Heterocyclic compounds and their uses

Abstract

This disclosure provides compounds that specifically target PDGFRα and thereby reduce and / or inhibit the activation of the receptor ("PDGFRα inhibitors"). This disclosure also provides methods for treating demyelinating diseases using the disclosed PDGFRα inhibitors.

Description

[Technical Field]

[0001] Cross-reference of related applications This PCT application claims priority to U.S. Provisional Patent Application No. 63 / 501,005, filed on 9 May 2023, which is incorporated herein by reference in its entirety.

[0002] Reference to sequence listings submitted electronically via EFS-WEB The contents of the electronically submitted sequence listing (title: 5411_003PC01_Seqlisting_ST26, size: 29,305 bytes, and creation date: May 8, 2024) submitted in this application are incorporated herein by reference in their entirety.

[0003] This disclosure relates to compounds that can promote remyelination. Specifically, this disclosure relates to PDGFRα inhibitors and the use of such inhibitors to treat diseases associated with myelinating disorders (e.g., multiple sclerosis). [Background technology]

[0004] Proper myelination by oligodendrocytes is essential for the development and function of the central nervous system (CNS). Gacem et al., Life 11(4):327 (Apr. 2021). Myelin maintains the conduction velocity of axonal potentials of nerve signals and provides metabolic support to axons, thereby supporting their survival. During normal development, oligodendrocyte progenitor cells (OPCs) undergo morphological and molecular changes as they differentiate into oligodendrocytes capable of myelination of axons. Therefore, any abnormal developmental process or pathogenic immune activation, as well as failure of oligodendrocytes to myelinate axons or loss of myelin, can lead to neurodegenerative diseases such as multiple sclerosis.

[0005] Multiple sclerosis (MS) is generally characterized by inflammation and demyelination of nerve axons. Patients with early MS often suffer from a single immune attack, accompanied by effective remyelination and recovery between attacks. However, as the disease progresses, MS patients have a reduced ability to remyelinate effectively, leading to permanent neuropathy. Current treatments for MS and other neurodegenerative diseases mainly focus on reducing the immune attack against myelin, but do not restore myelin on damaged nerve axons.

[0006] Therefore, there remains a need for new and more effective treatments for such demyelinating diseases, particularly treatments that can induce remyelination of demyelinated vulnerable nerve axons. SUMMARY OF THE INVENTION

[0007] This disclosure describes the discovery that compounds that inhibit PDGFRα can promote remyelination. Accordingly, the compounds and methods of this disclosure are useful for treating diseases associated with demyelination (e.g., hypomyelination).

[0008] Herein, Formula I:

Chemical formula

[0009] or pharmaceutically acceptable salts or solvates thereof are provided, wherein

Chemical formula

[0010] In some embodiments, Y1 is N, Y 2 is C.

[0011] In some embodiments, Y 1 is C, Y 2 is N.

[0012] In some embodiments, X 1 is N, X 2 is CR a and X 3 is CR a and X 4 is CR a and X

[0013] In some embodiments, X 1 is CR a and X 2 is N, X 3 is CR a and X 4 is CR a and X

[0014] In some embodiments, X 1 is CR a and X 2 is CR a and X 3 is N, X 4 is CR a and X

[0015] In some embodiments, X 1 is CR a and X 2 is CR a and X 3 is CR a and X 4 is N.

[0016] In some embodiments, X 1 is CR a and X 2 is CR a and X 3 is CR a and X 4 is CR a and X

[0017] In some embodiments, R 1 It is a 5-membered or 6-membered heteroaryl that is optionally substituted with a C1-C4 alkyl or oxo molecule.

[0018] In some embodiments, R 2 is one R 3 It will be replaced by this.

[0019] In some embodiments, R 1 This is pyrazolyl, which is substituted by choice.

[0020] In some embodiments, R 1 teeth, [ka] Selected from,

[0021] R 10 However, it is selected from H, C1-C4 alkyl, C1-C4 alkoxy, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, and C1-C4 alkylsulfonyl, with C1-C4 alkyl, C1-C4 alkoxy, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, and C1-C4 alkylsulfonyl being optional, and hydroxyl, C1-C4 alkoxy, NR 10a R 10b R may be substituted with one or more substituents selected from halo and deuterium, where R 10a and R 10b However, it is selected from hydrogen and C1-C4 alkyl, or R 10a and R 10b However, together with the nitrogen atoms bonded to them, they form 4- to 8-membered rings.

