmol) in THF (5 mL) from Step B above was added NaH (60% mineral oil, 0.14 g, 3.6 mmol) at 0°C and stirred for 30 min . Then add p-toluenesulfonyl chloride (342 mg, 1.8 mmol) and stir for 45 min. After the reaction was completed (monitored by TLC), the reaction mixture was quenched with water and extracted with EtOAc (20 mL×3). The combined organic extracts were washed with water, brine, and dried over Na ₂ SO ₄ . Filtration and evaporation of the solvent under reduced pressure yielded 7-methoxy-5-tosyl-1,3,4,5-tetrahydro-2H-pyrido[4,3-b] as an off-white solid Tert-butyl indole-2-carboxylic acid (550 mg, 66%). The product was used as is in the next step. MS: 357.0 (M+H) ⁺ . Step D To the solution of the title compound (550 mg, 1.204 mmol) in anhydrous DCM (5 mL) from Step C above was slowly added HCl (g) (2 M, 5 mL in dioxane) at 0°C. ), and stirred at 25 °C for 12 h. After the reaction was completed (monitored by TLC), the reaction mixture was evaporated under reduced pressure to produce a crude product. The crude product was washed with diethyl ether to obtain 7-methoxy-5-tosyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole as an off-white solid ( 350 mg, 81%). MS: 357.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.73 (d, J = 8.40 Hz, 1H), 7.56 (s, 1H), 7.37 (d, J = 8.00 Hz, 2H), 7.27 (d, J = 8.40 Hz, 1H), 6.85-6.86 (m, 1H), 3.82 (s, 3H), 3.72 (s, 2H), 2.90-2.97 (m, 4H), 2.32 (s, 3H).

步驟 A 在0℃下向8-甲基-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(1 g, 5.37 mmol)於DCM (20 mL)中之攪拌溶液添加TEA (2.25 mL, 16.1 mmol)及BOC酸酐(1.85 mL, 8.05 mol)，然後在25℃下攪拌12 h。根據TLC在反應完成後，向反應混合物添加水，之後使用DCM進行萃取。將有機層分離並用鹽水溶液洗滌，濃縮並用己烷洗滌，得到呈灰白色固體之標題化合物(1.1 g, 71%)。 MS: 287.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 10.74 (s, 1H), 7.16 (d,J = 8.44 Hz, 1H), 6.85 (d,J = 8.28 Hz, 1H), 4.49 (s, 2H), 3.69 (t,J = 5.64 Hz, 2H), 2.74 (t,J = 5.28 Hz, 2H), 2.35 (s, 3H), 1.47 (s, 9H)。步驟 B 在0℃下向氫化鈉(0.125 g, 3.13 mmol)於THF (10 mL)中之懸浮液逐滴添加來自上文步驟A之標題化合物(0.6 g, 2.08 mmol) (溶解於20 mL THF中)，然後在室溫下攪拌30 min。之後，在0℃下逐滴添加甲苯磺醯氯(1.19 g, 6.25 mmol) (溶解於20 mL THF中)，且然後在室溫下攪拌3 h。根據TLC在反應完成後，利用冰水將反應混合物淬滅，過濾所形成之固體且用水洗滌並乾燥，得到呈白色固體之標題化合物(0.550 g, 57%)。 MS: 341.1 (M+H)⁺ -Boc。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.91 (d,J = 8.52 Hz, 1H), 7.73 (d,J = 8.20 Hz, 2H), 7.34 (d,J = 8.40 Hz, 2H), 7.26 (s, 1H), 7.15 (d, J = 8.56 Hz, 1H), 4.41 (s, 2H), 3.68 (t,J = 5.60 Hz, 2H), 3.06 (s, 2H), 2.29-2.30 (m, 6H), 1.43 (s, 9H),步驟 C 在0℃下向來自上文步驟B之標題化合物(0.55 g, 1.20 mmol)於DCM中之溶液添加於二噁烷中之4 M HCl (4 mL)。將反應混合物攪拌2 h。在反應完成後，使該反應混合物蒸發以去除溶劑並利用二乙醚過濾，得到呈白色固體之標題化合物(0.40 g, 85.9%)。 Preparation Example 64

Step A To 8-methyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole (1 g, 5.37 mmol) in DCM (20 mL) at 0°C To the stirred solution, TEA (2.25 mL, 16.1 mmol) and BOC anhydride (1.85 mL, 8.05 mol) were added, and then stirred at 25°C for 12 h. After completion of the reaction according to TLC, water was added to the reaction mixture, followed by extraction with DCM. The organic layer was separated and washed with brine solution, concentrated and washed with hexane to give the title compound (1.1 g, 71%) as an off-white solid. MS: 287.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.74 (s, 1H), 7.16 (d, J = 8.44 Hz, 1H), 6.85 (d, J = 8.28 Hz, 1H), 4.49 (s, 2H), 3.69 (t, J = 5.64 Hz, 2H), 2.74 (t, J = 5.28 Hz, 2H), 2.35 (s, 3H), 1.47 (s, 9H). Step B To the suspension of sodium hydride (0.125 g, 3.13 mmol) in THF (10 mL) was added dropwise the title compound (0.6 g, 2.08 mmol) from step A above (dissolved in 20 mL THF at 0°C Medium), and then stirred at room temperature for 30 min. After that, tosyl chloride (1.19 g, 6.25 mmol) (dissolved in 20 mL of THF) was added dropwise at 0°C, and then stirred at room temperature for 3 h. After completion of the reaction according to TLC, the reaction mixture was quenched with ice water, the solid formed was filtered and washed with water and dried to give the title compound (0.550 g, 57%) as a white solid. MS: 341.1 (M+H) ⁺ -Boc. ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.91 (d, J = 8.52 Hz, 1H), 7.73 (d, J = 8.20 Hz, 2H), 7.34 (d, J = 8.40 Hz, 2H) , 7.26 (s, 1H), 7.15 (d, J = 8.56 Hz, 1H), 4.41 (s, 2H), 3.68 (t, J = 5.60 Hz, 2H), 3.06 (s, 2H), 2.29-2.30 ( m, 6H), 1.43 (s, 9H), Step C. To a solution of the title compound (0.55 g, 1.20 mmol) from Step B above in DCM at 0°C was added 4 M HCl in dioxane ( 4 mL). The reaction mixture was stirred for 2 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and filtered with diethyl ether to obtain the title compound (0.40 g, 85.9%) as a white solid.

步驟 A 在0℃下向8-氯-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(1 g, 4.84 mmol)於DCM (20 mL)中之攪拌溶液添加TEA (2.02 mL, 14.5 mmol)及BOC酸酐(1.67 mL, 7.26 mmol)，然後在25℃下攪拌12 h。根據TLC在反應完成後，向反應混合物添加水，之後使用DCM進行萃取。將有機層分離並用鹽水溶液洗滌，濃縮得到呈白色固體之8-氯-1,3,4,5-四氫吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(1 g, 66.5%)。 MS: 305.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 11.12 (bs, 1H), 7.46 (d,J = 1.68 Hz, 1H), 7.30 (d,J = 8.56 Hz, 1H), 7.03 (dd,J = 1.96, 8.52 Hz, 1H), 4.51 (s, 2H), 3.70 (t, J = 5.64 Hz, 2H), 2.77 (t,J = 5.32 Hz, 2H), 1.47 (s, 9H)。步驟 B 在0℃下向冷卻至0℃之氫化鈉(0.096 g, 2.42 mmol)於THF (10 mL)中之懸浮液逐滴添加來自上文步驟A之標題化合物(0.5 g, 1.61 mol) (溶解於20 mL THF中)，然後在室溫下攪拌30 min。之後，在0℃下逐滴添加甲苯磺醯氯(0.921 g, 48.3 mol) (溶解於20 mL THF中)，且然後在室溫下攪拌3 h。根據TLC在反應完成後，利用冰水將反應混合物淬滅，之後使用乙酸乙酯進行萃取(100 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮。藉由矽膠管柱層析使用石油醚:乙酸乙酯(75:25)純化產物，得到呈白色固體之8-氯-5-(對甲苯基磺醯基)-3,4-二氫-1H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(0.450 g, 60%)。 MS: 361.1 (M+H)⁺ -Boc。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.05 (d,J = 8.88 Hz, 1H), 7.78 (d,J = 8.16 Hz, 2H), 7.64 (d,J = 1.96 Hz, 1H), 7.35-7.36 (m, 3H), 4.44 (s, 2H), 3.68 (t,J = 5.60 Hz, 2H), 3.08 (bs, 2H), 2.32 (s, 3H), 1.43 (s, 9H)。步驟 C 在0℃下向來自上文步驟B之標題化合物(0.45 g, 0.97 mol)於DCM中之溶液添加於二噁烷中之4 M HCl (3 mL)。將反應混合物攪拌2 h。在反應完成後，使該反應混合物蒸發以去除溶劑並利用二乙醚濃縮，得到呈白色固體之標題化合物 (0.32 g, 91%)。產物原樣用於下一步驟。 MS: 361.1 (M+H)⁺ 。 Preparation Example 65

Step A To 8-chloro-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole (1 g, 4.84 mmol) in DCM (20 mL) at 0 °C To the stirred solution, TEA (2.02 mL, 14.5 mmol) and BOC anhydride (1.67 mL, 7.26 mmol) were added, and then stirred at 25°C for 12 h. After completion of the reaction according to TLC, water was added to the reaction mixture, followed by extraction with DCM. The organic layer was separated and washed with brine solution, and concentrated to give 8-chloro-1,3,4,5-tetrahydropyrido[4,3-b]indole-2-carboxylic acid tert-butyl ester as a white solid ( 1 g, 66.5%). MS: 305.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.12 (bs, 1H), 7.46 (d, J = 1.68 Hz, 1H), 7.30 (d, J = 8.56 Hz, 1H), 7.03 (dd, J = 1.96, 8.52 Hz, 1H), 4.51 (s, 2H), 3.70 (t, J = 5.64 Hz, 2H), 2.77 (t, J = 5.32 Hz, 2H), 1.47 (s, 9H). Step B To a suspension of sodium hydride (0.096 g, 2.42 mmol) in THF (10 mL) cooled to 0°C was added dropwise at 0°C the title compound (0.5 g, 1.61 mol) from step A above ( Dissolved in 20 mL THF), then stirred at room temperature for 30 min. After that, tosyl chloride (0.921 g, 48.3 mol) (dissolved in 20 mL of THF) was added dropwise at 0°C, and then stirred at room temperature for 3 h. After completion of the reaction according to TLC, the reaction mixture was quenched with ice water, followed by extraction with ethyl acetate (100 mL). The organic layer was separated, dried over sodium sulfate, filtered and then concentrated. The product was purified by silica gel column chromatography using petroleum ether: ethyl acetate (75:25) to obtain 8-chloro-5-(p-tolylsulfonyl)-3,4-dihydro-1H as a white solid -Pyrido[4,3-b]indole-2-carboxylic acid tert-butyl ester (0.450 g, 60%). MS: 361.1 (M+H) ⁺ -Boc. ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.05 (d, J = 8.88 Hz, 1H), 7.78 (d, J = 8.16 Hz, 2H), 7.64 (d, J = 1.96 Hz, 1H) , 7.35-7.36 (m, 3H), 4.44 (s, 2H), 3.68 (t, J = 5.60 Hz, 2H), 3.08 (bs, 2H), 2.32 (s, 3H), 1.43 (s, 9H). Step C To a solution of the title compound (0.45 g, 0.97 mol) from DCM in DCM at 0 °C was added 4 M HCl in dioxane (3 mL). The reaction mixture was stirred for 2 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and concentrated with diethyl ether to obtain the title compound (0.32 g, 91%) as a white solid. The product was used as is in the next step. MS: 361.1 (M+H) ⁺ .

步驟 A 在0℃下向5-溴-2-氯苯并[d]噁唑(1 g, 4.30 mmol)於無水DCM (10 mL)中之溶液添加嗎啉(0.56 g, 6.42 mmol)及Et₃ N (1.7 mL, 12.9 mmol)，且在25℃下攪拌4 h。在反應完成後(藉由TLC監測)，用H₂ O (10 mL)稀釋反應混合物並用DCM (10 mL × 2)萃取。將合併之有機萃取物經Na₂ SO₄ 乾燥，過濾並在減壓下蒸發以產生粗產物。使粗產物與二乙醚(100 mL)一起研磨，過濾，用二乙醚(5 mL)洗滌並乾燥，得到呈灰白色固體之標題化合物(0.85 g, 71%)。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.48 (d,J = 2.40 Hz, 1H), 7.34-7.38 (m, 1H), 7.16-7.17 (m, 1H), 3.70-3.72 (m, 4H), 3.58-3.59 (m, 4H)。 Preparation Example 66

Step A To a solution of 5-bromo-2-chlorobenzo[d]oxazole (1 g, 4.30 mmol) in anhydrous DCM (10 mL) was added morpholine (0.56 g, 6.42 mmol) and Et at 0°C ₃ N (1.7 mL, 12.9 mmol), and stirred at 25 °C for 4 h. After the reaction was completed (monitored by TLC), the reaction mixture was diluted with H ₂ O (10 mL) and extracted with DCM (10 mL×2). The combined organic extracts were dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to produce crude product. The crude product was triturated with diethyl ether (100 mL), filtered, washed with diethyl ether (5 mL) and dried to give the title compound (0.85 g, 71%) as an off-white solid. ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.48 (d, J = 2.40 Hz, 1H), 7.34-7.38 (m, 1H), 7.16-7.17 (m, 1H), 3.70-3.72 (m , 4H), 3.58-3.59 (m, 4H).

