Heterocyclic compounds for imaging aggregated tau proteins of tauopathies

Abstract

Heterocyclic compounds of Formula I or Formula II or pharmaceutically acceptable salts, solvates, hydrates, tautomers or stereoisomers thereof for imaging tau protein aggregates are provided. Compounds of Formula I or Formula II may be used to detect tau protein aggregates in the diagnosis or monitoring of the progression of a disease or disorder such as Alzheimer's disease, corticobasal degeneration and progressive suprakaryotic paralysis.

Description

Cross Reference to Related Applications

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 465,817, filed May 11, 2023. The contents of the application are incorporated herein by reference in their entirety. Statement as to Federally Sponsored Research

[0002] This invention was made with government support under U19 NS110456 awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND

[0003] Development of biomarkers to assess tau accumulation in the living human brain is critical to better understand the pathophysiology of tauopathies such as Alzheimer’s disease, progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD). Over the past 20 years, there has been great progress in developing positron emission tomography (PET) imaging biomarkers for AD. Despite three FDA-approved amyloid-β plaque PET radiopharmaceuticals becoming available for widespread imaging of amyloid-β plaques in AD over the past decade, first- and second-generation AD tau-PET tracers are still emerging. AD aggregates are composed of a mixture of both 3R-tau and 4R-tau, whereas PSP and CBD tau aggregates are primarily composed of 4R-tau, and PET imaging agents developed for imaging of tau deposits in AD such as AV-1451, PI-2620, MK-6240, and PM-PBB3 can not bind with high affinity to tau deposits in PSP and CBD patients. Thus, development of high-affinity 4R-tau selective PET imaging agents is one of the biggest challenges in PSP and CBD research.

[0004] There is a need in the art for 4R-tau selective binding agents, and the present disclosure meets this need. SUMMARY

[0005] According to one aspect of the present disclosure, there is provided a compound having the structure of Formula I or Formula II, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof: Formula I Formula II wherein: X1is N, O, or S; X2, X3, X4, X5are each independently N or CH; X6is CH or O; R1is H, halogen, CN, Me3N + X7 - , Et3N + X7 - , Pr3N + X7 - , Bu3N + X7 - , or other suitable ammonium salts, OH (with the proviso that when R1is OH, R1does not substitute at position 5 of the indole ring), OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate species, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin species, I(O-acyl)2, ArI + X7 - , IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, B-pinacol, B-neopentyl glycol, or other suitable boron species; wherein (1) X7 - is a suitable counterion; (2) ArI + X7 - is a suitable iodonium species known in the art; and (3) IR7is a suitable iodonium ylide known in the art; R2is H, alkyl, t-butyloxycarbonyl, tosyl, benzyl, p-methoxybenzyl, CBz, Fmoc, or other suitable nitrogen protecting groups; R3is H, halogen, NO2, Me3N + X7 - , Et3N + X7 - , Pr3N + X7 - , Bu3N + X7 - , or other suitable ammonium salts, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate species, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin species, I(O-acyl)2, ArI + X7 - , IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, B-pinacol, B-neopentyl glycol, or other suitable boron species; wherein (1) X7 - is a suitable counterion; (2) ArI+ X7 - (2) is a suitable iodonium substance known in the art; and (3) IR7 is a suitable iodonium ylide known in the art; R4 is H, halogen, OH, alkoxy, carbonyl, or OEtOEtF; R5 is H, halogen, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substance; R6 is H, halogen, OH, O-acyl, O-alkyl, OBoc, or other suitable oxygen-protecting substance; or R5 and R6 form an epoxide: m and n are each independently 0 or 1; and o is 0, 1 or 2.

[0006] In some embodiments, X1 is N; R1 is H or a halogen; R2 is H; R3 is H or a halogen; R4 is an alkoxy group; R5 is a halogen; and R6 is OH. In some embodiments, the halogen is F, Cl, I, or Br. In some embodiments, the halogen is F.

[0007] In some embodiments, the compound has the following structure: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

[0008] In some embodiments, the compound has the following structure: , , , , , , , , , , or .

[0009] In some implementations, any of the compounds described herein selectively binds to the 4Rτ protein.

[0010] In some implementations, any of the compounds described herein contains one or more detection markers.

[0011] In some implementations, the detection marker is selected from radionuclides, positron emitters, gamma emitters, and fluorescent markers.

[0012] In some implementations, the detection marker is selected from... 2 H, 3 H, 18 F, 123 I, 124 I, 125 I, 131 I, 11 C 13 C 14 C 13 N、 15 N、 15 O、 75 Br、 76 Br and 77 Br.

[0013] In some implementations, the detection marker is 18 F.

[0014] In some implementations, any of the compounds described herein are used to detect disorders or diseases associated with τ protein aggregates.

[0015] In some implementations, the disease or disorder is a 4R-τ protein disorder or a disease or disorder associated with 4R-τ protein aggregates.

[0016] In some implementations, the disease or disorder is a neurodegenerative disease or disorder.

[0017] In some implementations, the disorder or disease is selected from Alzheimer's disease, progressive supranuclear palsy, Pick's disease, frontotemporal dementia (FTLD), frontotemporal dementia-17 with Parkinson's syndrome (FTDP-17), primary age-related tau protein disease (PART) dementia, aurophilic granuloma, glioglobulin tau protein disease, chronic traumatic encephalopathy (CTE), corticobasal syndrome (CBS), and corticobasal degeneration.

[0018] In some implementations, detection is performed using one or more of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging, or autoradiography.

[0019] In some implementations, detection is performed using positron emission tomography (PET) imaging.

[0020] In one aspect, a method is provided for diagnosing a subject suffering from a disease or disorder, or for monitoring the progression of a subject's disease or disorder. The method comprises administering to the subject a detectable amount of an isotopically labeled compound of formula I or II, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof.

[0021] In some implementations, the disease or disorder is associated with τ protein aggregates.

[0022] In some implementations, the disease or disorder is a 4R-τ protein disorder or a disease or disorder associated with 4R-τ protein aggregates.

[0023] In some implementations, the disease or disorder is a neurodegenerative disease or disorder.

[0024] In some implementations, the disease or disorder is selected from Alzheimer's disease, progressive supranuclear palsy, Pick's disease, frontotemporal dementia (FTLD), frontotemporal dementia-17 with Parkinson's syndrome (FTDP-17), primary age-related tau protein disease (PART) dementia, aurophilic granuloma, glioglobulin tau protein disease, chronic traumatic encephalopathy (CTE), corticobasal syndrome (CBS), and corticobasal degeneration.

[0025] In some embodiments, the compound is 2 H, 3 H, 18 F, 123 I, 124 I, 125 I, 131 I, 11 C 13 C 14 C 13 N、15 N、 15 O、 75 Br、 76 Br or 77 Br isotope labeling.

[0026] In some implementations, detection is performed using one or more of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging, or autoradiography.

[0027] In all the methods described herein, the subject may be a human being.

[0028] The foregoing description of the invention, along with the following accompanying drawings and detailed descriptions, are exemplary and illustrative. They are intended to provide further details of this disclosure but should not be construed as limiting. Other objects, advantages, and novel features will readily become apparent to those skilled in the art from the following detailed descriptions of this disclosure. Attached Figure Description

[0029] Figure 1 shows the X-ray crystal structure of compound 38.

[0030] Figure 2 shows the X-ray crystal structure of compound 65.

[0031] Figure 3 shows the X-ray crystal structure of compound 66.

[0032] Figure 4 shows the X-ray crystal structure of compound 69 (JSS20-183A).

[0033] Figure 5 shows the contents of a male rhesus monkey (12 kg) [ 18 F]68 brain PET images and brain time-activity curves.

[0034] Figure 6 shows the contents of a male rhesus monkey (13 kg) [ 18 F]69([ 18 Brain PET images and brain time-activity curves (F]JSS20-183A).

[0035] Figure 7 shows post-mortem medial frontal cortex sections from PSP, CBD, and AD subjects, as well as the cingulate cortex from a 60-year-old control subject, using 5 nM [ 3 H]69([ 3 H]JSS20-183A) and self-blocking in adjacent slices with 1 mM unlabeled 69 (JSS20-183A). Detailed Implementation I. Overview

[0036] This invention relates to the discovery of novel compounds that selectively bind to tau proteins, particularly 4R-tau proteins. These compounds can be used, for example, in methods for imaging aggregated tau proteins in tau protein disorders.

[0037] Tau protein is a protein that helps stabilize the internal framework of nerve cells (neurons) in the brain. This internal framework has a tubular shape, through which nutrients and other essential substances reach different parts of the neuron. Tau proteinopathies are age-related neurodegenerative diseases characterized by the presence of aggregates of abnormally phosphorylated tau protein. In these neurodegenerative diseases, tau protein separates from microtubules to form insoluble aggregates, thus leading to tau proteinopathies.

[0038] Six distinct isoforms of microtubule-associated tau protein exist in the adult brain: ON3R, ON4R, 1N3R, 1N4R, 2N3R, and 2N4R. These isoforms differ in the number of N-terminal inserts (ON, 1N, 2N) and C-terminal repeat domains (3R or 4R) and exhibit differential expression depending on brain region and developmental stage. Adult human tau protein exhibits roughly equal expression of the 3R and 4R tau protein isoforms, with 1N3R and 1N4R being the most abundant forms.

[0039] As used in this article, "τ protein" refers to a highly soluble microtubule-binding protein that is primarily found in neurons and includes six main isoforms; cleaved or truncated forms; and other modified forms, such as those produced by phosphorylation, glycosylation, glycation, prolyl isomerization, nitration, acetylation, polyamineization, ubiquitination, SUMOylation, and oxidation.

[0040] Although all tau protein isoforms can aggregate and form neurofibrillary tangles, some tau protein diseases (such as Pick's disease and progressive supranuclear palsy) are characterized by the accumulation of specific tau protein isoforms. AD aggregates are composed of a mixture of 3R-tau and 4R-tau proteins, while PSP and CBD tau aggregates are primarily composed of 4R-tau proteins (Buee L; Delacourte A, “Comparative biochemistry of tau in progressive supranuclear palsy, corticobasal degeneration, FTDP-17 and Pick's disease,” Brain Pathology 1999, 9(4), 681-693). Expression of the 3R isoform causes more severe axonal transport defects and motor impairment, ultimately leading to a shorter lifespan than the 4R isoform. In contrast, the 4R isoform causes greater neurodegeneration and impairment of learning and memory. The expression of τ protein isoforms is directly related to brain development: during neurogenesis, only the shortest τ protein isoform, ON3R, is expressed, while in the adult brain, all six isoforms are present in roughly equal amounts as 3R and 4R isoforms.

[0041] The diagnosis of 4R-τ protein-specific neurological disorders has numerous benefits. Developing biomarkers to assess τ protein accumulation in the living human brain is crucial for a better understanding of the pathophysiology of Alzheimer's disease (AD) and non-AD τ protein disorders such as progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). 4R-τ protein biomarkers can assist clinicians in making more definitive disease diagnoses and confirming target binding, and can also help determine dosing regimens for novel anti-τ protein therapies currently under development.

[0042] Significant progress has been made in the development of positron emission tomography (PET) imaging biomarkers for Alzheimer's disease (AD) over the past 20 years. While several FDA-approved PET radiopharmaceuticals for amyloid-β plaques (AV-1451, PI-2620, MK-6240, and PM-PBB3) have been developed in the last decade for extensive imaging of AD amyloid-β plaques, first- and second-generation AD τ protein PET tracers continue to emerge. AD aggregates are composed of a mixture of 3R-τ and 4R-τ proteins, while PSP and CBD τ protein aggregates are primarily composed of 4R-τ protein, and PET imaging agents developed for imaging τ protein deposits in AD (AV-1451, PI-2620, MK-6240, PM-PBB3) may not bind with high affinity to τ protein deposits in PSP and CBD patients. Therefore, developing high-affinity 4R-τ protein-selective PET imaging agents is one of the biggest challenges in PSP and CBD research (Betthauser TJ., “In vitro evidence for a nonselective 4R-τ PET tracer,” Mol Psychiatry. January 19, 2023. doi: 10.1038 / s41380-023-01950-2. Electronic version published before print). For example, […] has been used in PSP and CBD patients. 18 F]PI-2620 and [ 18 PET imaging studies using 18F-APN-1607. (Li et al., “Progressive Supranuclear Palsy Neuroimage Initiative (PSPNI). Clinical Utility of 18F-APN-1607 Tau PET Imaging in Patients with Progressive Supranuclear Palsy,” Mov Disord. Oct 2021; 36(10):2314-2323; Tagai et al., “An optimized reference tissue method for quantification of tau protein depositions in diverse neurodegenerative disorders by PET with 18F-PM-PBB3 (18F-APN-1607),” Neuroimage. Dec 1, 2022; 264:119763).

[0043] Previously reported tau protein ligands include the following: 18 F-flortaucipir (AV-1451) is a tau protein PET ligand that binds to paired helical filaments of the tau protein associated with aging and AD, but its applicability in detecting aggregates of frontotemporal dementia remains uncertain (Tsai et al.). 18 F-flortaucipir (AV-1451) tau PET in frontotemporal dementia syndromes, “Alzheimer's Research and Therapy, 11(13) (2019)”. PI-2620 and MK-6240 are second-generation tau protein PET tracers (Malarte et al., “Discriminative binding of tau PET tracers PI2620, MK6240, and RO948 in Alzheimer's Disease, corticobasal degeneration and progressive supranuclear palsy brains,” Molecular Psychiatry, 28:1272-1283(2023)) as well as PM-PBB3 and CBD-2115 (Zhou et al., “Dissecting the Binding Profile of PETTracers to Corticobasal Degeneration Tau Fibrils,” ACS Chem. Neurosci., 12:3487-3496 (2021)).

[0044] However, the in vivo specificity of existing 4R-τ protein tracers is quite low, and there is likely a need to improve the binding affinity of 4R-τ protein PET tracers to improve the imaging signal of 4R-τ protein biomarkers in PSP and CBD patients. This invention surprisingly and unexpectedly solves this problem present in the prior art.

[0045] In one aspect of this disclosure, the compounds of formulas I and II described selectively bind to the 4Rτ protein, as defined by whether said compound binds to other protein binding sites (e.g., aggregated amyloid-β and aggregated α-synuclein). The binding affinity (Kb) of the compound at other protein binding sites is determined. d or Ki ), and at other binding sites, K is greater than about 50 nm. d and / or K i The values ​​indicate that the compound did not exhibit significant in vivo-specific imaging signals.

[0046] Non-limiting examples of disorders or diseases associated with tau protein aggregates (including, for example, 4R-tau protein aggregates) include Alzheimer's disease, progressive supranuclear palsy, Pick's disease, frontotemporal dementia (FTLD), frontotemporal dementia-17 with Parkinson's syndrome (FTDP-17), primary age-related tau protein disease (PART) dementia, aurophilic granuloma, glioglobulinosis, chronic traumatic encephalopathy (CTE), corticobasal syndrome (CBS), and corticobasal degeneration. II. Experimental results

[0047] In one exemplary aspect, Enamine Z3777013540 (Z-3540 or compound 1) was identified by a molecular similarity fingerprint search of an Enamine compound library based on CBD-2115. Z-3540 is an exemplary compound according to the present invention. Synthesis [ 3 H]Z-3540 was evaluated in human CBD, PSP and AD tissue binding assays. III. Compounds

[0048] This article presents heterocyclic compounds for imaging aggregated tau protein in tau protein diseases. To date, no high-affinity 4R-tau protein-selective PET tracer suitable for neuroimaging exists.

[0049] The heterocyclic compounds described herein have a structure according to Formula I or Formula II, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof: Formula I Formula II in: X1 is N, O, or S; X2, X3, X4, and X5 are each independently N or CH; X6 is CH or O; R1 is H, halogen, CN, Me3N + X7 - Et3N + X7 - Pr3N + X7 - Bu3N+ X7 - Or other suitable ammonium salts, OH (provided that R1 is not substituted at position 5 of the indole ring when R1 is OH), OMs, OTs, OTf, O-nitrobenzenesulfonates, O-bromobenzenesulfonates, or other suitable sulfonates, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin compounds, I(O-acyl)2, ArI + X7 - IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, β-pinacol, β-neopentyl glycol, or other suitable boron substances; of which (1) X7 - It is a suitable counterion; (2) ArI + X7 - (2) is a suitable iodonium substance known in the art; and (3) IR7 is a suitable iodonium ylide known in the art; R2 is H, alkyl, tert-butoxycarbonyl, toluenesulfonyl, benzyl, p-methoxybenzyl, CBz, Fmoc, or other suitable nitrogen protecting group; R3 is H, halogen, NO2, Me3N + X7 - Et3N + X7 - Pr3N + X7 - Bu3N + X7 - Or other suitable ammonium salts, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin substances, I(O-acyl)2, ArI + X7 - IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, β-pinacol, β-neopentyl glycol, or other suitable boron substances; of which (1) X7 - It is a suitable counterion; (2) ArI + X7 - (2) is a suitable iodonium substance known in the art; and (3) IR7 is a suitable iodonium ylide known in the art; R4 is H, halogen, OH, alkoxy, carbonyl, or OEtOEtF; R5 is H, halogen, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances; R6 is H, halogen, OH, O-acyl, O-alkyl, OBoc, or other suitable oxygen-protecting substance; or R5 and R6 form an epoxide: m and n are each independently 0 or 1; and o is 0, 1 or 2.

[0050] In some embodiments, X1 is N; R1 is H or a halogen; R2 is H; R3 is H or a halogen; R4 is an alkoxy group; R5 is a halogen; and R6 is OH. In some embodiments, the halogen is F, Cl, I, or Br. In some embodiments, the halogen is F.

[0051] Compounds of formula I can have one of the following structures: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

[0052] Compounds of formula II may have one of the following structures:

[0053] , , , , , , , , , , or .

[0054] In some embodiments, the compound of formula I has one of the following structures:

[0055] , , , , or .

[0056] In some embodiments, the compound of formula II has the following structure: .

[0057] Salts of compounds of Formula I or II may be pharmaceutically acceptable salts. Other salts may be used to prepare compounds according to this disclosure or pharmaceutically acceptable salts thereof.

[0058] When the compounds in this disclosure are acidic, a suitable "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic alkali, including inorganic and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, ferric salts, ferrous salts, lithium salts, magnesium salts, ferric salts, ferrous salts, potassium salts, sodium salts, zinc salts, etc. Particularly preferred are ammonium salts, calcium salts, magnesium salts, potassium salts, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic alkaloids include salts of the following substances: primary amines, secondary amines, and tertiary amines; substituted amines (including naturally occurring substituted amines); cyclic amines; and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

[0059] When the compounds of the present invention are basic, the salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucoic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. Particularly preferred are citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, and tartaric acid.

[0060] "Amine protecting group" is any protecting group suitable for protecting an amine group in a contemplated chemical reaction. Examples of suitable protecting groups are well known to those skilled in the art. These examples include, but are not limited to, urethanes, amides, imides, N-alkylamines, N-arylamines, imides, and enamines. Suitable protecting groups are discussed, for example, in the textbook *Greene and Wuts, Protecting groups in Organic Synthesis*, 3rd edition, pp. 494-653, which is incorporated herein by reference. Specific preferred examples of amine protecting groups include tert-butyloxycarbonyl (Boc), trimethylsilylethoxycarbonyl (Teoc), carbabenzyloxy (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), 9-fluorenylmethoxycarbonyl (Fmoc), acetyl (Ac), or trifluoroacetyl.

[0061] Compounds of Formula I or II can be labeled. A “label” (which may be a radiolabel or other detectable label, or tag, marker, detectable marker, tracer, radiotracer, or equivalent) is any atom or group suitable for in vivo or in vitro imaging and / or determination (e.g., identification, imaging, diagnosis, evaluation, detection, and / or quantification), and particularly imaging and diagnosis. Suitable labels include, for example, radioisotopes (also referred to as “radiolabeled atoms”), radionuclides, isotopes, positron emitters, gamma emitters, fluorescent groups, luminescent groups, chromophores, biotin (complexed with streptoavidin), or photoaffinity groups. The type of label chosen will depend on the desired detection method. There are no particular limitations on the location where the label is integrated or attached to the compound of the present invention.

[0062] Examples of isotopes (such as radioactive isotopes, radionuclides, positron emitters, and gamma emitters) that can be used to label the compounds of the present invention include, but are not limited to: 2 H, 3 H, 18 F, 123 I, 124 I, 125 I, 131 I, 11 C 13 C 14 C 13 N、 15 N、 15 O、 75 Br、 76 Br and 77 Br. In some implementations, the detection marker is 18 F.

[0063] In some embodiments, any of the compounds disclosed herein contains one or more detection tags. In some embodiments, the R1 and R3 substituents are detectably labeled. In some embodiments, in any of the compounds described herein, one or more H, one or more C, one or more N, one or more O, one or more F, one or more Br, and / or one or more I may be present. 3 H; 11 C 13 C or 14 C; 13 N; 15 O; 18 F; 75 Br or 76 Br; or 123 I, 124 I, 125 I, 131 I is an alternative. In some implementations, the detection marker is... 18 F.

[0064] In some embodiments, the compounds of this disclosure selectively bind to the 4R-τ protein. In some embodiments, the compounds of this disclosure can bind to 4R-τ protein aggregates both in vivo and in vitro. In some embodiments, the compounds of this disclosure can be used as diagnostic agents (for in vivo and / or in vitro diagnostic purposes) to diagnose diseases or disorders associated with 4R-τ protein aggregates (i.e., τ proteinopathies). In some embodiments, the compounds of this disclosure are used to detect disorders or diseases associated with τ protein aggregates. In some embodiments, the disease or disorder is a 4R-τ proteinopathy or a disease or disorder associated with 4R-τ protein aggregates. In some embodiments, the disease or disorder is a neurodegenerative disease or disorder. IV. Method

[0065] This article also provides methods for diagnosing a subject suffering from a disease or disorder, or for monitoring the progression of a subject's disease or disorder. In one aspect, the disease or disorder is a neurological disorder characterized by the accumulation of 4Rτ protein aggregates in the subject's body.

[0066] In another aspect, it covers a method for diagnosing subjects with non-AD τ protein disorders characterized by the accumulation of 4R τ protein aggregates in the subject's body.

[0067] In another aspect, it covers a method for diagnosing subjects with non-AD diseases or disorders, such as eliminating the diagnosis of AD.

[0068] In one aspect, the method covers diagnosing a subject with a neurological disorder and subsequently administering treatment to the disorder or impairment, said neurological disorder being characterized by the accumulation of 4R τ protein aggregates in the subject's body. The disorder or impairment may be, for example, progressive supranuclear palsy, Pick's disease, frontotemporal dementia (FTLD), frontotemporal dementia-17 with Parkinson's syndrome (FTDP-17), primary age-related τ proteinopathies (PART) dementia, aerophilic granulomatosis, glioglobulinosis, chronic traumatic encephalopathy (CTE), corticobasal syndrome (CBS), or corticobasal degeneration.

[0069] The methods disclosed herein may include (a) administering to a subject in need a detectable amount of an isotopically labeled compound of formula I or II or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof; and (b) detecting the binding of said compound to τ protein aggregates in the patient’s body, and particularly to 4R-τ protein aggregates present in the patient’s body.

[0070] In some implementations, detection is performed using one or more of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging, or autoradiography. In some implementations, detection is performed using positron emission tomography (PET). V. Definition

[0071] As used herein, the term "alkyl" refers to both straight-chain and branched saturated hydrocarbon groups. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, pentyl, and hexyl. Examples of unbranched alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, and n-butyl. Examples of branched alkyl groups include, but are not limited to, isopropyl, tert-butyl, isobutyl, 1-ethylpropyl, and 1-ethylbutyl.

[0072] As used herein, the term "halogen" or "halogenated group" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0073] As used herein, the term "alkoxy" refers to an alkyl group having a specified number of carbon atoms attached by an oxygen bridge.

[0074] As used herein, the terms "heterocyclic group," "heterocycle," or "heterocyclic" refer to stable 5- to 7-membered monocyclic or stable 8- to 11-membered bicyclic heterocycles that are saturated or unsaturated and consist of a carbon atom and one to four heteroatoms selected from N, O, and S, and include any bicyclic group in which any heterocycle as defined above is fused to a benzene ring. Heterocycles can be attached to any heteroatom or carbon atom, resulting in a stable structure. The terms heterocyclic group, heterocycle, or heterocyclic include a heteroaryl moiety.

[0075] As used herein, “pharmaceutical acceptable” means compounds, materials, compositions, and / or dosage forms that, to the extent of reasonable medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0076] As used herein, “about” will be understood by those skilled in the art and will vary to some extent depending on the context in which it is used. If the use of the term is not apparent to those skilled in the art in light of the context in which it is used, “about” will mean up to 10% positive or negative of a particular term.

[0077] Unless otherwise indicated herein or clearly contradicted by the context, the use of the terms “a / an” and “described” and similar indicative terms in the context of describing elements (especially in the context of the following claims) should be interpreted as covering both the singular and plural. Unless otherwise indicated herein, the numerical ranges listed herein are intended only as a shorthand method for individually referring to each individual value falling within that range, and each individual value is incorporated into the specification as if it were individually listed herein. Unless otherwise indicated herein or clearly contradicted by the context, all methods described herein may be performed in any suitable order. Unless otherwise stated, the use of any and all examples or exemplary language (e.g., “such as”) provided herein is intended only to better illustrate the embodiments and does not impose any limitation on the scope of the claims. The language in the specification should not be construed as indicating that any unclaimed element is necessary.

[0078] Various implementation schemes are described below. It should be noted that a particular implementation scheme is not intended as an exhaustive description or as a limitation on the broader aspects discussed herein. An aspect described in conjunction with a particular implementation scheme is not necessarily limited to that scheme and can be practiced in conjunction with any other one or more other implementation schemes.

[0079] The invention, as thus generally described, will be more readily understood by referring to the following embodiments, which are provided by way of illustration and are not intended to limit the invention. Example

[0080] Example 1. Identification of compounds with high affinity for aggregated 4R-τ proteins

[0081] The purpose of this embodiment is to identify compounds with high affinity for aggregated 4R-τ proteins.

[0082] Three 6-fluoroindole-based compounds 1–3 (shown below) available from commercial sources (Enamine) were identified as having high affinity for aggregated 4R-τ protein in human brain tissue samples from patients with AD, PSP, and CBD (Table 1).

[0083] These compounds were identified by combining ultra-high-throughput computer simulation screening with in vitro binding assays for “sprout confirmation.” Subsequent chemical synthesis and structure-activity relationship studies identified numerous 4R-τ protein ligands (shown below) that showed high affinity in human brain tissue samples from patients with AD, PSP, and CBD (Table 2 below).

[0084] Example 2. Synthesis of 4R-τ protein ligands

[0085] The purpose of this embodiment is to synthesize various 4R-τ protein ligands.

[0086] NMR spectra were obtained at specified frequencies using a Bruker Avance III spectrometer. Spectra in CDCl3 were referenced to the internal TMS, while spectra in other deuterated solvents were referenced to solvent residual protons.

[0087] Compounds 4 and 5 (Scheme 1) were prepared according to the previously described procedure (Kumar, A. et al.; Synthesis 2008, 707-710); while compound 6 was commercially available. Compounds 4-6 were coupled with 5-bromo-2-chloropyrimidine under Pd catalysis to give compounds 7-11, wherein the N-Boc protected (7, 9, and 11) and N-deprotected (8 and 10) compounds were isolated. Compound 7 was reacted with piperidine, 4-fluoropiperidine hydrochloride, or 4,4-difluoropiperidine hydrochloride to give compounds 12, 13, and 14, respectively, followed by deprotection with TFA to give compounds 15, 16, and 17, respectively. Compound 10 and the Boc protected compounds 7 and 11 were reacted with trans / racemic-4-fluoro-3-hydroxypyrrolidine hydrochloride to give compounds 18, 19, and 20, respectively, followed by removal of the N-Boc protecting group from 19 and 20 with TFA to give compounds 21 and 22, respectively. 5-Bromo-2-chloropyrimidine was reacted with various cyclic amines (Scheme 2) to give compounds 23, 24, (R)-25, and 26. Compound 26 was then oxidized with Dess-Martin periodinane to give compound 27. 2,5-Dibromopyridine was reacted with piperidine or morpholine (Scheme 3) to give compounds 28 and 29, respectively. 2,5-Dibromopyrazine was reacted with morpholine (Scheme 3) to give compound 30. Pd-catalyzed coupling of 4 (Scheme 4) with compounds 23, (R)-25, 27, 28, and 29 yielded N-Boc protected compounds 31, (R)-32, 33, 34, and 35, and N-deprotected compound 36, respectively. Compounds 31-35 were then deprotected with TFA to give compounds 36, (R)-37, 38, 39, and 40 (the structure of 38 was confirmed by X-ray crystallography, Figure 1). Commercially available compounds 41, 6, and 42 (Scheme 5) were coupled with compound 26 to obtain compounds 43, 44, and 45, respectively. Compounds 44 and 45 were then deprotected with TFA to obtain compounds 46 and 47, respectively.