[0022] In some embodiments, R 1 teeth, [ka] That is the case.

[0023] In some embodiments, R10 This is CH3.

[0024] In some embodiments, R 2 It is a heterocycline.

[0025] In some embodiments, R 2 teeth, [ka] [ka] Selected from,

[0026] [ka] However, it exhibits single or double bonds such that all valencies are satisfied.

[0027] m is selected from 0, 1, 2, 3, 4, 5, and 6.

[0028] Z 1 , Z 2 , and Z 3 However, N and CR a Selected from.

[0029] In some embodiments, R 2 teeth, [ka] Selected from.

[0030] In some embodiments, the compound, or a pharmaceutically acceptable salt or solvate thereof, is of formula Ia: [ka] It has,

[0031] In the formula, a and b are each independently selected from 1, 2, and 3.

[0032] Q is selected from -CH- and -N-, provided that if Q is -N-, then a and b are not 1.

[0033] In some embodiments, the compound, or a pharmaceutically acceptable salt or solvate thereof, is of formula II: [ka] It holds.

[0034] In some embodiments, the compound, or a pharmaceutically acceptable salt or solvate thereof, is of formula IIa: [ka] It holds.

[0035] In some embodiments, R 3 is 1, 2, 3, 4, or 5 R 30 It is a heteroaryl that is optionally substituted by [the specified method].

[0036] In some embodiments, R 3 teeth, [ka] [ka] Selected from,

[0037] In the formula, A 1 However, O, S, and NR 37 Selected from, R 36 However, R is selected from hydrogen, optionally substituted C1-C6 alkyls, and optionally substituted C1-C6 alkylaryls. 37 However, it is selected from hydrogen and C1-C6 alkyl groups.

[0038] In some embodiments, R 3 teeth, [ka] [ka] Selected from.

[0039] In some embodiments, R 30 teeth, [ka] That is the case.

[0040] In some embodiments, R 300 teeth, [ka] Selected from.

[0041] In some embodiments, R 300 teeth, [ka] That is the case.

[0042] In some embodiments, R 301 H is R 302 It is either -OH or CH3.

[0043] In some embodiments, R 3 teeth, [ka] Selected from.

[0044] In some embodiments, R 3 teeth, [ka] Selected from.

[0045] In some embodiments, the compound, or a pharmaceutically acceptable salt or solvate thereof, is of formula IV: [ka] It has, in the formula, L1 However, it is either NH or O.

[0046] In some embodiments, the compound, or a pharmaceutically acceptable salt or solvate thereof, is of formula V: [ka] That is the case.

[0047] In some embodiments, R 31 teeth, [ka] [ka] [ka] Selected from,

[0048] In the formula, R 31a However, it is selected from H, D, alkylamino, optionally substituted C1-C4 alkyl, C1-C4 alkoxy, amino, and C1-C4 haloalkyl,

[0049] Each R 31b The elements are independently selected from H, D, halo, hydroxy, amino, cyano, alkylamino, optionally substituted C1-C4 alkyl, C1-C4 haloalkyl, optionally substituted C1-C4 alkoxy, and C3-C6 cycloalkyl, where q is 1, 2, or 3.

[0050] In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, is selected from any one of the compounds in Table 1.

[0051] In some embodiments, compounds of formula I may exhibit one or more of the following properties: (i) promoting differentiation from OPCs to oligodendrocytes, (ii) promoting the expression of proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17 (GPR17), myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or combinations thereof), (iii) promoting axon myelination, (iv) promoting remyelination of demyelinated axons, (v) inhibiting PDGFRα kinase activity, (vi) achieving a brain-to-plasma ratio greater than 0.1 when systemically administered to a subject, and (vii) any combination thereof.

[0052] In some embodiments, the compound of formula I can inhibit PDGFRα kinase activity.

[0053] In some embodiments, the compound of formula I has PDGFRα kinase activity of less than 500 nM, less than 400 nM, less than 300 nM, less than 200 nM, less than 100 nM, less than 75 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, less than 2.5 nM, less than 1 nM, less than 0.5 nM, or less than 0.2 nM IC 50 This can be inhibited.

[0054] In some embodiments, IC of a PDGFRα inhibitor of formula I 50 This is determined using an enzymatic PDGFRα kinase assay (e.g., the Promega kinase assay described in Example 131).