步驟 A 向4,6-二氯吡啶-3-胺(2.5 g, 15.3 mmol)於THF (50 mL)中之攪拌溶液逐滴添加於THF中之三光氣(4.55 g, 15.3 mol)，之後添加TEA (4.28 mL, 30.7 mol)且加熱至回流2 h。將反應混合物在真空下濃縮。將殘餘物溶解於乙腈(50 mL)及甲苯(50 mL)中，且添加嗎啉(1.34 g, 15.3 mmol)並加熱至110℃持續12 h。之後，檢查TLC，將粗製物濃縮並藉由矽膠管柱層析使用石油醚:乙酸乙酯(20:80)純化，得到呈白色固體之N-(4,6-二氯-3-吡啶基)嗎啉-4-甲醯胺(3.0 g, 70.1%)。 MS: 276.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.55 (s, 1H), 8.38-8.40 (m, 1H), 7.78-7.79 (m, 1H), 3.57-3.58 (m, 4H), 3.40-3.42 (m, 4H)。步驟 B 向來自上文步驟A之標題化合物(3.0 g, 10.8 mmol)於1,4-二噁烷(5 mL)中之溶液添加Cs₂ CO₃ (10.5 g, 32.4 mol)、1,10-菲咯啉(0.972 g, 5.40 mol)及碘化銅(1.03 g, 5.40 mol)，然後加熱至120℃持續12 h。經由矽藻土過濾反應混合物並用DCM/MeOH洗滌，濃縮且使用Biotage Isolera One純化系統，採用EtOAc/己烷梯度(40/60)，在矽膠管柱上純化粗製物，得到呈灰白色固體之標題化合物(0.150 g，粗製)。粗產物原樣用於下一步驟。 MS: 240.1 (M+H)⁺ 。 Preparation Example 67

Step A To a stirred solution of 4,6-dichloropyridin-3-amine (2.5 g, 15.3 mmol) in THF (50 mL) was added dropwise phosgene (4.55 g, 15.3 mol) in THF, then added TEA (4.28 mL, 30.7 mol) and heated to reflux for 2 h. The reaction mixture was concentrated under vacuum. The residue was dissolved in acetonitrile (50 mL) and toluene (50 mL), and morpholine (1.34 g, 15.3 mmol) was added and heated to 110° C. for 12 h. After that, the TLC was checked, and the crude product was concentrated and purified by silica gel column chromatography using petroleum ether: ethyl acetate (20:80) to obtain N-(4,6-dichloro-3-pyridyl) as a white solid ) Morpholine-4-carboxamide (3.0 g, 70.1%). MS: 276.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.55 (s, 1H), 8.38-8.40 (m, 1H), 7.78-7.79 (m, 1H), 3.57-3.58 (m, 4H), 3.40 -3.42 (m, 4H). Step B To the solution of the title compound (3.0 g, 10.8 mmol) from 1, A in 1,4-dioxane (5 mL) was added Cs ₂ CO ₃ (10.5 g, 32.4 mol), 1,10- Phenanthroline (0.972 g, 5.40 mol) and copper iodide (1.03 g, 5.40 mol) were then heated to 120°C for 12 h. The reaction mixture was filtered through diatomaceous earth and washed with DCM/MeOH, concentrated and the crude material was purified on a silica gel column using a Biotage Isolera One purification system using an EtOAc/hexane gradient (40/60) to give the title compound as an off-white solid (0.150 g, crude). The crude product was used as is in the next step. MS: 240.1 (M+H) ⁺ .

步驟 A 在0℃下向8-乙氧基-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(1 g, 4.62 mmol)於DCM (20 mL)中之攪拌溶液添加TEA (1.97 mL, 13.87 mmol)及BOC酸酐(1.5 mL, 6.93 mol)，然後在25℃下攪拌12 h。根據TLC在反應完成後，向反應混合物添加水，之後使用DCM進行萃取。將有機層分離並用鹽水溶液洗滌，濃縮並用己烷洗滌，獲得呈棕色固體之8-乙氧基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(0.8 g, 54%)。 MS: 317.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 10.69 (bs, 1H), 7.16 (d,J = 8.68 Hz, 1H), 6.87 (s, 1H), 6.66 (t,J = 6.76 Hz, 1H), 4.48 (s, 1H), 3.97-3.99 (m, 2H), 3.69 (t,J = 5.48 Hz, 2H), 2.74 (bs, 2H), 1.44 (s, 9H), 1.23 (t,J = 6.84 Hz, 3H)。步驟 B 在0℃下向氫化鈉(0.136 g, 2.84 mmol)於THF (5 mL)中之懸浮液逐滴添加來自上文步驟A之標題化合物(0.3 g, 0.95 mmol) (溶解於10 mL THF中)，然後在室溫下攪拌30 min。之後，在0℃下逐滴添加甲苯磺醯氯(0.27 g, 1.42 mmol) (溶解於10 mL THF中)，且然後在室溫下攪拌3 h。根據TLC在反應完成後，利用冰水將反應混合物淬滅，過濾所形成之固體且用水洗滌並乾燥，得到呈白色固體之8-乙氧基-5-甲苯磺醯基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-甲酸第三丁基酯(0.32 g, 72%)。產物原樣用於下一步驟。 MS: 371.2 (M+H)⁺ -Boc。步驟 C 在0℃下向來自上文步驟B之標題化合物(0.32 g, 0.68 mmol)於DCM中之溶液添加於二噁烷中之4 M HCl (4 mL)。將反應混合物攪拌2 h。在反應完成後，使該反應混合物蒸發以去除溶劑並利用二乙醚過濾，得到呈棕色固體之標題化合物(0.13 g, 56% )。產物原樣用於下一步驟。 MS: 371.2 (M+H)⁺ 。 Preparation Example 68

Step A To 8-ethoxy-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole (1 g, 4.62 mmol) in DCM (20 mL) at 0°C To the stirred solution, TEA (1.97 mL, 13.87 mmol) and BOC anhydride (1.5 mL, 6.93 mol) were added, and then stirred at 25°C for 12 h. After completion of the reaction according to TLC, water was added to the reaction mixture, followed by extraction with DCM. The organic layer was separated and washed with brine solution, concentrated and washed with hexane to obtain 8-ethoxy-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indole as a brown solid Tert-Butyl-2-carboxylic acid (0.8 g, 54%). MS: 317.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.69 (bs, 1H), 7.16 (d, J = 8.68 Hz, 1H), 6.87 (s, 1H), 6.66 (t, J = 6.76 Hz, 1H), 4.48 (s, 1H), 3.97-3.99 (m, 2H), 3.69 (t, J = 5.48 Hz, 2H), 2.74 (bs, 2H), 1.44 (s, 9H), 1.23 (t, J = 6.84 Hz, 3H). Step B To the suspension of sodium hydride (0.136 g, 2.84 mmol) in THF (5 mL) was added dropwise the title compound (0.3 g, 0.95 mmol) from step A above (dissolved in 10 mL THF at 0°C Medium), and then stirred at room temperature for 30 min. After that, tosyl chloride (0.27 g, 1.42 mmol) (dissolved in 10 mL of THF) was added dropwise at 0°C, and then stirred at room temperature for 3 h. After completion of the reaction according to TLC, the reaction mixture was quenched with ice water, the solid formed was filtered and washed with water and dried to give 8-ethoxy-5-tosyl-1,3,4 as a white solid ,5-Tetrahydro-2H-pyrido[4,3-b]indole-2-carboxylic acid tert-butyl ester (0.32 g, 72%). The product was used as is in the next step. MS: 371.2 (M+H) ⁺ -Boc. Step C To a solution of the title compound (0.32 g, 0.68 mmol) in DCM from Step B above was added 4 M HCl in dioxane (4 mL) at 0°C. The reaction mixture was stirred for 2 h. After the reaction was completed, the reaction mixture was evaporated to remove the solvent and filtered with diethyl ether to obtain the title compound (0.13 g, 56%) as a brown solid. The product was used as is in the next step. MS: 371.2 (M+H) ⁺ .

步驟 A 向4-側氧基六氫吡啶-1-甲酸第三丁基酯(10.00 g, 0.0502 mol)於乙醇(100 mL)中之攪拌溶液添加羥胺鹽酸鹽(6.98 g, 0.100 mol)及CH₃ COONa (8.23 g, 0.100 mol)，然後在氮氣氛下加熱至90℃持續12 h。根據LCMS在反應完成後，將反應混合物濃縮，且向粗製材料添加水(100 mL)，之後藉由使用二氯甲烷(250 mL)進行萃取。將有機層濃縮且藉由矽膠管柱(Biotage)使用於石油醚中之18%-30%乙酸乙酯純化粗產物，獲得呈白色固體之4-(羥基亞胺基)六氫吡啶-1-甲酸第三丁基酯(5 g, 46.4%)。 MS: 159.1 (M+H)⁺ -第三丁基¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 10.45 (s, 1H), 3.33-3.36 (m, 4H), 2.42-2.44 (m, 2H), 2.20-2.22 (m, 2H), 1.41 (s, 9H)。步驟 B 在0℃下向氫化鈉(0.268 g, 7.00 mmol)於DMF (3 mL)中之懸浮液逐滴添加來自上文步驟A之標題化合物(0.500 g, 2.33 mmol) (溶解於5 mL DMF中)，然後在室溫下攪拌60 min。之後，在0℃下逐滴添加2-氟吡啶(0.340 g, 3.50 mmol) (溶解於2 mL DMF中)，且然後在室溫下攪拌3 h。根據TLC在反應完成後，利用冰水將反應混合物淬滅，之後使用乙酸乙酯(30 mL)進行萃取。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，獲得呈淡棕色固體之4-側氧基-3-(2-側氧基-1,2-二氫吡啶-3-基)六氫吡啶-1-甲酸第三丁基酯(300 mg，粗製)。粗產物原樣用於下一步驟。 MS: 293.2 (M+H)⁺ 。步驟 C 將來自上文步驟B之標題化合物(0.5 g, 1.71 Mmol)於濃H₂ SO₄ (2.0 mL)中之懸浮液在氮氣氛下在室溫下攪拌16 h。之後檢查LCMS，其僅指示起始材料，然後在氮氣氛下將反應混合物加熱至60℃持續16 h。之後檢查LCMS，其指示80%之產物質量。將反應混合物冷卻至室溫，向此添加於水中之10%乙腈(20.0 mL)，且藉由使用固體K₂ CO₃ 使反應混合物鹼化，然後將固體過濾。將濾液濃縮，得到呈棕色黏性油狀物之標題化合物(250 mg，粗製)。產物原樣用於下一步驟。 MS: 175.1 (M+H)⁺ 。步驟 D 在0℃下向來自上文步驟C之標題化合物(0.6 g, 3.15 mmol)於四氫呋喃(10.0 mL)中之攪拌溶液添加TEA (1.32 mL, 9.46 m0mol)及BOC酸酐(0.869 mL, 3.78 mmol)，然後在氮氣氛下在25℃下攪拌12 h。根據TLC在反應完成後，將反應混合物濃縮且藉由矽膠管柱(Biotage)使用於石油醚中之15%-20%乙酸乙酯純化粗產物，得到呈灰白色固體之-7,8-二氫呋喃并[2,3-b:4,5-c']二吡啶-6(5H)-甲酸第三丁基酯(500 mg, 53%)。 MS: 275.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.21-8.22 (m, 1H), 8.05-8.06 (m, 1H), 7.30-7.31 (m, 1H), 4.51 (s, 2H), 3.76 (t,J = 5.60 Hz, 2H), 2.86 (t,J = 5.60 Hz, 2H), 1.44 (s, 9H)。步驟 E 在0℃下向來自上文步驟D之標題化合物(0.5 g, 1.67 mmol)於二氯甲烷(5 mL)中之攪拌溶液添加於二噁烷中之4.0 M HCl (2 mL)，然後在0℃-20℃下攪拌2 h。根據TLC及LCMS在反應完成後，將反應混合物濃縮，得到呈灰白色固體之標題化合物(350 mg，定量)。 MS: 175.1 (M+H)⁺ 。 Preparation Example 69

Step A To a stirred solution of 4-pentoxyhexahydropyridine-1-carboxylic acid tert-butyl ester (10.00 g, 0.0502 mol) in ethanol (100 mL) was added hydroxylamine hydrochloride (6.98 g, 0.100 mol) and CH ₃ COONa (8.23 g, 0.100 mol), then heated to 90°C for 12 h under a nitrogen atmosphere. After the reaction was completed according to LCMS, the reaction mixture was concentrated, and water (100 mL) was added to the crude material, followed by extraction by using dichloromethane (250 mL). The organic layer was concentrated and the crude product was purified by silica gel column (Biotage) using 18%-30% ethyl acetate in petroleum ether to obtain 4-(hydroxyimino)hexahydropyridine-1-as a white solid Tert-butyl formate (5 g, 46.4%). MS: 159.1 (M+H) ⁺ -tert-butyl ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.45 (s, 1H), 3.33-3.36 (m, 4H), 2.42-2.44 (m , 2H), 2.20-2.22 (m, 2H), 1.41 (s, 9H). Step B To the suspension of sodium hydride (0.268 g, 7.00 mmol) in DMF (3 mL) was added dropwise the title compound from step A above (0.500 g, 2.33 mmol) (dissolved in 5 mL DMF Medium), and then stirred at room temperature for 60 min. After that, 2-fluoropyridine (0.340 g, 3.50 mmol) (dissolved in 2 mL DMF) was added dropwise at 0°C, and then stirred at room temperature for 3 h. After completion of the reaction according to TLC, the reaction mixture was quenched with ice water, and then extracted with ethyl acetate (30 mL). The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to obtain 4-oxo-3-(2-oxo-1,2-dihydropyridin-3-yl)hexahydro as a light brown solid Pyridine-1-carboxylic acid tert-butyl ester (300 mg, crude). The crude product was used as is in the next step. MS: 293.2 (M+H) ⁺ . Step C A suspension of the title compound (0.5 g, 1.71 Mmol) in concentrated H ₂ SO ₄ (2.0 mL) from Step B above was stirred at room temperature under a nitrogen atmosphere for 16 h. The LCMS was then checked, which indicated only the starting material, and then the reaction mixture was heated to 60° C. under a nitrogen atmosphere for 16 h. Then check the LCMS, which indicates 80% product quality. The reaction mixture was cooled to room temperature, 10% acetonitrile (20.0 mL) in water was added thereto, and the reaction mixture was basified by using solid K ₂ CO ₃ and then the solid was filtered. The filtrate was concentrated to give the title compound (250 mg, crude) as a brown viscous oil. The product was used as is in the next step. MS: 175.1 (M+H) ⁺ . Step D To a stirred solution of the title compound (0.6 g, 3.15 mmol) in tetrahydrofuran (10.0 mL) from Step C above was added TEA (1.32 mL, 9.46 m0mol) and BOC anhydride (0.869 mL, 3.78 mmol) at 0°C ), and then stirred at 25°C for 12 h under a nitrogen atmosphere. After completion of the reaction according to TLC, the reaction mixture was concentrated and the crude product was purified by silica gel column (Biotage) using 15%-20% ethyl acetate in petroleum ether to give -7,8-dihydrogen as off-white solid Furo[2,3-b:4,5-c']dipyridin-6(5H)-carboxylic acid tert-butyl ester (500 mg, 53%). MS: 275.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.21-8.22 (m, 1H), 8.05-8.06 (m, 1H), 7.30-7.31 (m, 1H), 4.51 (s, 2H), 3.76 (t, J = 5.60 Hz, 2H), 2.86 (t, J = 5.60 Hz, 2H), 1.44 (s, 9H). Step E To the stirred solution of the title compound (0.5 g, 1.67 mmol) in dichloromethane (5 mL) from Step D above was added 4.0 M HCl (2 mL) in dioxane at 0°C, then Stir at 0℃-20℃ for 2 h. After completion of the reaction according to TLC and LCMS, the reaction mixture was concentrated to give the title compound (350 mg, quantitative) as an off-white solid. MS: 175.1 (M+H) ⁺ .