[0088] Option 1

[0089] Option 2

[0090] Option 3

[0091] Option 4

[0092] Option 5

[0093] 2,6-Difluoropyridine and 2-chloro-6-nitropyridine were each reacted with piperidine or morpholine (Scheme 6) to give compounds 48, 49, 50, and 51, which were then brominated with NBS to give compounds 52, 53, 54, and 55 (Smith, AC et al., Organic Process Research & Development 2018, 22, 681-696). 2-Bromo-4-fluoropyridine was reacted with morpholine (Scheme 7) to give compound 56, which was then brominated to give compound 57. 2,5-Dibromo-3-fluoropyridine, 5-bromo-2-chloro-3-fluoropyridine, and 5-bromo-2-chloro-4-fluoropyridine (Scheme 8) were each coupled with piperidine or morpholine under Pd catalysis (Ji, J. et al., Organic Letters 2003, 5, 4611-4614; Stroup, B. W et al., Organic Letters 2007, 9, 2039-2042) to give compounds 58, 59, 60, 61, and 62.

[0094] Option 6

[0095] Option 7

[0096] Option 8

[0097] Compounds 52-55 were coupled with compound 6 under Pd catalysis (Scheme 9) to obtain N-Boc protected compounds 63, 64, 65, and 66, and N-deprotected compound 67, respectively. Compounds 63 and 64 were then deprotected with TFA to obtain compounds 68 and 69 (JSS20-183A), respectively. X-ray crystallography confirmed the structures of compounds 65 (Figure 2), 66 (Figure 3), and 69 (Figure 4).

[0098] Option 9

[0099] Compound 42 was coupled with compounds 24 and 29 (Scheme 10) to give compounds 70 and 71, respectively, and then deprotected with TFA to give compounds 72 and 73, respectively. Compound 4 was coupled with 52 (Scheme 11) to give compound 74, and then deprotected with TFA to give compound 75. Compound 76 was coupled with compounds 30 and 57-59 (Scheme 11) to give compounds 77, 78, 79, and 80, respectively, and then deprotected with TFA to give compounds 81, 82, 83, and 84, respectively. Compound 6 was coupled with compounds 57-59 (Scheme 11) to give compounds 85, 86, and 87, respectively, and then deprotected with TFA to give compounds 88, 89, and 90, respectively.

[0100] Option 10

[0101] Option 11

[0102] Compound 6 was coupled with compound 60 (Scheme 12) to give compound 91, which was then deprotected with TFA to give compound 92. Compounds 6 and 76 were coupled with compounds 61 and 62 respectively (Scheme 12) to give compounds 93, 94, 95 and 96, which were then deprotected with TFA to give compounds 97, 98, 99 and 100, respectively.

[0103] Option 12

[0104] Compound 44 was deprotonated with NaH (Scheme 13), then O-alkylated with 2-(2-fluoroethoxy)ethyl 4-methylbenzenesulfonic acid to give compound 101, which was then deprotected with TFA to give compound 102. Compound 2 was deprotonated with NaH (Scheme 14), then N-methylated with methyl toluenesulfonate to give compound 103.

[0105] Option 13

[0106] Option 14

[0107] method

[0108] Overview

[0109] Solvents and reagents were used exactly as received. NMR spectra were obtained at the specified frequencies using a Bruker Avance III spectrometer. In CDCl3... 1H NMR spectra refer to the internal TMS, while spectra in DMSO-d6, CD3OD, or acetone-d6 refer to the solvent residual protons. 13 C10 NMR spectroscopy reference solvent resonance. 19 The F NMR spectrum is unreferenced. A silica rotor from Miles Scientific was used at Harrison Research Chromatotron. TM Radial chromatography was performed. The silica gel used was Silicycle SiliaFlash® P60 40-63 μm (230-400 mesh).

[0110] Chemical

[0111] 6-Fluoro-2-(trimethylstanyl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 4). Compound 104 (2.23 g, 9.48 mmol) and Me3SnCl (2.09 g, 10.49 mmol, 1.1 equivalents) were mixed with N... 2(g) Rinse for 40 min, then dissolve in freshly distilled THF (100 mL) and cool in a CH3CN / dry ice bath. Add LDA solution (2.0 M THF / heptane / ethylbenzene, 6 mL, 12 mmol, 1.3 equivalence) dropwise over 3 min intervals, stirring the reaction mixture at CH3CN / dry ice temperature for 5 min, then warm to ambient temperature and stirring for 4 h. Add H2O (0.5 mL), stir the mixture for 5 min, and then concentrate to a dark brown oil. Add CH2Cl2 and hexane and remove them to obtain a dark green / brown slurry / residue, which is dissolved in CH2Cl2 and purified by vacuum rapid chromatography on silica (15 cm h x 4 cm id): % CH2Cl2 / hexane - 25% (200 mL), 50% (100 mL) to obtain a colorless slurry (3.64 g) that slowly solidifies. The crude product was purified twice by radial chromatography (4 mm silica):hexane (100 mL) to give compound 4 (3.52 g, 93%) as a white crystalline solid. 1 H NMR (500 MHz, CDCl3) δ 7.64 (dd, 1 H, J =10.5 Hz, J = 2.0 Hz), 7.41 (dd, 1 H, J = 8.5 Hz, J = 5.5 Hz), 6.94 (td, 1 H,J = 9.0 Hz, J = 2.0 Hz), 6.68 (t, 1H, 3 J SnH= 9.0 Hz), 1.70 (s, 9 H), 0.30(t, 9 H, 2 J SnH = 28.0 Hz).

[0112] 2-(2-chloropyrimidin-5-yl)-6-fluoro-1H-indole-1-carboxylic acid tert-butyl ester (compound 7) and 2-(2-chloropyrimidin-5-yl)-6-fluoro-1H-indole (compound 8). Compound 4 (0.38 g, 9.55 x 10⁻⁶ g) -4 mol), 5-bromo-2-chloropyrimidine (0.25 g, 1.29 mmol, 1.4 equivalents), Pd(PPh3)4 (80 mg, 6.92 x 10⁻⁶ ... -5 mol (0.07 equivalent) and toluene (25 mL) were refluxed in N2O. 2(g) The mixture was stirred for 15 h, then cooled to ambient temperature and stirred for 3 h. The reaction mixture was poured onto dry silica (55 mm h x 45 mm id) and eluted under vacuum: hexane (50 mL); CH2Cl2 (100 mL); %MeOH / CH2Cl2 - 1% (100 mL), 2.5% (200 mL), 5% (100 mL), 10% (50 mL) to give an orange / brown residue (0.33 g). Purification was performed by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL), 50 : 45 : 5 (50 mL) to give compound 7 (57 mg, 17%) as a grayish-white solid and compound 8 (30 mg, 13%) as a light brown solid.

[0113] Compound 7: 1 H NMR (300 MHz, CDCl3) δ 8.68 (s, 2 H), 7.94 (dd, 1 H, J =10.5 Hz, J = 2.4 Hz), 7.53 (dd, 1 H, J = 8.4 Hz, J = 5.4 Hz), 7.06 (td, 1 H,J = 8.7 Hz, J = 2.4 Hz), 6.68 (d, 1 H, J = 0.6 Hz), 1.48 (s, 9 H); HRMS (ESI)[M+H] + C 17 H 16 Calculated value of ClFN3O2: 348.0910, measured value: 348.0902.

[0114] Compound 8: 1 ¹H NMR (300 MHz, acetone-d6) δ 11.10 (br s, ¹H), 9.15 (s, ²H), 7.64 (dd, ¹H, J = 8.7 Hz, J = 5.4 Hz), 7.19 (m-overlap resonance, ²H), 6.91 (ddd, ¹H, J = 9.9 Hz, J = 8.7 Hz, J = 2.1 Hz); HRMS (ESI) [M+H] + C 12 Calculated value of H8ClFN3: 248.0385, measured value: 248.0382.

[0115] 2-(2-chloropyrimidin-5-yl)-6-cyano-1H-indole-1-carboxylic acid tert-butyl ester (compound 9) and 2-(2-chloropyrimidin-5-yl)-1H-indole-6-carboxynitrile (compound 10). 1,4-Dioxane was treated with N... 2(g) Purge for 40 min. Compound 5 (0.46 g, 1.14 mmol), 5-bromo-2-chloropyrimidine (0.23 g, 1.19 mmol), and Pd(dppf)Cl2 (78 mg, 1.07 x 10⁻⁶ mmol) were then added. -4 0.14 g (1.32 mmol, 1.2 equivalent) of Na₂CO₃ (0.14 g, 1.32 mmol, 1.2 equivalent) were used with N 2(g) Rinse for 15 min, then add 1,4-dioxane (20 mL). Recirculate the reaction mixture under reflux in N2O2. 2(g) Stir for 4 h, cool, and remove solvent to obtain a brown residue, which is dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (200 mL), 4% (100 mL) to obtain an orange / brown residue (0.16 g, crude compound 9) and an orange / brown solid (62 mg, crude compound 10).

[0116] Compound 9: The residue was dissolved in CH2Cl2 / MeOH, poured onto dry silica (33 mm h x 33 mm i.d.), and eluted under vacuum: % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL) to give a deep orange residue, which was briefly dried under vacuum. The residue was dissolved / suspended in CHCl3 (1 mL), filtered, and the precipitate was washed with CHCl3 (1 mL x 2). The filtrate was purified by radial chromatography (1 mm silica): 75 : 20 : 5 v / v / v hexane / EtOAc / NEt3 (100 mL) to give compound 9 (38 mg, 9%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ 8.71 (s, 2 H), 8.58 (d, 1 H, J = 0.6 Hz), 7.69 (d, 1 H, J = 8.1 Hz), 7.56 (dd, 1 H, J = 8.1 Hz, J = 1.5 Hz), 6.77 (s, 1 H), 1.49 (s, 9 H); HRMS(ESI) [M+H] + C 18 H 16 Calculated value of O2N4Cl: 355.0956, measured value: 355.0958.

[0117] Compound 10: The solid was dissolved / suspended in CHCl3 (2.5 mL), filtered, and the precipitate was washed with CHCl3 (2.5 mL x 3) and then dried under vacuum to give Compound 10 (29 mg, 10%) as a yellow / orange solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.44 (s, 1 H), 9.31 (s, 2 H), 7.96 (s, 1 H), 7.79 (d, 1H, J = 8.0 Hz), 7.40 (d, 1 H, J = 8.0 Hz), 7.36 (s, 1 H).

[0118] 2-(2-chloropyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 11). 1-Boc-indole-2-boronic acid (compound 6) (0.25 g, 9.58 x 10⁻⁶) -4 mol), 5-bromo-2-chloropyrimidine (0.18 g, 9.31 x 10⁻⁶ ... -4 mol) and Pd(dppf)Cl2 (52 mg, 7.11 x 10 -5mol (0.08 equivalents) with N 2(g) Rinse for 10 min, then add 1,4-dioxane (10 mL), followed by a solution of K₂CO₃ in H₂O (1.5 mL, 2 M, 3 mmol, 3.2 equivalence). Recycle the reaction mixture under reflux in N₂O. 2(g) Stir for 7 h, cool to ambient temperature, stir overnight, then filter through diatomaceous earth and wash the diatomaceous earth with 1,4-dioxane. Concentrate the filtrate into a brown oil, then add CHCl3 and hexane and remove it to obtain a brown slurry, dissolve it in CH2Cl2, pour it onto dry silica (45 mm h x 45 mm id) and elute under vacuum: CH2Cl2 (200 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (50 mL), 3% (50 mL) to give a brown solid (0.19 g). Purification by radial chromatography (2 mm silica): CH2Cl2 (100 mL) yielded a grayish-white solid (0.11 g), which was further purified by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (35 mL), 85 : 12 : 3 (100 mL), 75 : 20 : 5 (50 mL) to give compound 11 (82 mg, 27%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.69 (s, 2 H), 8.20 (d, 1 H, J = 8.4Hz), 7.60 (d, 1 H, J = 7.6 Hz), 7.40 (ddd, 1 H, J = 8.4 Hz, J = 7.2 Hz, J =1.2 Hz), 7.30 (ddd, 1 H, J = 7.8 Hz, J = 7.2 Hz, J = 0.8 Hz), 6.71 (s, 1 H), 1.49 (s, 9 H).

[0119] 6-Fluoro-2-(2-(piperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 12). Compound 7 (42 mg, 1.21 x 10⁻⁶) -4 mol), piperidine (0.05 mL, 5.1 x 10⁻⁶ ...). -4 mol, 4.2 equivalents), K2CO3 (24 mg, 1.74 x 10⁻⁶ ...) -4 mol (1.4 equivalents) and CH3CN (10 mL) were refluxed in N2O. 2(g)Stir for 75 min, then cool to ambient temperature. Remove CH3CN to obtain the residue, dissolve it in CH2Cl2, pour onto dry silica (33 mm h x 33 mm id) and elute under vacuum: CH2Cl2 (25 mL); % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (50 mL) to give compound 12 (47 mg, 98%) as a light brown solid. 1 H NMR (500 MHz, CDCl3) δ8.33 (s, 2 H), 7.93 (dd, 1 H, J = 10.5 Hz, J = 2.5 Hz), 7.45 (dd, 1 H, J =8.5 Hz, J = 5.5 Hz), 7.01 (td, 1 H, J = 8.5 Hz, J = 2.5 Hz), 6.48 (s, 1 H), 3.84 (t, 4 H, J = 5.0 Hz), 1.71 (m, 2 H), 1.62 (m, 4 H), 1.48 (s, 9 H).

[0120] 6-Fluoro-2-(2-(4-fluoropiperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 13). Compound 7 (60 mg, 1.73 x 10⁻⁶) -4 mol), 4-Fluoroperidol•HCl (34 mg, 2.44 x 10⁻⁶ ...) -4 mol (1.4 equivalents), K2CO3 (88 mg, 6.37 x 10⁻⁶ mol ... -4 mol (3.7 equivalents) and CH3CN (5 mL) were refluxed in N2O. 2(g) Stir for 5 h, then cool. Remove the solvent to obtain the residue, dissolve it in CH2Cl2, pour it onto dry silica (33 mm h x 33 mm i.d.), and elute with CH2Cl2 (125 mL) under vacuum to give compound 13 (64 mg, 89%) as a white foam: 1H NMR (300 MHz, CDCl3) δ 8.35 (s, 2 H), 7.93 (dd, 1 H, J = 10.8 Hz, J = 2.4Hz), 7.46 (dd, 1 H, J = 8.7 Hz, J = 5.7 Hz), 7.01 (td, 1 H, J = 8.7 Hz, J =2.4 Hz), 6.50 (s, 1 H), 4.99 (tt, 0.5 H, J = 6.3 Hz, J = 3.3 Hz) and 4.83 (dt,0.5 H, J = 9.3 Hz, J = 4.8 Hz) ( 2 J HF = 48.3 Hz), 3.97 (t, 4 H, J = 5.7 Hz),1.93 (m, 4 H), 1.48 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of F2N4O2: 415.1940, measured value: 415.1944.

[0121] 2-(2-(4,4-difluoropiperidin-1-yl)pyrimidin-5-yl)-6-fluoro-1H-indole-1-carboxylic acid tert-butyl ester (compound 14). Compound 7 (94.5 mg, 2.72 x 10⁻⁶) -4 mol), 4,4-difluoropiperidine•HCl (60 mg, 3.81 x 10⁻⁶ ...) -4 mol (1.4 equivalents), K2CO3 (186 mg, 1.35 mmol, 5 equivalents) and CH3CN (20 mL) were refluxed in N2O. 2(g)The mixture was stirred for 21 h, then cooled to ambient temperature, filtered, and the precipitate was washed with CH3CN. CH3CN was removed to give an orange residue, which was then removed by adding CH2Cl2 and hexane to give a pale orange solid, which was dried under vacuum. The solid was dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id), and eluted under vacuum with CH2Cl2 (350 mL) to give a colorless residue. Purification was performed by radial chromatography (2 mm silica): % CH2Cl2 / hexane - 25% (50 mL), 50% (125 mL) to give a white solid (46 mg), which was further purified by radial chromatography (1 mm silica): % CH2Cl2 / hexane - 25% (100 mL), 35% (100 mL), 50% (50 mL) to give compound 14 (32 mg, 27%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.37 (s, 2 H), 7.93 (dd, 1 H, J = 10.8Hz, J = 2.0 Hz), 7.46 (dd, 1 H, J = 8.4 Hz, J = 5.6 Hz), 7.02 (td, 1 H, J =8.8 Hz, J = 2.0 Hz), 6.51 (s, 1 H), 4.03 (t, 4 H, J = 6.0 Hz), 2.02 (septet, 4 H, J = 6.0 Hz), 1.48 (s, 9 H) HRMS (ESI) [M+H] + C 22 H 24 Calculated value of O2N4F3: 433.1846, measured value: 433.1836.

[0122] 6-Fluoro-2-(2-(piperidin-1-yl)pyrimidin-5-yl)-1H-indole (compound 15). Compound 12 (41 mg, 1.03 x 10⁻⁶) -4The solution was dissolved in TFA (1 mL, 13 mmol, 126 equivalents) and stirred at ambient temperature for 15 min. The solution was then poured into a mixture of NaHCO3 (1.23 g, 14.64 mmol, 1.1 equivalents of TFA), H2O (30 mL), and CH2Cl2 (30 mL). The mixture was stirred, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (10 mL). The combined CH2Cl2 layers were washed with brine (15 mL) and dried over MgSO4. The solution was concentrated, poured onto dry silica (33 mm h x 33 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (50 mL), 75 : 20 : 5 (50 mL), 50: 45 : 5 (75 mL) to give compound 15 (27 mg, 88%) as a pale yellow solid. 1 H NMR (300 MHz, CDCl3) δ 8.56 (s, 2 H), 8.20 (br s, 1 H), 7.49 (dd, 1 H, J = 8.7 Hz, J = 5.4Hz), 7.08 (dd, 1 H, J = 9.6 Hz, J = 2.1 Hz), 6.88 (ddd, 1 H, J = 9.6 Hz, J =8.7 Hz, J = 2.4 Hz), 6.62 (dd, 1 H, J = 2.1 Hz, J = 0.9 Hz), 3.84 (t, 4 H, J= 5.4 Hz), 1.68 (m, 6 H); 13 C NMR (125 MHz, DMSO-d6) δ 160.08, 158.71 (d, 1 J FC = 234.4 Hz), 154.49, 136.68 (d, J FC = 13.0 Hz), 134.17 (d, J FC =3.4 Hz), 125.44, 120.46 (d, J FC = 10.2 Hz), 114.48, 107.73 (d, J FC = 24.1 Hz), 97.05(d, J FC = 25.7 Hz), 96.91, 44.34, 25.25, 24.27; 19F NMR (470.6 MHz, DMSO-d6) δ-121.61 (m); HRMS (ESI) [M+H] + C 17 H 18 Calculated value of FN4: 297.1510, measured value: 297.1515.

[0123] 6-Fluoro-2-(2-(4-fluoropiperidin-1-yl)pyrimidin-5-yl)-1H-indole (compound 16). Compound 13 (59 mg, 1.42 x 10⁻⁶) -4 The aqueous layer was dissolved in TFA (1.5 mL, 19.5 mmol, 137 equivalents) and stirred at ambient temperature for 20 min. Then, it was poured into a mixture of NaHCO3 (1.82 g, 21.67 mmol, 1.1 equivalents of TFA), H2O (45 mL), and CH2Cl2 (45 mL). The mixture was stirred until the color disappeared, then the layers were separated, and the aqueous layer was extracted with CH2Cl2 (10 mL). The combined CH2Cl2 layers were washed with brine (20 mL) and dried over MgSO4. The solution was concentrated, poured onto dry silica (33 mmh x 33 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (50 mL), 75: 20 : 5 (50 mL), 50 : 45 : 5 (100 mL) to give compound 16 (44 mg, 99%) as a light brown solid. 1 H NMR (300 MHz, CDCl3) δ 8.58 (s, 2 H), 8.21 (br s, 1 H), 7.50 (dd, 1 H, J =8.7 Hz, J = 5.4 Hz), 7.08 (dd, 1 H, J = 9.6 Hz, J = 2.1 Hz), 6.89 (ddd, 1 H,J = 9.6 Hz, J = 8.7 Hz, J = 2.1 Hz), 6.64 (dd, 1 H, J = 2.1 Hz, J = 0.9 Hz), 5.00 (tt, 0.5 H, J = 6.0 Hz, J = 3.3 Hz) and 4.83 (dt, 0.5 H, J = 9.6 Hz, J =4.8 Hz) ( 2 J HF = 48.3 Hz), 3.97 (m, 4 H), 1.96 (m, 4 H); 1H NMR (300 MHz, DMSO-d6) δ 11.58 (s, 1 H), 8.83 (s, 2 H), 7.49 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.13 (dd, 1 H, J = 9.9 Hz, J = 2.1 Hz), 6.85 (ddd, 1 H, J = 9.9 Hz, J = 8.7Hz, J = 2.1 Hz), 6.82 (d, 1 H, J = 1.8 Hz), 4.93 (dtt, 1 H, 2 J HF = 48.6 Hz, J= 7.2 Hz, J = 3.6 Hz), 3.97 (m, 2 H), 3.78 (m, 2 H), 1.93 (m, 2 H), 1.72 (m,2 H); HRMS (ESI) [M+H] + C 17 H 17 Calculated value of F2N4: 315.1416, measured value: 315.1420.

[0124] 2-(2-(4,4-difluoropiperidin-1-yl)pyrimidin-5-yl)-6-fluoro-1H-indole (compound 17). Compound 14 (27 mg, 6.24 x 10⁻⁶) was used. -5 The solution was dissolved in TFA (0.6 mL, 7.8 mmol, 125 equivalents) and stirred at ambient temperature for 20 min. The solution was then poured into a mixture of NaHCO3 (0.856 g, 10.19 mmol, 1.3 equivalents of TFA), H2O (20 mL), and CH2Cl2 (20 mL). The mixture was stirred for 15 min, and the layers were separated. The aqueous layer was extracted with CH2Cl2 (5 mL x 2). The combined CH2Cl2 layers were washed with brine (15 mL) and dried over MgSO4. The solution was poured onto dry silica (33 mm h x 33 mm id) and eluted under vacuum: CH2Cl2 (50 mL) % MeOH / CH2Cl2 - 1% (50 mL), 2% (25 mL), 4% (50 mL) to give compound 17 (18 mg, 87%) as a grayish-white solid. 1H NMR (300 MHz, CDCl3)δ 8.60 (s, 2 H), 8.23 ​​(br s, 1 H), 7.51 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.08 (dd, 1 H, J = 9.6 Hz, J = 2.1 Hz), 6.90 (ddd, 1 H, J = 9.6 Hz, J = 8.7Hz, J = 2.1 Hz), 6.66 (d, 1 H, J = 1.5 Hz), 4.03 (t, 4 H, J = 5.7 Hz), 2.04(tt, 4 H, 3 J HF = 13.5 Hz, J = 5.7 Hz); HRMS (ESI) [M+H] + C 17 H 16 Calculated value of F3N4: 333.1322, measured value: 333.1314.

[0125] 2-(2-(3-fluoro-4-hydroxypyrrolidone-1-yl)pyrimidin-5-yl)-1H-indol-6-carboxynitrile (compound 18). Compound 10 (29 mg, 1.14 x 10⁻⁶) -4 mol), trans / racemic-4-fluoro-3-hydroxypyrrolidine•HCl (33 mg, 2.33 x 10⁻⁶ mol) -4 mol, 2 equivalents), K2CO3 (120 mg, 8.68 x 10⁻⁶ ...) -4 mol (7.6 equivalents) and CH3CN (10 mL) were refluxed in N2O. 2(g) Stir for 3 h, cool, and remove CH3CN to obtain a deep orange residue. Dissolve / suspend the residue in CH2Cl2 / MeOH, filter, and wash the precipitate with CH2Cl2. Remove the solvent from the filtrate to obtain an orange residue, which is briefly dried under vacuum (56 mg). Dissolve the residue in CH2Cl2 / MeOH, pour onto dry silica (33 mm h x 33 mm id), and elute under vacuum: % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (100 mL) to give compound 18 (9 mg, 24%) as a light brown solid. 1H NMR (300 MHz, DMSO-d6) δ 12.08 (s, 1H), 8.92 (s, 2 H), 7.82 (s, 1 H), 7.67 (d, 1 H, J = 8.1 Hz), 7.33 (dd, 1 H, J= 8.1 Hz, J = 1.5 Hz), 6.98 (d, 1 H, J = 1.5 Hz), 5.61 (d, 1 H, J = 3.9 Hz), 5.10 (dm, 1 H, 2 J HF = 49.8 Hz), 4.37 (m, 1 H), 3.88 (s, 1 H), 3.77 (m, 1 H), 3.69 (s, 2 H), HRMS (ESI) [MH] + C 17 H 13 Calculated value of FN5O: 322.1099, measured value: 322.1093.

[0126] 6-Fluoro-2-(2-(3-Fluoro-4-hydroxypyrrolidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 19). Compound 7 (56 mg, 1.61 x 10⁻⁶) -4 mol), trans / racemic-4-fluoro-3-hydroxypyrrolidine•HCl (52 mg, 3.67 x 10⁻⁶ mol) -4 mol (2.3 equivalents), K2CO3 (168 mg, 1.22 mmol, 7.5 equivalents) and CH3CN (15 mL) were refluxed in N2O. 2(g) Stir for 3 h, then cool to ambient temperature. Remove CH3CN to obtain a grayish-white solid, dissolve / suspend it in CH2Cl2, filter, and wash the precipitate with CH2Cl2. Remove CH2Cl2 from the filtrate to obtain a yellow paste, dry it under vacuum (89 mg), then dissolve it in CH2Cl2 / MeOH, pour it onto dry silica (45 mm h x 45 mm id) and elute under vacuum: % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (100 mL), 4% (50 mL). Combine the desired fractions and purify by radial chromatography (1 mm silica): % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (25 mL) to give compound 19 (41 mg, 61%) as a grayish-white solid. 1HNMR (300 MHz, CDCl3) δ 8.39 (s, 2 H), 7.91 (dd, 1 H, J = 10.8 Hz, J = 2.4Hz), 7.46 (dd, 1 H, J = 8.7 Hz, J = 5.7 Hz), 7.01 (td, 1 H, J = 8.7 Hz, J =2.4 Hz), 5.11 (dm, 1 H, 2 J HF = 51.0 Hz), 4.59 (dm, 1 H, J = 6.9 Hz), 4.03 (m,1 H), 3.90 (m, 3 H), 1.92 (br d, 1 H, J = 2.7 Hz), 1.50 (s, 9 H).