[0055] In some embodiments, the enzymatic PDGFRα kinase assay comprises 20 ng of purified PDGFRα protein, 150 μM of ATP, and 1 μg of the substrate poly(Glu4Tyr1) in a volume of 15 μl.

[0056] In some embodiments, the compound of formula I reduces PDGFRα kinase activity to approximately 500 nM to approximately 0.001 nM, approximately 400 nM to approximately 0.001 nM, approximately 300 nM to approximately 0.001 nM, approximately 200 nM to approximately 0.001 nM, approximately 100 nM to approximately 0.001 nM, approximately 75 nM to approximately 0.001 nM, approximately 50 nM to approximately 0.001 nM, and approximately ICs with a capacitance of approximately 40nM to 0.001nM, 30nM to 0.001nM, 20nM to 0.001nM, 10nM to 0.001nM, 5nM to 0.001nM, 2.5nM to 0.001nM, 1nM to 0.001nM, 0.5nM to 0.001nM, or 0.2nM to 0.001nM 50 This can be inhibited.

[0057] In some embodiments, the present disclosure provides compounds that can inhibit cellular PDGFRα activity and may further exhibit one or more of the following properties: (i) promote differentiation from OPCs to oligodendrocytes; (ii) promote the expression of proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17 (GPR17), myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof); (iii) promote axon myelination; (iv) promote remyelination of demyelinated axons; (v) inhibit PDGFRα kinase activity; (vii) achieve a brain-to-plasma ratio greater than 0.1 when systemically administered to a subject; or (viii) further exhibit one or more of any combination thereof.

[0058] In some embodiments, the compound has an IC of less than 500 nM, less than 400 nM, less than 300 nM, less than 200 nM, less than 100 nM, less than 75 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, or less than 2 nM of PDGFRα kinase activity. 50 This can be inhibited.

[0059] In some embodiments, IC of PDGFRα inhibitors 50This is determined using an enzymatic PDGFRα kinase assay (e.g., the Promega® kinase assay described in Example 131).

[0060] In some embodiments, the enzymatic PDGFRα kinase assay comprises 20 ng of purified PDGFRα protein, 150 μM of ATP, and 1 μg of the substrate poly(Glu4Tyr1) in a volume of 15 μl.

[0061] This disclosure also provides pharmaceutical compositions comprising a compound disclosed herein, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable additive.

[0062] This disclosure also provides a kit comprising the compounds disclosed herein or their pharmaceutically acceptable salts or solvates, or the compositions disclosed herein, along with instructions for use.

[0063] This disclosure also provides a method for producing PDGFRα inhibitors, which includes synthesizing the compounds disclosed herein.

[0064] This disclosure also provides compounds or pharmaceutical compositions disclosed herein for use as pharmaceuticals.

[0065] This disclosure also provides compounds or pharmaceutical compositions disclosed herein for use in therapeutic purposes.

[0066] The disclosure also provides a method for treating demyelinating diseases in subjects requiring treatment for such diseases, the method comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition disclosed herein.

[0067] The disclosure also provides a method for improving the performance of a subject in tests for evaluating one or more symptoms associated with demyelinating disease, the method comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition disclosed herein, wherein, after administration, the subject's performance in the tests is improved compared to a reference subject (e.g., the subject before administration).

[0068] In some embodiments, the examination may be selected from one or more of the following: visual evoked potential (VEP) testing, multifocal visual evoked potential (mfVEP) testing, low-contrast visual acuity (LC-VA) testing, magnetic resonance imaging (MRI) (e.g., magnetization transport rate (MTR), myelin water fraction (MWF), quantitative susceptibility mapping (QSM), and T2 imaging), electromyography (EMG), nerve conduction velocity (NCV) testing, expanded disability status scale (EDSS), gait time measurement (e.g., 25-foot walk time measurement), 9-hole peg test (9HPT), optical coherence tomography (OCT), quality of life measurement tests (e.g., quality of life in multiple sclerosis - 54 and visual quality of life), cognitive function assessments (e.g., code-number modality tests or Montreal cognitive function assessment), or a combination thereof.