步驟 A 向2,5-二氯-1,3-苯并噁唑(0.25 g, 0.0013 mol)於乙腈(15 mL)中之攪拌溶液添加K₂ CO₃ (0.55 g, 0.0040 mol)及(2S,6R)-2,6-二甲基嗎啉(0.17 g, 0.0014 mol)。之後將反應混合物加熱至70℃持續12 h。 根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮至呈白色固體之標題化合物(0.2 g, 56%)。 MS: 267.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.43 (d,J = 8.44 Hz, 1H), 7.34 (d,J = 1.96 Hz, 1H), 7.04-7.04 (m, 1H), 3.99 (d,J = 13.16 Hz, 2H), 3.66-3.67 (m, 2H), 2.82 (t,J = 10.84 Hz, 2H), 1.00 (d,J = 6.28 Hz, 6H)。 Preparation Example 70

Step A To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.25 g, 0.0013 mol) in acetonitrile (15 mL), add K ₂ CO ₃ (0.55 g, 0.0040 mol) and (2S ,6R)-2,6-dimethylmorpholine (0.17 g, 0.0014 mol). The reaction mixture was then heated to 70 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to the title compound (0.2 g, 56%) as a white solid. MS: 267.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.43 (d, J = 8.44 Hz, 1H), 7.34 (d, J = 1.96 Hz, 1H), 7.04-7.04 (m, 1H), 3.99 ( d, J = 13.16 Hz, 2H), 3.66-3.67 (m, 2H), 2.82 (t, J = 10.84 Hz, 2H), 1.00 (d, J = 6.28 Hz, 6H).

步驟 A 向2,5-二氯-1,3-苯并噁唑(0.6 g, 0.0032 mol)於乙腈(15 mL)中之攪拌溶液添加K₂ CO₃ (1.32 g, 0.0096 mol)及第三丁基胺(0.255 g, 0.0035 mol)。之後將反應混合物加熱至70℃持續12 h。 根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，得到呈棕色固體之標題化合物(0.4 g, 55%)。 MS: 225.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 7.97 (s, 1H), 7.31-7.32 (m, 2H), 6.96-6.97 (m, 1H), 1.40 (s, 9H)。 Preparation Example 71

Step A To a stirred solution of 2,5-dichloro-1,3-benzoxazole (0.6 g, 0.0032 mol) in acetonitrile (15 mL), add K ₂ CO ₃ (1.32 g, 0.0096 mol) and the third Butylamine (0.255 g, 0.0035 mol). The reaction mixture was then heated to 70 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to give the title compound (0.4 g, 55%) as a brown solid. MS: 225.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 7.97 (s, 1H), 7.31-7.32 (m, 2H), 6.96-6.97 (m, 1H), 1.40 (s, 9H).

步驟 A 向6-溴-2-氯喹唑啉(0.3 g, 1.234 mmol)於乙腈(10 mL)中之攪拌溶液添加K₂ CO₃ (0.34 g, 2.467 mmol)及嗎啉(0.17 g, 1.85 mmol)。之後將反應混合物加熱至70℃持續12 h。 根據TLC在反應完成後，向反應混合物添加DCM及水(50 mL)。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮，得到呈淡黃色固體之標題化合物(0.25 g, 69%)。產物原樣用於下一步驟。 MS: 294.0 (M+H)⁺ 。 Preparation Example 72

Step A To a stirred solution of 6-bromo-2-chloroquinazoline (0.3 g, 1.234 mmol) in acetonitrile (10 mL) was added K ₂ CO ₃ (0.34 g, 2.467 mmol) and morpholine (0.17 g, 1.85 mmol ). The reaction mixture was then heated to 70 °C for 12 h. After completion of the reaction according to TLC, DCM and water (50 mL) were added to the reaction mixture. The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to give the title compound (0.25 g, 69%) as a light yellow solid. The product was used as is in the next step. MS: 294.0 (M+H) ⁺ .

步驟 A 向2,6-二氯-1,5-萘啶(100 mg, 0.502 mmol)於二噁烷(3 mL)中之攪拌溶液添加三乙胺(0.210 ml, 1.507 mmol)及嗎啉(0.052 ml, 0.603 mmol)。然後在110℃下攪拌反應混合物。4 h後，反應未完成，因此添加三乙胺(0.210 ml, 1.507 mmol)，且將反應混合物進一步在110℃下攪拌過夜。將反應混合物濃縮至乾燥且然後溶解於二氯甲烷中，並用飽和NH₄ Cl水溶液洗滌。用二氯甲烷將水層萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下蒸發，得到呈米色固體之標題產物(93 mg, 74%)。 MS: 250.02 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.05 (d,J = 9.4 Hz, 1H), 7.98 (d,J = 8.8 Hz, 1H), 7.59 (d,J = 8.7 Hz, 1H), 7.53 (d,J = 9.5 Hz, 1H), 3.71 (hept,J = 3.3, 2.6 Hz, 8H)。 Preparation Example 73

Step A To a stirred solution of 2,6-dichloro-1,5-naphthyridine (100 mg, 0.502 mmol) in dioxane (3 mL) was added triethylamine (0.210 ml, 1.507 mmol) and morpholine ( 0.052 ml, 0.603 mmol). The reaction mixture was then stirred at 110°C. After 4 h, the reaction was not completed, so triethylamine (0.210 ml, 1.507 mmol) was added, and the reaction mixture was further stirred at 110° C. overnight. The reaction mixture was concentrated to dryness and then dissolved in dichloromethane, and washed with saturated aqueous NH ₄ Cl. The aqueous layer was extracted twice with dichloromethane. The combined organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give the title product (93 mg, 74%) as a beige solid. MS: 250.02 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.05 (d, J = 9.4 Hz, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.59 (d, J = 8.7 Hz, 1H) , 7.53 (d, J = 9.5 Hz, 1H), 3.71 (hept, J = 3.3, 2.6 Hz, 8H).

步驟 A 將2-溴-5-氯吡啶(200 mg, 1.039 mmol)溶解於乙腈(2.5 mL)中，向此添加(1S,4S)-2-氧雜-5-氮雜二環[2.2.1]庚烷鹽酸鹽(211 mg, 1.559 mmol)及三乙胺(0.362 mL, 2.60 mmol)，且將懸浮液於微波中輻照至160℃持續1h20。然後將樣品在水(20 mL)與二氯甲烷(20 mL)之間進行萃取。將水層用二氯甲烷洗滌兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統利用二氯甲烷/甲醇梯度(100/0 -＞ 90/10)在矽膠管柱上純化粗製物，得到呈米色固體之產物(57 mg, 26%)。 MS: 211.03 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.06 (d,J = 2.6 Hz, 1H), 7.56 (dd,J = 9.0, 2.7 Hz, 1H), 6.57 (d, J = 9.0 Hz, 1H), 4.80 (d,J = 2.3 Hz, 1H), 4.65 (d,J = 2.4 Hz, 1H), 3.76 (dd,J = 7.3, 1.5 Hz, 1H), 3.61 (d,J = 7.3 Hz, 1H), 3.43 (dd,J = 10.1, 1.5 Hz, 1H), 3.20 (d,J = 10.3 Hz, 1H), 1.90 (dd,J = 9.7, 2.3 Hz, 1H), 1.87 - 1.81 (m, 1H)。 Preparation Example 74

Step A Dissolve 2-bromo-5-chloropyridine (200 mg, 1.039 mmol) in acetonitrile (2.5 mL), and add (1S,4S)-2-oxa-5-azabicyclo[2.2. 1] Heptane hydrochloride (211 mg, 1.559 mmol) and triethylamine (0.362 mL, 2.60 mmol), and the suspension was irradiated in the microwave to 160°C for 1 h20. The sample was then extracted between water (20 mL) and dichloromethane (20 mL). The aqueous layer was washed twice with dichloromethane. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified on a silica gel column using a Biotage Isolera One purification system using a dichloromethane/methanol gradient (100/0 -> 90/10) to give the product as a beige solid (57 mg, 26%). MS: 211.03 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.06 (d, J = 2.6 Hz, 1H), 7.56 (dd, J = 9.0, 2.7 Hz, 1H), 6.57 (d, J = 9.0 Hz, 1H), 4.80 (d, J = 2.3 Hz, 1H), 4.65 (d, J = 2.4 Hz, 1H), 3.76 (dd, J = 7.3, 1.5 Hz, 1H), 3.61 (d, J = 7.3 Hz, 1H), 3.43 (dd, J = 10.1, 1.5 Hz, 1H), 3.20 (d, J = 10.3 Hz, 1H), 1.90 (dd, J = 9.7, 2.3 Hz, 1H), 1.87-1.81 (m, 1H ).

步驟 A 將2-溴-5-氯吡啶(200 mg, 1.039 mmol)溶解於乙腈(2.5 mL)中，向此添加(1R,4R)-2-氧雜-5-氮雜二環[2.2.1]庚烷鹽酸鹽(211 mg, 1.559 mmol)及三乙胺(0.362 mL, 2.60 mmol)，且將懸浮液於微波中輻照至160℃持續1h20。然後將樣品在水(20 mL)與二氯甲烷(20 mL)之間進行萃取。將水層用二氯甲烷洗滌兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並在減壓下濃縮。使用Biotage Isolera One純化系統利用二氯甲烷/甲醇梯度(100/0 -＞ 90/10)在矽膠管柱上純化粗製物，得到呈米色固體之產物(51 mg, 23%)。 MS: 211.04 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d₆ ) δ = 8.06 (dd,J = 2.7, 0.7 Hz, 1H), 7.56 (dd,J = 9.0, 2.7 Hz, 1H), 6.63 - 6.50 (m, 1H), 4.80 (s, 1H), 4.64 (s, 1H), 3.75 (dd,J = 7.3, 1.5 Hz, 1H), 3.61 (d,J = 7.3, 0.9 Hz, 1H), 3.43 (dd,J = 10.1, 1.6 Hz, 1H), 3.20 (d,J = 10.1, 1.1 Hz, 1H), 1.94 - 1.78 (m, 2H)。 Preparation Example 75

Step A Dissolve 2-bromo-5-chloropyridine (200 mg, 1.039 mmol) in acetonitrile (2.5 mL), and add (1R,4R)-2-oxa-5-azabicyclo[2.2. 1] Heptane hydrochloride (211 mg, 1.559 mmol) and triethylamine (0.362 mL, 2.60 mmol), and the suspension was irradiated in the microwave to 160°C for 1 h20. The sample was then extracted between water (20 mL) and dichloromethane (20 mL). The aqueous layer was washed twice with dichloromethane. The combined organic layer was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The crude product was purified on a silica gel column using a Biotage Isolera One purification system using a dichloromethane/methanol gradient (100/0 -> 90/10) to give the product as a beige solid (51 mg, 23%). MS: 211.04 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 8.06 (dd, J = 2.7, 0.7 Hz, 1H), 7.56 (dd, J = 9.0, 2.7 Hz, 1H), 6.63-6.50 (m, 1H ), 4.80 (s, 1H), 4.64 (s, 1H), 3.75 (dd, J = 7.3, 1.5 Hz, 1H), 3.61 (d, J = 7.3, 0.9 Hz, 1H), 3.43 (dd, J = 10.1, 1.6 Hz, 1H), 3.20 (d, J = 10.1, 1.1 Hz, 1H), 1.94-1.78 (m, 2H).

步驟 A 向經脫氣之四氫呋喃(5 mL)添加氯-(2-二環己基膦基-2′,6′-二異丙氧基-1,1′-聯苯)[2-(2-胺基乙基)苯基]鈀(II)-甲基-第三丁基醚加成物(PdRuPhos G1) (0.017 g, 0.024 mmol)、2-二環己基膦基-2′,6′-二異丙氧基聯苯(RuPho) (0.011 g, 0.024 mmol)、來自製備實例2 之標題化合物(0.05 g, 0.024 mmol)及市售4-(6-溴苯并[d]噻唑-2-基)嗎啉(0.073 g, 0.029 mmol)。然後，添加雙(三甲基矽基)胺基鋰(LiHMDS)於四氫呋喃(1 mL, 1 mmol)中之1 M溶液。將所得反應混合物在回流下加熱2小時。將反應混合物冷卻至室溫，溶解於二氯甲烷(100 mL)中。將有機相用水及鹽水洗滌且經Na₂ SO₄ 乾燥。將溶劑在減壓下去除。使用Biotage Isolera One純化系統，採用乙酸乙酯/正庚烷梯度(80/20 =＞ 100/0)，在矽膠管柱上純化粗產物，得到標題化合物(0.070 g, 69%)。¹ H-NMR (400 MHz，氯仿-d ) δ = 7.51 (d,J = 8.8 Hz, 1H), 7.31 (d,J = 2.4 Hz, 1H), 7.27 (s, 1H), 7.20 (dd,J = 8.8, 4.2 Hz, 1H), 7.14 (td,J = 8.6, 2.4 Hz, 2H), 6.94 (td,J = 9.1, 2.5 Hz, 1H), 4.40 (s, 2H), 3.88 - 3.81 (m, 4H), 3.70 (t,J = 5.7 Hz, 2H), 3.64 (s, 3H), 3.59 (t,J = 4.9 Hz, 4H), 2.92 (t,J = 5.7 Hz, 2H)。 Example 1

Step A To degassed tetrahydrofuran (5 mL), add chloro-(2-dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl) (2-(2- (Aminoethyl)phenyl)palladium(II)-methyl-third butyl ether adduct (PdRuPhos G1) (0.017 g, 0.024 mmol), 2-dicyclohexylphosphino-2′,6′- Diisopropoxybiphenyl (RuPho) (0.011 g, 0.024 mmol), the title compound from Preparation Example 2 (0.05 g, 0.024 mmol) and commercially available 4-(6-bromobenzo[d]thiazole-2- Base) morpholine (0.073 g, 0.029 mmol). Then, a 1 M solution of lithium bis(trimethylsilyl)amide (LiHMDS) in tetrahydrofuran (1 mL, 1 mmol) was added. The resulting reaction mixture was heated at reflux for 2 hours. The reaction mixture was cooled to room temperature and dissolved in dichloromethane (100 mL). The organic phase was washed with water and brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure. Using the Biotage Isolera One purification system using ethyl acetate/n-heptane gradient (80/20 => 100/0), the crude product was purified on a silica gel column to obtain the title compound (0.070 g, 69%). ¹ H-NMR (400 MHz, chloroform- d ) δ = 7.51 (d, J = 8.8 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 7.27 (s, 1H), 7.20 (dd, J = 8.8, 4.2 Hz, 1H), 7.14 (td, J = 8.6, 2.4 Hz, 2H), 6.94 (td, J = 9.1, 2.5 Hz, 1H), 4.40 (s, 2H), 3.88-3.81 (m, 4H), 3.70 (t, J = 5.7 Hz, 2H), 3.64 (s, 3H), 3.59 (t, J = 4.9 Hz, 4H), 2.92 (t, J = 5.7 Hz, 2H).