[0127] 2-(2-(3-fluoro-4-hydroxypyrrolidone-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 20). Compound 11 (79 mg, 2.40 x 10⁻⁶) -4 mol), trans / racemic-4-fluoro-3-hydroxypyrrolidine•HCl (97 mg, 6.85 x 10⁻⁶) -4 mol (2.9 equivalents), K2CO3 (158 mg, 1.14 mmol, 4.7 equivalents) and CH3CN (15 mL) were refluxed in N2O. 2(g) Stir for 5 h, then cool to ambient temperature. Remove CH3CN to obtain a residue, briefly dry it under vacuum, then dissolve it in CH2Cl2, pour it onto dry silica (33 mm h x 33 mm id) and elute under vacuum: CH2Cl2 (25 mL); % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (50 mL) to give compound 20 (85 mg, 89%) as a grayish-white foam: 1 ¹H NMR (300 MHz, CDCl₃) δ 8.41 (s, 2 H), 8.17 (d, 1 H, J = 8.4 Hz), 7.55 (d, 1 H, J = 7.5 Hz), 7.33 (partially resolved ddd, 1 H, J = 7.8 Hz, J = 1.2 Hz), 7.25 (m- partially blocked by CHCl₃ resonance, 1 H), 6.55 (s, 1 H), 5.10 (dm, 1 H, 2 J HF= 50.7 Hz), 4.59 (m, 1 H), 4.03 (m, 1 H), 3.91 (m, 3 H), 1.89 (d, 1 H, J = 3.9 Hz), 1.50 (s, 9 H).

[0128] 4-Fluoro-1-(5-(6-fluoro-1H-indol-2-yl)pyrimidin-2-yl)pyrrolidine-3-ol (compound 21). Compound 19 (27 mg, 6.48 x 10⁻⁶) -5 The aqueous layer was dissolved in TFA (0.75 mL, 9.7 mmol, 150 equivalents) and stirred at ambient temperature for 15 min. Then, it was poured into a mixture of NaHCO3 (0.91 g, 10.83 mmol, 1.1 equivalents of TFA), H2O (25 mL), and CH2Cl2 (25 mL). The mixture was stirred for 5 min, and then the layers were separated. The aqueous layer was extracted with CH2Cl2 (5 mL x 2). The combined CH2Cl2 layers were dried over MgSO4 to remove the solvent, yielding a grayish-white solid, which was dried under vacuum to give compound 21 (17 mg, 83%). 1 H NMR (300 MHz, acetone-d6) δ 10.71 (br s, 1 H), 8.80 (s, 2H), 7.52 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.11 (dd, 1 H, J = 9.9 Hz, J =2.1 Hz), 6.84 (ddd, 1 H, J = 9.9 Hz, J = 8.7 Hz, J = 2.4 Hz), 6.79 (m, 1 H),5.12 (dm, 1 H, 2 J HF = 51.0 Hz), 4.67 (d, 1 H, J = 3.9 Hz), 4.51 (m, 1 H), 3.96 (d, 1 H, J = 2.1 Hz), 3.85 (m, 1 H), 3.79 (m, 2 H); 13 C NMR (125 MHz, DMSO-d6)δ 158.74 (d, 1 J FC = 234.6 Hz), 159.10, 154,57, 136.75 (d, J FC = 12.6 Hz), 134.13 (d, J FC= 3.5 Hz), 125.43, 120.51, 115.14, 107.76 (d, J FC = 24.3 Hz), 97.09 (d, J FC = 25.5 Hz), 97.07, 95.16 (d, J FC = 177.0 Hz), 71.40 (d, J FC =27.2 Hz), 52.55, 50.71 (d, J FC = 21.9 Hz); 19 F NMR (470.6 MHz, DMSO-d6) δ -121.54 (m), -181.85 (m); HRMS (ESI) [MH] + C 16 H 13 Calculated value of F2N4O: 315.1052, measured value: 315.1048.

[0129] 1-(5-(1H-indol-2-yl)pyrimidin-2-yl)-4-fluoropyrrolidine-3-ol (compound 22). Compound 20 (79 mg, 1.98 x 10⁻⁶) -4 A mol) solution was dissolved in CH2Cl2 (5 mL), and HCl / 1,4-dioxane (4 M, 1 mL, 4 mmol) was added. The mixture was stirred for 5 h, then filtered, and the precipitate was washed with EtOEt (2 mL x 3). The solvent was removed from the filtrate to give a pale yellow solid, which was briefly dried under vacuum and then dissolved in CH2Cl2 (10 mL). H2O (10 mL) was added, followed by concentrated NH4OH. (aq)(5 drops), and the mixture was stirred for 5 min (pH 11-12). Brine (5 mL) was added, the layers were mixed and separated, and the CH2Cl2 layer was dried over MgSO4. TLC (3% MeOH / CH2Cl2) analysis showed residual N-Boc protected compound 20. The solvent was removed to obtain a white foam, which was dried under vacuum (50 mg). The foam was dissolved in TFA (2 mL, 26 mmol), stirred for 15 min, and then poured into a mixture of CH2Cl2 (40 mL), H2O (40 mL), and NaHCO3 (2.42 g, 28.81 mmol, 1.1 equivalent TFA). The mixture was stirred for 15 min, and the layers were separated, and the H2O layer was extracted with CH2Cl2 (10 mL). The combined CH2Cl2 layers were washed with brine (25 mL), dried over MgSO4, and the solvent was removed to obtain a grayish-white solid (24 mg). Compound 22 (21 mg, 36%) was purified by rapid column chromatography on silica (3% MeOH / CH2Cl2) to obtain a grayish-white solid. 1 ¹H NMR (400 MHz, DMSO-d⁶) δ 11.47 (s, 1 H), 8.86 (s, 2 H), 7.50 (d, 1 H, J = 8.0 Hz), 7.38 (d, 1 H, J = 8.0 Hz), 7.08 (partially resolved ddd, 1 H, J = 7.2 Hz, J = 1.2 Hz), 6.99 (partially resolved ddd, 1 H, J = 7.2 Hz, J = 1.2 Hz), 6.81 (d 1 H, J = 1.2 Hz), 5.60 (d, 1 H, J = 3.6 Hz), 5.09 (dm, 1 H, 2 J HF = 50.8 Hz), 4.37 (m, 1 H), 3.86 (s, 1 H), 3.79 (m, 1 H), 3.67 (s, 2 H);HRMS (ESI) [MH] + C 16 H 14 Calculated value of FN4O: 297.1152, measured value: 297.1145.

[0130] 5-Bromo-2-(4-methoxypiperidin-1-yl)pyrimidine (compound 23). 5-Bromo-2-chloropyrimidine (0.88 g, 4.55 mmol), 4-methoxypiperidin (0.65 mL, 5.25 mmol, 1.2 equivalents), K₂CO₃ (0.89 g, 6.44 mmol, 1.4 equivalents), and CH₃CN (25 mL) were refluxed in an N₂ atmosphere. 2(g) Stir for 1 h, then cool to ambient temperature, filter, and wash the precipitate with CH3CN. Remove CH3CN to obtain a pale yellow residue, dissolve it in CH2Cl2, pour onto dry silica (55 mm h x 45 mm id) and elute under vacuum: CH2Cl2 (50 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (100 mL), 4% (100 mL) to give compound 23 (1.21 g, 98%) as a white solid. 1 HNMR (300 MHz, CDCl3) δ 8.27 (s, 2 H), 4.19 (m, 2 H), 3.44 (m, 3 H), 3.39 (s, 3 H), 1.92 (m, 2 H), 1.57 (m, 2 H).

[0131] 4-(5-bromopyrimidin-2-yl)morpholine (compound 24). 5-Bromo-2-chloropyrimidin (1.48 g, 7.65 mmol), morpholine (1 mL, 11.6 mmol, 1.5 equivalents), K₂CO₃ (1.30 g, 9.41 mmol, 1.2 equivalents) and CH₃CN (55 mL) were refluxed in N₂O₃. 2(g) The mixture was stirred for 3 h, then cooled to ambient temperature, filtered, and the precipitate was washed with CH3CN. The solvent was removed from the filtrate to give a white solid, which was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (100 mL), 4% (100 mL) to give compound 24 (1.74 g, 93%) as a white solid. 1 H NMR (300 MHz, CDCl3)δ 8.31 (s, 2 H), 3.76 (s, 8 H); 13C NMR (125 MHz, CDCl3) δ 160.15, 158.09,106.39, 66.85, 44.52; HRMS (ESI) [M+H] + C8H 11 Calculated value of BrN3O: 244.0080, measured value: 244.0084.

[0132] (R)-5-bromo-2-(3-fluoropyrrolidine-1-yl)pyrimidine ((R)-compound 25). 5-bromo-2-chloropyrimidine (0.51 g, 2.64 mmol), (R)-3-fluoropyrrolidine•HCl (0.49 g, 3.90 mmol, 1.5 equivalents), K₂CO₃ (1.13 g, 8.18 mmol, 3.1 equivalents), and CH₃CN (30 mL) were refluxed in N₂O₃ solution. 2(g) Stir for 90 min, then cool to ambient temperature, filter, and wash the precipitate with EtOAc 3x. Remove the solvent from the filtrate to obtain a white solid, which is dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: % CH2Cl2 / hexane - 50% (50 mL), 75% (100 mL), CH2Cl2 (200 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (100 mL), 5% (100 mL), 10% (50 mL) to give (R)-compound 25 (0.50 g, 77%) as a grayish-white solid. 1 H NMR (300 MHz, CDCl3) δ 8.32 (s, 2 H), 5.36 (dt, 1 H, 2 J HF = 52.8, J = 3.3Hz), 3.99 – 3.57 (m, 4 H), 2.45 – 1.99 (m, 2 H).

[0133] 1-(5-bromopyrimidin-2-yl)piperidin-4-ol (compound 26). 5-Bromo-2-chloropyrimidin (0.97 g, 5.01 mmol), 4-hydroxy-piperidin (0.56 g, 5.54 mmol, 1.1 equivalents), K₂CO₃ (0.92 g, 6.66 mmol, 1.3 equivalents), and CH₃CN (30 mL) were refluxed in N₂O₃. 2(g)Stir for 1 h, then cool to ambient temperature, filter, and wash the precipitate with CH3CN. Remove CH3CN from the filtrate to obtain a solid, dissolve / suspend it in CH2Cl2, pour it onto dry silica (55 mm h x 45 mm id) and elute under vacuum: % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (200 mL), 5% (150 mL) to give compound 26 (1.27 g, 98%) as a white solid. 1 ¹H NMR (300MHz, CDCl₃) δ 8.27 (s, 2H), 4.33 (dtd, 2H, J = 13.8 Hz, J = 4.8 Hz, J = 0.9Hz), 3.96 (apparent octet, 1H, J = 4.2 Hz), 3.33 (ddd, 2H, J = 13.5 Hz, J = 9.6 Hz, J = 3.3 Hz), 1.94 (m, 2H), 1.53 (m, 3H).

[0134] 1-(5-bromopyrimidin-2-yl)piperidin-4-one (compound 27). Compound 26 (0.54 g, 2.09 mmol) was dissolved in CH2Cl2 (25 mL), followed by the addition of H2O (0.05 mL), and then Dysmartin periodane (1.03 g, 2.43 mmol, 1.2 equivalence). The reaction mixture was stirred under reflux for 7 h, then cooled to ambient temperature and stirred for 17 h, and poured into a mixture of CH2Cl2 (25 mL), H2O (50 mL), Na2S2O3 • 5 H2O (2.17 g, 8.74 mmol), and NaHCO3 (2.25 g, 26.78 mmol). The mixture was stirred for 2 h, and then the layers were separated, and the H2O layer was extracted with CH2Cl2 (10 mL x 2). The combined CH2Cl2 layers were washed with brine (25 mL) and dried over MgSO4. The solution was concentrated, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (200 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (200 mL) to give a grayish-white solid (0.55 g). Purification was performed by radial chromatography (4 mm silica): % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 4% (50 mL), 5% (100 mL) to give the recovered white solid compounds 26 (0.11 g, 20%) and 27 (0.38 g, 71%). 1 H NMR (300MHz, CDCl3) δ 8.35 (s, 2 H), 4.10 (t, 4 H, J = 6.3 Hz), 2.50 (t, 4 H, J = 6.3Hz).

[0135] 5-Bromo-2-(piperidin-1-yl)pyridine (compound 28). 2,5-Dibromopyridine (0.87 g, 3.67 mmol) and piperidine (5 mL, 50.6 mmol, 13.8 equivalents) were mixed in an N2 atmosphere. 2(g)The mixture was stirred in a hot sand bath (105ºC) for 15 h and then cooled to ambient temperature. EtOAc (5 mL) was added, the mixture was filtered, and the precipitate was washed with EtOAc (5 mL x 2). The filtrate was concentrated to an oily / residue mixture, then dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane (50 mL), hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL) to give compound 28 (0.86 g, 97%) as a colorless oily substance. 1 H NMR (300 MHz, CDCl3) δ8.16 (d, 1 H, J = 2.4 Hz), 7.48 (dd, 1 H, J = 9.3 Hz, J = 2.4 Hz), 6.54 (d, 1H, J = 9.3 Hz), 3.50 (m, 4 H), 1.63 (m, 6 H).

[0136] 4-(5-bromopyridin-2-yl)morpholine (compound 29). 2,5-Dibromopyridine (0.87 g, 3.67 mmol) and morpholine (5 mL, 58 mmol, 16 equivalents) were mixed in an N2 atmosphere. 2(g) The mixture was stirred in a hot sand bath (approximately 110ºC) for 15 h, then cooled to ambient temperature. EtOAc (5 mL) was added, the mixture was filtered, and the precipitate was washed with EtOAc (5 mL x 2). The filtrate was concentrated, diluted with hexane, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (200 mL) to give compound 29 (0.86 g, 96%) as a grayish-white solid. 1 H NMR (300 MHz, CDCl3) δ 8.21 (d, 1 H, J = 2.4 Hz), 7.56 (dd, 1 H, J = 9.0 Hz, J = 2.4 Hz), 6.53 (d, 1 H, J = 9.0 Hz), 3.81 (t, 4 H, J= 4.8 Hz), 3.47 (t, 4 H, J = 4.8 Hz).

[0137] 4-(5-bromopyrazin-2-yl)morpholine (compound 30). At 0ºC in N 2(g)Morpholine (0.3 mL, 3.5 mmol) was added to a suspension of NaH (87 mg, 90%, 3.26 mmol) in THF (6 mL), and the mixture was stirred for 15 min. Then, 2,5-dibromopyrazine (0.81 g, 3.41 mmol) in THF (6 mL) was added, and the reaction mixture was stirred under reflux overnight. The solvent was evaporated, and the residue was dissolved in EtOAc (20 mL), washed with H2O (10 mL), and dried over MgSO4. The solvent was removed, and the residue was purified by rapid column chromatography (4:1 v / v hexane / EtOAc) to give compound 30 (0.86 g, quantified) as a white solid. 1 ¹H NMR (CDCl₃, 500 MHz): δ 8.16 (d, ¹H, J = 2.0 Hz), 7.86 (d, ¹H, J = 2.5 Hz), 3.83 (apparent t, ⁴H, J = 8.0 Hz), 3.53 (apparent t, ⁴H, J = 8.0 Hz). HRMS (ESI) [M+H] + C8H 11 Calculated value of ON3Br: 244.0080, measured value: 244.0076. 6-Fluoro-2-(2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 31). Compound 4 (0.65 g, 1.63 mmol), compound 23 (0.49 g, 1.80 mmol, 1.1 equivalents), and Pd(dppf)Cl2 (0.11 g, 1.50 x 10⁻⁶ mmol) were added. -4 0.56 g (0.56 g, 5.28 mmol, 3.2 equivalents) and Na2CO3 (0.56 g, 5.28 mmol, 3.2 equivalents) were treated with N 2(g) Rinse for 10 min, then add 1,4-dioxane (45 mL). Recirculate the reaction mixture under reflux in N2O. 2(g)The mixture was stirred for 15 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with 1,4-dioxane. The filtrate was concentrated into a brown slurry, dried under vacuum, and then dissolved in CH2Cl2. The slurry was poured onto dry silica (55 mm h x 45 mm i.d.) and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (150 mL) to give a brown residue (0.65 g). The residue was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane (25 mL); hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (100 mL), 50 : 45 : 5 (100 mL), 20 : 75 : 5 (150 mL) to give a deep orange solid (0.52 g). Purification was performed by radial chromatography (2 mm silica): CH2Cl2 (100 mL); 1% MeOH / CH2Cl2 (50 mL) to give crude compound 31 (0.30 g, light brown paste) and crude compound 36 (91 mg, light brown residue - further purified below).

[0138] The crude compound 31 was purified by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 95 : 4 : 1 (100 mL), 90 : 8 : 2 (25 mL) to obtain a viscous, slightly yellow residue, which was further purified by radial chromatography (2 mm silica): CHCl3 (50 mL); %EtOH / CHCl3 - 1% (50 mL), 2% (25 mL) to obtain compound 31 (0.26 g, 37%) as a slightly yellow, viscous paste that slowly solidified. 1 H NMR (300 MHz, CDCl3) δ 8.34(s, 2 H), 7.93 (dd, 1 H, J = 10.8 Hz, J = 2.4 Hz), 7.45 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.01 (td, 1 H, J = 8.7 Hz, J = 2.4 Hz), 6.49 (s, 1 H), 4.34 (m,2 H), 3.48 (m, 3 H), 3.41 (s, 3 H), 1.96 (m, 2 H), 1.59 (m, 2 H), 1.47 (s, 9H); HRMS (ESI) [M+H] +C 23 H 28 Calculated value of FN4O3: 427.2140, measured value: 427.2131.

[0139] (R)-6-fluoro-2-(2-(3-fluoropyrrolidone-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester ((R)-compound 32). Compound 4 (0.40 g, 1.00 mmol), (R)-compound 25 (0.30 g, 1.22 mmol, 1.2 equivalents), and Pd(dppf)Cl2 (76 mg, 1.04 x 10⁻⁶) were added. -4 mol (0.1 equivalent), Na₂CO₃ (0.17 g, 1.60 mmol, 1.6 equivalent) and 1,4-dioxane (25 mL) were refluxed in N₂CO₃. 2(g) The mixture was stirred for 6 h and then cooled to ambient temperature. The reaction mixture was filtered through diatomaceous earth, which was then washed with EtOAC. The filtrate was concentrated into a brown slurry, which was then removed by adding CH2Cl2 and hexane to obtain a brown residue, which was dried under vacuum. The residue was dissolved in CH2Cl2 (+ a few drops of MeOH), poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (200 mL) to obtain a deep orange / brown residue (0.37 g). The residue was dissolved in CH2Cl2 / MeOH and purified by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL) to give a pale yellow solid (0.17 g), which was further purified by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL) to give a pale yellow solid (0.14 g). A third purification was performed by radial chromatography (2 mm silica): CHCl3 (150 mL); 1% EtOH / CHCl3 (50 mL) to give (R)-compound 32 (0.13 g, 32%) as a grayish-white solid. 1H NMR (400 MHz, CDCl3) δ 8.39 (s, 2 H), 7.92 (dd, 1 H, J = 10.6 Hz, J = 2.4 Hz), 7.46 (dd, 1 H, J = 8.8 Hz, J = 5.4 Hz), 7.01 (td, 1 H, J = 8.8 Hz, J = 2.4Hz), 6.51 (s, 1 H), 5.40 (dt, 1 H, 2 J HF = 52.8 Hz, J = 3.2 Hz), 4.03 (ddd, 1H, 3 J HF = 25.2 Hz, J = 13.6 Hz, J = 1.6 Hz), 3.93 (t, 1 H, J = 10.0 Hz), 3.81 (dd, 0.5 H, J = 13.6 Hz, J = 3.6 Hz), 3.76 – 3.69 (m - overlap resonance, 1.5 H), 2.46– 2.37 (m, 1 H), 2.26 – 2.06 (m, 1 H), 1.49 (s, 9 H); HRMS (ESI) [M+H] + C 21 H 23 Calculated value of O2N4F2: 401.1784, measured value: 401.1774.

[0140] 6-Fluoro-2-(2-(4-oxopiperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 33). 1,4-Dioxane was treated with N... 2(g) Bubbling for 30 min. Compound 4 (0.24 g, 6.03 x 10⁻⁶) -4 mol), compound 27 (0.18 g, 7.03 x 10⁻⁶ ... -4 mol, 1.2 equivalents), Pd(dppf)Cl2 (46.5 mg, 6.36 x 10⁻⁶ mol, 1.2 equivalents), Pd(dppf)Cl� ... -5 0.1 mol (0.1 equivalent) and Na₂CO₃ (0.14 g, 1.32 mmol, 2.2 equivalent) were treated with N₂CO₃. 2(g) Rinse for 10 min, then add 1,4-dioxane (25 mL), and reflux the mixture under N2O conditions. 2(g)Stir for 5 h, then cool to ambient temperature. Filter the mixture through diatomaceous earth and wash the diatomaceous earth with EtOAc. Concentrate the filtrate into a deep orange oil, then add CH2Cl2 and hexane and remove it to obtain a deep orange residue, which is dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL), 50 : 45 : 5 (200 mL), 20:75:5 (100 mL) to give a pale orange solid (0.134 g). Purification by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL) yielded a grayish-white solid (79 mg), which was further purified by radial chromatography (2 mm silica): % MeOH / CH2Cl2 -1% (50 mL), 2% (50 mL) to give compound 33 (70 mg, 28%) as a grayish-white solid. 1 H NMR (300 MHz, CDCl3) δ 8.42 (s, 2 H), 7.93 (dd, 1 H, J = 10.8 Hz, J = 2.4 Hz), 7.47 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.02 (td, 1 H, J = 8.7 Hz, J = 2.4Hz), 6.53 (s, 1 H), 4.20 (t, 4 H, J = 6.3 Hz), 2.54 (t, 4 H, J = 6.3 Hz), 1.49 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 24 Calculated value of O3N4F: 411.1827, measured value: 411.1815.

[0141] 6-Fluoro-2-(6-(piperidin-1-yl)pyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 34). Add 1,4-dioxane (30 mL) to N... 2(g)Purge for 30 min. Compound 4 (0.44 g, 1.11 mmol), compound 28 (0.29 g, 1.20 mmol, 1.1 equivalents), Pd(dppf)Cl2 (0.10 g, 0.14 mmol, 0.1 equivalents), and Na2CO3 (0.54 g, 5.09 mmol, 4.6 equivalents) were then treated with N2O2. 2(g) Rinse for 10 min, then add 1,4-dioxane. Recycle the reaction mixture under reflux in N2O. 2(g) The mixture was stirred for 5 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with EtOAc. The solvent was removed from the filtrate to obtain a brown oily substance, which was then treated with CH2Cl2 and hexane to obtain a brown residue, which was dried under vacuum. The residue was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: % CH2Cl2 / hexane - 50% (100 mL), 75% (100 mL), CH2Cl2 (200 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (100 mL) to obtain recovered compound 4 (0.22 g crude, subsequently purified to 94 mg, 21% recovery) and brown residue (0.31 g). The brown residue was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (200 mL), 5% (200 mL) to give a brown residue (0.27 g), which was purified by radial chromatography (2 mm silica): CHCl3 (100 mL) to give a light brown solid (0.12 g). Purification by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75:20:5 (25 mL) gave compound 34 (89 mg, 20%) as a grayish-white solid. 1H NMR (300 MHz, CDCl3) δ8.22 (d, 1 H, J = 2.4 Hz), 7.92 (dd, 1 H, J = 10.8 Hz, J = 2.4 Hz), 7.45 (m,2 H), 6.99 (td, 1 H, J = 8.7 Hz, 2.4 Hz), 6.67 (d, 1 H, J = 8.7 Hz), 6.46 (s,1 H), 3.59 (m, 4 H), 1.67 (m, 6 H), 1.43 (s, 9 H); HRMS (ESI) [M+H] + C 23 H 27 Calculated value of FN3O2: 396.2082, measured value: 396.2081.

[0142] 6-Fluoro-2-(6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 35). Compound 4 (0.41 g, 1.03 mmol), compound 29 (0.26 g, 1.07 mmol, 1 equivalent), Pd(dppf)Cl2 (81 mg, 0.11 mmol, 0.1 equivalent), Na2CO3 (0.32 g, 3.02 mmol, 2.9 equivalent) and 1,4-dioxane (25 mL) were refluxed in N2O. 2(g) The mixture was stirred for 6 h and then cooled to ambient temperature. The reaction mixture was filtered through diatomaceous earth, which was then washed with EtOAC. The filtrate was concentrated into a brown oil, which was then removed by adding CH2Cl2 and hexane to obtain a brown residue, which was dried under vacuum. The residue was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum in the following ratios: hexane / EtOAc / NEt3 v / v / v 90:8:2 (125 mL), 75:20:5 (100 mL), 50:45:(150 mL), 20:75:5 (200 mL) to give a yellow paste (0.14 g). Purification was performed by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (125 mL), 75 : 20 : 5 (100 mL) to give compound 35 (66 mg, 16%) as a pale yellow foam. 1H NMR (400 MHz, CDCl3) δ 8.26(d, 1 H, J = 2.0 Hz), 7.91 (dd, 1 H, J = 10.6 Hz, J = 2.0 Hz), 7.54 (dd, 1 H,J = 8.8 Hz, J = 2.4 Hz), 7.45 (dd, 1 H, J = 8.8 Hz, J = 5.4 Hz), 7.00 (td, 1H, J = 8.8 Hz, J = 2.4 Hz), 6.67 (d, 1 H, J = 8.8 Hz), 6.48 (s, 1 H), 3.85(t, 4 H, J = 4.8 Hz), 3.56 (t, 4 H, J = 4.8 Hz), 1.44 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of FN3O3: 398.1875, measured value: 398.1866.

[0143] 6-Fluoro-2-(2-(4-methoxypiperidin-1-yl)pyrimidin-5-yl)-1H-indole (compound 36). Crude compound 36 (91 mg – from the coupling reaction of compound 31 above) was purified by radial chromatography (1 mm silica): CH2Cl2 (50 mL); % MeOH / CH2Cl2 – 1% (50 mL), 2% (75 mL) to give compound 36 (46 mg, 9%) as a brownish-brown solid. 1 H NMR (300 MHz, CDCl3) δ 8.57 (s, 2 H), 8.23 ​​(br s, 1 H), 7.50 (dd, 1H, J = 8.7 Hz, J = 5.4 Hz), 7.08 (dd, 1 H, J = 9.6 Hz, J = 2.1 Hz), 6.89(ddd, 1 H, J = 9.7 Hz, J = 8.7 Hz, J = 2.4 Hz), 6.63 (d, 1 H, J = 1.5 Hz), 4.31 (m, 2 H), 3.50 (m, 3 H), 3.41 (s, 3 H), 1.97 (m, 2 H), 1.62 (m, 2 H); 13 CNMR (125 MHz, DMSO-d6) δ 160.04, 158.74 (d, 1 J FC= 234.4 Hz), 154.52, 136.70(d, J FC = 12.7 Hz), 134.06 (d, J FC = 3.5 Hz), 125.42, 120.51 (d, J FC = 10.2Hz), 114.81, 107.76 (d, J FC = 24.4 Hz), 97.08 (d, J FC = 25.7 Hz), 97.05,75.49, 54.97, 40.97, 30.20; 19 F NMR (470.6 MHz, DMSO-d6) δ -121.53 (m).

[0144] N-Boc deprotection of compound 31 to compound 36: Compound 31 (64.5 mg, 1.51 x 10⁻⁶) -4 The mixture of NaHCO3 (1.84 g, 21.90 mmol, 1.1 TFA equivalents) and TFA (1.5 mL, 19.5 mmol, 129 equivalents) was stirred at ambient temperature for 15 min, then poured into a mixture of NaHCO3 (1.84 g, 21.90 mmol, 1.1 TFA equivalents), H2O (45 mL), and CH2Cl2 (45 mL). The mixture was stirred, and the layers were separated. The H2O layer was extracted with CH2Cl2 (10 mL). The combined CH2Cl2 layers were washed with brine (20 mL) and dried over MgSO4. The solution was concentrated, poured onto dry silica (33 mm h x 33 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (50 mL), 75 : 20 : 5 (50 mL), 50 : 45 : 5 (100 mL) to give compound 36 (30 mg, 61%) as a pale yellow solid. 1¹H NMR (300 MHz, CDCl₃) δ 8.57 (s, 2 H), 8.24 (br s, 1 H), 7.49 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.07 (dd, 1 H, J = 9.3 Hz, J = 2.1 Hz), 6.89 (ddd, 1 H, J = 9.6 Hz, J = 8.7 Hz, J = 2.4 Hz), 6.62 (partially resolved dd, 1 H, J = 2.1 Hz, J = 0.6 Hz), 4.31 (m, 2 H), 3.50 (m, 3 H), 3.41 (s, 3 H), 1.97 (m, 2 H), 1.62 (m, 2 H); HRMS (ESI) [M+H] + C 18 H 20 Calculated value of FN4O: 327.1616, measured value: 327.1620.