[0069] In some cases, demyelinating diseases include acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis, acute transverse myelitis, adrenoleukodystrophy, adrenal spinal neuropathy, Alexander disease, Alzheimer's disease, aminoaciduria, amyotrophic lateral sclerosis, anti-MAG peripheral neuropathy, anti-MOG related spectrum, Barlow concentric sclerosis, brain injury, CAMFAK syndrome, Canavan disease, carbon monoxide poisoning, central pontine myelinolysis, cerebral hypoxia, cerebral ischemia, Charcot-Marie-Tooth disease, and chronic inflammatory demyelinating disease. Myelopolyneuropathy, chronic traumatic encephalopathy, syndrome consisting of the first episode (CIS), congenital cataract, copper deficiency-related conditions, delayed hypoxic leukoencephalopathy, Schilder's generalized encephalosclerosis, generalized myelin-destructive sclerosis, extrapontine myelinolysis, Gaucher disease, Guillain-Barré syndrome, hereditary neuropathy, hereditary pressure-fragility neuropathy, HTLV-1-associated myelopathy, Hurler syndrome, myelin hypoplasia, hypoxic brain injury, Krabbe disease, Leber's hereditary optic atrophy and associated mitochondrial disorders, leukodystrophy Fee's disease, Marquiafava-Bignami disease, metachromatic leukodystrophy, multiple sclerosis (e.g., primary progressive multiple sclerosis (PPMS), relapsing-remitting multiple sclerosis (RRMS), secondary progressive multiple sclerosis (SPMS), progressive relapsing multiple sclerosis, Marburg multiple sclerosis, tumor-like multiple sclerosis, and optic and spinal cord multiple sclerosis), multiple system atrophy, myelin-destroying disorders, myelopathy, nerve injury, neuromyelitis optica (NMO), Niemann-Pick disease, optic neuropathy, optic neuritis (e.g., acute optic nerve This includes one or more of the following conditions: optic neuritis and chronic relapsing inflammatory optic neuritis (CRION), osmotic demyelinating syndrome, Parkinson's disease, Pelizaeus-Merzbach disease, peripheral neuropathy, phenylketonuria, progressive inflammatory neuropathy, progressive multifocal leukoencephalopathy, progressive subcortical ischemic demyelination, reperfusion injury, Schilder's disease, isolated sclerosis, spinal cord injury, subacute sclerosing panencephalitis, tabes dorsalis, Tay-Sachs disease, transverse myelitis, traumatic brain injury, tropical spastic paraplegia, vitamin B12 deficiency, and cerebral palsy.

[0070] In some embodiments, demyelinating diseases are characterized by the demyelination of one or more cells within the affected central nervous system (CNS).

[0071] In some cases, demyelinating diseases are multiple sclerosis.

[0072] In some manifestations, multiple sclerosis includes the syndrome consisting of a first episode ("CIS"), relapsing-remitting MS ("RRMS"), secondary progressive MS ("SPMS"), primary progressive MS ("PPMS"), optic neuritis, or transverse myelitis.

[0073] In some cases, demyelinating diseases are optic neuritis.

[0074] In some embodiments, treatment for demyelinating diseases includes reducing one or more symptoms associated with the demyelinating disease.

[0075] In some embodiments, one or more symptoms include one or more of the following: fatigue, dizziness, malaise, high fever and hyperthermia, extreme coldness of the hands and feet, weakness and stiffness of muscles and joints, weight changes, digestive or gastrointestinal disorders, hypotension, hypertension, irritability, anxiety, depression, visual impairment (e.g., blurred vision, diplopia, decreased low-contrast visual acuity (LC-VA)), ataxia, clonus, seizures, dysarthria, weakness, clumsiness, paralysis of the hands, hemiplegia, loss of genital sensation, sexual dysfunction, incoordination, paresthesia, ophthalmoplegia, muscular incoordination, loss of sensation, tingling, anesthesia, pain, neurological symptoms, cognitive impairment, unsteady gait, balance problems, dizziness, spastic paraplegia, incontinence, hearing impairment, speech impairment, loss of smell, and anosmia.

[0076] The disclosure also provides a method for promoting axonal myelination in subjects requiring enhanced axonal myelination, the method comprising administering to the subject an effective amount of a compound or pharmaceutical composition disclosed herein.

[0077] In some embodiments, enhanced axonal myelination results in increased expression of one or more of the following markers within the subject: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

[0078] In some embodiments, axonal myelination can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

[0079] The disclosure also provides a method for promoting the remyelination of demyelinated axons in subjects requiring the promotion of remyelination of demyelinated axons, the method comprising administering to the subject an effective amount of a compound or pharmaceutical composition disclosed herein.