步驟 A 向製備實例1之標題化合物(0.150 g, 1 eq)於無水1,4-二噁烷(5 mL)中之攪拌溶液添加市售4-(6-溴苯并[d]噻唑-2-基)嗎啉(1 eq)、第三丁醇鈉(3 eq)，且將混合物在N₂ 氣氛下脫氣10分鐘。向此反應混合物添加Pd₂ (dba)₃ (0.05 eq)及Ru-Phos (0.1 eq)，且將混合物加熱至100℃直至反應完成為止。在反應完成後，經由矽藻土床過濾該反應混合物，並用EtOAc洗滌。將濾液濃縮且藉由管柱層析或製備型HPLC純化粗產物，得到如表2中所指示之標題化合物6 。 Example 2

Step A To a stirred solution of the title compound of Preparation Example 1 (0.150 g, 1 eq) in anhydrous 1,4-dioxane (5 mL) was added commercially available 4-(6-bromobenzo[d]thiazole-2 -Yl) morpholine (1 eq), sodium tert-butoxide (3 eq), and the mixture was degassed under N ₂ atmosphere for 10 minutes. To this reaction mixture, Pd ₂ (dba) ₃ (0.05 eq) and Ru-Phos (0.1 eq) were added, and the mixture was heated to 100° C. until the reaction was completed. After the reaction was completed, the reaction mixture was filtered through a bed of celite and washed with EtOAc. The filtrate was concentrated and the crude product was purified by column chromatography or preparative HPLC to obtain the title compound 6 as indicated in Table 2.

Examples 3 to 96e In addition to using the tricyclic amino derivatives and bromine/chlorine derivatives indicated in the table below, following the palladium coupling procedure as described in Examples 1 and 2 , the following compounds were prepared. Examples 71 and 72 were prepared following the procedures described in Preparation Examples 42 and 43 , respectively, followed by the deprotection procedures described in Example 97 . Table 2

將來自製備實例33之標題化合物(0.072 g, 0.142 mmol)及碳酸銫(0.107 g, 0.328 mmol)置入至微波管中，之後置入MeOH (1.5 mL)及THF (3 mL)。將反應混合物於微波中輻照至110℃持續30分鐘，且然後使其冷卻至室溫。將溶劑在減壓下去除，且使用Biotage Isolera One純化系統，利用正庚烷/乙酸乙酯梯度(80/20-＞ 0/100)在矽膠管柱上純化殘餘物，得到呈黃色固體之標題化合物(0.027 g, 53%)。 MS: 354.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 10.96 (s, 1H), 7.39 (d,J = 9.9 Hz, 1H), 7.28 (s, 1H), 7.26 (t,J = 4.4 Hz, 1H), 7.20 (dd,J = 10.0, 2.6 Hz, 1H), 6.85 (td,J = 9.2, 2.6 Hz, 1H), 4.62 (s, 2H), 3.91 (t,J = 5.7 Hz, 2H), 3.79 - 3.67 (m, 4H), 3.37 - 3.32 (m, 4H), 2.88 (t,J = 5.6 Hz, 2H)。 Example 97

The title compound from Preparation Example 33 (0.072 g, 0.142 mmol) and cesium carbonate (0.107 g, 0.328 mmol) were placed in a microwave tube, followed by MeOH (1.5 mL) and THF (3 mL). The reaction mixture was irradiated in the microwave to 110°C for 30 minutes, and then allowed to cool to room temperature. The solvent was removed under reduced pressure and the residue was purified on a silica gel column using a Biotage Isolera One purification system using a n-heptane/ethyl acetate gradient (80/20->0/100) to give the title as a yellow solid Compound (0.027 g, 53%). MS: 354.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 10.96 (s, 1H), 7.39 (d, J = 9.9 Hz, 1H), 7.28 (s, 1H), 7.26 (t, J = 4.4 Hz, 1H), 7.20 (dd, J = 10.0, 2.6 Hz, 1H), 6.85 (td, J = 9.2, 2.6 Hz, 1H), 4.62 (s, 2H), 3.91 (t, J = 5.7 Hz, 2H), 3.79-3.67 (m, 4H), 3.37-3.32 (m, 4H), 2.88 (t, J = 5.6 Hz, 2H).

Examples 98 to 105f The following compounds were prepared by following the tosylate cracking procedure as described in Example 97 except that the tosylate protection precursor indicated in the table below was used: Table 3

步驟 A 向來自製備實例7之標題化合物(500 mg, 1.31 mmol)於DMF (5 mL)中之溶液添加2,5-二氯苯并噁唑(327 mg, 1.74 mmol)及碳酸鉀(602 mg, 4.36 mmol)，且加熱至100℃持續8小時。在反應完成後，將反應混合物傾倒至冰冷卻之水中；將沈澱出之固體過濾並乾燥，得到標題化合物。產物不經進一步純化即原樣用於下一步驟(0.500 g, 77%)。 MS: 496.1 (M+H)⁺ 。步驟 B 於50 mL雙頸圓底燒瓶中添加無水1,4-二噁烷(5 mL)並脫氣20 min。向此添加乙酸鈀(67 mg, 0.1008 mmol)及2-二環己基膦基-2′,4′,6′-三異丙基聯苯(144 mg, 0.302 mmol)。所得溶液呈橙色且再繼續脫氣10 min。將懸浮液在預先加熱之油浴上在100℃下加熱數秒(20- 30秒)。色彩變為深綠色。將油浴移除並脫氣5-10 min。向此添加來自上文步驟A之標題化合物(500 mg, 1.008 mmol)、嗎啉(88 mg, 1.008 mmol)及碳酸銫(0.98 g, 3.024 mmol)，且將反應混合物加熱至100℃持續3小時。3 h後，LC-MS顯示產物為主要峰。經由矽藻土墊過濾反應混合物並在真空下濃縮，得到標題化合物。產物不經進一步純化即原樣用於下一步驟(0.450 g, 82%)。 MS: 547.3 (M+H)⁺ 。步驟 C 向來自上文步驟B之標題化合物(450 mg, 0.82 mmol)於1,4-二噁烷:甲醇(1:1；5 mL)中之溶液添加第三丁醇鈉(230 mg, 2.4 mmol)，且加熱至70℃持續16小時。將反應混合物在真空下濃縮。使用Biotage Isolera One純化系統，採用石油醚/ EtOAc梯度(0 - 100%)，在矽膠管柱上實施純化，得到呈固體之標題化合物(0.270 g, 84%)。 MS: 393.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.10 (s, 1H), 7.47-7.48 (m, 1H), 7.28 (d,J = 8.72 Hz, 1H), 7.08-7.09 (m, 1H), 6.92 (d,J = 2.20 Hz, 1H), 6.83-6.84 (m, 1H), 6.62-6.63 (m, 1H), 4.81 (s, 2H), 3.98-4.00 (m, 2H), 3.73-3.74 (m, 4H), 3.04-3.05 (m, 4H), 2.94-2.95 (m, 2H)。 Example 106

Step A To a solution of the title compound (500 mg, 1.31 mmol) in DMF (5 mL) from Preparation Example 7 was added 2,5-dichlorobenzoxazole (327 mg, 1.74 mmol) and potassium carbonate (602 mg , 4.36 mmol), and heated to 100 ℃ for 8 hours. After the reaction was completed, the reaction mixture was poured into ice-cooled water; the precipitated solid was filtered and dried to obtain the title compound. The product was used as such in the next step without further purification (0.500 g, 77%). MS: 496.1 (M+H) ⁺ . Step B Anhydrous 1,4-dioxane (5 mL) was added to a 50 mL double-necked round bottom flask and degassed for 20 min. To this, palladium acetate (67 mg, 0.1008 mmol) and 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (144 mg, 0.302 mmol) were added. The resulting solution was orange and degassed for another 10 minutes. The suspension was heated on a preheated oil bath at 100°C for several seconds (20-30 seconds). The color becomes dark green. Remove the oil bath and degas for 5-10 min. To this was added the title compound (500 mg, 1.008 mmol), morpholine (88 mg, 1.008 mmol) and cesium carbonate (0.98 g, 3.024 mmol) from Step A above, and the reaction mixture was heated to 100° C. for 3 hours . After 3 h, LC-MS showed that the product was the main peak. The reaction mixture was filtered through a pad of celite and concentrated under vacuum to obtain the title compound. The product was used as such in the next step without further purification (0.450 g, 82%). MS: 547.3 (M+H) ⁺ . Step C To the solution of the title compound (450 mg, 0.82 mmol) in 1,4-dioxane: methanol (1:1; 5 mL) from step B above was added sodium tert-butoxide (230 mg, 2.4 mmol) and heated to 70°C for 16 hours. The reaction mixture was concentrated under vacuum. Using the Biotage Isolera One purification system with a petroleum ether/EtOAc gradient (0-100%), purification was performed on a silica gel column to obtain the title compound as a solid (0.270 g, 84%). MS: 393.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.10 (s, 1H), 7.47-7.48 (m, 1H), 7.28 (d, J = 8.72 Hz, 1H), 7.08-7.09 (m, 1H ), 6.92 (d, J = 2.20 Hz, 1H), 6.83-6.84 (m, 1H), 6.62-6.63 (m, 1H), 4.81 (s, 2H), 3.98-4.00 (m, 2H), 3.73- 3.74 (m, 4H), 3.04-3.05 (m, 4H), 2.94-2.95 (m, 2H).

向來自製備實例7之標題化合物(500 mg, 1.318 mmol)於DMF (5 mL)中之溶液添加2,5-二氯苯并噻唑(268 mg, 1.32 mmol)及碳酸鉀(545 mg, 3.95 mmol)，且加熱至100℃持續8小時。在反應完成後，將反應混合物傾倒至冰冷卻之水中，將沈澱出之固體過濾並乾燥，得到標題化合物(500 mg, 74%)。產物不經進一步純化即原樣用於下一步驟。 MS: 513.1 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(200 mg, 0.39 mmol)於無水二噁烷(5 mL)中之攪拌溶液添加嗎啉(51 mg, 0.58 mmol)、第三丁醇鈉(112 mg, 1.17 mmol)，且在N₂ 氣氛下脫氣10 min。向此反應混合物添加參(二亞苄基丙酮)二鈀(0) (17.9 mg, 0.019 mmol)及2-二環己基膦基-2′,6′-二異丙氧基聯苯(18.2 mg, 0.039 mmol)，且加熱至100℃直至反應完成為止。在反應完成後(藉由LCMS監測)，經由矽藻土過濾反應混合物並用EtOAc洗滌。將濾液在減壓下濃縮以產生粗產物。使用Biotage Isolera One純化系統，採用石油醚/ EtOAc梯度(0 - 100%)，在矽膠管柱上實施純化，得到呈固體之標題化合物(50 mg, 33%)。 MS: 409.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.11 (s, 1H), 7.60 (d,J = 8.68 Hz, 1H), 7.48-7.49 (m, 1H), 7.07-7.08 (m, 2H), 6.86-6.88 (m, 2H), 4.77 (s, 2H), 3.97-3.98 (m, 2H), 3.74-3.76 (m, 4H), 3.11-3.12 (m, 4H), 2.95-2.96 (m, 2H)。 Example 107

To a solution of the title compound (500 mg, 1.318 mmol) in DMF (5 mL) from Preparation Example 7, 2,5-dichlorobenzothiazole (268 mg, 1.32 mmol) and potassium carbonate (545 mg, 3.95 mmol) were added ), and heated to 100 ℃ for 8 hours. After the reaction was completed, the reaction mixture was poured into ice-cooled water, and the precipitated solid was filtered and dried to obtain the title compound (500 mg, 74%). The product was used as is in the next step without further purification. MS: 513.1 (M+H) ⁺ . Step B To a stirred solution of the title compound (200 mg, 0.39 mmol) from anhydrous dioxane (5 mL) from step A above, add morpholine (51 mg, 0.58 mmol) and sodium tert-butoxide (112 mg , 1.17 mmol), and degassed under N ₂ atmosphere for 10 min. To this reaction mixture was added ginseng (dibenzylideneacetone) dipalladium (0) (17.9 mg, 0.019 mmol) and 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl (18.2 mg , 0.039 mmol), and heated to 100 ℃ until the reaction was completed. After the reaction was completed (monitored by LCMS), the reaction mixture was filtered through celite and washed with EtOAc. The filtrate was concentrated under reduced pressure to produce crude product. Using the Biotage Isolera One purification system, using a petroleum ether/EtOAc gradient (0-100%), purification was performed on a silica gel column to obtain the title compound (50 mg, 33%) as a solid. MS: 409.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.11 (s, 1H), 7.60 (d, J = 8.68 Hz, 1H), 7.48-7.49 (m, 1H), 7.07-7.08 (m, 2H ), 6.86-6.88 (m, 2H), 4.77 (s, 2H), 3.97-3.98 (m, 2H), 3.74-3.76 (m, 4H), 3.11-3.12 (m, 4H), 2.95-2.96 (m , 2H).

步驟 A 向來自製備實例7之標題化合物(1 g, 2.63 mmol)於DMF (10 mL)中之溶液添加2,6-二氯苯并噻唑(0.537 g, 2.63 mmol)及碳酸鉀(1.09 g, 7.89 mmol)，且加熱至100℃持續8小時。在反應完成後，將反應混合物傾倒至冰冷卻之水中，將沈澱出之固體過濾並乾燥，得到標題化合物(1.0 g, 74%)。產物不經進一步純化即原樣用於下一步驟。 MS: 513.2 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(150 mg, 0.293 mmol)於無水二噁烷(5 mL)中之攪拌溶液添加嗎啉(39 mg, 0.44 mmol)、第三丁醇鈉(85 mg, 0.87 mmol)，且在N₂ 氣氛下脫氣10 min。向此反應混合物添加參(二亞苄基丙酮)二鈀(0) (13.4 mg, 0.0015 mmol)及2-二環己基膦基-2′,6′-二異丙氧基聯苯(13.65 mg, 0.029 mmol)，且加熱至100℃直至反應完成為止。在反應完成後(藉由LCMS監測)，經由矽藻土過濾反應混合物並用EtOAc洗滌。將濾液在減壓下濃縮以產生粗產物。使用Biotage Isolera One純化系統，採用石油醚/ EtOAc梯度(0 - 100%)，在矽膠管柱上實施純化，得到呈固體之標題化合物(60 mg, 50%)。 MS: 409.1 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.10 (s, 1H), 7.48-7.50 (m, 1H), 7.36-7.38 (m, 2H), 7.08-7.09 (m, 1H), 6.96-6.97 (m, 1H), 6.85-6.86 (m, 1H), 4.74 (s, 2H), 3.94-3.95 (m, 2H), 3.74-3.75 (m, 4H), 3.06-3.07 (m, 4H), 2.94-2.95 (m, 2H)。 Example 108

Step A To a solution of the title compound from Preparation Example 7 (1 g, 2.63 mmol) in DMF (10 mL) was added 2,6-dichlorobenzothiazole (0.537 g, 2.63 mmol) and potassium carbonate (1.09 g, 7.89 mmol) and heated to 100°C for 8 hours. After the reaction was completed, the reaction mixture was poured into ice-cooled water, and the precipitated solid was filtered and dried to obtain the title compound (1.0 g, 74%). The product was used as is in the next step without further purification. MS: 513.2 (M+H) ⁺ . Step B To a stirred solution of the title compound (150 mg, 0.293 mmol) in anhydrous dioxane (5 mL) from step A above, add morpholine (39 mg, 0.44 mmol) and sodium tert-butoxide (85 mg , 0.87 mmol), and degassed under N ₂ atmosphere for 10 min. To this reaction mixture was added ginseng (dibenzylideneacetone) dipalladium (0) (13.4 mg, 0.0015 mmol) and 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl (13.65 mg , 0.029 mmol), and heated to 100 ℃ until the reaction was completed. After the reaction was completed (monitored by LCMS), the reaction mixture was filtered through celite and washed with EtOAc. The filtrate was concentrated under reduced pressure to produce crude product. Using the Biotage Isolera One purification system, using a petroleum ether/EtOAc gradient (0-100%), purification was performed on a silica gel column to obtain the title compound (60 mg, 50%) as a solid. MS: 409.1 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.10 (s, 1H), 7.48-7.50 (m, 1H), 7.36-7.38 (m, 2H), 7.08-7.09 (m, 1H), 6.96 -6.97 (m, 1H), 6.85-6.86 (m, 1H), 4.74 (s, 2H), 3.94-3.95 (m, 2H), 3.74-3.75 (m, 4H), 3.06-3.07 (m, 4H) , 2.94-2.95 (m, 2H).