[0145] (R)-6-fluoro-2-(2-(3-fluoropyrrolidone-1-yl)pyrimidin-5-yl)-1H-indole ((R)-compound 37). (R)-compound 32 (0.12 g, 3.00 x 10⁻⁶ g) -4 The aqueous layer was dissolved in TFA (3 mL, 38.9 mmol, 130 equivalents) and stirred at ambient temperature for 20 min. The solution was then poured into a mixture of NaHCO3 (3.95 g, 47.02 mmol, 1.2 equivalents of TFA), H2O (75 mL), and CH2Cl2 (75 mL). The mixture was stirred for 20 min, and the layers were separated. The aqueous layer was extracted with CH2Cl2 (25 mL x 2). The combined CH2Cl2 layers were washed with brine (25 mL), dried over MgSO4, and concentrated (forming a precipitate). MeOH was added to redissolve the precipitate, and the solution was poured onto dry silica (55 mm h x 45 mm id) and eluted under vacuum: % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (200 mL), 10% (50 mL) to give a yellow solid (85 mg). The solid was placed in a medium-grain glass filter, washed with 1:1 v / v CH2Cl2 / hexane (1 mL x 7), and dried under vacuum to give (R)-compound 37 (67 mg, 74%) as a pale yellow solid. 1HNMR (300 MHz, acetone-d6) δ 10.71 (br s, 1 H), 8.80 (s, 2 H), 7.52 (dd, 1 H, J =8.4 Hz, J = 5.4 Hz), 7.10 (dd, 1 H, J = 9.9 Hz, J = 2.1 Hz), 6.84 (ddd, 1 H,J = 9.9 Hz, J = 8.7 Hz, J = 2.4 Hz), 6.79 (s, 1 H), 5.45 (dm, 1 H, 2 J HF = 52.8Hz), 4.01 – 3.58 (m, 4 H), 2.37 – 2.12 (m, 2 H); HRMS (ESI) [M+H] + C 16 H 15 Calculated value of N4F2: 301.1259, measured value: 301.1253.

[0146] 1-(5-(6-fluoro-1H-indol-2-yl)pyrimidin-2-yl)piperidin-4-one (compound 38). Compound 33 (65 mg, 1.58 x 10⁻⁶) -4 The aqueous layer was dissolved in TFA (1.6 mL, 20.8 mmol, 131 equivalents) and stirred at ambient temperature for 20 min. Then, it was poured into a mixture of NaHCO3 (2.13 g, 25.36 mmol, 1.2 equivalents of TFA), H2O (40 mL), and CH2Cl2 (40 mL). The mixture was stirred for 10 min, and the layers were separated. The aqueous layer was extracted with CH2Cl2 (15 mL x 2). The combined CH2Cl2 layers were washed with brine (25 mL), dried over MgSO4, concentrated, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (200 mL), 10% (50 mL) to give a grayish-white solid (43 mg). Purification by radial chromatography (1 mm silica): % MeOH / CH2Cl2- 1% (50 mL), 2.5% (25 mL) yielded compound 38 (38 mg, 77%) as a light brown solid. 1H NMR (300MHz, CDCl3) δ 8.65 (s, 2 H), 8.27 (br s, 1 H), 7.52 (dd, 1 H, J = 8.7 Hz, J =5.4 Hz), 7.10 (dd, 1 H, J = 9.3 Hz, J = 2.1 Hz), 6.91 (ddd, 1 H, HRMS (ESI) [M+H] + C 17 H 16 Calculated value of FN4O: 311.1303, measured value: 311.1296. X-ray quality crystals were grown by slow evaporation of acetone.

[0147] 6-Fluoro-2-(6-(piperidin-1-yl)pyridin-3-yl)-1H-indole (compound 39). Compound 34 (82 mg, 2.07 x 10⁻⁶) -4 The aqueous layer was dissolved in TFA (2 mL, 26 mmol, 126 equivalents) and stirred at ambient temperature for 20 min. The solution was then poured into a mixture of Na2CO3 (2.43 g, 28.93 mmol, 1.1 equivalents of TFA), H2O (60 mL), and CH2Cl2 (60 mL). The mixture was stirred for 10 min, and the layers were separated. The aqueous layer was extracted with CH2Cl2 (10 mL x 3). The combined CH2Cl2 layers were washed with brine (30 mL), dried over MgSO4, and then poured onto dry silica (33 mm h x 33 mm id) and eluted under vacuum: % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (50 mL) to give a grayish-white solid (58 mg). Purification by radial chromatography (1 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (40 mL), 75 : 20 : 5 (50 mL) yielded compound 39 (50 mg, 82%) as a grayish-white solid. 1H NMR (300MHz, CDCl3) δ 8.46 (d, 1 H, J = 2.4 Hz), 8.28 (br s, 1 H), 7.68 (dd, 1 H, J =9.0 Hz, J = 2.4 Hz), 7.48 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.06 (dd, 1 H, J= 9.6 Hz, J = 1.8 Hz), 6.86 (ddd, 1 H, J = 9.6 Hz, J = 8.7 Hz, J = 2.1 Hz), 6.70 (d, 1 H, J = 8.7 Hz), 6.61 (d, 1 H, J = 1.5 Hz), 3.59 (m, 4 H), 1.67 (m, 6 H); 13 C NMR (125 MHz, DMSO-d6) δ 158.57 (d, 1 J FC = 233.9 Hz), 157.98, 144.42,136.86 (d, J FC = 3.5 Hz), 136.66 (d, J FC = 12.6 Hz), 134.13, 125.61, 120.25(d, J FC = 9.9 Hz), 116.64, 107.49 (d, J FC = 24.0 Hz), 106.98, 96.95 (d, J FC =25.5 Hz), 96.51, 45.50, 24.99, 24.29; 19 F NMR (470.6 MHz, DMSO-d6) δ -122.04(m); HRMS (ESI) [M+H] + C 18 H 19 Calculated value of FN3: 296.1558, measured value: 296.1568.

[0148] 4-(5-(6-fluoro-1H-indol-2-yl)pyridin-2-yl)morpholine (compound 40). Compound 35 (61 mg, 1.53 x 10⁻⁶) was used. -4The aqueous layer was dissolved in TFA (1.5 mL, 19.5 mmol, 127 equivalents) and stirred at ambient temperature for 20 min. Then, it was poured into a mixture of Na2CO3 (1.82 g, 21.67 mmol, 1.1 equivalents of TFA), H2O (40 mL), and CH2Cl2 (40 mL). The mixture was stirred for 15 min, and then the layers were separated. The aqueous layer was extracted with CH2Cl2 (15 mL x 2). The combined CH2Cl2 layers were washed with brine (25 mL), dried over MgSO4, poured onto dry silica (33 mm h x 33 mm i.d.), and eluted under vacuum: % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (50 mL) to give compound 40 (42 mg, 92%) as a light brown solid. 1 H NMR (300 MHz, CDCl3) δ 8.50 (d, 1 H, J =2.1 Hz), 8.28 (br s, 1 H), 7.75 (dd, 1 H, J = 8.7 Hz, J = 2.4 Hz), 7.49 (dd,1 H, J = 8.7 Hz, J = 5.4 Hz), 7.07 (dd, 1 H, J = 9.3 Hz, J = 2.1 Hz), 6.88 (ddd, 1 H, J = 9.6 Hz, J = 8.7 Hz, J = 2.1 Hz), 6.71 (d, 1 H, J = 8.7 Hz), 6.65 (dd, 1 H, J = 2.1 Hz, J = 0.6 Hz), 3.85 (t, 4 H, J = 4.8 Hz), 3.57 (t, 4H, J = 4.8 Hz); 13 C NMR (125 MHz, DMSO-d6) δ 158.66 (d, 1 J FC = 234.3 Hz),158.19, 144.28, 136.72 (d, J FC = 12.7 Hz), 136.54 (d, J FC = 3.4 Hz), 134.19,125.55, 120.41 (d, J FC = 10.1 Hz), 117.95, 107.60 (d, J FC = 24.3 Hz), 106.92,97.00 (d, J FC= 25.5 Hz), 96.99, 65.91, 45.04; 19 F NMR (470.6 MHz, DMSO-d6) δ -121.79; HRMS (ESI) [M+H] + C 17 H 17 Calculated value of FN3O: 298.1350, measured value: 298.1345.

[0149] 1-(5-(5-fluorobenzofuran-2-yl)pyrimidin-2-yl)piperidin-4-ol (compound 43). Combine 1,4-dioxane (30 mL) and H₂O (6 mL), and use N₂O. 2(g) Purge for 45 min. Add 5-fluorobenzofuran-2-boronic acid (compound 41) (0.25 g, 1.39 mmol), compound 26 (0.42 g, 1.63 mmol, 1.2 equivalents), and Pd(dppf)Cl2 • CH2Cl2 (97 mg, 1.19 x 10⁻⁶ mmol / L). -4 0.71 g (6.70 mmol, 4.8 equivalents) and Na2CO3 (0.71 g, 6.70 mmol, 4.8 equivalents) were treated with N 2(g) Rinse for 10 min, then add the H2O / 1,4-dioxane mixture (17 mL). Transfer the reaction mixture to an N2O container. 2(g) The mixture was stirred in a hot sand bath (approximately 95ºC) for 14 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with EtOAc. The filtrate was concentrated, and EtOAc was added and removed to obtain a brown residue, which was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (100 mL), 50 : 45 : 5 (150 mL), 20 : 75 : 5 (250 mL), EtOAc (50 mL). The desired fractions were combined and purified by radial chromatography (2 mm silica): % EtOH / CHCl3 - 1% (50 mL), 2% (75 mL), 3% (100 mL) to give a brown solid (77 mg). The solid was suspended in CHCl3 (2 mL), filtered, washed with CHCl3 (1.5 mL x 2), and dried under vacuum to give compound 43 (47 mg, 11%) as a white solid. 1¹H NMR (300 MHz, DMSO-d⁶) δ 8.85 (s, 2 H), 7.60 (dd, 1 H, J = 9.0 Hz, J = 4.2 Hz), 7.43 (dd, 1 H, J = 9.0 Hz, J = 2.4 Hz), 7.26 (s, 1 H), 7.10 (td, 1 H, J = 9.0 Hz, J = 2.4 Hz), 4.77 (d, 1 H, J = 4.2 Hz), 4.30 (partially resolved dddd, 2 H, J = 13.5 Hz, J = 4.5 Hz), 3.77 (apparent octet, 1 H, J = 4.2 Hz), 3.39 (ddd - partially blocked by H₂O resonance, 2 H, J = 9.6 Hz, J = 3.3 Hz), 1.80 (m, 2 H), 1.36 (m, 2 H); HRMS (ESI) [M+H] + C 17 H 17 Calculated value of FN3O2: 314.1299, measured value: 314.1296.

[0150] 2-(2-(4-hydroxypiperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 44). Combine 1,4-dioxane (10 mL) and H₂O (2 mL), and use N₂O. 2(g) Purge for 10 min. Compound 26 (0.24 g, 9.30 x 10⁻⁶ mcg) was then placed in the purge tank. -4 mol), Boc-indole-2-boronic acid (compound 6) (0.26 g, 9.96 x 10⁻⁶ ... -4 mol, 1.1 equivalent), Pd(dppf)Cl2 (94 mg, 1.15 x 10⁻⁶ mol), -4 0.45 g (0.45 g, 4.25 mmol, 4.6 equivalents) and Na2CO3 (0.45 g, 4.25 mmol, 4.6 equivalents) were treated with N 2(g) Rinse for 15 min, then add the H2O / 1,4-dioxane mixture. Place the reaction mixture in an N2O atmosphere. 2(g)The mixture was stirred in a hot sand bath (85ºC) for 16 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with EtOAc. The solvent was removed to obtain a black slurry, which was dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (100 mL). The desired fractions were combined, concentrated, poured onto dry silica (45 mm h x 45 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90:8:2 (50 mL), 75:20:5 (100 mL), 50:45:5 (100 mL), 20:75:5 (200 mL) to give orange / brown foam (0.25 g). Purified by radial chromatography (2 mm silica): CHCl3 (50 mL); % EtOH / CHCl3 - 1% (50 mL), 2% (75 mL), 3% (50 mL) to give compound 44 (0.23 g, 63%) as pale yellow foam. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.36 (s, 2 H), 8.19 (d, 1 H, J = 8.4 Hz), 7.54 (d, 1 H, J = 6.0 Hz), 7.32 (apparent t, 1 H, J = 7.6 Hz), 7.25 (apparent t - partially blocked by CHCl₃ resonance, 1 H, J = 7.6 Hz), 6.53 (s, 1 H), 4.46 (partially resolved dddd, 2 H, J = 13.6 Hz, J = 4.4 Hz), 3.99 (br s, 1 H), 3.40 (partially resolved ddd, 2 H, J = 11.4 Hz, J = 2.8 Hz), 1.98 (m, 2 H), 1.57 (m, 2 H) 1.48 (s, 10 H).

[0151] 5-Fluoro-2-(2-(4-hydroxypiperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 45). Combine 1,4-dioxane (30 mL) and H₂O (6 mL), and use N₂O. 2(g) Purge for 45 min. Add N-Boc-5-fluoroindole-2-boronic acid (compound 42) (0.25 g, 8.96 x 10⁻⁶ g) to the solution. -4mol), compound 26 (0.25 g, 9.69 x 10⁻⁶ ...). -4 mol, 1.1 equivalent), Pd(dppf)Cl2 • CH2Cl2 (79 mg, 9.67 x 10⁻⁶ ... -5 0.47 g (0.43 mmol, 0.1 equivalent) and Na2CO3 (0.47 g, 4.43 mmol, 5 equivalent) were treated with N 2(g) Rinse for 10 min, then add a mixture of H₂O / 1,4-dioxane (16 mL). Transfer the reaction mixture to an N₂O container. 2(g) The mixture was stirred in a hot sand bath (approximately 95ºC) for 14 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with EtOAc. The filtrate was concentrated into an oil, then EtOAc was added and removed to obtain a brown residue, which was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (100 mL), 50 : 45 : 5 (150 mL), 20 : 75 : 5 (200 mL) to give a brown foam (0.23 g). Purification was performed by radial chromatography (2 mm silica): % EtOH / CHCl3 - 1% (50 mL), 2% (75 mL) to give compound 45 (0.17 g, 46%) as a grayish-white foam. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.35 (s, 2 H), 8.14 (dd, 1 H, J = 8.8 Hz, J = 4.8 Hz), 7.19 (dd, 1 H, J = 8.8 Hz, J = 2.4 Hz), 7.04 (td, 1 H, J = 9.2 Hz, J = 2.4 Hz), 6.49 (s, 1 H), 4.46 (partially resolved dddd, 2 H, J = 13.6 Hz, J = 4.4 Hz), 4.00 (apparent octet, 1 H, J = 4.4 Hz), 3.40 (ddd, 1 H, J = 10.0 Hz, J = 3.2 Hz), 1.98 (m, 2 H), 1.56 (m, 3 H) 1.47 (s, 9 H).

[0152] 1-(5-(1H-indol-2-yl)pyrimidin-2-yl)piperidin-4-ol (compound 46). Compound 44 (83 mg, 2.10 x 10⁻⁶) -4The aqueous layer was dissolved in TFA (2 mL, 26 mmol, 124 equivalents) and stirred at ambient temperature for 20 min. The solution was then poured into a mixture of NaHCO3 (2.40 g, 28.57 mmol, 1.1 equivalents of TFA), H2O (60 mL), and CH2Cl2 (60 mL). The mixture was stirred until gas escape ceased, then the layers were separated, and the aqueous layer was extracted with CH2Cl2 (10 mL). The combined CH2Cl2 layers were washed with brine (25 mL), dried over MgSO4, and concentrated to produce a precipitate. MeOH (a few drops) was added until the precipitate dissolved, and the solution was poured onto dry silica (55 mm h x 45 mm id) and eluted under vacuum: %MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (150 mL), 10% (50 mL) to give a grayish-white solid. The solid was dissolved / suspended in CHCl3 (approximately 4 mL), and then MeOH (approximately 0.5 mL) was added to dissolve the remaining solid. The solution was purified by radial chromatography (2 mm silica): CHCl3 (25 mL); % EtOH / CHCl3 - 1% (50 mL), 2% (75 mL), 3% (50 mL), 4% (50 mL), 10% (60 mL) to give compound 46 (58 mg, 94%) as a grayish-white solid. 1 ¹H NMR (400 MHz, acetone-d6) δ 10.59 (br s, 1 H), 8.80 (s, 2 H), 7.54 (d, 1 H, J = 7.6 Hz), 7.37 (d, 1 H, J = 8.4 Hz), 7.08 (td, 1 H, J = 7.6 Hz, J = 1.2 Hz), 7.01 (td, 1 H, J = 7.6 Hz, J = 1.2 Hz), 6.78 (d, 1 H, J = 1.2 Hz), 4.42 (partially resolved dddd, 2 H, J = 13.6 Hz, J = 4.4 Hz), 3.91 (apparent octet, 1 H, J = 4.4 Hz), 3.84 (d, 1 H, J = 4.4 Hz). Hz), 3.42 (ddd, 2 H, J = 13.4 Hz, J = 9.8 Hz,J = 3.2 Hz), 1.90 (m, 2 H), 1.48 (m, 2 H); HRMS (ESI) [M+H] + C 17 H 19Calculated value of N4O: 295.1553, measured value: 295.1553.

[0153] 1-(5-(5-fluoro-1H-indol-2-yl)pyrimidin-2-yl)piperidin-4-ol (compound 47). Compound 45 (90 mg, 2.18 x 10⁻⁶) -4 The aqueous layer was dissolved in TFA (2 mL, 26 mmol, 119 equivalents) and stirred at ambient temperature for 20 min. The solution was then poured into a mixture of NaHCO3 (2.45 g, 29.16 mmol, 1.1 equivalents of TFA), H2O (60 mL), and CH2Cl2 (60 mL). The mixture was stirred for 20 min, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (10 mL). The combined CH2Cl2 layers were washed with brine (25 mL) and dried over MgSO4. The solution was concentrated, poured onto dry silica (55 mm h x 45 mm i.d.), and eluted under vacuum: CH2Cl2 (50 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (100 mL), 10% (100 mL) to give a pale yellow solid (54 mg). Purification was performed by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (100 mL), 50 : 45 : 5 (100 mL), 20 : 75 : 5 (250 mL) to give compound 47 (47 mg, 69%) as a light brown solid. 1¹H NMR (300 MHz, acetone-d6) δ 10.67 (br s, 1 H), 8.79 (s, 2 H), 7.36 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.23 (dd, 1 H, J = 9.6 Hz, J = 2.7 Hz), 6.87 (ddd, 1 H, J = 9.6 Hz, J = 8.7 Hz, J = 2.7 Hz), 6.77 (m, 1 H), 4.41 (partially resolved dddd, 2 H, J = 13.5 Hz, J = 4.5 Hz), 3.92 (apparent octet, 1 H, J = 4.2 Hz), 3.83 (d, 1 H, J = 4.2 Hz), 3.43 (ddd, 2 H, J = 4.2 Hz), 2 H, J = 5.4 Hz. H, J = 13.2 Hz, J = 9.6 Hz, J = 3.3 Hz), 1.90 (m, 2 H), 1.48 (dddd, 2H, J = 13.2 Hz, J = 9.0 Hz, J = 4.2 Hz); HRMS (ESI) [M+H] + C 17 H 18 Calculated value of FN4O: 313.1459, measured value: 313.1456.

[0154] 2-Fluoro-6-(piperidin-1-yl)pyridine (compound 48). 2,6-Difluoropyridine (1.34 g, 11.64 mmol), piperidine (1.2 mL, 12.1 mmol, 1 equivalent), i-Pr2NEt (3.2 mL, 18.4 mmol, 1.6 equivalent), and 1,4-dioxane (10 mL) were refluxed in N2O. 2(g) Stir for 90 min, then cool to ambient temperature. Concentrate the reaction mixture into a yellow oil, add CH2Cl2 and hexane, pour the solution onto dry silica (55 mm h x 45 mm id) and elute under vacuum: hexane (100 mL); % CH2Cl2 / hexane - 10% (100 mL), 25% (200 mL), 50% (100 mL), 75% (100 mL), CH2Cl2 (100 mL) to give compound 48 (1.50 g, 71%) as a colorless oil. 1 H NMR (300 MHz, CDCl3) δ 7.48 (dd, 1 H, J HF= 16.7 Hz, J = 8.1 Hz), 6.40 (dd, 1 H, J = 8.1 Hz, J = 2.7 Hz), 6.09 (dd, 1 H, J = 7.8 Hz, J = 3.0 Hz), 3.51 (m, 4 H), 1.64 (m,6 H); 19 F NMR (282.5 MHz, CDCl3) δ -68.60 (d, J = 6.5 Hz); 13 C NMR (125 MHz, CDCl3) δ 162.96 (d, 1 J CF = 233.0 Hz), 158.58 (d, 3 J CF = 15.9 Hz), 141.73 (d, 3 J CF = 8.4 Hz), 102.69 (d, 4 J CF = 4.0 Hz), 94.86 (d, 2 J CF = 37.6 Hz), 46.12,25.46, 24.70); HRMS (ESI) [M+H] + C 10 H 14 Calculated value of FN2: 181.1136, measured value: 181.1133.

[0155] 4-(6-Fluoropyridin-2-yl)morpholine (compound 49). 2,6-Difluoropyridine (1.34 g, 11.64 mmol), morpholine (1 mL, 11.6 mmol, 1 equivalent), i-Pr2NEt (3 mL, 17.2 mmol, 1.5 equivalent), and 1,4-dioxane (10 mL) were refluxed in N2O. 2(g)Stir for 90 min, cool to ambient temperature, and concentrate into an oily substance. Dry under vacuum to obtain a grayish-white solid. Dissolve the solid in CH2Cl2, pour onto dry silica (55 mm h x 45 mm id), and elute under vacuum: hexane (50 mL); % CH2Cl2 / hexane - 25% (50 mL), 50% (75 mL), 75% (100 mL), CH2Cl2 (500 mL); % MeOH / CH2Cl2 - 2% (75 mL), 5% (50 mL), 10% (100 mL) to obtain compound 49 (0.72 g, 34%) as a colorless oily substance, which slowly turns into a white solid. 1 H NMR (300 MHz, CDCl3) δ 7.55(dd, 1 H, J CF = 16.5 Hz, J = 8.1 Hz), 6.41 (dd, 1 H, J = 8.1 Hz, J = 2.7 Hz), 6.21 (dd, 1 H, J = 7.8 Hz, J = 2.7 Hz), 3.80 (t, 4 H, J = 4.8 Hz), 3.50 (t, 4H, J = 4.8 Hz); 19 F NMR (282.5 MHz, CDCl3) δ -68.47 (d, J = 7.1 Hz); 13 C NMR (125 MHz, CDCl3) δ 162.87 (d, 1 J CF = 234.6 Hz), 158.64 (d, 3 J CF = 15.5 Hz), 142.07 (d, 3 J CF = 8.3 Hz), 102.74 (d, 4 J CF = 4.0 Hz), 96.74 (d, 2 J CF = 37.1 Hz),66.65, 45.40; HRMS (ESI) [M+H] + C9H 12 Calculated value of FN2O: 183.0928, measured value: 183.0926.

[0156] 2-Nitro-6-(piperidin-1-yl)pyridine (compound 50). 2-Chloro-6-nitropyridine (0.28 g, 1.77 mmol), piperidine (0.2 mL, 2.0 mmol, 1.1 equivalent), i-Pr2NEt (0.5 mL, 2.9 mmol, 1.6 equivalent), and 1,4-dioxane (5 mL) were refluxed in N2O. 2(g) Stir for 2 h, then cool to ambient temperature. Pour the reaction mixture onto dry silica (55 mm h x 45 mm id) and elute under vacuum: hexane (50 mL); hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (150 mL) to give a yellow / orange slurry (0.27 g). Purify by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (25 mL) to give compound 50 (0.16 g, 44%) as an orange slurry. 1 H NMR (300 MHz, CDCl3) δ7.64 (dd, 1 H, J = 8.4 Hz, J = 7.5 Hz), 7.37 (d, 1 H, J = 7.5 Hz), 6.89 (d, 1H, J = 8.4 Hz), 3.63 (m, 4 H), 1.67 (m, 6 H).

[0157] 4-(6-nitropyridin-2-yl)morpholine (compound 51). 2-chloro-6-nitropyridine (0.28 g, 1.77 mmol), morpholine (0.2 mL, 2.3 mmol, 1.3 equivalents), i-Pr2NEt (0.5 mL, 2.9 mmol, 1.6 equivalents), and 1,4-dioxane (5 mL) were refluxed in N2O. 2(g)The mixture was stirred for 4 h, then cooled to ambient temperature, filtered, and the precipitate was washed with EtOAc. The solvent was removed from the filtrate to obtain an orange residue, which was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (200 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (150 mL) to give a yellow / orange solid. Purification was achieved by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (75 mL) to give compound 51 (0.14 g, 38%) as a yellow / orange solid. 1 H NMR (300 MHz, CDCl3) δ 7.73 (dd, 1 H, J = 8.4Hz, J = 7.5 Hz), 7.49 (d, 1 H, J = 7.5 Hz), 6.90 (d, 1 H, J = 8.4 Hz), 3.83(t, 4 H, J = 5.1 Hz), J = 3.63 (t, 4 H, J = 5.1 Hz); 13 C NMR (125 MHz, CDCl3)δ 158.17, 156.06, 140.49, 111.79, 106.21, 66.67, 45.27; HRMS (ESI) [M+H] + C9H 12 Calculated value of N3O3: 210.0873, measured value: 210.0871.

[0158] 3-Bromo-2-fluoro-6-(piperidin-1-yl)pyridine (compound 52). In N 2(g)Compound 48 (0.77 g, 4.27 mmol) was dissolved in CH3CN (25 mL) and cooled to 0ºC. NBS (0.86 g, 4.83 mmol, 1.1 equivalents) was added, and the reaction mixture was stirred at 0ºC for 5 min, then warmed to ambient temperature and stirred for 6 h. CH3CN was removed to give a light brown residue, which was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted with CH2Cl2 (150 mL) under vacuum to give a colorless oil (1.26 g). Purification by radial chromatography (2 mm silica): %CH2Cl2 / hexane - 10% (100 mL), 25% (50 mL) gave compound 52 (0.78 g, 70%) as a colorless oil. 1 ¹H NMR (300 MHz, CDCl₃) δ 7.57 (apparent t, ¹H, J = 8.7 Hz), 6.33 (dd, ¹H, J = 8.7 Hz, J = 1.5 Hz), 3.49 (m, ⁴H), 1.62 (m, ⁶H); 19 F NMR (470.6 MHz, CDCl3) δ -66.52 (d, J = 8.5 Hz); 13 C NMR (125 MHz, CD3OD) δ 159.43 (d, 1 J CF =229.3 Hz), 158.53 (d, 3 J CF = 14.9 Hz), 145.54 (d, 3 J CF = 2.4 Hz), 106.08 (d, 4 J CF = 4.4 Hz), 87.26 (d, 2 J CF = 38.4 Hz), 47.17, 26.48, 25.66; HRMS (ESI) [M+H] + C 10 H 13 79 Calculated value of BrFN2: 259.0241, measured value: 259.0238.