[0080] In some embodiments, enhanced remyelination of demyelinated axons results in increased expression of one or more of the following markers within the subject: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

[0081] In some embodiments, remyelination of demyelinated axons can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

[0082] This disclosure also provides a method for reducing myelinated nerve axon demyelination in subjects requiring a reduction in myelinated nerve axon demyelination, the method comprising administering to a subject an effective amount of a compound or pharmaceutical composition disclosed herein.

[0083] In some embodiments, reduced demyelination of myelinated nerve axons results in increased expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

[0084] In some embodiments, the reduction of demyelination of myelinated nerve axons can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

[0085] This disclosure also provides a method for activating oligodendrocyte progenitor cells (OPCs) in a subject requiring activation of such OPCs in the central nervous system (CNS), the method comprising administering to the subject an effective amount of one of the compounds or pharmaceutical compositions disclosed herein.

[0086] In some embodiments, the subjects have or are at risk of developing demyelinating diseases, such as those disclosed herein.

[0087] In some embodiments, the method is a method for treating or preventing demyelinating diseases, such as those disclosed herein.

[0088] In some embodiments, the compound or pharmaceutical composition is administered to the subject in a single dose.

[0089] In some embodiments, the compound or pharmaceutical composition is administered to the subject two or more times using intermittent dosing.

[0090] In some embodiments, intermittent dosing includes administering a compound or pharmaceutical composition every other day, every three days, every four days, every five days, every six days, once a week, every eight days, every nine days, every ten days, every eleven days, every twelve days, every thirteen days, once every two weeks, once every three weeks, once a month, once every two months, once every three months, once every four months, once every five months, once every six months, or once every twelve months.

[0091] In some embodiments, intermittent dosing comprises administering a first dose and a second dose of a compound or pharmaceutical composition to a subject, the second dose being administered at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or at least 12 months after the administration of the first dose.

[0092] In some embodiments, the second dose is administered to the subject 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 days, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or 12 months after the first dose.

[0093] In some embodiments, after administration, the compound or pharmaceutical composition may achieve a brain-to-plasma ratio greater than 0.1, greater than 0.2, greater than 0.3, greater than 0.4, greater than 0.5, greater than 0.6, greater than 0.7, greater than 0.8, greater than 0.9, greater than 1.0, greater than 1.1, greater than 1.2, greater than 1.3, greater than 1.4, greater than 1.5, greater than 1.6, greater than 1.7, greater than 1.8, greater than 1.9, or greater than 2.0. In some embodiments, the brain-to-plasma ratio may be evaluated in preclinical species using bioanalytical gases or liquid chromatography and mass spectrometry.

[0094] In some embodiments, the method further includes administering an additional therapeutic agent to the subject.

[0095] In some embodiments, additional therapeutic agents include standard therapeutic agents.

[0096] In some embodiments, the additional therapeutic agent includes an immunomodulator.

[0097] In some embodiments, additional therapeutic agents are selected from interferon beta-1b, interferon beta-1a, pegylated interferon beta-1a, alemtuzumab, natalizumab, ocrelizumab, ofatumumab, ubrituximab-xiiy, glatiramer acetate, teriflunomide, dimethyl fumarate, monomethyl fumarate, diloximel fumarate, fingolimod hydrochloride, siponimod fumarate, ozanimod hydrochloride, ponesimod, cladribine, mitoxantrone, BTK inhibitors (e.g., masitinib, evobrutinib, or trebrutinib), statins (e.g., simvastatin), or pharmaceutically acceptable salts thereof.

[0098] In some embodiments, the additional therapeutic agent is administered to the subject before, simultaneously with, or after the administration of the compound or pharmaceutical composition.

[0099] This disclosure also provides a method for inducing the differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes, the method comprising contacting the OPCs with an effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0100] In some embodiments, induction of differentiation from OPCs to oligodendrocytes results in increased expression of the following markers in the subjects: GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or combinations thereof.

[0101] In some embodiments, differentiation from OPCs to oligodendrocytes is measured by determining the expression of GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

[0102] This disclosure also provides a method for inhibiting PDGFRα activity in cells, the method comprising contacting cells with an effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0103] In some embodiments, inhibition of PDGFRα activity is measured by one or more of the following: an in vitro OPC differentiation assay (e.g., as described in Example 132), a cuprizone model for demyelination, an in vivo OPC differentiation assay (e.g., as described in Example 134), an enzymatic PDGFRα kinase assay (e.g., as described in Example 131), or any combination thereof.

[0104] In some embodiments, contact occurs ex vivo or in vivo.