步驟 A 向來自製備實例7之標題化合物(250 mg, 0.725 mmol)於DMF (5 mL)中之溶液添加2,6-二氯苯并噁唑(166 mg, 0.88 mmol)及碳酸鉀(326 mg, 2.36 mmol)，且加熱至100℃持續8小時。在反應完成後，將反應混合物傾倒至冰冷卻之水中，將沈澱出之固體過濾並乾燥，得到標題化合物(250 mg, 70%)。產物不經進一步純化即原樣用於下一步驟。 MS: 496.1 (M+H)⁺ 。步驟 B 向來自上文步驟A之標題化合物(250 mg, 0.505 mmol)於無水二噁烷(5 mL)中之攪拌溶液添加嗎啉(53 mg, 0.60 mmol)、第三丁醇鈉(145 mg, 1.52 mmol)，且在N₂ 氣氛下脫氣10 min。向此反應混合物添加參(二亞苄基丙酮)二鈀(0) (23.1 mg, 0.0252 mmol)及2-二環己基膦基-2′,6′-二異丙氧基聯苯(23.53 mg, 0.051 mmol)，且加熱至100℃直至反應完成為止。在反應完成後(藉由LCMS監測)，經由矽藻土過濾反應混合物並用EtOAc洗滌。將濾液在減壓下濃縮以產生粗產物。使用Biotage Isolera One純化系統，採用石油醚/ EtOAc梯度(0 - 100%)，在矽膠管柱上實施純化，得到呈固體之標題化合物(50 mg, 25%)。 MS: 393.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ = 11.09 (s, 1H), 7.45-7.46 (m, 1H), 7.17 (d,J = 8.56 Hz, 1H), 7.08-7.09 (m, 2H), 6.87-6.79 (m, 2H), 4.77 (s, 2H), 3.95-3.96 (m, 2H), 3.72-3.74 (m, 4H), 3.03-3.04 (m, 4H), 2.91-2.93 (m, 2H)。 Example 109

Step A To a solution of the title compound (250 mg, 0.725 mmol) in DMF (5 mL) from Preparation Example 7 was added 2,6-dichlorobenzoxazole (166 mg, 0.88 mmol) and potassium carbonate (326 mg , 2.36 mmol), and heated to 100 ℃ for 8 hours. After the reaction was completed, the reaction mixture was poured into ice-cooled water, and the precipitated solid was filtered and dried to obtain the title compound (250 mg, 70%). The product was used as is in the next step without further purification. MS: 496.1 (M+H) ⁺ . Step B To a stirred solution of the title compound (250 mg, 0.505 mmol) in anhydrous dioxane (5 mL) from Step A above, add morpholine (53 mg, 0.60 mmol), sodium tert-butoxide (145 mg , 1.52 mmol), and degassed under N ₂ atmosphere for 10 min. To this reaction mixture was added ginseng (dibenzylideneacetone) dipalladium (0) (23.1 mg, 0.0252 mmol) and 2-dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl (23.53 mg , 0.051 mmol), and heated to 100 ℃ until the reaction was completed. After the reaction was completed (monitored by LCMS), the reaction mixture was filtered through celite and washed with EtOAc. The filtrate was concentrated under reduced pressure to produce crude product. Using the Biotage Isolera One purification system with a petroleum ether/EtOAc gradient (0-100%), purification was performed on a silica gel column to obtain the title compound (50 mg, 25%) as a solid. MS: 393.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ = 11.09 (s, 1H), 7.45-7.46 (m, 1H), 7.17 (d, J = 8.56 Hz, 1H), 7.08-7.09 (m, 2H ), 6.87-6.79 (m, 2H), 4.77 (s, 2H), 3.95-3.96 (m, 2H), 3.72-3.74 (m, 4H), 3.03-3.04 (m, 4H), 2.91-2.93 (m , 2H).

步驟 A 向來自製備實例1 之標題化合物(0.15 g, 0.43 mmol)於無水1,4-二噁烷(5 mL)中之攪拌溶液添加來自製備實例50 之標題化合物(0.17 g, 0.43 mmol)及Cs₂ CO₃ (0.420 g, 1.29 mmol)。將反應混合物在N₂ 氣氛下脫氣10 min。然後添加Pd(OAc)₂ (0.009 g, 0.043 mmol)及2-二環己基膦基-2′,4′,6′-三異丙基聯苯‎(0.062 g; 0.129 mmol)，且將反應混合物加熱至100℃直至反應完成為止。在反應完成後(藉由LCMS監測)，經由矽藻土過濾反應混合物並用乙酸乙酯洗滌。將濾液在減壓下濃縮以產生粗產物。藉由急速管柱層析或製備型HPLC純化粗製材料，得到經甲苯磺醯基保護之化合物。向甲苯磺醯基化合物(1.0 eq)於二噁烷:MeOH (1:1, 10 vol)中之溶液添加NaOtBu (3 eq)，且加熱至70℃持續6小時。將反應混合物在真空下濃縮，且使粗產物進行管柱純化，得到期望產物。藉由急速管柱層析或製備型HPLC純化粗製材料，得到標題化合物(0.042 g, 24%)。¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 11.00 (bs, 1H), 8.70 (s, 1H), 7.96 (d,J = 9.60 Hz, 1H), 7.45-7.46 (m, 1H), 7.39 (d,J = 9.20 Hz, 1H), 7.07-7.08 (m, 2H), 6.82-6.83 (m, 1H), 4.64 (s, 2H), 4.02-4.04 (m, 2H), 3.73 (d,J = 4.80 Hz, 4H), 3.59 (d,J = 4.40 Hz, 4H), 2.90 (bs, 2H)。 MS: 404.2 (M+H)⁺ 。 Example 110

Step A To a stirred solution of the title compound from Preparation Example 1 (0.15 g, 0.43 mmol) in anhydrous 1,4-dioxane (5 mL) was added the title compound (0.17 g, 0.43 mmol) from Preparation Example 50 and Cs ₂ CO ₃ (0.420 g, 1.29 mmol). The reaction mixture was degassed under N ₂ atmosphere for 10 min. Then add Pd(OAc) ₂ (0.009 g, 0.043 mmol) and 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl‎ (0.062 g; 0.129 mmol), and react The mixture was heated to 100°C until the reaction was completed. After the reaction was completed (monitored by LCMS), the reaction mixture was filtered through celite and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to produce crude product. The crude material was purified by flash column chromatography or preparative HPLC to obtain the compound protected by tosylate. To a solution of the tosylate compound (1.0 eq) in dioxane:MeOH (1:1, 10 vol) was added NaOtBu (3 eq) and heated to 70°C for 6 hours. The reaction mixture was concentrated under vacuum, and the crude product was subjected to column purification to obtain the desired product. The crude material was purified by flash column chromatography or preparative HPLC to obtain the title compound (0.042 g, 24%). ¹ H-NMR (400 MHz, DMSO- d ₆ ): δ 11.00 (bs, 1H), 8.70 (s, 1H), 7.96 (d, J = 9.60 Hz, 1H), 7.45-7.46 (m, 1H), 7.39 (d, J = 9.20 Hz, 1H), 7.07-7.08 (m, 2H), 6.82-6.83 (m, 1H), 4.64 (s, 2H), 4.02-4.04 (m, 2H), 3.73 (d, J = 4.80 Hz, 4H), 3.59 (d, J = 4.40 Hz, 4H), 2.90 (bs, 2H). MS: 404.2 (M+H) ⁺ .

實例 113

步驟 A 將乙酸鈀(II)(0.013 g, 0.058 mmol)及4,5-雙(二苯基膦基)-9,9-二甲基𠮿

(XANTPHOS) (0.101 g, 0.174 mmol)置於反應小瓶中並脫氣。添加1,4-二噁烷(6 mL)。將所得溶液短暫脫氣。將懸浮液在100℃下(在預加熱之加熱塊上)加熱少於1分鐘，直至溶液之色彩自橙色變為深粉色為止。然後，將小瓶自加熱塊移除，且添加來自製備實例52之標題化合物(148 mg, 0.581 mmol)、來自製備實例1之標題化合物(0.200 g, 0.581 mmol)及碳酸銫(0.662 g, 2.033 mmol)。將反應小瓶填充氬，之後將其關閉。將反應混合物在100℃下加熱18 h。用乙酸乙酯及水稀釋該反應混合物。分離有機層，且將水層用乙酸乙酯再萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯(100/00 --＞ 50/50)梯度，在矽膠管柱上純化粗產物。將所獲得之粗製反應混合物(0.261 g, 0.469 mmol)及碳酸銫(0.214 g, 0.658 mmol)置入至微波管中，之後置入MeOH (4 mL)及THF (8 mL)。將反應混合物於微波中輻照至110℃持續30分鐘，且然後使其冷卻至室溫。將溶劑在減壓下去除，且使用Biotage Isolera One純化系統，利用二氯甲烷/甲醇梯度(100/00 --＞ 95/05)在矽膠管柱上純化殘餘物，得到呈黃色固體之標題化合物(0.020 g, 9%)。¹ H NMR (400 MHz, DMSO-d₆ ) δ 10.97 (s, 1H), 7.95 (d, 1H), 7.54 (d, 2H), 7.33 - 7.10 (m, 4H), 6.86 (td, 1H), 4.41 (s, 2H), 3.72 (dt, 6H), 3.55 (t, 4H), 2.95 (d, 2H)。 MS: 403.19 (M+H)⁺ 。 Examples 111 and 112 After deprotection of tosylate, following the palladium coupling procedure as described in Example 110 , the following compounds were prepared. Table 4

Example 113

Step A: Palladium(II) acetate (0.013 g, 0.058 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethyl 𠮿

(XANTPHOS) (0.101 g, 0.174 mmol) was placed in the reaction vial and degassed. Add 1,4-dioxane (6 mL). The resulting solution was briefly degassed. The suspension was heated at 100°C (on the preheated heating block) for less than 1 minute until the color of the solution changed from orange to dark pink. Then, the vial was removed from the heating block, and the title compound from Preparation Example 52 (148 mg, 0.581 mmol), the title compound from Preparation Example 1 (0.200 g, 0.581 mmol), and cesium carbonate (0.662 g, 2.033 mmol) were added ). The reaction vial was filled with argon and then closed. The reaction mixture was heated at 100 °C for 18 h. The reaction mixture was diluted with ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice more with ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. Using the Biotage Isolera One purification system, the crude product was purified on a silica gel column using a n-heptane/ethyl acetate (100/00 --> 50/50) gradient. The obtained crude reaction mixture (0.261 g, 0.469 mmol) and cesium carbonate (0.214 g, 0.658 mmol) were placed in a microwave tube, followed by MeOH (4 mL) and THF (8 mL). The reaction mixture was irradiated in the microwave to 110°C for 30 minutes, and then allowed to cool to room temperature. The solvent was removed under reduced pressure, and the residue was purified on a silica gel column using a Biotage Isolera One purification system using a dichloromethane/methanol gradient (100/00 --> 95/05) to obtain the title compound as a yellow solid (0.020 g, 9%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.97 (s, 1H), 7.95 (d, 1H), 7.54 (d, 2H), 7.33-7.10 (m, 4H), 6.86 (td, 1H), 4.41 (s, 2H), 3.72 (dt, 6H), 3.55 (t, 4H), 2.95 (d, 2H). MS: 403.19 (M+H) ⁺ .

步驟 A 將乙酸鈀(II)(0.013 g, 0.058 mmol)及4,5-雙(二苯基膦基)-9,9-二甲基𠮿

(XANTPHOS) (0.101 g, 0.174 mmol)置於反應小瓶中並脫氣。添加1,4-二噁烷(6 mL)。將所得溶液短暫脫氣。將懸浮液在100℃下(在預加熱之加熱塊上)加熱少於1分鐘，直至溶液之色彩自橙色變為深粉色為止。然後，將小瓶自加熱塊移除，且添加來自製備實例51之標題化合物(144 mg, 0.581 mmol)、來自製備實例1之標題化合物(0.200 g, 0.581 mmol)及碳酸銫(0.662 g, 2.033 mmol)。將反應小瓶填充氬，之後將其關閉。將反應混合物在100℃下加熱18 h。用乙酸乙酯及水稀釋該反應混合物。分離有機層，且將水層用乙酸乙酯再萃取兩次。使合併之有機層經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發。使用Biotage Isolera One純化系統，採用正庚烷/乙酸乙酯梯度(100/00 --＞ 50/50)，在矽膠管柱上純化粗產物。將所獲得之粗製反應混合物(0.122 g, 0.219 mmol)及碳酸銫(0.214 g, 0.658 mmol)置入至微波管中，之後置入MeOH (4 mL)及THF (8 mL)。將反應混合物於微波中輻照至110℃持續30分鐘，且然後使其冷卻至室溫。將溶劑在減壓下去除，且使用Biotage Isolera One純化系統，利用二氯甲烷/甲醇梯度(100/00 --＞ 95/05)在矽膠管柱上純化殘餘物，得到呈黃色固體之標題化合物(0.081 g, 35%)。¹ H NMR (400 MHz, DMSO-d₆ ) δ 10.97 (s, 1H), 7.88 (d, 1H), 7.56 (d, 1H), 7.40 - 7.13 (m, 3H), 7.02 (d, 1H), 6.97 - 6.79 (m, 2H), 4.49 (s, 2H), 3.80 (t, 2H), 3.72 (t, 4H), 3.60 (t, 4H), 2.93 (d, 2H)。 MS: 403.20 (M+H)⁺ 。 Example 114

Step A: Palladium(II) acetate (0.013 g, 0.058 mmol) and 4,5-bis(diphenylphosphino)-9,9-dimethyl 𠮿

(XANTPHOS) (0.101 g, 0.174 mmol) was placed in the reaction vial and degassed. Add 1,4-dioxane (6 mL). The resulting solution was briefly degassed. The suspension was heated at 100°C (on the preheated heating block) for less than 1 minute until the color of the solution changed from orange to dark pink. Then, the vial was removed from the heating block, and the title compound from Preparation Example 51 (144 mg, 0.581 mmol), the title compound from Preparation Example 1 (0.200 g, 0.581 mmol), and cesium carbonate (0.662 g, 2.033 mmol) were added ). The reaction vial was filled with argon and then closed. The reaction mixture was heated at 100 °C for 18 h. The reaction mixture was diluted with ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted twice more with ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. Using the Biotage Isolera One purification system, the crude product was purified on a silica gel column using a n-heptane/ethyl acetate gradient (100/00 --> 50/50). The obtained crude reaction mixture (0.122 g, 0.219 mmol) and cesium carbonate (0.214 g, 0.658 mmol) were placed in a microwave tube, followed by MeOH (4 mL) and THF (8 mL). The reaction mixture was irradiated in the microwave to 110°C for 30 minutes, and then allowed to cool to room temperature. The solvent was removed under reduced pressure, and the residue was purified on a silica gel column using a Biotage Isolera One purification system using a dichloromethane/methanol gradient (100/00 --> 95/05) to obtain the title compound as a yellow solid (0.081 g, 35%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.97 (s, 1H), 7.88 (d, 1H), 7.56 (d, 1H), 7.40-7.13 (m, 3H), 7.02 (d, 1H), 6.97-6.79 (m, 2H), 4.49 (s, 2H), 3.80 (t, 2H), 3.72 (t, 4H), 3.60 (t, 4H), 2.93 (d, 2H). MS: 403.20 (M+H) ⁺ .