[0159] 4-(5-bromo-6-fluoropyridin-2-yl)morpholine (compound 53). In N 2(g)Compound 49 (0.36 g, 1.98 mmol) was dissolved in CH3CN (12 mL) and cooled to 0ºC. NBS (0.39 g, 2.19 mmol, 1.1 equivalence) was added, and the reaction mixture was stirred at 0ºC for 5 min, then warmed to ambient temperature and stirred for 6 h. CH3CN was removed to give a red / brown slurry, which was dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id), and eluted under vacuum with CH2Cl2 (300 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL) to give a white solid (0.54 g). Purification by radial chromatography (2 mm silica): % CH2Cl2 / hexane - 25% (100 mL), 50% (100 mL), 75% (100 mL) yielded compound 53 (0.28 g, 54%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ7.65 (apparent t, 1 H, J = 8.7 Hz), 6.34 (dd, 1 H, J = 8.7 Hz, J = 1.5 Hz), 3.79 (t, 4 H, J = 4.8 Hz), 3.48 (t, 4 H, J = 4.8 Hz); 19 F NMR (470.6 MHz, CDCl3) δ-66.22 (d, J = 8.5 Hz); 13 C NMR (125 MHz, CDCl3) δ 158.35 (d, 1 J CF = 241.5 Hz), 157.37 (d, 3 J CF = 23.8 Hz), 144.64 (d, 3 J CF = 2.6 Hz), 104.63 (d, 4 J CF = 4.5Hz), 89.09 (d, 2 J CF = 38.5 Hz), 66.60, 45.44; HRMS (ESI) [M+H] + C9H 11 79 Calculated value of BrFN2O: 261.0033, measured value: 261.0063.

[0160] 3-Bromo-2-nitro-6-(piperidin-1-yl)pyridine (compound 54). Compound 50 (0.16 g, 7.72 x 10⁻⁶) -4 mol) with N 2(g) Rinse, then dissolve in CH3CN (10 mL) and cool to 0ºC. Add NBS (0.14 g, 7.87 x 10⁻⁶ ppm). -4 (mol), the reaction mixture was heated to ambient temperature, and in N 2(g) Stir for 5.5 h. Remove CH3CN to obtain an orange slurry, then add CH2Cl2 and hexane and remove them to obtain an orange residue, which is dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id) and eluted under vacuum: hexane (50 mL); % CH2Cl2 / hexane - 25% (100 mL), 50% (100 mL), CH2Cl2 (150 mL) to obtain a deep yellow slurry. Purify by radial chromatography (2 mm silica): % CH2Cl2 / hexane - 25% (100 mL), 50% (50 mL) to give compound 54 (0.15 g, 68%) as a yellow slurry: 1 H NMR (500 MHz, CDCl3) δ 7.64 (d, 1 H, J = 9.0 Hz), 6.66 (d,1 H, J = 9.0 Hz), 3.54 (t, 4 H, J = 5.5 Hz), 1.66 (m, 2 H), 1.62 (m, 4 H); 13 CNMR (125 MHz, CDCl3) δ 156.45, 156.14, 144.12, 111.10, 92.85, 46.17, 25.49,24.52; HRMS (ESI) [M+H] + C 10 H 13 79 Calculated value of BrN3O2: 286.0186, measured value: 286.0172.

[0161] 4-(5-bromo-6-nitropyridin-2-yl)morpholine (compound 55). Compound 51 (0.12 g, 5.74 x 10⁻⁶) -4 Dissolve 0.15 g of NBS (8.43 x 10 mol) in CH3CN (10 mL), cool to 0ºC, and add NBS (0.15 g, 8.43 x 10 mol). -4mol, 1.5 equivalents). The reaction mixture was stirred in a capped flask at ambient temperature for 6 h, concentrated into an orange oil, then CH2Cl2 and hexane were added and removed, repeated twice to give an orange solid, which was dried under vacuum. The solid was dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id) and eluted under vacuum: CH2Cl2 (400 mL), 1% MeOH / CH2Cl2 (100 mL) to give a yellow solid (0.13 g). Purified by radial chromatography (2 mm silica) with CHCl3 (100 mL) to give compound 55 (82 mg, 50%) as a yellow solid: 1 H NMR (400 MHz, CDCl3) δ 7.76 (d, 1 H, J = 9.2 Hz), 6.68 (d, 1 H, J = 9.2 Hz), 3.80 (t, 4 H, J = 4.8 Hz), 3.54 (t, 4 H, J = 4.8 Hz); 13 C NMR (125 MHz, CDCl3) δ 156.65,156.07, 144.61, 110.99, 94.88, 66.51, 45.21; HRMS (ESI) [M+H] + C9H 11 79 Calculated value of BrN3O3: 287.9978, measured value: 287.9981.

[0162] 4-(4-Fluoropyridin-2-yl)morpholine (compound 56). 2-Bromo-4-fluoropyridine (1.0 g, 5.68 mmol), morpholine (0.50 g, 5.74 mmol), sodium tert-butoxide (0.58 g, 6.04 mmol), toluene (50 mL), and Pd2 (dba) were added. 3（ 73 mg, 7.97 x 10 -5 mol) and XantPhos (0.14 g, 2.42 x 10 -4 mol) with N 2(g) Rinse for 10 min, then heat overnight at 100ºC. Cool the reaction mixture to ambient temperature, filter through a short silica gel pad, and wash with EtOAc. Evaporate the filtrate to dryness and purify the residue by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 56 (0.88 g, 85%) as a white solid. 1H NMR (CDCl3, 300 MHz): δ 8.13 (dd, 1 H, J = 9.4 Hz, J = 5.4 Hz), 6.41 (ddd, 1 H, J = 8.1 Hz, J = 5.4 Hz, J = 2.1 Hz), 6.28 (dd, 1 H, J = 12.3 Hz, J = 2.1 Hz), 3.81 (apparent t, 4 H, J = 4.8 Hz), 3.49 (apparent t, 4 H, J = 4.8 Hz); HRMS (ESI) [M+H] + C9H 12 Calculated value of ON2F: 183.0928, measured value: 183.0931.

[0163] 4-(5-bromo-4-fluoropyridin-2-yl)morpholine (compound 57). Compound 56 (0.87 g, 4.78 mmol) was dissolved in CH3CN (50 mL), cooled to 0ºC, and then NBS (0.85 g, 4.78 mmol) was added. The reaction mixture was stirred at 0ºC for 90 min to remove the solvent, and the residue was purified by rapid column chromatography (4:1 v / v hexane / EtOAc) to give compound 57 (1.20 g, 96%) as a grayish-white solid. 1 ¹H NMR (300 MHz, CDCl₃) δ 8.21 (d, ¹H, J = 9.9 Hz), 6.36 (d, ¹H, J = 11.5 Hz), 3.80 (apparent t, ⁴H, J = 4.8 Hz), 3.48 (apparent t, ⁴H, J = 4.8 Hz). HRMS (ESI) [M+H] + C9H 11 Calculated value of ON2BrF: 261.0033, measured value: 261.0029.

[0164] 5-Bromo-3-fluoro-2-(piperidin-1-yl)pyridine (compound 58). 2,5-Dibromo-3-fluoropyridine (0.27 g, 1.06 mmol), piperidine (85 mg, 1.00 mmol), sodium tert-butoxide (144 mg, 1.50 mmol), toluene (20 mL), and Pd2(dba)3 were added. （ 18 mg, 1.97 x 10 -5 mol) and XantPhos (35 mg, 6.05 x 10) -5 mol) with N 2(g)Rinse for 5 min, then heat overnight at 100ºC. Cool the reaction mixture to ambient temperature, filter through a short silica gel pad, and wash with EtOAc. Evaporate the filtrate to dryness and purify the residue by radial chromatography (7:3 v / v hexane / EtOAc) to give compound 58 (0.15 g, 58%) as a white solid. 1 H NMR (300 MHz, CDCl3): δ 8.02 (dd, 1 H, J = 2.1Hz, J = 0.9 Hz), 7.33 (dd, 1 H, J = 12.0 Hz, J = 2.1 Hz), 3.41 (m, 4 H), 1.65(m, 6 H); HRMS (ESI) [M+H] + C 10 H 13 Calculated value of N2BrF: 259.0241, measured value: 259.0237.

[0165] 4-(5-bromo-3-fluoropyridin-2-yl)morpholine (compound 59). 2,5-Dibromo-3-fluoropyridinium (0.27 g, 1.06 mmol), morpholine (87 mg, 1.00 mmol), sodium tert-butoxide (144 mg, 1.50 mmol), toluene (20 mL), and Pd2(dba)3 (18 mg, 1.97 x 10⁻⁶ mmol) were added. -5 mol) and XantPhos (35 mg, 6.05 x 10) -5 mol) with N 2(g) Rinse for 5 min, then heat overnight at 100ºC. Cool the reaction mixture to ambient temperature, filter through a short silica gel pad, and wash with EtOAc (4 x 10 mL). Evaporate the filtrate to dryness and purify the residue by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 59 (0.16 g, 61%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ8.05 (dd, 1 H, J = 1.8 Hz, J = 0.9 Hz), 7.39 (dd, 1 H, J = 12.0 Hz, J = 1.8 Hz), 3.82 (apparent t, 4 H, J = 4.8 Hz), 3.46 (apparent t, 4 H, J = 4.8 Hz); HRMS (ESI)[M+H] + C9H 11Calculated value of ON2BrF: 261.0033, measured value: 261.0044.

[0166] 2-Chloro-3-fluoro-5-(piperidin-1-yl)pyridine (compound 60). 5-Bromo-2-chloro-3-fluoropyridine (0.27 g, 1.28 mmol), piperidine (88 mg, 1.03 mmol), sodium tert-butoxide (144 mg, 1.50 mmol), toluene (20 mL), and Pd2(dba)3 (18 mg, 1.97 x 10⁻⁶) were added. -5 mol) and XantPhos (35 mg, 6.05 x 10) -5 mol) with N 2(g) Rinse for 5 min, then heat overnight at 100ºC. Cool the reaction mixture to ambient temperature, filter through a short silica gel pad, and wash with EtOAc. Evaporate the filtrate to dryness and purify the residue by radial chromatography (7:3 v / v hexane / EtOAc) to give compound 60 (0.22 g, 99%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ 7.84 (d, 1 H, J =2.7 Hz), 6.95 (dd, 1 H, J = 11.4 Hz, J = 2.7 Hz), 3.20 (m, 4H), 1.67 (m, 6H); HRMS (ESI) [M+H] + C 10 H 13 Calculated value of N2ClF: 215.0746, measured value: 215.0743.

[0167] 4-(6-chloro-5-fluoropyridin-3-yl)morpholine (compound 61). 5-bromo-2-chloro-3-fluoropyridine (0.27 g, 1.28 mmol), morpholine (0.11 g, 1.26 mmol), sodium tert-butoxide (144 mg, 1.50 mmol), toluene (20 mL), and Pd2(dba)3 (18 mg, 1.97 x 10⁻⁶ mmol) were added. -5 mol) and XantPhos (35 mg, 6.05 x 10) -5 mol) with N 2(g) Rinse for 5 min, then heat overnight at 100ºC. Cool the reaction mixture to ambient temperature, filter through a short silica gel pad, and wash with EtOAc. Evaporate the filtrate to dryness and purify the residue by radial chromatography (7:3 v / v hexane / EtOAc) to give compound 61 (0.25 g, 92%) as a white solid. 1H NMR (300 MHz, CDCl3) δ 7.85 (d, 1 H, J = 2.7Hz), 6.97 (dd, 1 H, J = 10.8 Hz, J = 2.7 Hz), 3.87 (m – AA'XX', 4 H, J = 4.8Hz, J = 1.8 Hz), 3.19 (m – AA'XX', 4 H, J = 4.8 Hz, J = 1.8 Hz); HRMS (ESI)[M+H] + C9H 11 Calculated value of ON2ClF: 217.0539, measured value: 217.0536.

[0168] 4-(6-Chloro-4-fluoropyridin-3-yl)morpholine (compound 62). 2-Chloro-4-fluoro-5-bromopyridine (0.27 g, 1.28 mmol), morpholine (87 mg, 1.00 mmol), sodium tert-butoxide (144 mg, 1.50 mmol), toluene (20 mL), and Pd2(dba)3 (18 mg, 1.97 x 10⁻⁶ mmol) were added. -5 mol) and XantPhos (35 mg, 6.05 x 10) -5 mol) with N 2(g) Rinse for 5 min, then heat overnight at 100ºC. Cool the reaction mixture to ambient temperature, filter through a short silica gel pad, and wash with EtOAc. Evaporate the filtrate to dryness and purify the residue by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 62 (97 mg, 45%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ 7.98 (d, 1H, J= 10.5 Hz), 7.04 (d, 1 H, J = 11.1 Hz), 3.86 (m – AA'XX', 4 H, J = 4.8 Hz, J= 1.8 Hz), 3.12 (m – AA'XX', 4 H, J = 4.8 Hz, J = 1.8 Hz); HRMS (ESI) [M+H] + C9H 11 Calculated value of ON2ClF: 217.0539, measured value: 217.0536.

[0169] 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 63). Combine 1,4-dioxane (30 mL) and H₂O (6 mL), and use N₂O. 2(g) Purge for 30 min. Add N-Boc-indole-2-boronic acid (compound 6) (0.25 g, 9.58 x 10⁻⁶ ppm) to the solution. -4 mol), compound 52 (0.31 g, 1.20 mmol, 1.2 equivalents), Pd(dppf)Cl2 (95 mg, 1.30 x 10⁻⁶ mol), compound 52 (0.31 g, 1.20 mmol, 1.2 equivalents), Pd(dppf)Cl� ... -4 0.47 g (0.14 mmol, 4.43 mmol, 4.6 mmol) of Na₂CO₃ and N₂CO₃ were used with N₂CO₃. 2(g) Rinse for 10 min, then add the H2O / 1,4-dioxane mixture (15 mL). Reflux the reaction mixture under N2O conditions. 2(g) The mixture was stirred for 4 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with 1,4-dioxane, followed by washing with EtOAc. The solvent was removed from the filtrate to obtain a brown oily substance, which was dried under vacuum to obtain a brown residue. The residue was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum with CH2Cl2 (250 mL) to obtain a slightly brown slurry (0.19 g). Purification by radial chromatography (2 mm silica) with 90:8:2 v / v / v hexane / EtOAc / NEt3 (100 mL) yielded compound 63 (87 mg, 23%) as a white foam. 1 H NMR (300MHz, CDCl3) δ 8.20 (d, 1 H, J = 8.4 Hz), 7.55 (m, 2 H), 7.31 (ddd, 1 H, J =8.4 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.22 (td, 1 H, J = 7.2 Hz, J = 0.9 Hz), 6.53(s, 1 H), 6.49 (dd, 1 H, J = 8.4 Hz, J = 1.8 Hz), 3.58 (m, 4 H), 1.66 (m, 6H), 1.45 (s, 9 H); 19 F NMR (282.5 MHz, CDCl3) δ -67.96 (d, J = 9.6 Hz); HRMS(ESI) [M+H] + C 23 H 27Calculated value of FN3O2: 396.2082, measured value: 396.2074.

[0170] 2-(2-fluoro-6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 64). Combine 1,4-dioxane (30 mL) and H₂O (6 mL), and use N₂O. 2(g) Purge for 30 min. Add N-Boc-indole-2-boronic acid (6) (0.24 g, 9.19 x 10⁻⁶ g) to the solution. -4 mol), compound 53 (0.25 g, 9.58 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (0.11 g, 1.50 x 10 -4 0.30 g (2.83 mmol, 3.1 equivalents) and Na2CO3 (0.30 g, 2.83 mmol, 3.1 equivalents) were treated with N 2(g) Rinse for 15 min, then add the H2O / 1,4-dioxane mixture (15 mL). Reflux the reaction mixture under N2O conditions. 2(g) The mixture was stirred for 4 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with 1,4-dioxane and then with EtOAc. The solvent was removed from the filtrate to obtain a brown oily substance, which was dried under vacuum to obtain a brown residue. The residue was dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2.5% (100 mL), 5% (150 mL). The desired fractions were combined, concentrated, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (300 mL), 50 : 45 : 5 (100 mL) to obtain a brown residue (0.23 g). Purification by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL) yielded compound 64 (0.20 g, 55%) as a white foam. 1H NMR (300 MHz, CDCl3) δ 8.20 (d, 1 H, J =8.1 Hz), 7.62 (dd, 1 H, J = 9.6 Hz, J = 8.1 Hz), 7.54 (d, 1 H, J = 7.2 Hz), 7.32 (ddd, 1 H, J = 8.4 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.23 (td, 1 H, J = 7.5Hz, J = 0.9 Hz), 6.55 (s, 1 H), 6.48 (dd, 1 H, J = 8.1 Hz, J = 1.8 Hz), 3.82(t, 4 H, J = 4.8 Hz), 3.54 (t, 4 H, J = 4.8 Hz), 1.46 (s, 9 H); 13 C NMR (125MHz, CDCl3) δ 159.39 (d, 1 J FC = 237.7 Hz), 158.10 (d, J FC = 15.6 Hz), 150.26,142.03 (d, J FC = 4.8 Hz), 137.27, 133.70 (d, J FC = 4.3 Hz), 129.22, 124.62,123.01, 120.57, 115.69, 110.61, 105.72 (d, J FC = 31.2 Hz), 102.33 (d, J FC =3.8 Hz), 83.81, 66.73, 45.64, 28.01; 19 F NMR (282.5 MHz, CDCl3) δ -67.85 (d, J= 9.9 Hz); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of FN3O3: 398.1875, measured value: 398.1867.

[0171] 2-(2-nitro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 65). Combine 1,4-dioxane (15 mL) and H₂O (3 mL), and use N₂O. 2(g)Purge for 30 min. Add N-Boc-indole-2-boronic acid (6) (92 mg, 3.52 x 10⁻⁶ mg) to the solution. -4 mol), compound 54 (110 mg, 3.84 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (32 mg, 4.37 x10 -5 mol (0.1 equivalent) and Na2CO3 (196 mg, 1.85 mmol, 5.2 equivalent) were used with N 2(g) Rinse for 10 min, then add the H2O / 1,4-dioxane mixture (10 mL). Reflux the reaction mixture under N2O conditions. 2(g) The mixture was stirred for 5 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with EtOAc. The solvent was removed from the filtrate to obtain a dark green / black residue, which was dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id), and eluted under vacuum with 75:20:5 v / v / v hexane / EtOAc / NEt3 (200 mL) to give a dark orange slurry (0.14 g). Purification was performed by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90:8:2 (100 mL), 75:20:5 (100 mL) to give a yellow solid (65 mg), which was further purified by radial chromatography (2 mm silica) with CHCl3 (50 mL) to give an orange foam (56 mg). Purification by radial chromatography (1 mm silica): % CH2Cl2 / hexane - 25% (25 mL), 50% (50 mL), CH2Cl2 (25 mL) yielded compound 65 (48 mg, 32%), which presented as yellow / orange foam. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.20 (d, 1 H, J = 8.4 Hz), 7.58 (d, 1 H, J = 8.8 Hz), 7.53 (d, 1 H, J = 8.0 Hz), 7.33 (apparent t, 1 H, J = 7.6 Hz), 7.24 (apparent t - partially blocked by CHCl₃ resonance, 1 H, J = 7.2 Hz), 6.86 (d, 1 H, J = 8.8 Hz), 6.50 (s, 1 H), 3.66 (m, 4 H), 1.67 (m, 6 H), 1.41 (s, 9 H); 13C NMR (125 MHz, CDCl3) δ 157.10, 154.90, 150.07, 142.88, 136.97, 134.21, 129.21, 124.83,123.08, 120.72, 116.06, 111.29, 110.65, 109.53, 83.90, 46.24, 27.97, 25.60,24.74; HRMS (ESI) [M+H] + C 23 H 27 Calculated value of N4O4: 423.2027, measured value: 423.2030. X-ray quality crystals were grown by slow evaporation of CH2Cl2 / hexane.

[0172] 2-(6-morpholino-2-nitropyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 66). Combine 1,4-dioxane (15 mL) and H₂O (3 mL), and use N₂O. 2(g) Purge for 30 min. Add N-Boc-indole-2-boronic acid (60 mg, 2.30 x 10⁻⁶ ppm) to the solution. -4 mol), compound 55 (66 mg, 2.30 x 10⁻⁶) -4 mol), Pd(dppf)Cl2 (19 mg, 2.60 x 10 -5 mol (0.1 equivalent) and Na2CO3 (117 mg, 1.10 mmol, 4.8 equivalent) were used with N 2(g) Rinse, then add the H2O / 1,4-dioxane mixture (10 mL). Reflux the reaction mixture under N2O conditions. 2(g) The mixture was stirred for 4 h, then cooled to ambient temperature, filtered through diatomaceous earth, and the diatomaceous earth was washed with EtOAc. The filtrate was concentrated, and EtOAc was added and removed to give a dark green / brown oily substance. CH2Cl2 and hexane were added and removed to give a residue, which was dissolved in CH2Cl2, poured onto dry silica (45 mm h x 45 mm id), and eluted under vacuum: CH2Cl2 (200 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (150 mL) to give a dark yellow / brown residue (69 mg). Purified by radial chromatography (1 mm silica): CH2Cl2 (100 mL) to give compound 66 (36 mg, 37%) as a yellow / orange foam: 1¹H NMR (400 MHz, CDCl₃) δ 8.18 (d, 1H, J = 8.1 Hz), 7.67 (d, 1H, J = 8.1 Hz), 7.54 (d, 1H, J = 7.6 Hz), 7.34 (ddd, 1H, J = 8.4 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.25 (m - partially blocked by CHCl₃ resonance, 1H), 6.88 (d, 1H, J = 8.4 Hz), 6.52 (s, 1H), 3.84 (t, 4H, J = 4.8 Hz), 3.65 (t, 4H, J = 4.8 Hz), 1.43 (s, 9H); 13 CNMR (100 MHz, CDCl3) δ 157.25, 154.70, 150.06, 143.25, 136.89, 133.83,129.17, 125.00, 123.17, 120.83, 116.09, 113.03, 110.87, 109.56, 84.12, 66.67,45.35, 28.01; HRMS (ESI) [M+H] + C 22 H 25 Calculated value of N4O5: 425.1820, measured value: 425.1815. X-ray quality crystals were grown by slow evaporation of CH2Cl2 / hexane.

[0173] 4-(5-(1H-indol-2-yl)-6-nitropyridin-2-yl)morpholine (compound 67). Separated as a byproduct from the reaction of compound 66: 1 H NMR (400 MHz, acetone-d6) δ 10.53 (br s, 1 H), 8.09 (d, 1 H, J =8.8 Hz), 7.56 (d, 1 H, J = 7.6 Hz), 7.41 (d, 1 H, J = 8.4 Hz), 7.20 (d, 1 H,J = 8.8 Hz), 7.13 (t, 1 H, J = 7.6 Hz), 7.04 (t, 1 H, J = 7.6 Hz), 6.49 (s, 1H), 3.77 (t, 4 H, J = 4.8 Hz), 3.62 (t, 4 H, J = 4.8 Hz); 13C NMR (125 MHz, DMSO-d6) δ 156.31, 154.79, 140.95, 136.77, 130.23, 128.21, 121.94, 120.14,119.51, 111.30, 110.32, 107.21, 100.09, 65.66, 44.65; HRMS (ESI) [M+H] + C 17 H 17 Calculated value of N4O3: 325.1295, measured value: 325.1295.

[0174] 2-(2-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole (compound 68). Compound 63 (71 mg, 1.80 x 10⁻⁶) -4 1 mol) and TFA (1.8 mL, 23.4 mmol, 130 equivalents) were stirred for 20 min, then poured into a mixture of NaHCO3 (2.29 g, 27.26 mmol, 1.2 equivalents of TFA), H2O (45 mL), and CH2Cl2 (45 mL). The mixture was stirred for 10 min, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (20 mL). The combined CH2Cl2 layer was washed with brine (25 mL), dried over MgSO4, concentrated, poured onto dry silica (33 mm h x 33 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (25 mL), 75 : 20 : 5 (200 mL), 50 : 45 : 5 (50 mL) to give compound 68 (25 mg, 47%) as a grayish-white solid. 1 H NMR (400 MHz, CDCl3) δ 8.77 (br s,1 H), 7.92 (dd, 1H, J = 10.6 Hz, J = 8.4 Hz), 7.59 (d, 1 H, J = 8.0 Hz), 7.39(d, 1 H, J = 7.6 Hz), 7.17 (td, 1 H, J = 7.6 Hz, J = 0.8 Hz), 7.10 (td, 1 H,J = 7.6 Hz, J = 0.8 Hz), 6.72 (d, 1 H, J = 1.6 Hz), 6.54 (dd, 1 H, J = 8.4Hz, J = 2.4 Hz), 3.59 (m, 4 H), 1.67 (m, 6 H); 13C NMR (125 MHz, DMSO-d6) δ158.00 (d, 1 J FC = 236.7 Hz), 156.04 (d, J FC = 16.5 Hz), 139.16 (d, J FC = 4.4Hz), 136.42, 131.47 (d, J FC = 7.3 Hz), 128.57, 121.11, 119.60, 119.21, 111.00,103.91 (d, J FC = 3.5 Hz), 100.65 (d, J FC = 28.0 Hz), 99.81 (d, J FC = 8.3 Hz),45.44, 24.93, 24.09; 19 F NMR (470.6 MHz, DMSO-d6) δ -67.52; 19 F NMR (376.5 MHz, CDCl3) δ -70.32; HRMS (ESI) [M+H] + C 18 H 19 Calculated value of FN3: 296.1558, measured value: 296.1604.

[0175] 4-(6-fluoro-5-(1H-indol-2-yl)pyridin-2-yl)morpholine (compound 69, JSS20-183A). Compound 64 (76 mg, 1.91 x 10⁻⁶ mg) was used. -4 1 mol) and TFA (1.8 mL, 23.4 mmol, 123 equivalents) were stirred for 20 min, then poured into a mixture of NaHCO3 (2.22 g, 26.43 mmol, 1.1 equivalents of TFA), H2O (45 mL), and CH2Cl2 (45 mL). The mixture was stirred for 10 min, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (20 mL). The combined CH2Cl2 layer was washed with brine (25 mL), dried over MgSO4, concentrated, poured onto dry silica (33 mm h x 33 mm id), and eluted under vacuum: hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (100 mL), 50 : 45 : 5 (200 mL), 20:75:5 (75 mL) to give compound 69 (54 mg, 95%) as a light brown solid. 1H NMR (400 MHz, CDCl3) δ8.78 (br s, 1 H), 7.99 (dd, 1 H, J = 10.4 Hz, J = 8.8 Hz), 7.60 (d, 1 H, J =7.6 Hz), 7.40 (d, 1 H, J = 8.0 Hz), 7.18 (td, 1 H, J = 7.6 Hz, J = 0.8 Hz),7.11 (td, 1 H, J = 7.6 Hz, J = 0.8 Hz), 6.77 (d, 1 H, J = 1.2 Hz), 6.55 (dd,1 H, J = 8.4 Hz, J = 2.0 Hz), 3.82 (t, 4 H, J = 4.8 Hz), 3.57 (t, 4 H, J =4.8 Hz); 13 C NMR (125 MHz, DMSO-d6) δ 157.86 (d, 1 J FC = 237.2 Hz), 156.31 (d,J FC = 16.1 Hz), 139.26 (d, J FC = 4.4 Hz), 136.47, 131.13 (d, J FC = 7.2 Hz),128.50, 121.30, 119.71, 119.27, 111.06, 104.14 (d, J FC = 3.6 Hz), 102.11 (d,J FC = 27.5 Hz), 100.23 (d, J FC = 8.6 Hz), 65.71, 44.83; 19 F NMR (376.5 MHz, CDCl3) δ -70.37; HRMS (ESI) [M+H] + C 17 H 17 Calculated value of FN3O: 298.1350, measured value: 298.1396. X-ray quality crystals were grown by slow evaporation of CDCl3.

[0176] 5-Fluoro-2-(2-morpholinopyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 70). Compound 42 (0.21 g, 7.52 x 10⁻⁶) -4 mol), compound 24 (0.10 g, 4.10 x 10 ...)-4 mol) and Pd(dppf)Cl 2（ 20 mg, 2.73 x 10 -5 Dissolve mol) in 1,4-dioxane (10 mL), then add K2CO 3(aq) (2 M, 0.5 mL). The reaction mixture was then heated with N2. 2(g) Rinse for 10 minutes, then at 100ºC under N2O 2(g) Heat overnight. Cool the reaction mixture to ambient temperature, evaporate to dryness, dissolve the residue in EtOAc (30 ml), wash with H2O (10 mL), dry over MgSO4, and filter. Pass the filtrate through a silica gel pad, wash the silica with EtOAc, and concentrate the filtrate to dryness. Purify the residue by radial chromatography (hexane / EtOAc v / v 90:10 to 80:20) to give compound 70 as a solid (0.15 g, 92%). 1 HNMR (300 MHz, CDCl3) δ 8.38 (s, 2 H), 8.13 (dd, 1 H, J = 9.0 Hz, J = 4.5 Hz), 7.20 (dd, 1 H, J = 8.7 Hz, J = 2.7 Hz), 7.05 (td, 1 H, J = 9.0 Hz, J = 2.7Hz), 6.50 (d, 1 H, J = 0.3 Hz), 3.88 (m, 4 H), 3.79 (m, 4 H), 1.49 (s, 9 H);HRMS (ESI) [M+H] + C 21 H 24 Calculated value of O3N4F: 399.1827, measured value: 399.1837.