[0105] In some aspects, the method is a therapeutic procedure.

[0106] The disclosure also provides a method for treating PDGF-related tumors in subjects requiring treatment of PDGF-related tumors, the method comprising administering to a subject a therapeutically effective amount of a compound or pharmaceutical composition disclosed herein, wherein PDGFRα activity is reduced in the subject after administration.

[0107] In some embodiments, PDGF-related tumors include oligodendrogliomas.

[0108] In some aspects, the method is a therapeutic procedure. [Modes for carrying out the invention]

[0109] This disclosure describes compounds that can induce the differentiation of OPCs into cells possessing the characteristics of mature oligodendrocytes, including morphological properties and protein expression patterns related to myelination, and that can also influence remyelination. Therefore, as described herein, the compounds of this disclosure may be useful in treating various diseases, including demyelination-related diseases. Additional embodiments of this disclosure are provided throughout this application.

[0110] Before describing this disclosure in more detail, it should be understood that this disclosure is not limited to the specific compositions or process steps described, which can naturally vary. As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has separate components and features that can be readily separated from or combined with any of the features of any of the other embodiments without departing from the scope or spirit of this disclosure. Any described method may be performed in the order of the described events or in any other logically possible order.

[0111] The headings provided herein are not limitations on the various aspects of this disclosure that may be defined by reference to the specification as a whole. It should also be understood that the scope of this disclosure is limited only by the appended claims, and therefore the terminology used herein is intended to describe, and not limit, specific aspects.

[0112] I. Terminology To facilitate understanding of this disclosure, certain terms are defined first. Where used in this application, each of the following terms shall have the meanings set forth below, unless otherwise expressly provided herein. Additional definitions are provided throughout this application.

[0113] When the term "a" or "an" refers to an entity, it means one or more of those entities; for example, "a nucleotide sequence" is understood to mean one or more nucleotide sequences. Therefore, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably herein.

[0114] Furthermore, as used herein, “and / or” shall be considered a specific disclosure of each of two designated features or components, whether one is accompanied by the other or not. Thus, as used herein in phrases such as “A and / or B,” the term “and / or” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Similarly, as used in expressions such as “A, B, and / or C,” the term “and / or” is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0115] Where an aspect is described herein using the word “including,” it will be understood that other similar aspects described using the expressions “consisting of” and / or “essentially consisting of” are also provided.

[0116] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art in the field to which this disclosure relates. For example, *Concise Dictionary of Biomedicine and Molecular Biology*, Juo, Pei-Show, 2nd ed., 2002, CRC Press, *The Dictionary of Cell and Molecular Biology*, 5th ed., 2013, Academic Press, and *Oxford Dictionary of Biochemistry and Molecular Biology*, 2nd ed., 2008, Oxford University Press provide general dictionaries for many of the terms used herein.

[0117] Units, prefixes, and symbols shall be written in the form recognized by the International System of Units (SI). Numerical ranges shall include the number that defines the range. Where ranges of values ​​are enumerated, each integer value interposing between the upper and lower limits enumerated for that range, and each fractional part thereof, shall be understood to be explicitly disclosed, along with each subrange between such values. The upper and lower limits of any range may be independently included in or excluded from a range, and each range that includes either an upper or lower limit, each range that does not include either, or each range that includes both, is also included in this disclosure. Therefore, ranges enumerated herein shall be understood to be all abbreviated representations of the values ​​within the range that include the enumerated endpoint. For example, the range 1 to 10 shall be understood to include any number, combination of numbers, or subrange from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.

[0118] Where a value is explicitly stated, it should be understood that values ​​that are approximately the same number or amount as the stated value (e.g., 10) (e.g., ±10%) are also included within the scope of this disclosure. Where a combination is disclosed, each of the partial combinations of the elements of that combination is also specifically disclosed and included within the scope of this disclosure. Conversely, where different elements or groups of elements are disclosed individually, their combinations are also disclosed. Where any element of this disclosure is disclosed as having multiple substitutes, examples of disclosures in which each substitute is excluded, either alone or in any combination with other substitutes, are also disclosed herein, and two or more elements of the disclosure may have such exclusions, and all combinations of elements having such exclusions are disclosed herein.

[0119] As used herein, the term “halo,” either by itself or as part of another group, refers to Cl, F, Br, or I.

[0120] As used herein, the term "nitro," either by itself or as part of another group, refers to NO2.