步驟 A 將Pd(OAc)₂ (16 mg,0.072 mmol)及2-二環己基膦基-2′,4′,6′-三異丙基聯苯‎(XPhos；103 mg, 0.2177 mmol)添加至反應小瓶，且添加經脫氣之二噁烷(5 ml)。將小瓶填充氬氣且密封。將懸浮液在100℃下加熱1分鐘，然後添加8-氟-5-甲苯磺醯基-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(製備實例1 ) (250 mg, 0.725 mmol)、4-(4-溴苯基)嗎啉(210 mg, 0.87 mmol)及Cs₂ CO₃ (707 mg, 2.177 mmol)，且將溶液在100℃下加熱18小時。用乙酸乙酯(30 mL)及水(30 mL)稀釋反應混合物。分離有機相，且將水相用乙酸乙酯再萃取兩次。使合併之有機相經Na₂ SO₄ 乾燥，過濾並將溶劑在減壓下蒸發。藉由採用DCM/MeOH梯度(100/0 -＞ 95/05)，在HP-Sil管柱(Biotage)上純化粗製物，得到呈黃色固體之4-(4-(8-氟-5-甲苯磺醯基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-基)苯基)嗎啉(235 mg, 64%)。 向4-(4-(8-氟-5-甲苯磺醯基-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-基)苯基)嗎啉(0.235 mg, 0.465 mmol)於二噁烷:MeOH (1:1, 5 vol)中之溶液添加NaOtBu (133 mg,1.39 mmol)，且加熱至70℃持續6小時。將反應混合物在真空下濃縮，且藉由採用DCM/MeOH梯度(100/0 -＞ 95/05)，在HP-Sil管柱(Biotage)上管柱純化粗產物，得到呈灰白色固體之標題化合物(57 mg, 35%) MS: 352.0 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.93 (bs, 1H), 7.21-7.24 (m, 2H), 6.99-7.01 (m, 2H), 6.85-6.86 (m, 3H), 4.24 (s, 2H), 3.73 (bs, 4H), 3.52-3.53 (m, 2H), 2.98 (bs, 4H), 2.86 (bs, 2H)。 Example 115

Step A Add Pd(OAc) ₂ (16 mg, 0.072 mmol) and 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl‎ (XPhos; 103 mg, 0.2177 mmol) To the reaction vial, and degassed dioxane (5 ml) was added. The vial was filled with argon and sealed. The suspension was heated at 100°C for 1 minute, and then 8-fluoro-5-tosyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole (preparation was added Example 1 ) (250 mg, 0.725 mmol), 4-(4-bromophenyl)morpholine (210 mg, 0.87 mmol) and Cs ₂ CO ₃ (707 mg, 2.177 mmol), and the solution was heated at 100°C 18 hours. The reaction mixture was diluted with ethyl acetate (30 mL) and water (30 mL). The organic phase was separated and the aqueous phase was extracted twice more with ethyl acetate. The combined organic phases were dried over Na ₂ SO ₄ , filtered and the solvent was evaporated under reduced pressure. By using a DCM/MeOH gradient (100/0 -> 95/05), the crude material was purified on an HP-Sil column (Biotage) to give 4-(4-(8-fluoro-5-toluene as a yellow solid Sulfonyl-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indol-2-yl)phenyl)morpholine (235 mg, 64%). To 4-(4-(8-fluoro-5-toluenesulfonyl-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indol-2-yl)phenyl) A solution of morpholine (0.235 mg, 0.465 mmol) in dioxane:MeOH (1:1, 5 vol) was added NaOtBu (133 mg, 1.39 mmol) and heated to 70°C for 6 hours. The reaction mixture was concentrated under vacuum, and the crude product was purified by using a DCM/MeOH gradient (100/0 -> 95/05) on an HP-Sil column (Biotage) to give the title compound as an off-white solid (57 mg, 35%) MS: 352.0 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ 10.93 (bs, 1H), 7.21-7.24 (m, 2H), 6.99-7.01 (m, 2H), 6.85-6.86 (m, 3H), 4.24 ( s, 2H), 3.73 (bs, 4H), 3.52-3.53 (m, 2H), 2.98 (bs, 4H), 2.86 (bs, 2H).

向7-氟-5-甲苯磺醯基-2,3,4,5-四氫-1H-吡啶并[4,3-b]吲哚(製備實例7 ) (250 mg, 0.7267 mmol)於無水二噁烷(5 mL)中之攪拌溶液添加5-氯-N-(2-甲氧基乙基)-N-甲基苯并[d]噁唑-2-胺(製備實例60 ) (190 mg, 0.7994 mmol)、第三丁醇鈉(202 mg, 2.180 mmol)，且在N₂ 氣氛下脫氣10 min。向此反應混合物添加Ruphos G4 Pd (60 mg, 0.0726 mmol)，且加熱至100℃直至反應完成為止。在反應完成後，經由矽藻土床過濾該反應混合物，用EtOAc洗滌。將濾液濃縮，且藉由採用DCM/MeOH梯度(100/0 -＞ 95/05)在HP-Sil管柱(Biotage)上藉由管柱層析純化粗製物，得到標題化合物(37 mg, 13%) MS: 395.0 (M+H)⁺ 。 1H-NMR (400 MHz, DMSO-d ₆ ) δ 10.94 (bs, 1H), 7.43-7.44 (m, 1H), 7.23 (d,J = 8.80 Hz, 1H), 7.06-7.06 (m, 1H), 6.99 (d,J = 2.40 Hz, 1H), 6.80-6.81 (m, 1H), 6.71-6.72 (m, 1H), 4.32 (s, 2H), 3.64-3.65 (m, 2H), 3.56-3.58 (m, 4H), 3.27 (s, 3H), 3.13 (s, 3H), 2.86 (t,J = 5.20 Hz, 2H)。 Example 116

To 7-fluoro-5-tosyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole (Preparation Example 7 ) (250 mg, 0.7267 mmol) in anhydrous The stirred solution in dioxane (5 mL) was added 5-chloro-N-(2-methoxyethyl)-N-methylbenzo[d]oxazol-2-amine (Preparation Example 60 ) (190 mg, 0.7994 mmol), sodium tert-butoxide (202 mg, 2.180 mmol), and degassed under N ₂ atmosphere for 10 min. To this reaction mixture was added Ruphos G4 Pd (60 mg, 0.0726 mmol), and heated to 100°C until the reaction was completed. After the reaction was completed, the reaction mixture was filtered through a bed of celite and washed with EtOAc. The filtrate was concentrated, and the crude material was purified by column chromatography on an HP-Sil column (Biotage) using a DCM/MeOH gradient (100/0 -> 95/05) to obtain the title compound (37 mg, 13 %) MS: 395.0 (M+H) ⁺ . 1H-NMR (400 MHz, DMSO- d ₆ ) δ 10.94 (bs, 1H), 7.43-7.44 (m, 1H), 7.23 (d, J = 8.80 Hz, 1H), 7.06-7.06 (m, 1H), 6.99 (d, J = 2.40 Hz, 1H), 6.80-6.81 (m, 1H), 6.71-6.72 (m, 1H), 4.32 (s, 2H), 3.64-3.65 (m, 2H), 3.56-3.58 ( m, 4H), 3.27 (s, 3H), 3.13 (s, 3H), 2.86 (t, J = 5.20 Hz, 2H).

Examples 117 to 145 Following the procedures reported in Examples 115 and 116 , the following compounds were prepared. Table 5

步驟 A 在0℃下向氫化鈉(0.0293 g, 0.764 mmol)於DMF (3 mL)中之懸浮液逐滴添加5-(7-氟-1,3,4,5-四氫吡啶并[4,3-b]吲哚-2-基)-2-嗎啉基-1,3-苯并噁唑(實例45 ) (0.100 g, 0.255 mmol) (溶解於3 mL DMF中)，然後在室溫下攪拌60 min。之後，在0℃下逐滴添加碘乙烷(0.0596 g, 0.382 mmol) (溶解於2 mL DMF中)，且然後在室溫下攪拌3 h。根據TLC在反應完成後，利用冰水將反應混合物淬滅，之後使用乙酸乙酯(20 mL)進行萃取。將有機層分離，經硫酸鈉乾燥，過濾且然後濃縮以得到粗製物，且然後藉由矽膠管柱(Biotage)使用於石油醚中之40%-50%乙酸乙酯純化為呈灰白色固體之5-(5-乙基-7-氟-3,4-二氫-1H-吡啶并[4,3-b]吲哚-2-基)-2-嗎啉基-1,3-苯并噁唑。(15 mg, 14%) MS: 421.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 7.47-7.48 (m, 1H), 7.26-7.28 (m, 2H), 7.06 (d,J = 2.40 Hz, 1H), 6.84-6.85 (m, 1H), 6.78-6.79 (m, 1H), 4.33 (s, 2H), 4.10 (q,J = 7.20 Hz, 2H), 3.55-3.57 (m, 10H), 2.90 (t,J = 4.80 Hz, 2H), 1.22 (t,J = 7.20 Hz, 3H)。 Example 146

Step A To a suspension of sodium hydride (0.0293 g, 0.764 mmol) in DMF (3 mL) was added 5-(7-fluoro-1,3,4,5-tetrahydropyrido[4 ,3-b]indol-2-yl)-2-morpholinyl-1,3-benzoxazole (Example 45 ) (0.100 g, 0.255 mmol) (dissolved in 3 mL DMF), then in the room Stir at a temperature of 60 min. After that, iodoethane (0.0596 g, 0.382 mmol) (dissolved in 2 mL DMF) was added dropwise at 0°C, and then stirred at room temperature for 3 h. After completion of the reaction according to TLC, the reaction mixture was quenched with ice water, and then extracted with ethyl acetate (20 mL). The organic layer was separated, dried over sodium sulfate, filtered and then concentrated to obtain a crude product, and then purified by silica gel column (Biotage) using 40%-50% ethyl acetate in petroleum ether to 5 as an off-white solid. -(5-ethyl-7-fluoro-3,4-dihydro-1H-pyrido[4,3-b]indol-2-yl)-2-morpholinyl-1,3-benzox Azole. (15 mg, 14%) MS: 421.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ 7.47-7.48 (m, 1H), 7.26-7.28 (m, 2H), 7.06 (d, J = 2.40 Hz, 1H), 6.84-6.85 (m, 1H), 6.78-6.79 (m, 1H), 4.33 (s, 2H), 4.10 (q, J = 7.20 Hz, 2H), 3.55-3.57 (m, 10H), 2.90 (t, J = 4.80 Hz, 2H ), 1.22 (t, J = 7.20 Hz, 3H).

Examples 147 to 153 In addition to using the tricyclic amino derivatives and bromine/chlorine derivatives indicated in the following table, following the palladium coupling procedure as described in Examples 1 , 2 and 115 , the following compounds were prepared. Table 5a

步驟 A 向冷卻至0℃之4-(6-(7-氟-1,3,4,5-四氫-2H-吡啶并[4,3-b]吲哚-2-基)噻唑并[4,5-c]吡啶-2-基)嗎啉(實例89 ) (0.1 g, 0.244 mmol)於無水DCM (5 mL)中之溶液添加4 M HCl於二噁烷(0.2 mL)並攪拌1小時。將反應混合物在真空下濃縮且與二乙醚一起研磨，得到呈棕色固體之標題化合物(0.09 g, 83%)。 MS: 410.2 (M+H)⁺ 。¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.14 (bs, 1H), 8.28 (s, 1H), 7.98 (s, 1H), 7.38-7.40 (m, 1H), 7.11-7.12 (m, 1H), 6.85-6.86 (m, 1H), 4.75 (s, 2H), 4.02 (t,J = 5.64 Hz, 2H), 3.73-3.75 (m, 8H), 2.97 (s, 2H)。 Examples of HCl salts of compounds of the present invention 89 HCl

Step A To 4-(6-(7-fluoro-1,3,4,5-tetrahydro-2H-pyrido[4,3-b]indol-2-yl)thiazolo[0, 4,5-c)pyridin-2-yl)morpholine (Example 89 ) (0.1 g, 0.244 mmol) in anhydrous DCM (5 mL) was added 4 M HCl in dioxane (0.2 mL) and stirred for 1 hour. The reaction mixture was concentrated under vacuum and triturated with diethyl ether to give the title compound (0.09 g, 83%) as a brown solid. MS: 410.2 (M+H) ⁺ . ¹ H-NMR (400 MHz, DMSO- d ₆ ) δ 11.14 (bs, 1H), 8.28 (s, 1H), 7.98 (s, 1H), 7.38-7.40 (m, 1H), 7.11-7.12 (m, 1H), 6.85-6.86 (m, 1H), 4.75 (s, 2H), 4.02 (t, J = 5.64 Hz, 2H), 3.73-3.75 (m, 8H), 2.97 (s, 2H).

Examples Following the hydrochloride procedure as described in Example 89 HCl , the following compounds were prepared. Table 6

將觸媒添加至氚反應容器，之後添加實例12 起始材料(1.0 mg, 0.003 mmol)於二氯甲烷中之溶液。將容器附接至氚管線並在-200℃下對氚氣加壓。將溶液攪拌8小時，冷卻至-200℃且去除過量氣體。用4 × 1 mL甲醇沖洗反應燒瓶，將其各自轉移至100 mL回收燒瓶。使合併之有機相在真空下蒸發(粗產率：124 mCi)。藉由HPLC純化粗製材料，使移動相在真空下蒸發且將產物重新溶解於無水乙醇中(產率：17 mCi，純度＞ 97%)。藉由質譜測定比活性為80.6 Ci/mmol。 Synthesis Example ³ of Radioactive Ligand of the Compound of the Invention ³ H-12

The catalyst was added to the tritium reaction vessel, after which a solution of the starting material of Example 12 (1.0 mg, 0.003 mmol) in dichloromethane was added. Attach the vessel to the tritium line and pressurize the tritium gas at -200°C. The solution was stirred for 8 hours, cooled to -200°C and excess gas was removed. Rinse the reaction flask with 4 × 1 mL methanol and transfer each to a 100 mL recovery flask. The combined organic phase was evaporated under vacuum (crude yield: 124 mCi). The crude material was purified by HPLC, the mobile phase was evaporated under vacuum and the product was redissolved in absolute ethanol (yield: 17 mCi, purity >97%). The specific activity measured by mass spectrometry was 80.6 Ci/mmol.