[0177] 5-Fluoro-2-(6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 71). Compound 42 (0.21 g, 7.52 x 10⁻⁶) -4 mol), compound 29 (0.10 g, 4.11 x 10⁻⁶ ... -4 mol), Pd(dppf)Cl2 (20 mg, 2.73 x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 10 minutes, then at 100ºC under N2O 2(g)Stir overnight. Cool the reaction mixture to ambient temperature, evaporate to dryness, dissolve the residue in EtOAc (30 ml), wash with H2O (10 mL), dry over MgSO4, and filter. Pass the filtrate through a silica gel pad, wash the silica with EtOAc, and concentrate the filtrate to dryness. Purify the residue by radial chromatography (hexane / EtOAc v / v 90:10 to 80:20) to give compound 71 as a solid (0.12 g, 73%). 1 ¹H NMR (300 MHz, CDCl₃) δ 8.26 (d, ¹H, J = 2.1 Hz), 8.12 (dd, ¹H, J = 9.0 Hz, J = 4.5 Hz), 7.55 (dd, ¹H, J = 8.7 Hz, J = 1.8 Hz), 7.19 (dd, ¹H, J = 8.7 Hz, J = 2.4 Hz), 7.03 (td, ¹H, J = 9.0 Hz, J = 2.4 Hz), 6.68 (d, ¹H, J = 8.7 Hz), 6.48 (s, ¹H), 3.85 (t, 4H, J = 4.8 Hz), 3.57 (partially resolved t, 4H, J = 4.8 Hz), 1.43 (s, 9H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of O3N3F: 398.1875, measured value: 398.1887.

[0178] 4-(5-(5-fluoro-1H-indol-2-yl)pyrimidin-2-yl)morpholine (compound 72). Compound 70 (0.14 g, 3.51 x 10⁻⁶) -4 Dissolve mol) in TFA (2 mL) and stir at ambient temperature for 2 h, then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 72 (58 mg, 55%) as a grayish-white solid. 1H NMR (300 MHz, acetone-d6) δ 10.80 (br s, 1 H), 8.83 (s, 2 H), 7.37 (dd, 1 H, J = 8.7 Hz, J = 4.5 Hz), 7.23 (dd, 1 H, J = 9.6 Hz, J = 2.4Hz), 6.88 (dd, 1 H, J = 9.6 Hz, J = 8.7 Hz, J = 2.4 Hz), 6.79 (m, 1 H), 3.82 (m, 4 H), 3.71 (m, 4 H); 13 C NMR (125 MHz, DMSO-d6): 160.33, 157.23 (d, 1 J FC =231.4 Hz), 154.69, 135.15, 133.59, 128.93 (d, J FC = 10.3 Hz), 115.41, 111.92(d, J FC = 9.9 Hz), 109.27 (d, J FC = 25.9 Hz), 104.31, 104.12, 97.50 (d, J FC =4.7 Hz), 65.95, 44.01; HRMS (ESI) [M+H] + C 16 H 16 Calculated value of ON4F: 299.1303, measured value: 299.1301.

[0179] 4-(5-(5-fluoro-1H-indol-2-yl)pyridin-2-yl)morpholine (compound 73). Compound 71 (0.12 g, 3.02 x 10⁻⁶) was used. -4 Dissolve mol) in TFA (2 mL) and stir at ambient temperature for 2 h, then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was then placed in a filter and washed with a small amount of acetone to give compound 73 (66 mg, 74%) as a grayish-white solid. 1H NMR (acetone-d6, 300 MHz): δ 10.62 (s, 1 H), 8.65 (dd, 1 H, J = 2.5 Hz, J = 0.7 Hz), 7.98 (dd, 1 H, J = 8.9 Hz, J = 2.6Hz), 7.34 (dd, 1 H, J = 8.8 Hz, J = 4.5 Hz), 7.21 (dd, 1 H, J = 9.9 Hz, J =2.5 Hz), 6.85 (m, 2 H), 6.74 (s, 1 H), 3.74 (m, 4 H), 3.56 (m, 4 H); 13 C NMR (125 MHz, DMSO-d6): 158.30, 157.17 (d, 1 J FC = 231.5 Hz), 144.57, 137.81,134.38, 133.54, 129.07 (d, J FC = 10.6 Hz), 117.78, 111.75 (d, J FC = 9.9 Hz), 108.88 (d, J FC = 26.0 Hz), 106.86, 104.05 (d, J FC = 23.1 Hz), 97.18 (d, J FC =4.5 Hz), 65.90, 45.01; HRMS (ESI) [M+H] + C 17 H 17 Calculated value of ON3F: 298.1350, measured value: 298.1348.

[0180] 6-Fluoro-2-(2-Fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 74). Add 1,4-dioxane (25 mL) to N... 2(g) Purge for 1 h. Compound 52 (0.25 g, 9.65 x 10⁻⁶) was then purged. -4 mol), compound 4 (0.41 g, 1.03 mmol, 1.1 equivalents), Na2CO3 (0.51 g, 4.81 mmol, 5 equivalents) and Pd(dppf)Cl2 (89 mg, 1.22 x 10⁻⁶ ... -4 mol (0.1 equivalent) with N 2(g)Rinse for 10 min, then add 1,4-dioxane. Recycle the reaction mixture under reflux in N2O. 2(g) Stir for 3 h, then cool to ambient temperature, filter through diatomaceous earth, and wash the diatomaceous earth with EtOAc. Concentrate the filtrate into a brown oil, then add hexane and remove it to obtain a brown residue, which is dissolved in CH2Cl2, poured onto dry silica (55 mm h x 45 mm id) and eluted under vacuum: hexane (50 mL), hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (100 mL), 75 : 20 : 5 (100 mL), 50 : 45 : 5 (50 mL) to obtain an orange slurry (0.43 g). Purification by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 95 : 4 : 1 (100 mL), 90 : 8 : 2 (50 mL) yielded a grayish-white foam (0.16 g), which was further purified by radial chromatography (2 mm silica): % CH2Cl2 / hexane - 25% (50 mL), 50% (75 mL), 75% (50 mL); CH2Cl2 (25 mL) to yield a grayish-white solid (0.14 g). Final purification by radial chromatography (2 mm silica): hexane / EtOAc / NEt3 v / v / v 90 : 8 : 2 (125 mL), 75 : 20 : 5 (25 mL) yielded compound 74 (0.13 g, 33%) as a white solid. 1 H NMR (300 MHz, CDCl3) δ 7.95 (dd, 1 H, J =10.8 Hz, J = 2.1 Hz), 7.54 (dd, 1 H, J = 9.6 Hz, J = 8.4 Hz), 7.44 (dd, 1 H,J = 8.4 Hz, J = 5.4 Hz), 6.99 (td, 1H, J = 8.7 Hz, J = 2.4 Hz), 6.54 (d, 1H, J = 7.5 Hz), 6.49 (s, 1 H), 3.57 (m, 4 H), 1.67 (s, 6 H), 1.44 (s, 9 H); 13 C NMR (100 MHz, CDCl3) δ 161.06 (d, J FC = 238.0 Hz), 159.46 (d, J FC = 234.5Hz), 158.04 (d, J FC= 16.2 Hz), 150.10, 141.80 (d, J FC = 4.8 Hz), 137.51 (d,J FC = 12.8 Hz), 134.52 (t, J FC = 4.3 Hz), 125.55 (d, J FC = 1.1 Hz), 120.95 (d,J FC = 9.8 Hz), 111.16 (d, J FC = 24.1 Hz), 109.84, 103.46 (d, J FC = 31.3 Hz), 103.11 (d, J FC = 28.6 Hz), 102.25 (d, J FC = 3.8 Hz), 84.08, 46.45, 27.93,25.55, 24.84; HRMS (ESI) [M+H] + C 23 H 26 Calculated value of F2N3O2: 414.1988, measured value: 414.1989.

[0181] 6-Fluoro-2-(2-Fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole (compound 75). Compound 74 (92 mg, 2.23 x 10⁻⁶) -4The solution was dissolved in TFA (2.2 mL, 28.6 mmol, 128 equivalents) and stirred at ambient temperature for 20 min. The solution was then poured into a mixture of NaHCO3 (2.68 g, 31.90 mmol, 1.1 equivalents of TFA), H2O (55 mL), and CH2Cl2 (55 mL). The mixture was stirred for 10 min, and the layers were separated. The H2O layer was extracted with CH2Cl2 (25 mL). The combined CH2Cl2 layers were washed with H2O (50 mL) and brine (50 mL) and dried over MgSO4. The solution was concentrated, poured onto dry silica (55 mm h x 45 mm id), and eluted under vacuum: hexane (50 mL); hexane / EtOAc / NEt3 v / v / v 75 : 20 : 5 (200 mL), 50 : 45 : 5 (150 mL) to give a grayish-white / brown solid (68 mg). The solid was suspended in CHCl3 (1 mL), filtered, washed with CHCl3 (1 mL x 3), then washed with hexane (5 mL), and dried under vacuum to give compound 75 (49 mg, 70%) as a white solid. 1 H NMR (300 MHz, DMSO-d6) δ 11.36(s, 1 H), 8.08 (dd, 1 H, J = 10.8 Hz, J = 8.7 Hz), 7.49 (dd, 1 H, J = 8.7 Hz,J = 5.7 Hz), 7.13 (dd, 1 H, J = 9.9 Hz, J = 1.8 Hz), 6.84 (m, 2 H), 6.69 (s, 1 H), 3.56 (m, 4 H), 1.59 (m, 6 H); 13 C NMR (125 MHz, DMSO-d6) δ 158.71 (d, 1 J FC = 232.9 Hz), 157.84 (d, 1 J FC = 234.8 Hz), 157.05 (d, J FC = 16.3 Hz), 139.06(d, J FC =4.4 Hz), 136.33 (d, J FC = 12.9 Hz), 132.23 (dd, J FC = 7.3 Hz, J FC = 3.6Hz), 125.34, 120.53 (d, JFC = 10.0 Hz), 107.61 (d, J FC = 24.3 Hz), 103.91 (d,J FC =3.5 Hz), 100.40 (d, J FC = 27.9 Hz), 99.70 (d, J FC = 8.0 Hz), 97.03 (d, J FC = 25.8 Hz), 45.41, 24.92, 24.06; HRMS (ESI) [M+H] + C 18 H 18 Calculated value of F2N3: 314.1463, measured value: 314.1454.

[0182] (1-(tert-butoxycarbonyl)-6-fluoro-1H-indol-2-yl)boronic acid (compound 76). In N 2(g) Compound 104 (2.35 g, 9.99 mmol) was dissolved in THF (100 mL) and cooled to 0ºC. Triisopropyl borate (2.82 g, 14.99 mmol) was then added. LDA (2 M THF / heptane / ethylbenzene, 6.5 mL, 13 mmol) was added dropwise over a 10-minute period. The reaction mixture was stirred at 0ºC for 2 h and then dissolved in 1 M HCl. (aq) Quenching. The reaction mixture was extracted with EtOAc (100 mL x 3), the combined extracts were washed with H2O (2 x 50 mL), dried over MgSO4, filtered, and evaporated to dryness. The residue was recrystallized from 1:2 v / v hexane / EtOAc to give compound 76 (1.78 g, 64%) as a white solid. 1 H NMR (300 MHz, CDCl3): δ 7.72 (dd, 1 H, J = 11.1 Hz, J = 2.4 Hz), 7.52 (dd, 1 H,J = 8.4 Hz, J = 5.7 Hz), 7.45 (d, 1 H, J = 0.6 Hz), 7.02 (td, 1 H, J = 8.7Hz, J = 2.4 Hz) 6.88 (br s, 2 H), 1.75 (s, 9 H).

[0183] 6-Fluoro-2-(5-morpholinopyrazin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 77). Compound 76 (0.21 g, 7.52 x 10⁻⁶)-4 mol), compound 30 (0.10 g, 4.10 x 10 ...) -4 mol), Pd(dppf)Cl2 (20 mg, 2.73 x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 10 minutes, then at 100ºC under N2O 2(g) Stir overnight. Cool the reaction mixture to ambient temperature, evaporate to dryness, dissolve the residue in EtOAc (30 ml), wash with H2O (10 mL), dry over MgSO4, and filter. Pass the filtrate through a silica gel pad, wash the silica with EtOAc, and concentrate the filtrate to dryness. Purify the residue by radial chromatography (hexane / EtOAc v / v 90:10 to 80:20) to give compound 77 (78 mg, 48%) as a solid. 1 ¹H NMR (300 MHz, CDCl₃) δ 8.27 (d, 1 H, J = 1.2 Hz), 8.17 (d, 1 H, J = 1.5 Hz), 7.88 (dd, 1 H, J = 10.8 Hz, J = 2.4 Hz), 7.48 (dd, 1 H, J = 8.7 Hz, J = 5.7 Hz), 7.01 (td, 1 H, J = 9.0 Hz, J = 2.4 Hz), 6.69 (s, 1 H), 3.86 (apparent t, 4 H, J = 4.8 Hz), 3.62 (apparent t, 4 H, J = 4.8 Hz), 1.44 (s, 9 H); HRMS (ESI) [M+H] + C 21 H 24 Calculated value of O3N4F: 399.1827, measured value: 399.1829.

[0184] 6-Fluoro-2-(4-Fluoro-6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 78). Compound 76 (0.34 g, 1.22 mmol) and compound 57 (0.15 g, 5.75 x 10⁻⁶ mmol) were used. -4 mol), Pd(dppf)Cl2 (28 mg, 3.83x 10 -5 mol), 1,4-dioxane (30 mL) and K2CO 3(aq) (2 M, 0.7 mL) with N 2(g)Rinse for 5 minutes, then in N 2(g) Stirred overnight at 100ºC (TLC indicated the remaining unreacted compound 57). Add the remaining compound 76 (0.15 g, 5.37 x 10⁻⁶). -4 mol) and the reaction mixture in N 2(g) The mixture was stirred overnight at 100ºC, then cooled to ambient temperature and evaporated to dryness. The residue was dissolved in H2O and extracted with EtOAc (10 ml x 3). The combined extracts were dried over MgSO4 and filtered. The filtrate was passed through a silica gel pad, the silica was washed with EtOAc, and the solvent was removed to obtain the residue, which was purified by radial chromatography (hexane / EtOAc v / v 99:1 to 80:20) to give compound 78 as a solid (0.15 g, 63%). 1 ¹H NMR (500 MHz, CDCl₃) δ 8.20 (d, 1H, J = 10.5 Hz), 7.92 (dd, 1H, J = 10.5 Hz, J = 1.5 Hz), 7.47 (dd, 1H, J = 9.0 Hz, J = 5.5 Hz), 7.01 (td, 1H, J = 9.0 Hz, J = 2.5 Hz), 6.53 (s, 1H), 6.36 (d, 1H, J = 12.5 Hz), 3.83 (t, 4H, J = 5.0 Hz), 3.56 (partially resolved m, 4H), 1.47 (s, 9H); HRMS (ESI) [M+H] + C 22 H 24 Calculated value of O3N3F2: 416.1780, measured value: 416.1793.

[0185] 6-Fluoro-2-(5-Fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 79). Compound 76 (60 mg, 2.15 x 10⁻⁶) -4 mol), compound 58 (53 mg, 2.05 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (10 mg, 1.37 x 10 -5 mol), 1,4-dioxane (6 mL) and K2CO 3(aq) (2 M, 0.3 mL) with N 2(g) Rinse for 5 minutes, then in N 2(g)The mixture was stirred overnight at 100ºC (TLC indicated the presence of unreacted compound 58). Additional compound 76 (64 mg, 2.29 x 10⁻⁶) was added. -4 mol) and the reaction mixture in N 2(g) The mixture was stirred overnight at 100ºC, then cooled to ambient temperature and evaporated to dryness. The residue was dissolved in H2O and extracted with EtOAc (10 ml x 3). The combined extracts were dried over MgSO4 and filtered. The filtrate was passed through a silica gel pad, the silica was washed with EtOAc, and the solvent was removed to obtain the residue, which was purified by radial chromatography (hexane / EtOAc v / v 99 : 1 to 80 : 20) to give compound 79 (59 mg, 70%) as a solid. 1 H NMR (300 MHz, CDCl3) δ 8.00 (m, 1 H), 7.89 (dd, 1 H, J= 10.8 Hz, J = 1.8 Hz), 7.41 (m, 1 H), 7.21 (dd, 1 H, J = 13.8 Hz, J = 1.8Hz), 6.96 (tt, 1 H, J = 9.0 Hz, J = 2.1 Hz), 6.47 (s, 1 H), 3.45 (m, 4 H),1.64 (m, 6 H), 1.39 (s, 9 H); HRMS (ESI) [M+H] + C 23 H 26 Calculated value of O2N3F2: 414.1988, measured value: 414.1988.

[0186] 6-Fluoro-2-(5-Fluoro-6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 80). Compound 76 (0.19 g, 6.81 x 10⁻⁶ g) -4 mol), compound 59 (77 mg, 2.95 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (12 mg, 1.64 x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 10 minutes, then at 100ºC under N2O 2(g)Stir overnight. Cool the reaction mixture to ambient temperature, evaporate to dryness, dissolve the residue in H2O, and extract with EtOAc (10 ml x 3). Dry the combined extracts in MgSO4 and filter, pass the filtrate through a silica gel pad, and wash the silica with EtOAc. Remove the solvent to obtain the residue, which is purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 80 (86 mg, 70%) as a solid. 1 ¹H NMR (400MHz, CDCl₃): δ 8.07 (s, 1 H), 7.92 (d, 1 H, J = 10.8 Hz), 7.46 (dd, 1 H, J = 8.8 Hz, J = 5.6 Hz), 7.30 (d, 1 H, J = 13.6 Hz), 7.02 (unresolved apparent td, 1 H, J = 8.8 Hz), 6.53 (s, 1 H), 3.86 (t, 4 H, J = 4.4 Hz), 3.54 (t, 4 H, J = 4.4 Hz), 1.45 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 24 Calculated value of O3N3F2: 416.1780, measured value: 416.1781.

[0187] 4-(5-(6-fluoro-1H-indol-2-yl)pyrazin-2-yl)morpholine (compound 81). Compound 77 (78 mg, 1.96 x 10⁻⁶) -4 mol) was dissolved in TFA (2 mL), stirred at ambient temperature for 2 h, and then N... 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 81 (58 mg, 99%) as a grayish-white solid. 1H NMR (500 MHz, acetone-d6) δ 10.75 (br s, 1 H), 8.72 (d, 1 H, J =1.0 Hz), 8.26 (d, 1 H, J = 1.5 Hz), 7.55 (dd, 1 H, J = 9.0 Hz, J = 5.5 Hz), 7.23 (dd, 1 H, J = 10.0 Hz, J = 2.0 Hz), 6.84 (ddd, 1 H, J = 10.0 Hz, J = 8.5Hz, J = 2.0 Hz), 3.79 (t, 4 H, J = 5.0 Hz), 3.62 (t, 4 H, J =5.0 Hz); 13 C NMR(125 MHz, DMSO-d6): 159.06 (d, 1 J FC = 234.3 Hz), 153.41, 138.40, 136.93 (d, J FC = 13.2 Hz), 136.29, 135.17, 129.82, 125.49, 121.08 (d, J FC = 9.6 Hz), 107.89(d, J FC = 24.9 Hz), 98.09, 97.51 (d, J FC = 26.0 Hz), 65.81, 44.54; HRMS (ESI)[M+H] + C 16 H 16 Calculated value of ON4F: 299.1303, measured value: 299.1302.

[0188] 4-(4-fluoro-5-(6-fluoro-1H-indol-2-yl)pyridin-2-yl)morpholine (compound 82). Compound 78 (0.15 g, 3.61 x 10⁻⁶ g) -4 Dissolve mol) in TFA (2 mL), stir at ambient temperature for 1.5 h, and then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 82 (0.11 g, 97%) as a grayish-white solid. 1¹H NMR (300 MHz, CDCl₃) δ 8.71 (br s, ¹H), 8.61 (d, ¹H, J = 11.1 Hz), 7.52 (dd, ¹H, J = 8.7 Hz, J = 5.4 Hz), 7.08 (dd, ¹H, J = 9.6 Hz, J = 2.1 Hz), 6.89 (ddd, ¹H, J = 9.9 Hz, J = 8.7 Hz, J = 2.1 Hz), 6.80 (dd, ¹H, J = 1.2 Hz), 6.40 (d, ¹H, J = 14.7 Hz), 3.85 (apparent t, ⁴H, J = 4.8 Hz), 3.57 (apparent t, ⁴H, J = 4.8 Hz); 13 C NMR (125 MHz, DMSO-d6): 166.26 (d, 1 J FC =257.9 Hz), 159.88 (d, J FC = 4.5 Hz), 158.99 (d, J FC = 241.6 Hz), 147.42 (d, J FC = 4.9 Hz), 136.47 (d, J FC = 12.7 Hz), 130.55, 125.23, 120.82 (d, J FC = 10.1Hz), 107.83 (d, J FC = 24.4 Hz), 107.36 (d, J FC = 10.9 Hz), 100.68 (d, J FC = 7.0Hz), 97.18 (d, J FC = 25.7 Hz), 93.74 (d, J FC = 22.6 Hz), 65.83, 45.03; HRMS(ESI) [M+H] + C 17 H 16 Calculated value of ON3F2: 316.1256, measured value: 316.1246.

[0189] 6-Fluoro-2-(5-Fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole (compound 83). Compound 79 (59 mg, 1.43 x 10⁻⁶) -4Dissolve mol) in TFA (1 mL), stir at ambient temperature for 2.5 h, and then use N 2(g) TFA was removed by flow. The residue was neutralized with aqueous NaHCO3 to give a solid, which was purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 83 (30 mg, 67%) as a grayish-white solid. 1 H NMR (300 MHz, acetone-d6) δ 10.74 (br s, 1 H), 8.47 (t, 1 H, J = 1.8 Hz), 7.78 (dd, 1 H, J = 14.7Hz, J = 2.1 Hz), 7.53 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.10 (dd, 1 H, J =9.9 Hz, J = 2.4 Hz), 6.86 (m, 1 H), 6.84 (ddd, 1 H, J = 9.9 Hz, J = 8.4 Hz, J= 2.4 Hz), 3.51 (m, 4 H), 1.67 (m, 6 H); 13 C NMR (125 MHz, DMSO-d6): 159.12(d, J FC = 235.1 Hz), 149.17 (d, J FC = 255.3 Hz), 148.54 (d, J FC = 6.4 Hz), 139.22 (d, J FC = 4.5 Hz), 137.06 (d, J FC =12.7 Hz), 135.17, 125.53, 120.91 (d,J FC = 10.1 Hz), 120.73, 119.83 (d, J FC = 20.8 Hz), 107.98 (d, J FC = 24.4 Hz), 98.56, 97.22 (d, J FC = 25.5 Hz), 48.37 (d, J FC = 5.7 Hz), 25.49, 24.31; HRMS(ESI) [M+H] + C 18 H 18 Calculated value of N3F2: 314.1463, measured value: 314.1455.

[0190] 4-(3-fluoro-5-(6-fluoro-1H-indol-2-yl)pyridin-2-yl)morpholine (compound 84). Compound 80 (80 mg, 1.93 x 10⁻⁶) -4 Dissolve mol) in TFA (2 mL), stir at ambient temperature for 1.5 h, and then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 84 (60 mg, 98%) as a grayish-white solid. 1 ¹H NMR (300 MHz, acetone-d6) δ 10.79 (br s, ¹H), 8.51 (apparent t, ¹H, J = 1.8 Hz), 7.84 (dd, ¹H, J = 14.4 Hz, J = 1.8 Hz), 7.54 (dd, ¹H, J = 8.7 Hz, J = 5.4 Hz), 7.11 (apparent dd, ¹H, J = 9.9 Hz, J = 2.4 Hz), 6.89 (m, ¹H), 6.85 (ddd, ¹H, J = 9.9 Hz, J = 8.7 Hz, J = 2.4 Hz), 3.78 (apparent t, ⁴H, J = 4.8 Hz), 3.50 (apparent t, ⁴H, J = 4.8 Hz). Hz); 13 C NMR (125 MHz, DMSO-d6): 159.01(d, J FC = 235.2 Hz), 149.21 (d, J FC = 255.4 Hz), 147.86 (d, J FC = 6.8 Hz), 139.13 (d, J FC = 4.7 Hz), 136.94 (d, J FC = 12.7 Hz), 134.79 (d, J FC = 3.2 Hz),125.33, 121.48 (d, J FC = 3.0 Hz), 120.90 (d, J FC = 10.1 Hz), 119.98 (d, J FC =20.6 Hz), 107.98 (d, J FC = 24.4 Hz), 98.82, 97.17 (d, J FC= 25.5 Hz), 65.97,47.67 (d, J FC = 5.4 Hz); HRMS (ESI) [M+H] + C 17 H 16 Calculated value of ON3F2: 316.1256, measured value: 316.1253.

[0191] 2-(4-fluoro-6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 85). Compound 6 (0.30 g, 1.15 mmol) and compound 57 (0.15 g, 5.75 x 10⁻⁶ mmol) were also tested. -4 mol), Pd(dppf)Cl2 (28 mg, 3.83 x10 -5 mol), 1,4-dioxane (30 mL) and K2CO 3(aq) (2 M, 0.7 mL) with N 2(g) Rinse for 10 minutes, in N 2(g) The mixture was stirred overnight at 100ºC, then cooled to ambient temperature and evaporated to dryness. The residue was dissolved in H2O and extracted with EtOAc (10 ml x 3). The extracts were combined, dried over MgSO4, and filtered. The filtrate was passed through a silica gel pad, and the silica was washed with EtOAc. The solvent was removed to obtain the residue, which was purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 85 (0.16 g, 70%) as a solid. 1 ¹H NMR (300 MHz, CDCl₃) δ 8.22 (d, ¹H, J = 10.5 Hz), 8.19 (d, ¹H, J = 7.5 Hz), 7.56 (dm, ¹H, J = 7.5 Hz), 7.33 (ddd, ¹H, J = 8.4 Hz, J = 7.2 Hz, J = 1.5 Hz), 7.24 (td, ¹H, J = 7.5 Hz, J = 1.2 Hz), 6.57 (s, ¹H), 6.36 (d, ¹H, J = 12.9 Hz), 3.83 (apparent t, ⁴H, J = 4.8 Hz), 3.56 (apparent t, ⁴H, J = 4.8 Hz), 1.48 (s, ⁹H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of O3N3F: 398.1875, measured value: 398.1887.

[0192] 2-(5-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 86). Compound 6 (90 mg, 3.45 x 10⁻⁶) -4 mol), compound 58 (90 mg, 3.47 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (19 mg, 2.60 x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2M, 0.5 mL) with N 2(g) Rinse for 5 minutes, then in N 2(g) The mixture was stirred overnight at 100ºC (TLC indicated the presence of unreacted compound 58). Additional compound 6 (85 mg, 3.26 x 10⁻⁶) was added. -4 mol), the reaction mixture in N 2(g) The mixture was stirred at 100ºC for 5 h, then cooled to ambient temperature and evaporated to dryness. The residue was dissolved in H2O and extracted with EtOAc (10 ml x 3). The extracts were combined, dried over MgSO4, and filtered. The filtrate was passed through a silica gel pad, and the silica was washed with EtOAc. The solvent was removed to obtain the residue, which was purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 86 (0.12 g, 87%) as a solid. 1 ¹H NMR (300 MHz, CDCl₃) δ 8.19 (d, 1 H, J = 8.4 Hz), 8.07 (t, 1 H, J = 1.5 Hz), 7.54 (unresolved apparent dd, 1 H, J = 7.8 Hz), 7.33 (partially resolved ddd, 1 H, J = 8.1 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.26 (m- masked by CHCl₃ resonance, 3 H) 6.55 (s, 1 H), 3.50 (m, 4 H), 1.69 (m, 6 H), 1.44 (s, 9 H); HRMS (ESI) [M+H] + C 23 H 27 Calculated value of O2N3F: 396.2082, measured value: 396.2082.