[0121] As used herein, either by itself or as part of another group, the term "cyano" refers to CN.

[0122] As used herein, the term "hydroxy," either by itself or as part of another group, refers to the OH group.

[0123] As used herein, the term "alkyl" refers to a group consisting of 1 to 12 carbon atoms (i.e., C1 to C12). 12 Alkyl refers to a linear or branched aliphatic hydrocarbon containing an alkyl group, or a specified number of carbon atoms (e.g., C1 alkyl groups such as methyl, C2 alkyl groups such as ethyl, etc.). In some embodiments, alkyl refers to C1-C 10It is alkyl. In some embodiments, the alkyl is C1-C6 alkyl. In some embodiments, the alkyl is C1-C4 alkyl. In some embodiments, the alkyl is C1-C3 alkyl, i.e., methyl, ethyl, propyl, or isopropyl. Non-limiting and exemplary C1-C 12 Alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, iso-butyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, and decyl. In some embodiments, one or more hydrogen atoms of the alkyl group are replaced by deuterium atoms, i.e., the alkyl group is isotopically labeled with deuterium. A non-limiting and exemplary deuterated alkyl group is -CD3.

[0124] As used herein, either by itself or as part of another group, the term “haloalkyl” refers to an alkyl group in which one or more hydrogen atoms of the alkyl group are replaced by a halo atom. In some embodiments, the haloalkyl group is a -CF3 group.

[0125] As used herein, either by itself or as part of another group, the term “alkoxy” refers to an alkyl group bonded to a terminal oxygen atom. In some embodiments, the alkyl is a C1-C8 alkyl group, and therefore the resulting alkoxy is referred to as a “C1-C8 alkoxy.” In some embodiments, the alkyl is a C1-C4 alkyl group. Non-limiting and exemplary alkoxy groups include methoxy, ethoxy, and tert-butoxy.

[0126] As used herein, either by itself or as part of another group, the term “alkyloxyalkyl” refers to an alkyl group substituted with an alkoxyl group. In some embodiments, the alkyl is a C1-C4 alkyl and the alkoxy is a C1-C4 alkoxyl, and therefore the resulting alkoxyalkyl is referred to as “C1-C4 alkoxyC1-C4 alkyl”.

[0127] As used herein, either by itself or as part of another group, the term "amino" refers to -NH2, which may be optionally substituted with one or two alkyl groups, two alkyl groups linked to form a ring, haloalkyl groups, (hydroxy)alkyl groups, (alkoxy)alkyl groups, (amino)alkyl groups, heteroalkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocyclo alkyl groups, optionally substituted aryl groups, optionally substituted heteroaryl groups, (aryl)alkyl groups, (cycloalkyl)alkyl groups, (heterocyclo)alkyl groups, or (heteroaryl)alkyl groups.

[0128] As used herein, either by itself or as part of another group, the term “heterocycylamino” refers to an amino group substituted with a heterocyclyl group.

[0129] As used herein, either by itself or as part of another group, the term “aminoalkyl” refers to an alkyl group substituted with an amino group. In some embodiments, the alkyl is a C1-C4 alkyl, and therefore the resulting aminoalkyl is referred to as “amino C1-C4 alkyl.”

[0130] As used herein, either by itself or as part of another group, the term “hydroxyalkyl” refers to an alkyl group substituted with a hydroxyl group. In some embodiments, the alkyl is a C1-C4 alkyl, and therefore the resulting hydroxyalkyl is referred to as a “hydroxy C1-C4 alkyl.”

[0131] As used herein, either by itself or as part of another group, the term "aminoalkylamino" refers to an amino group substituted with an aminoalkyl group. In some embodiments, the aminoalkylamino group is -NHCH2CH2NH2.

[0132] As used herein, the term "oxo" refers to an oxygen atom connected to a carbon atom by a double bond (i.e., forming a keto group).

[0133] As used herein, the term "cycloalkyl" refers to a group consisting of 3 to 12 carbon atoms (i.e., C3 to C2). 12 This refers to monocyclic, bicyclic, or tricyclic aliphatic hydrocarbons containing a specified number of carbon atoms (e.g., C3 cycloalkyls such as cyclopropyl, C4 cycloalkyls such as cyclobutyl, etc.), and saturated and partially unsaturated (e.g., containing one or two double bonds). In some embodiments, a cycloalkyl is bicyclic, i.e., it has two rings. In some embodiments, a cycloalkyl is monocyclic, i.e., it has one ring. In some embodiments, a cycloalkyl is C3-C8 cycloalkyl. In some embodiments, a cycloalkyl is C 3-6 A cycloalkyl group is a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl group. In some embodiments, the cycloalkyl group is a C5 cycloalkyl group, i.e., cyclopentyl or cyclopentenyl. In some embodiments, the cycloalkyl group is a C6 cycloalkyl group, i.e., cyclohexyl or cyclohexenyl. A cycloalkyl group containing one or two double bonds may also be referred to as a "cycloalkenyl" group.