將觸媒添加至氚反應容器，之後添加實例45 起始材料(1.0 mg, 0.003 mmol)於二氯甲烷中之溶液。將容器附接至氚管線並在-200℃下對氚氣加壓。將溶液攪拌8小時，冷卻至-200℃且去除過量氣體。用4 × 1 mL甲醇沖洗反應燒瓶，將其各自轉移至100 mL回收燒瓶。使合併之有機相在真空下蒸發(粗產率：124 mCi)。藉由HPLC純化粗製材料，使移動相在真空下蒸發且將產物重新溶解於無水乙醇中(產率：17 mCi，純度＞ 99%)。藉由質譜測定比活性為96.8 Ci/mmol。 Example ³ H-45

The catalyst was added to the tritium reaction vessel, after which a solution of the starting material of Example 45 (1.0 mg, 0.003 mmol) in dichloromethane was added. Attach the vessel to the tritium line and pressurize the tritium gas at -200°C. The solution was stirred for 8 hours, cooled to -200°C and excess gas was removed. Rinse the reaction flask with 4 × 1 mL methanol and transfer each to a 100 mL recovery flask. The combined organic phase was evaporated under vacuum (crude yield: 124 mCi). The crude material was purified by HPLC, the mobile phase was evaporated under vacuum and the product was redissolved in absolute ethanol (yield: 17 mCi, purity >99%). The specific activity measured by mass spectrometry was 96.8 Ci/mmol.

Bioanalysis Description The full-length Tau (flTau) depolymerization analysis of thioflavin T (ThT) was used to express and purify the longest isoform of human Tau (2N4R; 441 amino acids) in bacteria. For Tau depolymerization analysis by ThT, 35 μM recombinant full-length (fl) Tau in PBS was used at 37° C. in the presence of 50 μM heparin (Sigma-Aldrich) and 10 mM DTT (Sigma-Aldrich) at 750 RPM Gather for 24 hours with shaking. The compound was dissolved in anhydrous dimethyl sulfoxide (DMSO, Sigma-Aldrich) to reach a concentration of 10 mM. The serial dilutions of flTau aggregates and compounds were mixed together in PBS (50 μL volume) to a final concentration of 2 μM flTau aggregates and 160 μM to 0.04 μM compounds. The mixture was incubated at room temperature (RT) for 30 minutes, and then 40 μL of this mixture was transferred to a black 384-well plate analysis (Perkin-Elmer) and mixed with 10 μL of 100 μM ThT (250 mM glycan in PBS) Mix in amine acid (both from Sigma-Aldrich). Fluorescence (relative fluorescence unit; RFU) was measured in one or two copies on a Tecan reader (excitation: 440 nm; emission: 485 nm). The flTau disaggregation percentage was then calculated, and GraphPad Prism version 5 (GraphPad software) was used to determine a half-maximal effective concentration (EC ₅₀ ) assuming a binding site fitting model. Tau K18 depolymerization analysis by ThT The Tau K18 fragments covering amino acids 244 to 372 of the longest isoform (2N4R) of human Tau441 were expressed and purified in bacteria, or purchased from SignalChem. For K18 disaggregation analysis by ThT, 35 µM recombinant K18 in PBS was aggregated at 37°C in the presence of 50 µM heparin (Sigma-Aldrich) and 10 mM DTT (Sigma-Aldrich) under 750 RPM shaking 24 hour. The compound was dissolved in anhydrous dimethyl sulfoxide (DMSO, Sigma-Aldrich) to reach a concentration of 10 mM. K18 aggregates and serial dilutions of compounds were mixed together in PBS (50 µL volume) to a final concentration of 2 μM K18 aggregates and 160 µM to 0.04 µM compounds. The mixture was incubated at room temperature (RT) for 30 minutes, and then 40 μL of this mixture was transferred to a black 384-well plate analysis (Perkin-Elmer) and mixed with 10 μL of 100 μM ThT (250 mM glycan in PBS) Mix in amine acid (both from Sigma-Aldrich). Fluorescence (relative fluorescence unit; RFU) was measured in one or two copies on a Tecan reader (excitation: 440 nm; emission: 485 nm). K18 depolymerization percentage is then calculated, and using GraphPad Prism Version 5 (GraphPad Software) assuming one binding site to determine the half maximal effective concentration (EC ₅₀₎ fitted model.

圖例：+++ EC₅₀ ＜ 10 uM；++ EC₅₀ 10＜x＜25 uM；+ EC₅₀ 25＜x＜50 uM。The following example compounds were measured: Table 7

Legend: +++ EC ₅₀ <10 uM; ++ EC ₅₀ 10<x<25 uM; +EC ₅₀ 25<x<50 uM.

Intracellular Tau aggregation is reduced in complete medium [DMEM-F12 supplemented with 2.5 µg/ml G418 (Sigma-Aldrich) antibiotic selection 4.5 g/L Glutamax (Invitrogen), 15% FBS (Biochrom), 1% Peni/Strep ( Invitrogen)] have cultured human neuroblastoma cell lines expressing the full-length form of human Tau carrying the P301L mutation. On the day before the experiment, 5×10 ⁵ cells/well were plated in 3 mL complete medium in a 6-well plate. The next day, the cells were incubated with DMSO or 5 µM of the compound of the invention for 24 h at 37°C. After incubation, trypsinize the cells and resuspend in 100 µl of homogenization buffer [25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1 mM containing phosphatase inhibitors (30 mM NaF, 0.2 mM Na ₃ VO ₄ , 1 nM Okadaic acid, 1 mM PMSF, 5 mM Na ₄ P ₂ O ₇ ) EGTA and protease inhibitor mixture (Complete ^TM , Roche)], and then used three times for quick freezing and thawing Circulate for physical dissolution. Then test the sample directly in the AlphaLISA analysis.

AlphaLisa uses the following antibody pairs to quantify phosphorylation, aggregation, and total Tau: · HT7-Recipient Bead + Biotin (BT)-Tau13- Donor Bead: Total Tau · HT7-recipient beads + biotin (BT)-HT7-donor beads: gathering human Tau

Tau13 (Abcam) was biotinylated using EZ-Link® NHS-PEO solid phase biotinylation kit (Thermo Scientific), while the HT7-biotin system was from a commercial source (Thermo Scientific).

For each antibody pair, the concentration of recipient beads and biotinylated antibody is optimized. All samples are first tested in a dilution series in PBS to identify the linear range and optimal dilution of each sample and analysis. For the final protocol, add the following reagents to 384-well white OptiPlate (PerkinElmer): · 5 µL test diluted sample · Biotin-mAb recipient beads with a final concentration of less than 20 µL: · Combination of 1.25 nM HT7-BT and 10 µg/ml HT7-Acc beads · Combination of 5 nM Tau13-BT and 2.5 µg/ml HT7-Acc beads

After incubating this mixture at room temperature for 1 h, 25 µL of 25 µg/mL streptavidin (Streptavidin) donor beads (Perkin Elmer) was added in the dark. After incubation for 30 min, the plates were analyzed using EnSpire Alpha instrument and EnSpire workstation version 3.00. The data for aggregated Tau is normalized to total Tau and then expressed as a percentage of DMSO-treated cells.

圖例：+++%＞50；++% 50＜x＜25；+% 25＜x＜10。The following example compounds were measured: Table 8

Legend: +++%>50; ++% 50<x<25; +% 25<x<10.

In vivo efficacy test examples 12 and 45 of the compounds of the present invention are designed for in vivo studies. Double transgenic rTg4510 mice exhibit full-length human Tau carrying P301L (Tau4R0N-P301L) mutation under the control of the tet-inducible CaMKII promoter (Ramsden et al., J. Neurosci., 2005 • 25(46): 10637-10647). A single transgenic mouse expressing only tetracycline-controlled transactivator protein (tTA) was used as a genotype control. The study included 4 treatment groups (for the tTa group n=15 females and n=11 female mice/groups for treatment) with the following group assignments (see Table 9). Starting at 5 months of age, the compound or vehicle control was administered once a day or twice a day by gastric tube feeding for 1 month. All mice received 200 ppm deoxytetracycline in the feed for 3 weeks, plus a loading dose of 1.5 mg/mL in 4% sucrose in drinking water for the first two days. The administration of oxytetracycline at the time of administration started at the second week and continued throughout the administration period.

(a)             媒劑：0.5% w/v羥丙基甲基纖維素4000 cps；0.5% w/v Tween 80CSF analysis was performed on all animals, and histology of misfolded Tau (MC1), total microglia (Iba1), and phagocytic microglia (CD68) was performed on 10 animals/group. Table 9 : In vivo study design of Test Example 12 and Example 45

(a) Vehicle: 0.5% w/v hydroxypropyl methylcellulose 4000 cps; 0.5% w/v Tween 80

Cerebrospinal fluid (CSF) can be collected and analyzed by standard anesthetized mice were deeply anesthetized and perfused through the heart with cold PBS. Use surgical scissors to remove the fur and skin from the neck (the mice were not decapitated). The tissue surrounding the base of the skull and the brain stem was carefully removed with minimal bleed to minimize bleeding. Once the meninges are exposed in the occipital foramen region of the skull base, the 27ga butterfly needle is kept perpendicular to the skull and inserted laterally into the cerebrospinal fluid (CSF) space. The samples were quickly immersed in liquid nitrogen and stored at -80°C until biochemical analysis was performed by AlphaLISA. The following antibody pairs were used to quantify the total human Tau in CSF Tau: HT7-recipient beads + biotin (BT)-Tau13-donor beads. Tau13 (Abcam) antibody was biotinylated using EZ-Link® NHS-PEO solid phase biotinylation kit (Thermo Scientific). For the final protocol, add the following reagents to 384-well white OptiPlate (PerkinElmer): · 5 µL test dilution sample · 20 µL mixture biotin-mAb recipient beads, final concentration: 0.6 nM Tau13-BT and 2.5 µg/ ml HT7-Acc bead combination

After incubating the mixture at room temperature for 1 h, 25 µL of 25 µg/mL streptavidin donor beads (Perkin Elmer) was added in the dark. After a 30-minute incubation, the plate was analyzed using the EnSpire Alpha instrument and EnSpire workstation version 3.00. The data was analyzed by one-way ANOVA followed by post-hoc comparisons, and the statistics shown refer to the difference compared to vehicle-treated rTg4510 mice.

As shown in Figure 1, both Example 12 and Example 45 reduced the Tau content in the CSF.

Histological evaluation of rTg4510 mice treated with Example 12 and Example 45 After perfusion, the brain was then removed and half-sagittal. The right hemisphere was fixed in 4% paraformaldehyde in PBS at room temperature for 3 hours, refrigerated and stored for 3 days by immersion in 15% sucrose at 4°C, and prepared for frozen section. The fixed hemispheres were then frozen in dry ice on OCT medium in a low temperature mold, and sagittal frozen sections (10 mm thickness) were made with a Leica CM3050 cryostat. Sections from 10 mice/group were collected from approximately 12 medial and lateral levels. A systematic random slice group from seven sagittal levels/animals was used to quantify Iba1 and CD68 immunoreactivity, and one slice/animal from the medial-lateral level 4 was used to evaluate Tau misfolding (MC1 monoclonal antibody). The frozen sections were removed from -20°C and air-dried at room temperature for 25 minutes. Brain tissue was circled with a pap pen liquid blocker, and washed once in PBS at room temperature for 5 minutes and once for 10 minutes at room temperature. At room temperature, 10% normal goat serum (NGS) and 0.25% Triton X-100 in PBS were used for blocking and permeabilization treatment for 2 hours. The sections were then blotted dry and incubated with mouse monoclonal MC1 antibody diluted 1:1000 in PBS containing 5% NGS and 0.25% Triton X-100 at 4°C overnight in a humidity cabinet. The sections were washed three times in PBS at room temperature for 10 minutes, and avoided with a Cy3 labeled goat anti-mouse IgG (H+L) secondary antibody (Jackson) diluted 1:1000 in PBS at room temperature. Light incubate for 30 minutes. After washing three times in PBS for 10 minutes, the sections were incubated with a solution of 0.1% Sudan Black (Sigma) in 70% ethanol at room temperature for 30 seconds to reduce spontaneous fluorescence of the tissue. The sections were washed three times in PBS for 10 minutes, mounted with DAPI-containing ProLong Gold anti-fading reagent (Molecular Probes) and covered with coverslips. Sections were imaged using a digital slide scanner (Panoramic 250 Flash, 3D Histech Ltd.) and quantified using image visualization software Visiopharm.

MC1 staining was analyzed in the frontal cortex in the upper cortex. Draw a region of interest (ROI) around the frontal cortex corresponding to a 20x magnification field of view. This ROI was analyzed in Visiopharm using a predetermined threshold for MC1 staining, where the pixel intensity threshold is higher than 30 (8-bit image) and all detected objects smaller than 20 μm ^{2 are} excluded. The data was analyzed by single factor ANOVA followed by post-mortem testing.

As shown in Figure 2, both Example 12 and Example 45 reduced the MC1 positive area. These data indicate that treatment with both Example 12 and Example 45 in rTg4510 reduced the content of misfolded Tau in this aggressive Tau transgenic model.

In order to evaluate whether the above-mentioned effects of the tested compounds on Tau also have an effect on neuroinflammation markers, the sections of rat anti-mouse CD68 pure line FA-11 (BD Biosciences) and goat anti-Iba1 antibody (Abcam) were labeled, And counterstain with DAPI. Highly cross-absorbing fluorescently labeled secondary antibodies (Thermo Fisher) were used to visualize antibody binding. The antibody was diluted in antibody dilution (Dako), and M.O.M. serum (Vector) was used to block non-specific endogenous IgG binding before preliminary incubation. Using LED (Colibri2) illumination and a sensitive Orca Flash 4.0 monochrome camera, the mounted slices were imaged on an Axio Scan Z1 slide scanner driven by ZEN software at 20× magnification (flat-field apochromatic objectives). The data was analyzed by one-way ANOVA followed by post-hoc comparisons, and the statistics shown refer to the difference compared to vehicle-treated rTg4510 mice.

As shown in Figure 3A, both Example 12 and Example 45 reduced microglial hyperplasia, as measured by the Ibal immunoreactive area. In addition, both Examples 12 and 45 reduced highly activated phagocytic microglial cells positive for CD68 (Figure 3B). Taken together, these data indicate that treatment with both Example 12 and Example 45 in rTg4510 reduced the degree of nerve inflammation in this aggressive Tau transgenic model.