[0193] 2-(5-fluoro-6-morpholinopyridin-3-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 87). Compound 6 (0.17 g, 6.51 x 10⁻⁶) -4 mol), compound 59 (75 mg, 2.87 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (11 mg, 1.50x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 5 minutes, then at 100ºC in N 2(g) Stir overnight. Cool the reaction mixture to ambient temperature and evaporate to dryness. Dissolve the residue in H2O, extract with EtOAc (10 ml x 3), dry the extract with MgSO4, and filter. Pass the filtrate through a silica gel pad and wash the silica with EtOAc. Remove the solvent to obtain the residue, which is purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 87 (92 mg, 80%) as a grayish-white solid. 1 H NMR (400 MHz, CDCl3)δ 8.18 (d, 1 H, J = 8.4 Hz), 8.09 (s, 1 H), 7.55 (d, 1 H, J = 7.6 Hz), 7.33 (m, 2 H), 7.26 (m - shielded by CHCl3 resonance, 1 H), 6.57 (s, 1 H), 3.86 (t, 4 H, J = 4.4 Hz), 3.54 (t, 4 H, J = 4.4 Hz), 1.45 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of O3N3F: 398.1875, measured value: 398.1863.

[0194] 4-(4-fluoro-5-(1H-indol-2-yl)pyridin-2-yl)morpholine (compound 88). Compound 85 (54 mg, 1.36 x 10⁻⁶) -4 mol) was dissolved in TFA (2 mL), stirred at ambient temperature for 2 h, and then N... 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 solution to give a solid, which was washed with cold CH2Cl2 to give compound 88 (30 mg, 74%) as a grayish-white solid. 1¹H NMR (300 MHz, acetone-d6) δ 10.52 (br s, 1 H), 8.66 (d, 1 H, J = 11.4 Hz), 7.56 (d, 1 H, J = 8.1 Hz), 7.42 (d, 1 H, J = 8.1 Hz), 7.10 (partially resolved ddd, 1 H, J = 8.1 Hz, J = 6.9 Hz, J = 1.2 Hz), 7.02 (partially resolved ddd, 1 H, J = 7.8 Hz, J = 7.2 Hz, J = 1.2 Hz), 6.81 (m, 1 H), 6.71 (d, 1 H, J = 14.7 Hz), 3.76 (apparent t, 4 H, J = 4.8 Hz), 3.59 (apparent t, 4 H, J = 4.8 Hz); 13 C NMR (125 MHz, DMSO-d6) δ 166.33 (d, J FC = 257.8 Hz), 159.93 (d, J FC = 11.4 Hz), 147.61 (d, J FC = 3.8 Hz), 136.49, 129.80, 128.42, 121.48, 119.78, 119.32,111.12, 107.52 (d, J FC = 10.1 Hz), 100.69 (d, J FC = 6.3 Hz), 93.66 (d, J FC =22.5 Hz), 65.81, 45.00; HRMS (ESI) [M+H] + C 17 H 17 Calculated value of ON3F: 298.1350, measured value: 298.1343.

[0195] 2-(5-fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole (compound 89). Compound 86 (0.11 g, 2.78 x 10⁻⁶) -4 mol) was dissolved in TFA (2 mL), stirred at ambient temperature for 2 h, and then N... 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 89 (78 mg, 95%) as a grayish-white solid. 1¹H NMR (300 MHz, acetone-d6) δ 10.61 (br s, ¹H), 8.50 (t, ¹H, J = 1.8 Hz), 7.80 (dd, ¹H, J = 14.7 Hz, J = 1.8 Hz), 7.54 (apparent d, ¹H, J = 8.1 Hz), 7.38 (dd, ¹H, J = 8.7 Hz, J = 0.6 Hz), 7.10 (ddd, ¹H, J = 8.1 Hz, J = 6.9 Hz, J = 1.2 Hz), 7.02 (ddd, ¹H, J = 8.1 Hz, J = 6.9 Hz, J = 1.2 Hz), 6.85 (m, ¹H), 3.50 (m, ⁴H), 1.67 (m, 6 H); 13 C NMR (125 MHz, DMSO-d6):149.03 (d, J FC = 255.2 Hz), 148.34 (d, J FC = 6.4 Hz), 139.27 (d, J FC = 4.6 Hz),137.00, 134.25, 128.59, 121.49, 120.81 (d, J FC = 2.8 Hz), 119.82 (d, J FC =20.8 Hz), 119.80, 119.43, 111.08, 98.44, 48.25 (d, J FC = 5.8 Hz), 25.37,24.19; HRMS (ESI) [M+H] + C 18 H 19 Calculated value of N3F: 296.1558, measured value: 296.1550.

[0196] 4-(3-fluoro-5-(1H-indol-2-yl)pyridin-2-yl)morpholine (compound 90). Compound 87 (90 mg, 2.26 x 10⁻⁶ mg) -4 Dissolve mol) in TFA (2 mL), stir at ambient temperature for 1 h, and then use N 2(g) TFA was removed by flow. The residue was neutralized with aqueous NaHCO3 to give a solid, which was purified by radial chromatography to give compound 90 (60 mg, 89%) as a solid. 1¹H NMR (300 MHz, acetone-d6) δ 10.65 (br s, ¹H), 8.54 (apparent t, ¹H, J = 1.8 Hz), 7.86 (dd, ¹H, J = 14.4 Hz, J = 1.8 Hz), 7.55 (d, ¹H, J = 7.8 Hz), 7.39 (partially resolved dd, ¹H, J = 8.1 Hz, J = 0.9 Hz), 7.11 (ddd, ¹H, J = 8.7 Hz, J = 6.9 Hz, J = 1.2 Hz), 7.02 (partially resolved ddd, ¹H, J = 8.1 Hz, J = 7.8 Hz, J = 1.2 Hz), 6.88 (m, ¹H), 3.78 (apparent t, ¹H) (H, J = 4.8 Hz), 3.50 (apparent t, 4H, J = 4.8 Hz); 13 C NMR (125 MHz, DMSO-d6): 149.29 (d, J FC = 255.4 Hz), 147.87 (d, J FC = 6.9 Hz), 139.24 (d, J FC = 4.8 Hz), 137.09, 136.93, 133.83 (d,J FC = 10.2 Hz), 128.49 (d, J FC = 10.4 Hz), 121.65, 120.08 (d, J FC = 20.5 Hz),119.88, 119.49, 111.10, 98.78, 66.01, 47.73 (d, J FC = 5.4 Hz); HRMS (ESI) [M+H] + C 17 H 17 Calculated value of ON3F: 298.1350, measured value: 298.1348.

[0197] 2-(3-fluoro-5-(piperidin-1-yl)pyridin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 91). Compound 6 (0.22 g, 8.43 x 10⁻⁶ g) was used. -4 mol), compound 60 (90 mg, 4.19 x 10⁻⁶) -4 mol), Pd(dppf)Cl2 (19 mg, 2.60 x 10-5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 5 minutes, then in N 2(g) Stirred overnight at 100ºC (TLC indicated the remaining unreacted compound 60). Add the remaining compound 6 (90 mg, 3.45 x 10⁻⁶). -4 mol), and continue in N 2(g) The mixture was stirred overnight at 100ºC, then cooled to ambient temperature and evaporated to dryness. The residue was dissolved in H2O, extracted with EtOAc (10 ml x 3), dried over MgSO4, and filtered. The filtrate was passed through a silica gel pad, and the silica was washed with EtOAc. The solvent was removed to obtain the residue, which was purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give compound 91 (75 mg, 45%) as a solid. 1 ¹H NMR (300 MHz, CDCl₃) δ 8.19 (dd, ¹H, J = 8.4 Hz, J = 0.6 Hz), 8.15 (dd, ¹H, J = 2.7 Hz, J = 1.5 Hz), 7.57 (d, ¹H, J = 7.5 Hz), 7.33 (ddd, ¹H, J = 8.4 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.23 (apparent td - partially blocked by CHCl₃ resonance, ¹H, J = 7.5 Hz, J = 0.9 Hz), 6.91 (dd, ¹H, J = 12.6 Hz, J = 2.4 Hz), 6.78 (s, ¹H), 3.28 (m, 4H), 1.70 (m, 6H), 1.42 (s, 9 H); HRMS (ESI) [M+H] + C 23 H 27 Calculated value of O2N3F: 396.2082, measured value: 396.2082.

[0198] 2-(3-fluoro-5-(piperidin-1-yl)pyridin-2-yl)-1H-indole (compound 92). Compound 91 (75 mg, 1.90 x 10⁻⁶) -4 Dissolve mol) in TFA (1 mL), stir at ambient temperature for 1 h, and then use N 2(g)TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 92 (44 mg, 78%) as a grayish-white solid. 1 ¹H NMR (300 MHz, acetone-d6) δ 10.60 (br s, ¹H), 8.20 (t, ¹H, J = 2.4 Hz), 7.58 (partially resolved dd, ¹H, J = 8.1 Hz), 7.55 (partially resolved dd, ¹H, J = 8.1 Hz, J = 0.9 Hz), 7.19 (dd, ¹H, J = 14.7 Hz, J = 2.4 Hz), 7.12 (ddd, ¹H, J = 8.1 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.01 (ddd, ¹H, J = 7.8 Hz, J = 7.2 Hz, J = 1.2 Hz), 6.94 (m, ¹H), 3.37 (m, ¹H). 1.70 (m, 6 H); 13 C NMR (125 MHz, DMSO-d6): 157.07 (d, J FC = 258.7 Hz), 147.40 (d, J FC = 5.4 Hz), 136.20, 132.75(d, J FC = 7.2 Hz), 132.42, 128.70, 127.30 (d, J FC = 12.2 Hz), 121.70, 120.22,119.15, 111.69, 108.77 (d, J FC = 22.6 Hz), 100.67 (d, J FC = 11.9 Hz), 48.18,24.72, 23.64; HRMS (ESI) [M+H] + C 18 H 19 Calculated value of N3F: 296.1558, measured value: 296.1562.

[0199] 2-(3-fluoro-5-morpholinopyridin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 93). Compound 6 (0.17 g, 6.51 x 10⁻⁶) -4 mol), compound 61 (78 mg, 3.60 x 10⁻⁶ ...). -4mol), Pd(dppf)Cl2 (18 mg, 2.46x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 5 minutes, then at 100ºC in N 2(g) Stir overnight. Cool the reaction mixture to ambient temperature and evaporate to dryness. Dissolve the residue in H2O, extract with EtOAc (10 ml x 3), dry the extract with MgSO4, and filter. Pass the filtrate through a silica gel pad and wash the silica with EtOAc. Remove the solvent to obtain the residue, which is purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:2) to give the recovered compounds 61 (58 mg, 74%) and 93 (29 mg, 20%) as solids. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.18 (d, 1 H, J = 8.4 Hz), 8.15 (s, 1 H), 7.58 (d, 1 H, J = 7.6 Hz), 7.34 (apparent t, 1 H, J = 7.6 Hz), 7.24 (apparent t, 1 H, J = 7.2 Hz), 6.92 (dd, 1 H, J = 12.0 Hz, J = 2.0 Hz), 6.81 (s, 1 H), 3.90 (t, 4 H, J = 4.8 Hz), 3.26 (t, 4 H, J = 4.8 Hz), 1.43 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of O3N3F: 398.1875, measured value: 398.1865.

[0200] 6-Fluoro-2-(3-Fluoro-5-morpholinopyridin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 94). Compound 76 (0.19 g, 6.81 x 10⁻⁶ g) -4 mol), compound 61 (87 mg, 4.02 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (19 mg, 2.60 x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 5 minutes, then in N 2(g)The mixture was stirred overnight at 100ºC (TLC indicated the presence of unreacted compound 61). Additional compound 76 (93 mg, 3.33 x 10⁻⁶) was added. -4 mol), the reaction mixture in N 2(g) The mixture was stirred at 100ºC for 5 h, then cooled to ambient temperature and evaporated to dryness. The residue was dissolved in H2O and extracted with EtOAc (10 ml x 3). The extracts were combined, dried over MgSO4, and filtered. The filtrate was passed through a silica gel pad, and the silica was washed with EtOAc. The solvent was removed to obtain the residue, which was purified by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to obtain the recovered compounds 61 (23 mg, 26%) and 94 (96 mg, 57%) as solids. 1 ¹H NMR (400 MHz, CDCl₃) δ 8.15 (s, 1 H), 7.91 (partially resolved dd, 1 H, J = 10.8 Hz, J = 1.2 Hz), 7.49 (dd, 1 H, J = 8.4 Hz, J = 5.6 Hz), 7.00 (apparent td, 1 H, J = 9.6 Hz, J = 1.6 Hz), 6.92 (apparent dd, 1 H, J = 12.4 Hz, J = 1.6 Hz), 6.76 (s, 1 H), 3.90 (t, 4 H, J = 4.8 Hz), 3.26 (t, 4 H, J = 4.8 Hz), 1.43 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 24 Calculated value of O3N3F2: 416.1780, measured value: 416.1767.

[0201] 2-(4-fluoro-5-morpholinopyridin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 95). Compound 6 (85 mg, 3.26 x 10⁻⁶ mg) -4 mol), compound 62 (78 mg, 3.60 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (18 mg, 2.46x 10 -5 mol), 1,4-dioxane (5 mL) and K2CO 3(aq) (2 M, 0.5 mL) at 100ºC in N 2(g)Stir overnight. Cool the reaction mixture to ambient temperature, evaporate to dryness, dissolve the residue in H2O, and extract with EtOAc (10 ml x 3). Dry the combined extracts in MgSO4, filter, and pass the filtrate through a silica gel pad and wash with EtOAc. Concentrate the solution to dryness and purify the residue by radial chromatography (hexane / EtOAc v / v 95:5 to 70:30) to give recovered compound 62 (32 mg, 41%) and compound 95 (45 mg, 35%). 1 ¹H NMR (300 MHz, CDCl₃) δ 8.28 (d, 1 H, J = 11.1 Hz), 8.15 (d, 1 H, J = 8.1 Hz), 7.57 (d, 1 H, J = 7.8 Hz), 7.35 (partially resolved ddd, 1 H, J = 8.4 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.25 (m- partially blocked by CHCl₃ resonance, 1 H), 7.19 (d, 1 H, J = 12.9 Hz), 6.75 (s, 1 H), 3.90 (apparent t, 4 H, J = 4.5 Hz), 3.21 (apparent t, 4 H, J = 4.5 Hz), 1.42 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 25 Calculated value of O3N3F: 398.1875, measured value: 398.1865.

[0202] 6-Fluoro-2-(4-Fluoro-5-morpholinopyridin-2-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 96). Compound 76 (0.23 g, 8.24 x 10⁻⁶ g) -4 mol), compound 62 (97 mg, 4.48 x 10⁻⁶ ...). -4 mol), Pd(dppf)Cl2 (18 mg, 2.46 x 10 -5 mol), 1,4-dioxane (10 mL) and K2CO 3(aq) (2 M, 0.5 mL) with N 2(g) Rinse for 5 minutes, then in N 2(g)Stir and heat at 100ºC overnight. Cool the reaction mixture to ambient temperature, evaporate to dryness, dissolve the residue in H2O, and extract with EtOAc (10 ml x 3). Dry the combined extracts in MgSO4, filter, and pass the filtrate through a silica gel pad and wash with EtOAc. Concentrate the solution to dryness and purify the residue by radial chromatography (hexane / EtOAc v / v 95:5 to 80:20) to give recovered compounds 62 (68 mg, 70%) and 96 (29 mg, 16%) as solids. 1 H NMR (300 MHz, CDCl3) δ 8.27 (d, 1 H, J = 10.8 Hz), 7.88 (dd,1 H, J = 10.5 Hz, J = 2.4 Hz), 7.48 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.18(d, 1 H, J = 12.9 Hz), 7.01 (td, 1 H, J = 9.0 Hz, J = 2.4 Hz), 6.70 (s, 1 H), 3.90 (m – AA'XX', 4 H, J = 4.8 Hz, J = 1.8 Hz), 3.21 (m – AA'XX', 4 H, J =4.8 Hz, J = 1.8 Hz), 1.41 (s, 9 H); HRMS (ESI) [M+H] + C 22 H 24 Calculated value of O3N3F2: 416.1780, measured value: 416.1767.

[0203] 4-(5-fluoro-6-(1H-indol-2-yl)pyridin-3-yl)morpholine (compound 97). Compound 93 (29 mg, 7.30 x 10⁻⁶) was used. -5 Dissolve mol) in TFA (1 mL), stir at ambient temperature for 2 h, and then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 97 (17 mg, 78%) as a grayish-white solid. 1¹H NMR (400 MHz, acetone-d6) δ 10.65 (br s, ¹H), 8.22 (t, ¹H, J = 2.0 Hz), 7.60 (unresolved dq, ¹H, J = 8.0 Hz), 7.56 (partially resolved dq, ¹H, J = 8.0 Hz, J = 0.8 Hz), 7.24 (dd, ¹H, J = 14.4 Hz, J = 2.4 Hz), 7.14 (ddd, ¹H, J = 8.8 Hz, J = 7.2 Hz, J = 1.2 Hz), 7.02 (ddd, ¹H, J = 7.8 Hz, J = 7.2 Hz, J = 1.2 Hz), 6.98 (dm, ¹H, J = 4.4 Hz). 3.81 (apparent t, 4 H, J = 4.8 Hz), 3.32 (partially resolved m - AA'XX', 4 H, J = 4.8 Hz); 13 C NMR (125 MHz, DMSO-d6): 156.94 (d,J FC = 258.6 Hz), 147.20 (d, J FC = 4.3 Hz), 136.24, 132.53 (d, J FC = 6.4 Hz),132.14, 128.65, 128.46 (d, J FC = 10.9 Hz), 121.88, 120.33, 119.22, 111.74,108.95 (d, J FC = 22.1 Hz), 101.06 (d, J FC = 11.9 Hz), 65.69, 47.23; HRMS (ESI)[M+H] + C 17 H 17 Calculated value of ON3F: 298.1350, measured value: 298.1353.

[0204] 4-(5-fluoro-6-(6-fluoro-1H-indol-2-yl)pyridin-3-yl)morpholine (compound 98). Compound 94 (31 mg, 7.46 x 10⁻⁶) -5 Dissolve mol) in TFA (1 mL), stir at ambient temperature for 1 h, and then use N 2(g)TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 98 (23 mg, 98%) as a grayish-white solid. 1 ¹H NMR (300 MHz, acetone-d6) δ 10.77 (br s, 1 H), 8.21 (t, 1 H, J = 1.8 Hz), 7.58 (dd, 1 H, J = 8.7 Hz, J = 5.4 Hz), 7.30 (dd, 1 H, J = 10.2 Hz, J = 2.4 Hz), 7.23 (dd, 1 H, J = 14.4 Hz, J = 2.4 Hz), 6.96 (dm, 1 H, J = 4.2 Hz), 6.85 (ddd, 1 H, J = 9.9 Hz, J = 8.7 Hz, J = 2.4 Hz), 3.81 (apparent t, 4 H, J = 4.8 Hz), 3.32 (apparent t, 4 H, J = 4.8 Hz); 13 C NMR (125 MHz, DMSO-d6):159.18 (d, J FC = 235.3 Hz), 156.82 (d, J FC = 258.8 Hz), 147.26 (d, J FC = 5.5Hz), 136.24 (d, J FC = 13.2 Hz), 133.33 (unresolved m), 132.13 (d, J) FC = 2.8 Hz), 128.15 (d, J FC = 11.9 Hz), 125.49, 121.46 (d, J FC = 10.2 Hz), 108.93 (d, J FC =22.8 Hz), 107.83 (d, J FC = 24.5 Hz), 101.06 (d, J FC = 12.1 Hz), 97.51 (d, J FC =25.7 Hz), 65.69, 47.21; HRMS (ESI) [M+H] + C 17 H 16 Calculated value of ON3F2: 316.1256, measured value: 316.1258.

[0205] 4-(4-fluoro-6-(1H-indol-2-yl)pyridin-3-yl)morpholine (compound 99). Compound 95 (40 mg, 1.01 x 10⁻⁶) -4 Dissolve mol) in TFA (1.5 mL), stir at ambient temperature for 1.5 h, and then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with hexane and cold CH2Cl2 to give compound 99 (29 mg, 97%) as a grayish-white solid. 1 H NMR (300 MHz, acetone-d6) δ 10.73 (br s, 1 H), 8.28 (d,1 H, J = 11.1 Hz), 7.72 (d, 1 H, J = 13.8 Hz), 7.58 (d, 1 H, J = 8.1 Hz), 7.53 (d, 1 H, J = 8.1 Hz, J = 0.9 Hz), 7.14 (ddd, 1 H, J = 8.1 Hz, J = 7.2Hz, J = 1.2 Hz), 7.07 (m, 1 H), 7.02 (ddd, 1 H, J = 7.8 Hz, J = 7.2 Hz, J =0.9 Hz), 3.82 (m – AA'XX', 4 H, J = 4.8 Hz, J = 1.8 Hz), 3.19 (m – AA'XX', 4H, J = 4.8 Hz, J = 1.8 Hz); 13 C NMR (125 MHz, DMSO-d6): 160.55 (d, J FC = 258.7Hz), 146.18 (d, J FC = 8.0 Hz), 140.75 (unresolved d), 137.10, 136.37 (unresolved d), 134.62 (d, J FC = 5.2 Hz), 128.33, 122.15, 120.43, 119.43, 111.87, 107.55 (d,J FC = 18.4 Hz), 100.24, 65.99, 50.08; HRMS (ESI) [M+H] + C 17 H 17 Calculated value of ON3F: 298.1350, measured value: 298.1347.

[0206] 4-(4-fluoro-6-(6-fluoro-1H-indol-2-yl)pyridin-3-yl)morpholine (compound 100). Compound 96 (29 mg, 6.98 x 10⁻⁶) -5 Dissolve mol) in TFA (1 mL), stir at ambient temperature for 2 h, and then use N 2(g) TFA was removed by flushing. The residue was neutralized with aqueous NaHCO3 to give a solid, which was washed with cold CH2Cl2 to give compound 100 (21 mg, 95%) as a grayish-white solid. 1 H NMR (300 MHz, acetone-d6) δ 10.86 (br s, 1 H), 8.27 (d, 1 H,J = 11.1 Hz), 7.71 (d, 1 H, J = 13.8 Hz), 7.57 (dd, 1 H, J = 8.7 Hz, J = 5.4Hz), 7.26 (dd, 1 H, J = 9.9 Hz, J = 2.4 Hz), 7.08 (m, 1 H), 6.85 (ddd, 1 H, J= 9.9 Hz, J = 8.7 Hz, J = 2.4 Hz), 3.82 (m – AA'XX', 4 H, J = 4.8 Hz, J = 1.8Hz), 3.19 (m – AA'XX', 4 H, J = 4.8 Hz, J = 1.8 Hz); 13 C NMR (125 MHz, DMSO-d6): 160.56 (d, J FC = 258.8 Hz), 159.30 (d, J FC = 235.7 Hz), 145.85 (d, J FC =8.3 Hz), 140.71 (d, J FC = 3.0 Hz), 137.16 (dd, J FC = 3.6 Hz), 137.03 (d, J FC =13.2 Hz), 134.69 (d, J FC = 5.8 Hz), 125.17, 121.59 (d, J FC = 10.2 Hz), 108.11(d, J FC = 24.5 Hz), 107.46 (d, J FC = 18.5 Hz), 100.26, 97.63 (d, J FC= 25.8Hz), 65.99, 50.07 (d, J FC = 2.6 Hz); HRMS (ESI) [M+H] + C 17 H 16 Calculated value of ON3F2: 316.1256, measured value: 316.1245.

[0207] 2-(2-(4-(2-(2-fluoroethoxy)ethoxy)piperidin-1-yl)pyrimidin-5-yl)-1H-indole-1-carboxylic acid tert-butyl ester (compound 101). Compound 44 (79 mg, 2.00 x 10⁻⁶) -4 mol) with N 2(g) Rinse, dissolve in DMA (1 mL) and cool to 0ºC. Add NaH (60%, 26 mg, 6.50 x 10⁻⁶ ppm). -4 mol (3.2 equivalents), the mixture was incubated at 0ºC in N 2(g) Stir for 5 min, then add 2-(2-fluoroethoxy)ethyl 4-toluenesulfonic acid (110 mg, 4.19 x 10⁻⁶ oz). -4 A solution of 2.1 mol (2.1 equivalents) in DMA (0.5 mL). The reaction mixture was incubated at ambient temperature in N2O2. 2(g) Stir for 2 h, then add H2O (5 drops), stir the mixture for 10 min, and then pour it into a mixture of H2O (20 mL), EtOAc (20 mL), and hexane (10 mL). Mix the layers, separate them, and wash the organic layer with H2O (10 mL x 2), brine (10 mL), and dry with MgSO4. Remove the solvent to obtain a yellow residue (138 mg), which is dissolved in CH2Cl2, poured onto dry silica (55 cm h x 45 cm id), and eluted under vacuum: CH2Cl2 (100 mL); % MeOH / CH2Cl2 - 1% (100 mL), 2% (100 mL), 3% (100 mL), 5% (100 mL) to obtain a slightly brown paste (126 mg). Compound 101 (32 mg, 33%) was purified by radial chromatography (1 mm silica): CH2Cl2 (50 mL); % MeOH / CH2Cl2 - 1% (50 mL), 2% (25 mL) to obtain a colorless residue. 1¹H NMR (300 MHz, CDCl₃) δ 8.48 (s, 2 H), 7.62 (d, 1 H, J = 7.8 Hz), 7.41 (d, 1 H, J = 8.1 Hz), 7.24 (td, 1 H, J = 8.1 Hz, J = 0.9 Hz), 7.15 (partially resolved td, 1 H, J = 7.8 Hz, J = 0.9 Hz), 6.50 (s, 1 H), 4.86 (apparent septet, 1 H, J = 4.2 Hz), 4.52 (apparent t, 1 H, J = 4.2 Hz), 4.39 (m, 3 H), 4.30 (t, 2 H, J = 6.0 Hz), 3.80 (t, 2 H, J = 6.0 Hz). Hz), 3.55 (m, 4 H), 2.06 (m, 2H), 1.77 (m, 2 H), 1.52 (s, 9 H).

[0208] 2-(2-(4-(2-(2-fluoroethoxy)ethoxy)piperidin-1-yl)pyrimidin-5-yl)-1H-indole (compound 102). Compound 101 (31 mg, 6.40 x 10⁻⁶) -5 The mixture was stirred for 20 min with 0.65 mL of TFA (8.4 mmol, 132 equivalents) and then poured into a mixture of NaHCO3 (0.79 g, 9.40 mmol, 1.1 equivalents of TFA), H2O (25 mL), and CH2Cl2 (25 mL). The mixture was stirred for 5 min, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (5 mL x 2). The combined CH2Cl2 layers were washed with brine (20 mL), dried over MgSO4, concentrated, poured onto dry silica (33 mm h x 33 mm i.d.), and eluted under vacuum: % MeOH / CH2Cl2 - 1% (50 mL), 2% (75 mL), 3% (100 mL) to give compound 102 (16 mg, 65%) as a grayish-white solid. 1¹H NMR (MHz, CDCl₃) δ 8.48 (s, 2H), 7.62 (d, 1H, J = 7.6 Hz), 7.41 (d, 1H, J = 8.0 Hz), 7.24 (td, 1H, J = 8.0 Hz, J = 0.8 Hz), 7.15 (partially resolved td, 1H, J = 7.6 Hz, J = 0.8 Hz), 6.50 (s, 1H), 4.48 (dt, 2H, J = 13.4 Hz, J = 4.4 Hz), 4.44 (dt, 2H, 2 J HF = 48.0 Hz, J = 4.4 Hz), 4.30 (t, 2 H, J = 6.0 Hz), 4.01 (apparent septet, 1 H, J = 4.4 Hz), 3.80 (t, 2 H, J = 6.0 Hz), 3.55 (dt, 2 H, 3 J HF = 29.2 Hz, J = 4.4 Hz), 3.42 (ddd, 2H, J = 13.4 Hz, J = 10.0 Hz, J = 3.6 Hz), 2.01 (m, 2 H), 1.58 (m, 2 H); HRMS(ESI) [M+H] + C 21 H 26 Calculated value of FN4O2: 385.2034, measured value: 385.2041.

[0209] 4-(5-(6-fluoro-1-methyl-1H-indol-2-yl)pyrimidin-2-yl)morpholine (compound 103). Compound 2 (21 mg, 7.04 x 10⁻⁶) -5 mol) with N 2(g) Rinse, then dissolve in DMA (1 mL) and cool to 0ºC. Add NaH (60%, 9.6 mg, 2.40 x 10⁻⁶ ppm). -4 (mol, 3.4 equivalents), the reaction mixture was incubated at 0ºC in N2. 2(g) Stir for 18 min, then add MeOTs (26.5 mg, 1.42 x 10⁻⁶). -4mol, 2 equivalents). The reaction mixture was warmed to ambient temperature and stirred for 2 h, then H2O (1 drop) was added. The reaction mixture was combined with EtOAc (15 mL), hexane (5 mL) and H2O (10 mL), the layers were mixed and separated, and the organic layer was washed with H2O (10 mL x 2), brine (10 mL) and dried over MgSO4. The solvent was removed to give an off-white solid, which was dissolved in CH2Cl2, poured onto dry silica (33 mm h x 33 mm id) and eluted under vacuum: % MeOH / CH2Cl2 - 1% (100 mL), 2% (50 mL), 3% (50 mL) to give compound 103 (19 mg, 86%) as an off-white solid. 1 H NMR (400 MHz, CDCl3) δ 8.46 (s, 2 H), 7.52 (dd, 1 H,J = 8.4 Hz, J = 5.4 Hz), 7.02 (dd, 1 H, J = 10.0 Hz, J = 2.0 Hz), 6.91 (td, 1H, J = 9.0 Hz, J = 2.0 Hz), 6.49 (s, 1 H), 3.88 (m, 4 H), 3.81 (m, 4 H), 3.67(s, 3 H); HRMS (ESI) [M+H] + C 17 H 18 Calculated value of FN4O: 313.1459, measured value: 313.1455.

[0210] 6-Fluoro-1H-indole-1-carboxylic acid tert-butyl ester (compound 104). 6-Fluoro-indole (3.28 g, 24.27 mmol), Boc₂O (6.59 g, 30.19 mmol, 1.2 equivalents), i-Pr₂NEt (7 mL, 40.2 mmol, 1.7 equivalents), DMAP (0.31 g, 2.54 mmol, 0.1 equivalents), and CH₂Cl₂ (210 mL) were reacted at ambient temperature in an N₂ atmosphere. 2(g) The mixture was stirred for 16 h. The reaction mixture was concentrated into a yellow / orange oil, hexane was added, and the mixture was purified by vacuum rapid chromatography on silica (16 cm h x 4 cm id): hexane (100 mL); % CH2Cl2 / hexane - 10% (200 mL), 25% (400 mL) to give compound 104 (5.67 g, 99%) as a colorless oil. 1H NMR (300 MHz, CDCl3) δ 7.86 (br d, 1 H,J = 10.2 Hz), 7.56 (d, 1 H, J = 3.6 Hz), 7.46 (dd, 1 H, J = 8.4 Hz, J = 5.4Hz), 6.98 (td, 1 H, J = 9.0 Hz, J = 2.4 Hz), 6.53 (d, 1 H, J = 3.6 Hz), 1.67 (s, 9 H).

[0211] Example 3. In vitro binding assay

[0212] As determined by analytical HPLC, the purity of all screened compounds was >95%. Binding assays were performed as described above (Graham, TJA et al., Journal of Medicinal Chemistry 2023, 66, 10628-10638), and the results are shown in Table 2. a) n = 1. nd = Undetermined.

[0213] Example 4. In vitro competition (K i ) Measurement.

[0214] Using the published method (Klunk et al., “The binding of 2-(4'-methylaminophenyl)benzothiazole to postmortem brain homogenates is dominated by the amyloid component.” J Neurosci 2003, 23, 2086-2092), the equilibrium inhibition constant (K0) of the compound was determined relative to a tritium-labeled radioligand. iThe assay utilized autopsy-confirmed postmortem human brain tissue of AD, PSP, and CBD patients, obtained from the UCSF Neurodegenerative Disease Brain Bank, containing frequently autopsy-confirmed 3R / 4R-τ protein and amyloid-β aggregates (AD tissue) or containing only 4R-τ protein aggregates (PSP and CBD tissue) and no other detectable aggregated amyloid material. Briefly, the tissue was homogenized on ice at 300 mg / mL in ice-cold pH 7.0 phosphate-buffered saline (PBS), diluted 30-fold to 10 mg / mL with PBS, and a second homogenization was performed using a Brinkmann Polytron homogenizer before storage at -80ºC. Frozen brain tissue was thawed and diluted 10-fold to 1 mg / mL in PBS. The concentration of unlabeled competing agent compounds (approximately 400 μM in stock solution) was determined by quantitative NMR in DMSO (0.25% DMSO in the final assay vial). An appropriate concentration (range 0.1–1000 nM) of unlabeled competitive agent in 400 μL PBS buffer was combined with 500 μL of tritium-labeled radioligand in PBS (approximately 1 nM final concentration of radioligand). Assays were initiated by adding 100 μL of 1 mg / mL brain tissue homogenate to achieve a final concentration of 100 µg tissue / mL. After incubation at room temperature for 60 min, the binding mixture was filtered through a Whatman GF / B glass filter using a Brandel M-24R cell harvester (Gaithersburg, MD, USA) and rapidly washed four times with 3 mL PBS buffer. The filter membrane was counted in a Cytoscint-ES after thorough vortexing using a liquid scintillation counter. Complete (100%) inhibition of specific binding was defined as the number of counts displaced by 1 μM of unlabeled competitive agent. All assays were performed in triplicate at each concentration. 50% inhibition of radioligand binding (IC50) was determined by the concentration of the radioligand. 50 The Ki value is determined by the inhibitor concentration: K i = IC 50 / 2.

[0215] Example 5. Binding affinity (K) d ) Measurement

[0216] Tritium-labeled radioligand binding assays utilize brain AD, PSP, or CBD homogenates to determine the equilibrium dissociation constant (K0). dThe assay was performed using a slightly modified version of the procedure described in previous details (Klunk et al., “Uncharged thioflavin-T derivatives bind to amyloid-beta protein with high affinity and readily enter the brain.” Life Sciences 2001, 69(13), 1471-1484). In short, frozen aliquots (-80ºC) of homogenized cortical tissue from AD, PSP, or CBD brains (10 mg / mL in PBS (pH = 7.0)) were thawed and diluted 10-fold to 1 mg / mL in PBS. The unlabeled test compound was dissolved in DMSO at 400 µM and then diluted to 20 µM with PBS to produce a 5% DMSO / PBS solution. The remaining serial dilutions (typically from 6 µM to 4 nM) were prepared with 5% DMSO / PBS to maintain a constant DMSO concentration in the final assay. 50 µL of these solutions were combined with 50 µL of the tritium test compound and 800 µL of PBS to produce 0.25% DMSO, approximately 1 nM of the tritium compound, and 0.2 to 1000 nM of unlabeled compound in the final assay. Assays were initiated by adding 100 μL of 1 mg / mL brain homogenate to achieve a final concentration of 100 µg tissue / mL. After incubation at room temperature for 60 min, the binding mixture was filtered through a Whatman GF / B glass filter using a Brandel M-24R cell harvester (Gaithersburg, MD) and rapidly washed three times with 3 mL of PBS. After thorough vortexing and overnight incubation, the filter membrane was counted in a Cytoscint ES. All assays were performed in at least triplicate. The concentration of bound compounds was determined by the radioactivity retained on the filter after adjusting for non-displaceable radioactivity (defined as the radioactivity remaining in the presence of approximately 1 µM of unlabeled compound) and by the specific activity of the tritium compound after dilution with different concentrations of unlabeled compound. The slope of the Scatcherd plot of bound / free radioligand values ​​versus bound radioligand values ​​at different ligand concentrations (slope = -1 / K) was used. d To determine K d value.

[0217] Example 6. X-ray crystal structure data

[0218] Example 7. In tau protein disease tissue samples [ 3 H]1([ 3 H]Z-3540) and [ 3 H]2([ 3 Radiosynthesis and in vitro evaluation of H]Z-2340.

[0219] Z-3540 (Enamine Z3777013540, compound 1) and Z-2340 (Enamine Z4169252340, compound 2) were identified by molecular similarity fingerprinting of an Enamine compound library based on CBD-2115. Synthesis [ 3 H]Z-3540 and [ 3 H]Z-2340 was evaluated in binding assays in human CBD, PSP, and AD tissues.

[0220] In human organizations, [ 3 H]Z-3540's K d In AD, it is [ 3 H]CBD-2115's K d 1 / 3 and 1 / 7 of it in the CBD. [Found] 3 H]Z-3540 is a tau protein that selectively binds to and aggregates relative to β-amyloid and α-synuclein.

[0221] Example 8. Radiochemistry

[0222] The X-ray crystal structures of radiolabeled precursors 65 and 66 are shown in Figures 2 and 3, respectively. The process was performed manually using a manipulator in a lead-shielded hot chamber. 18 F]68 and [ 18 Radioactive labeling of F]69. [ 18 F] Fluoride was captured on WatersSep-Pak Plus Light QMA and then treated with Et4NHCO 3(aq) Elution to obtain Et4N 18 F, then azeotropically dry it with CH3CN. Dissolve nitro precursor 65 or 66 in DMSO and add it to the dried Et4N in a sealed 5-mL V-shaped vial. 18 In step F, the mixture was heated at approximately 140ºC for 30 minutes, and then the crude reaction mixture was purified by semi-preparative HPLC. The radioactive tracer was obtained in a 10% EtOH / 0.9% sterile saline solution (pH 5). 18 F]68, with a non-decay-corrected radiochemical yield of 4.4% (1.18 GBq (31.9 mCi)); radiochemical purity of 99.4%; and molar activity of A. m= 0.13 GBq / nmol (3.5 mCi / nmol). The radioactive tracer was obtained in a 10% EtOH / 0.9% sterile saline solution (pH 5). 18 F]69, with a non-decay-corrected radiochemical yield of 11.5% (3.49 GBq (94.3 mCi)); radiochemical purity of 98.9%; and molar activity of A. m = 0.05 GBq / nmol (1.4 mCi / nmol); logD = 1.96 ± 0.03 (n = 4). The radiochemical characterization of the radiotracer was confirmed by analytical HPLC co-injection with unlabeled standards.

[0223] Option 15

[0224] 2-(2-[ 18 F]Fluoro-6-(piperidin-1-yl)pyridin-3-yl)-1H-indole ([ 18 [F]68). Et4NHCO3 (70 mg) was dissolved in Milli-Q H2O (10 mL) to obtain a 7 mg / mL solution. Waters Sep-Pak Plus Light QMA (WAT023525) was dissolved in Et4NHCO3. 3(aq) The solution was applied (8 mL, 7 mg / mL), then rinsed with Milli-Q H2O (10 mL), followed by air (25 mL). Waters C 18 Sep-Pak (WAT020515) was rinsed with EtOH (10 mL) and then with Milli-Q H2O (10 mL). 18 O(p,n) 18 The F reaction (Siemens Eclipse HP cyclotron) produces carrier-free [ 18 F]F - , and in [ 18 Transferred from O]H2O to a lead-shielded hot chamber. 18 F]F - (26.6 GBq (719 mCi)) captured on QMA and with Et4NHCO 3(aq) The solution (1 mL, 7 mg / mL) was eluted, followed by elution with CH3CN (1 mL) (0.45 GBq (12.2 mCi) retained on QMA), and collected in 5-mL V-tubes. The solvent was evaporated at 110ºC under an argon flow, and then Et4N was collected. 18F was aliquoted with CH3CN (1 mL x 3) and azeotropically dried at 110ºC under an argon atmosphere. Compound 65 (3.8 mg) was dissolved in DMSO (0.6 mL) and added to the dried Et4N. 18 In step F, cap the V-shaped vial and gently mix for about 10 seconds until the solution turns a light pink color, then heat at about 140ºC for 30 min. Cool the V-shaped vial for about 1 min, dilute the solution with HPLC solvent (0.5 mL, 55:45 v / v CH3CN / 0.1 M NH4HCO2 pH 4.2), load it into the HPLC loop, rinse the V-shaped vial with HPLC solvent (0.5 mL), load the solvent into the HPLC loop, and purify the sample by preparative HPLC (Phenomenex Gemini, 250 x 10 mm + guard column; 5 mL / min for 6 min, then 10 mL / min). Collect the HPLC radioactive peak at about 24–25 min, dilute with Milli-Q H2O (50 mL), and purify by C 18 Sep-Pak was prepared and rinsed with Milli-Q H2O (10 mL). The radioactive tracer was eluted from Sep-Pak with EtOH (1 mL) and then with 0.9% sterile saline (9 mL), passed through a 0.2-micron filter, and collected in a sterile vial to obtain [ 18 F]68 (1.18 GBq (31.9 mCi); radiochemical yield 4.4% (uncorrected for decay); radiochemical purity 99.4% (Phenomenex Gemini, 5 μm, NX-C18 110 Å, 100 x 4.6 mm, 60:40 v / v CH3CN / 0.1 MNH4HCO2 pH 4.2; 1 mL / min; 254 nm); A m = 0.13 GBq / nmol (3.5 mCi / nmol); pH 5). Radiochemical characterization was confirmed by co-injection with 68. Endotoxin level of the final preparation < 2.00 EU / mL (CharlesRivers Endosafe®).

[0225] 4-(6-[ 18 F]Fluoro-5-(1H-indol-2-yl)pyridin-2-yl)morpholine ([ 18 F]69)([ 18[F]JSS20-183A). Dissolve 70 mg of Et4NHCO3 in 10 mL of Milli-Q H2O to obtain a 7 mg / mL solution. Dissolve Waters Sep-PakPlus Light QMA (WAT023525) in Et4NHCO3. 3(aq) The solution was applied (8 mL, 7 mg / mL), then rinsed with Milli-Q H2O (10 mL), followed by air (25 mL). Waters C 18 Sep-Pak (WAT020515) was rinsed with EtOH (10 mL) and then with Milli-Q H2O (10 mL). 18 O(p,n) 18 The F reaction (Siemens Eclipse HP cyclotron) produces carrier-free [ 18 F]F - , and in [ 18 Transferred from O]H2O to a lead-shielded hot chamber. 18 F]F - (30.3 GBq (818 mCi)) captured on QMA and with Et4NHCO 3(aq) The solution (1 mL, 7 mg / mL) was eluted, followed by elution with CH3CN (1 mL), then with air (1 mL) (0.3 GBq (8.0 mCi) retained on QMA), and collected in 5-mL V-tubes. The solvent was evaporated at 110ºC under an argon flow, and then Et4N was collected. 18 F was aliquoted with CH3CN (1 mL x 3) and azeotropically dried at 110ºC under an argon atmosphere. Compound 66 (2.5 mg) was dissolved in DMSO (0.5 mL) and added to the dried Et4N. 18 In F (the solution turns pale purple), cap the V-shaped vial and heat at approximately 140ºC for 30 min. Cool the V-shaped vial for approximately 1 min, dilute the solution with HPLC solvent (0.5 mL, 45:55 v / v CH3CN / 0.1 M NH4HCO2 pH 4.2), load it into the HPLC loop, rinse the V-shaped vial with HPLC solvent (0.5 mL), load the solvent into the HPLC loop, and purify the sample by preparative HPLC (Phenomenex Gemini, 250 x 10 mm + guard column; 5 mL / min for 6 min, then 8 mL / min). Collect the HPLC radioactive peak at approximately 20.4–22 min, dilute with Milli-Q H2O (50 mL), and purify by C 18Sep-Pak was prepared and rinsed with Milli-Q H2O (10 mL). The radioactive tracer was eluted from Sep-Pak with EtOH (1 mL) and then with 0.9% sterile saline (9 mL), passed through a 0.2-micron filter, and collected in a sterile vial to obtain [ 18 F]69 (3.49 GBq (94.3 mCi); radiochemical yield 11.5% (uncorrected for decay); radiochemical purity 98.9% (Phenomenex Gemini, 5 μm, NX-C18 110 Å, 100 x 4.6 mm, 45:55 v / v CH3CN / 0.1 M NH4HCO2 pH 4.2; 1 mL / min; 254 nm); A m = 0.05 GBq / nmol (1.4 mCi / nmol); pH 5; logD = 1.96 ± 0.03 (n = 4)). Radiochemical characterization was confirmed by co-injection with 69. Endotoxin level of the final preparation < 2.00 EU / mL (Charles Rivers Endosafe®).

[0226] Example 9. 3 H]69([ 3 Binding affinity of H]JSS20-183A)

[0227] Identify in post-mortem brain tissue [ 3 H]69([ 3 Binding affinity of H]JSS20-183A (Table 7). 3 H]69([ 3 H]JSS20-183A) showed high affinity in postmortem human AD, PSP, CBD, and Pick's disease brain tissue, as well as in P301L mouse brain tissue. 3 H]69([ 3 Compound 69 (H]JSS20-183A) showed low affinity in post-mortem brain tissue of Parkinson's disease (PD), thus demonstrating that 69 does not bind strongly to α-synuclein. In post-mortem human AD brain tissue, compound 69 binds to […]. 3 The poor competition from H]PiB demonstrates that 69 does indeed bind to the PiB binding site on amyloid-β.

[0228] Example 10. Brain PET Imaging Study

[0229] Evaluation through brain PET imaging studies of rhesus monkeys [ 18 F]68 (Figure 5) and [18 Brain uptake and rinsing of F]69 (Fig. 6). 18 The ratio of 2 minutes to 60 minutes for F]68 is 1.2 (whole brain), 1.3 (cortex), and 2.0 (cerebellum). 18 The ratio of 2 minutes to 60 minutes for F]69 was 3.0 (whole brain), 3.7 (cortex), and 3.8 (cerebellum).

[0230] Example 11. Autoradiography

[0231] Autoradiography has demonstrated the presence of PSP, CBD, and AD in postmortem human brain tissue. 3 Specific binding of H]69 was observed (Figure 7). Specific binding was almost entirely absent in post-mortem human brain control tissue. ***

[0232] While certain embodiments have been described and illustrated, it should be understood that changes and modifications may be made to the present invention in accordance with common art without departing from the broader aspects of the invention as defined in the appended claims.

[0233] The embodiments described herein may be suitably practiced in the absence of any one or more elements or limitations not expressly disclosed herein. Therefore, terms such as “comprising,” “including,” and “containing” should be interpreted broadly and without limitation. Furthermore, the terms and expressions used herein have been used as descriptive rather than restrictive terms and are not intended to exclude any equivalent forms of the features shown and described or portions thereof, but it should be recognized that various modifications are possible within the scope of the claimed technology. Additionally, the phrase “consistently composed of” will be understood to include those elements expressly stated and those additional elements that do not substantially affect the essential and novel features of the claimed technology. The phrase “consisting of” does not include any unspecified elements.

[0234] This disclosure is not limited to the specific embodiments described herein. As will be apparent to those skilled in the art, many modifications and alterations can be made without departing from the spirit and scope of this disclosure. It will be apparent from the foregoing description to those skilled in the art that functionally equivalent methods and compositions other than those listed herein exist within the scope of this disclosure. Such modifications and alterations are intended to fall within the scope of the appended claims. This disclosure is limited only by the claims of the appended claims and the full scope of equivalents referred to in such claims. It should be understood that this disclosure is not limited to specific methods, reagents, compounds, or compositions, which can, of course, vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be restrictive.

[0235] Furthermore, where features or aspects of this disclosure are described in accordance with the Markush group, those skilled in the art will recognize that this disclosure is therefore also described in accordance with any individual member or subgroup of the Markush group.

[0236] As those skilled in the art will understand, for any and all purposes, particularly for the purpose of providing a written description, all scopes disclosed herein also encompass any and all possible subscopes and combinations thereof. Any listed scope can be readily identified as adequately describing the same scope and capable of being decomposed into at least two, three, four, five, ten, etc., equal parts. As a non-limiting example, each scope discussed herein can be readily decomposed into a lower third, a middle third, and an upper third, etc. As those skilled in the art will also understand, all language such as “at most,” “at least,” “greater than,” “less than,” etc., includes the listed numbers and refers to a scope that can subsequently be decomposed into subscopes as discussed above. Finally, as those skilled in the art will understand, a scope includes each individual member.

[0237] All publications, patent applications, granted patents, and other documents mentioned in this specification are incorporated herein by reference as if each individual publication, patent application, granted patent, or other document were expressly and separately incorporated by reference in its entirety. Definitions contained in the text incorporated by reference are excluded to the extent that they contradict the definitions in this disclosure.

[0238] Other embodiments are set forth in the following claims.

Claims

1. A compound having the structure of Formula I or Formula II, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof: Formula I Formula II in: X1 is N, O, or S; X2, X3, X4, and X5 are each independently N or CH; X6 is CH or O; R1 is H, halogen, CN, Me3N + X7 - Et3N + X7 - Pr3N + X7 - Bu3N + X7 - Or other suitable ammonium salts, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin substances, I(O-acyl)2, ArI + X7 - IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, β-pinacol, β-neopentyl glycol, or other suitable boron substances, provided that when R1 is OH, R1 is not substituted at position 5 of the indole ring; Among them, X7 - It is a suitable counterion; among which ArI + X7 - It is a suitable iodonium substance known in the art; wherein IR7 is a suitable iodonium ylide known in the art; R2 is H, alkyl, tert-butoxycarbonyl, toluenesulfonyl, benzyl, p-methoxybenzyl, CBz, Fmoc, or other suitable nitrogen protecting group; R3 is H, halogen, NO2, Me3N + X7 - Et3N + X7 - Pr3N + X7 - Bu3N + X7 - Or other suitable ammonium salts, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin substances, I(O-acyl)2, ArI + X7 - IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, β-pinacol, β-neopentyl glycol, or other suitable boron substances; Among them, X7 - It is a suitable counterion; among which ArI + X7 - It is a suitable iodonium substance known in the art; wherein IR7 is a suitable iodonium ylide known in the art; R4 is H, halogen, OH, alkoxy, carbonyl, or OEtOEtF; R5 is H, halogen, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances; R6 is H, halogen, OH, O-acyl, O-alkyl, OBoc, or other suitable oxygen-protecting substance; Or R5 and R6 can form an epoxide: ; m and n are each independently 0 or 1; and o can be 0, 1, or 2.

2. The compound according to claim 1, wherein the compound has the following structure: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

3. The compound according to claim 1, wherein the compound has the following structure: , , , , , , , , , , or .

4. The compound according to any one of the preceding claims, wherein the compound selectively binds to the 4Rτ protein.

5. The compound according to any one of the preceding claims, wherein the compound comprises one or more detection markers.

6. The compound according to claim 5, wherein the detection label is selected from radionuclides, positron emitters, gamma emitters, and fluorescent labels.

7. The compound according to claim 5 or claim 6, wherein the detection marker is selected from... 2 H, 3 H, 18 F, 123 I, 124 I, 125 I, 131 I, 11 C 13 C 14 C 13 N、 15 N、 15 O、 75 Br、 76 Br and 77 Br.

8. The compound according to any one of claims 5 to 7, wherein the detection marker is 18 F.

9. The compound according to any one of the preceding claims, wherein the compound is used to detect disorders or diseases associated with τ protein aggregates.

10. The compound of claim 9 for the stated use, wherein the disease or disorder is a 4R-τ protein disease or a disease or disorder associated with 4R-τ protein aggregates.

11. The compound for the stated use according to claim 9 or 10, wherein the disease or disorder is a neurodegenerative disease or disorder.

12. The compound for the use according to claim 8 or 10, wherein the disorder or disease is selected from Alzheimer's disease, progressive supranuclear palsy, Pick's disease, frontotemporal dementia (FTLD), frontotemporal dementia-17 with Parkinson's syndrome (FTDP-17), primary age-related tau protein disease (PART) dementia, aerobatic granulopathy, glioglobulinosis, chronic traumatic encephalopathy (CTE), corticobasal syndrome (CBS), and corticobasal degeneration.

13. The compound for the purpose according to any one of claims 9-12, wherein detection is performed by one or more of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging, or autoradiography.

14. The compound for the said use according to claim 13, wherein detection is performed by positron emission tomography (PET) imaging.

15. A method for diagnosing a subject suffering from a disease or disorder, or for monitoring the progression of a subject's disease or disorder, the method comprising: (a) Administering to the subject a detectable amount of an isotopically labeled compound of formula I or II, or a pharmaceutically acceptable salt, solvate, hydrate, tautomer, or stereoisomer thereof: Formula I Formula II in: X1 is N, O, or S; X2, X3, X4, and X5 are each independently N or CH; X6 is CH or O; R1 is H, halogen, CN, Me3N + X7 - Et3N + X7 - Pr3N + X7 - Bu3N + X7 - Or other suitable ammonium salts, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin substances, I(O-acyl)2, ArI + X7 - IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, β-pinacol, β-neopentyl glycol, or other suitable boron substances, provided that when R1 is OH, R1 is not substituted at position 5 of the indole ring; Among them, X7 - It is a suitable counterion; among which ArI + X7 - It is a suitable iodonium substance known in the art; wherein IR7 is a suitable iodonium ylide known in the art; R2 is H, alkyl, tert-butoxycarbonyl, toluenesulfonyl, or other suitable nitrogen protecting group; R3 is H, halogen, NO2, Me3N + X7 - Et3N + X7 - Pr3N + X7 - Bu3N + X7 - Or other suitable ammonium salts, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances, SnMe3, SnEt3, SnPr3, SnBu3, or other suitable trialkyltin substances, I(O-acyl)2, ArI + X7 - IR7, S(O)Ar, SO2Ar, BF3K, B(OH)2, B(OMe)2, B(OEt)2, B(Opr)2, B(OiPr)2, B(OiPr)3Li, β-pinacol, β-neopentyl glycol, or other suitable boron substances; Among them, X7 - It is a suitable counterion; among which ArI + X7 - It is a suitable iodonium substance known in the art; wherein IR7 is a suitable iodonium ylide known in the art; R4 is H, halogen, OH, alkoxy, carbonyl, or OEtOEtF; R5 is H, halogen, OH, OMs, OTs, OTf, O-nitrobenzenesulfonate, O-bromobenzenesulfonate, or other suitable sulfonate substances; R6 is H, halogen, OH, O-acyl, O-alkyl, OBoc, or other suitable oxygen-protecting substance; Or R5 and R6 can form an epoxide: ; m and n are each independently 0 or 1; and o is 0, 1, or 2. as well as (b) Detect the binding of the compound to the τ protein aggregates in the patient.

16. The method of claim 15, wherein the disease or disorder is associated with τ protein aggregates.

17. The method of claim 16, wherein the disease or disorder is a 4R-τ protein disorder or a disease or disorder associated with 4R-τ protein aggregates.

18. The method of claim 16 or 17, wherein the disease or disorder is a neurodegenerative disease or disorder.

19. The method according to any one of claims 15-18, wherein the disease or disorder is selected from Alzheimer's disease, progressive supranuclear palsy, Pick's disease, frontotemporal dementia (FTLD), frontotemporal dementia-17 with Parkinson's syndrome (FTDP-17), primary age-related tau protein disease (PART) dementia, aurophilic granuloma, glioglobulinosis, chronic traumatic encephalopathy (CTE), corticobasal syndrome (CBS), and corticobasal degeneration.

20. The method according to any one of claims 15-19, wherein the compound is... 2 H, 3 H, 18 F, 123 I, 124 I, 125 I, 131 I, 11 C 13 C 14 C 13 N、 15 N、 15 O、 75 Br、 76 Br and 77 Br isotope labeling.

21. The method according to any one of claims 15-20, wherein the detection is performed by one or more of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging, or autoradiography.

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