Reducing Tau misfolding in neurofibrillary tangles (NFT) from human AD and PSP brain slices Fresh frozen tissue sections from one AD case and two PSP cases were obtained from a commercial distributor (Tissue Solutions, UK). Tissue sections were incubated with 100 µM Example 45 or DMSO in a humidity cabinet at room temperature for approximately 60 hours. After washing the incubation section three times in PBS, it was fixed with paraformaldehyde (PFA; Sigma) for 10 min at 4°C, and then washed three more times in PBS. The sections were then permeabilized in blocking buffer (PBS, 10% pure goat serum (NGS), 0.25% Triton X) for 1 hour at room temperature. After blocking, the sections were incubated with Tau conformation antibody (MC1 monoclonal antibody) in blocking buffer (5% NGS, 0.25% Triton X) at 4°C overnight. The next day, the sections were washed three times in PBS for 5 min, and incubated with Cy3-conjugated AffiniPure goat anti-mouse antibody (Jackson laboratories) at room temperature for 1 h. Wash in PBS three times for 5 min to wash out excess antibody. To reduce spontaneous fluorescence, the sections were incubated with 0.1% Sudan black (Sigma) dissolved in 70% ethanol at room temperature for 8 min. Finally, the sections were washed 5 times in PBS for 5 min, and mounted under a coverslip using ProLong Gold anti-fading reagent (Invitrogen) containing DAPI. The sections were then dried at room temperature for 24 hours, after which they were imaged using a digital slide scanner (Pannoramic P250 Flash III, 3D Histech Ltd.), and Visiopharm software was used to quantify Tau misfolding in NFT. As shown in Figure 4, Example 45 reduced Tau misfolding in NFT by approximately 30%, which had similar efficacy on AD and PSP human brain samples.

In vitro high-resolution autoradiography using ³ H- Example 45 in human AD brain slices Fresh frozen tissue slices from one AD case were obtained from a commercial distributor (Tissue Solutions, UK). For autoradiography, brain slices were fixed with 4% PFA (Sigma) for 15 minutes at 4°C. Sections were incubated with 20 nM [ ³ H]-Example 45 alone or together with 5 μM non-radioactive Example 45 at room temperature for 1 hour. The slices were then washed as follows: first, in ice-cold buffer for 1 minute, then in ice-cold 70% ethanol twice for 1 minute, in ice-cold buffer for 1 minute, and finally briefly rinsed in ice-cold distilled water. The sections were then dried under air flow for 1 hour, and then exposed to Ilford nuclear latex type K5 (Agar Scientific) at 4°C for 5 days in an opaque slide storage box. According to the manufacturer's instructions, it has been exposed to latex in a dark room illuminated with safety lights, and the latex fragments were melted in pre-heated water at an equal volume of 40°C. After 5 days, the sections were developed according to the manufacturer's instructions. The sections were mounted using ProLong Gold anti-fading reagent (Invitrogen) and imaged using a digital slide scanner (Pannoramic P250 Flash III, 3D Histech Ltd.).

As shown in Figure 5, Example 45 shows specific target engagement on Tau NFT in human AD brain samples.

Figure 1 : Tau in CSF quantified in rTg4510 mice treated with Examples 12 and 45. Figure 2 : Quantification of Tau misfolding in rTg4510 mice treated with Example 12 and Example 45. Figure 3 : Quantification of Iba1 (A) and CD68 (B) in rTg4510 mice treated with Example 12 and Example 45. Figure 4 Example 45 reduces Tau misfolding in NFT in human AD and PSP brain slices. Figure 5 High-resolution autoradiography of human AD brain slices using ³ H-Example 45 (A) and ³ H-Example 45 and Cold Example 45 (B).

Claims (30)

A compound of formula (I),

And all its stereoisomers, racemic mixtures, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates, and polymorphs; where A is selected from the group consisting of:

and

,among them

and

Can be connected to Q at any available location, where

and

Substituted by one or more substituents R ^j , and wherein

and

Optionally substituted with one or more substituents R ; B is selected from the group consisting of O and NR ^a ; E and V are independently selected from the group consisting of: N, NR ⁵ , O, and S; G is selected from the group consisting of Group consisting of: benzene ring, pyrimidine ring and pyridine ring; J is selected from the group consisting of O and NR ¹ ; Q is selected from the group consisting of N and CR ¹ ; Y is selected from the group consisting of CZ and N, conditional When Y is N and Y ¹ , Y ² and Y ³ are CZ, B is N-alkyl or O; Y ¹ is selected from the group consisting of CZ and N; Y ² is selected from the group consisting of CZ and N; Y ³ is selected from the group consisting of CZ and N; Z is independently selected from the group consisting of H, halogen, O-alkyl, alkyl and CN; R is independently selected from

And -NR ³ R ⁴ ; R ^a is selected from the group consisting of H and alkyl; R ^b , R ^c , R ^d , R ^e , R ^f and R ^{g are} independently selected from the group consisting of H and alkyl Group, or any two of R ^b , R ^c , R ^d , R ^e , R ^f and R ^g can be connected to form a 3 to 8 member ring; R ^{j is} independently selected from the group consisting of: -halogen,- O-alkyl, -CF ₃ , -CN, -NR ³ R ⁴ ,

and

, In which the bridge containing C _1-2 carbon atoms may exist between a carbon atom and c or d carbon atom or the bridge containing C _1-2 carbon atoms may exist between b carbon atom and c or d carbon atom ; R ¹ is selected from the group consisting of H and alkyl; R ^{2 is} independently selected from the group consisting of alkyl, F, and =O, where the alkyl is optionally substituted with halogen, -OH, or -O-alkyl, And if two R ² are twins, they can be connected to form a 3-member to 6-member ring; R ³ and R ^{4 are} independently selected from the group consisting of H and alkyl, where the alkyl may optionally be halogen, -OH or -O-alkyl substitution; R ⁵ is selected from the group consisting of H and alkyl; n is 0, 1, 2, 3, or 4, r and s are independently 0, 1, 2, or 3; and t and u are independently 1, 2, or 3. If the compound of claim 1, it is a compound of formula (Ia):

Wherein ^{A, B, R b, R} c, R d, R e, R f, R g, Y and Z line such as a request item defined. If the compound of claim 1, it is a compound of formula (Ib):

Wherein ^{A, B, R b, R} c, R d, R e, R f, R g and Z line such as a request item defined. The compound according to any one of claims 1 to 3, wherein A is

,among them

Can be connected to Q or N at any available location, and where

Optionally substituted with one or more substituents R. If the compound of any one of claims 1 to 3 is a compound of formula (Ic):

Where E , R , V and Z are as defined in claim 1. The compound of claim 1, wherein the compound is selected from the group consisting of:

. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 and optionally a pharmaceutically acceptable carrier or excipient. The compound according to any one of claims 1 to 3 and 6, which is used as a medicament. A compound of formula (I),

And all its stereoisomers, racemic mixtures, tautomers, pharmaceutically acceptable salts, prodrugs, hydrates, solvates and polymorphs; it is used to treat, improve or prevent aggregation with Tau protein Body-related diseases or abnormalities, where A is selected from the group consisting of:

and

,among them

and

Can be connected to Q at any available location, where

and

Substituted by one or more substituents R ^j , and wherein

and

Optionally substituted with one or more substituents R ; B is selected from the group consisting of O and NR ^a ; E and V are independently selected from the group consisting of: N, NR ⁵ , O, and S; G is selected from the group consisting of Group consisting of: benzene ring, pyrimidine ring and pyridine ring; J is selected from the group consisting of O and NR ¹ ; Q is selected from the group consisting of N and CR ¹ ; Y is selected from the group consisting of CZ and N; Y ¹ Is selected from the group consisting of CZ and N; Y ² is selected from the group consisting of CZ and N; Y ³ is selected from the group consisting of CZ and N; Z is independently selected from the group consisting of: H, halogen, O- Alkyl, alkyl and CN; R is independently selected from

And -NR ³ R ⁴ ; R ^a is selected from the group consisting of H and alkyl; R ^b , R ^c , R ^d , R ^e , R ^f and R ^{g are} independently selected from the group consisting of H and alkyl Group, or any two of R ^b , R ^c , R ^d , R ^e , R ^f and R ^g can be connected to form a 3 to 8 member ring; R ^{j is} independently selected from the group consisting of: -halogen,- O-alkyl, -CF ₃ , -CN, -NR ³ R ⁴ ,

and

, In which the bridge containing C _1-2 carbon atoms may exist between a carbon atom and c or d carbon atom or the bridge containing C _1-2 carbon atoms may exist between b carbon atom and c or d carbon atom ; R ¹ is selected from the group consisting of H and alkyl; R ^{2 is} independently selected from the group consisting of alkyl, F, and =O, where the alkyl is optionally substituted with halogen, -OH, or -O-alkyl, And if two R ² are twins, they can be connected to form a 3-member to 6-member ring; R ³ and R ^{4 are} independently selected from the group consisting of H and alkyl, where the alkyl may optionally be halogen, -OH or -O-alkyl substitution; R ⁵ is selected from the group consisting of H and alkyl; n is 0, 1, 2, 3, or 4; r and s are independently 0, 1, 2, or 3; and t and u are independently 1, 2, or 3. A compound as claimed in claim 9, wherein the compound of formula (I) is as defined in any one of claims 1 to 6. A use of the compound according to any one of claims 1 to 6 or 9 for the manufacture of a medicament for treating, preventing or ameliorating a disease or abnormality associated with Tau protein aggregates. The compound as claimed in claim 9 or the use as claimed in claim 12, wherein the disorder is selected from Alzheimer's disease (AD), familial AD, primary age-related Tau protein disease (PART), Creutzfeldt-Jacob disease, boxer-type dementia, Down's Syndrome, Gerstmann-Sträussler-Scheinker disease (GSS), inclusion Myositis, prion protein cerebral amyloid angiopathy, traumatic brain injury (TBI), amyotrophic lateral sclerosis (ALS), Guam Parkinson's disease-dementia complex (Parkinsonism- dementia complex of Guam), non-Guamanian motor neuron disease with neurofibrillary tangles, argyrophilic granulopathy, cortical basal nucleus degeneration (CBD), diffuse neurofibrillary tangles with calcification , Frontotemporal dementia associated with chromosome 17 with Parkinson's disease (FTDP-17), Hallervorden-Spatz disease, multiple system atrophy (MSA), type C Niemann Pick disease (Niemann-Pick disease type C), globus pallidus-pontine-melanosis, Pick's disease (PiD), progressive subcortical glioma disease, progressive supranuclear palsy (PSP), subcutaneous Acute sclerosing pan-encephalitis, tangled dominant dementia, post-encephalitis Parkinson's disease, myotonic dystrophy, subacute sclerosing pan-encephalopathy, LRRK2 mutation, chronic traumatic encephalopathy (CTE), family Familial British dementia, familial Danish dementia, other frontotemporal degeneration, Guadeloupean Parkinsonism, neurodegeneration with iron in the brain Cumulative, SLC9A6-related mental retardation, white matter tau protein lesions with globular inclusions, epilepsy, Lewy body dementia (LBD), mild cognitive impairment (MCI), multiple sclerosis, Parkinson's disease Parkinson's disease, HIV-related dementia, adult-onset diabetes, senile cardiac amyloidosis, glaucoma, ischemic stroke, psychiatric AD, and Huntington's disease disease), preferably Alzheimer's disease (AD), cortical basal nucleus degeneration (CBD), Pick's disease (PiD) and progressive supranuclear palsy (PSP). The compound of any one of claims 1 to 3, 6, 9, 10, and 12, which is used to reduce Tau aggregation. A compound as claimed in any one of claims 1 to 3, 6, 9, 10 and 12, which is used to prevent the formation of Tau aggregates and/or to inhibit Tau aggregation. The compound according to any one of claims 1 to 3, 6, 9, 10, and 12, which is used for intracellular interference with Tau aggregates. The compound of any one of claims 1 to 3, 6, 9, 10, and 12, which is used to reduce Tau misfolding and hyperphosphorylation in vivo. A compound as claimed in any one of claims 1 to 3, 6, 9, 10 and 12, which is used as a marker for reducing neuroinflammation. Use of a compound as claimed in any one of claims 1 to 6 or 9 for the manufacture of a medicament for reducing Tau aggregation. A use of the compound according to any one of claims 1 to 6 or 9 for the manufacture of an agent for preventing the formation of Tau aggregates and/or for inhibiting Tau aggregation. A use of the compound according to any one of claims 1 to 6 or 9 for the manufacture of a medicament for interfering with Tau aggregates in a cell. A use of the compound according to any one of claims 1 to 6 or 9 for the manufacture of a medicament for reducing Tau misfolding and hyperphosphorylation in vivo. Use of a compound as claimed in any one of claims 1 to 6 or 9 for the manufacture of a medicament for reducing neuroinflammation markers. A mixture comprising a compound as claimed in any one of claims 1 to 6 and at least one other biologically active compound selected from therapeutic agents different from the compound as claimed in any one of claims 1 to 6, Wherein the mixture further comprises at least one of a pharmaceutically acceptable carrier, diluent and excipient. The mixture of claim 23, wherein the compound and/or the other biologically active compound is present in a therapeutically effective amount. The mixture of claim 23 or 24, wherein the other biologically active compound is selected from the group consisting of: anti-oxidative stress compounds; anti-apoptotic compounds; metal chelating agents; DNA repair inhibitors, such as pirenzepine ) And metabolites; 3-amino-1-propanesulfonic acid (3APS); 1,3-propanedisulfonate (1,3PDS); α-secretase activator; β- and γ-secretase inhibitors ; Glycogen synthase kinase 3 inhibitor; O-linked N-acetylglucosamine hydrolase (O-glcnacase, OGA) inhibitor; neurotransmitters; β-sheet breakers (β-sheet breakers); class Starch β depleting/depleting cell component attractants; inhibitors of N-terminal truncated starch β (including pyroglutarated starch β 3-42), anti-inflammatory molecules, or cholinesterase inhibition Agents (ChEI), such as tacrine, rivastigmine, donepezil, and/or galantamine; M1 agonists; other drugs, including any Starch or Tau modified drugs and nutritional supplements; antibodies, including any functionally equivalent antibodies or functional parts thereof; or vaccines. The mixture of claim 25, wherein the other biologically active compound is a cholinesterase inhibitor (ChEI). A mixture as claimed in claim 25, wherein the other biologically active compound is selected from the group consisting of tacrine, rivastigmine, donepezil, galantamine, nicotinic acid, and memantine . The mixture of claim 25, wherein the other biologically active compound is an antibody, in particular a monoclonal antibody, including any functionally equivalent antibody or functional part thereof. The mixture of claim 23 or 24, wherein the compound and/or the other biologically active compound is present in a therapeutically effective amount. A use of the compound according to any one of claims 1 to 6 or 9 as an analytical reference or in vitro screening tool.

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