[0134] As used herein, either by itself or as part of another group, the term “heterocyclyl” refers to monocyclic, bicyclic, or tricyclic groups containing 1, 2, 3, or 4 heteroatoms and containing 3 to 14 ring members (i.e., 3 to 14 member heterocyclyls), which are saturated and partially unsaturated (e.g., containing one or two double bonds). Each heteroatom is independently oxygen, sulfur, or nitrogen. The term heterocyclyl includes groups in which one or more -CH2- groups are replaced by one or more -C(=O)- groups, including cyclic ureido groups such as imidazolidinyl-2-one, cyclic amide groups such as piperidine-2-one or piperazine-2-one, and cyclic carbamate groups such as oxazolidinyl-2-one.

[0135] As used herein, either by itself or as part of another group, the term “heterocyclylaryl” refers to an aryl group substituted with a heterocyclyl group.

[0136] As used herein, either by itself or as part of another group, the term “aryl C1-C4 alkyl” refers to a C1-C4 alkyl group substituted with an aryl group.

[0137] As used herein, either by itself or as part of another group, the term “heteroaryl C1-C4 alkyl” refers to a C1-C4 alkyl group substituted with a heteroaryl group.

[0138] As used herein, either by itself or as part of another group, the term “heterocyclyl C1-C4 alkyl” refers to a C1-C4 alkyl group substituted with a heterocyclyl group.

[0139] As used herein, either by itself or as part of another group, the term "cycloalkyl C1-C4 alkyl" refers to a C1-C4 alkyl group substituted with a cycloalkyl group.

[0140] As used herein, either by itself or as part of another group, the term "alkylsulfonyl" refers to a sulfonyl group substituted with an alkyl group, i.e., -SO2-. A non-limiting and exemplary alkylsulfonyl group is -SO2CH3.

[0141] As used herein, either by itself or as part of another group, the term "aryl" refers to an aromatic ring system having 6 to 14 carbon atoms, i.e., C6-C 14 This refers to aryl groups. Non-restrictive and exemplary aryl groups include phenyl (abbreviated as "Ph"), naphthyl, phenanthryl, anthrasyl, indenyl, azlenyl, biphenyl, biphenylenyl, and fluorenyl groups. In some embodiments, the aryl group is phenyl.

[0142] As used herein, either by itself or as part of another group, the term “heteroaryl” refers to monocyclic and bicyclic aromatic ring systems having 5 to 14 ring members (i.e., 5- to 14-membered heteroaryls) containing 1, 2, 3, or 4 heteroatoms. Each heteroatom is independently oxygen, sulfur, or nitrogen. In some embodiments, a heteroaryl has 3 heteroatoms. In some embodiments, a heteroaryl has 2 heteroatoms. In some embodiments, a heteroaryl has 1 heteroatom. In some embodiments, a heteroaryl is a 5- to 10-membered heteroaryl. In some embodiments, a heteroaryl has 5 ring atoms (e.g., thienyl, a 5-membered heteroaryl having 4 carbon atoms and 1 sulfur atom). In some embodiments, a heteroaryl has 6 ring atoms (e.g., pyridyl, a 6-membered heteroaryl having 5 carbon atoms and 1 nitrogen atom). Non-restrictive and exemplary heteroaryl groups include thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl, benzofuryl, pyranyl, isobenzofuranyl, benzoxazolyl, clomenyl, xanthenyl, 2H-pyrrolyl, pyrrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridadinyl, isoindolyl, 3H-indolyl, indolyl, This includes indazolyl, prinyl, isoquinolyl, quinolyl, phthalazinyl, naphthilidinyl, cinnolinyl, quinazolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl, β-carbolinyl, phenanthiazolyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenadinyl, thiazolyl, isothiazolyl, phenothiazolyl, isoxazolyl, flazanil, and phenoxadinyl. The term heteroaryl also encompasses N-oxides. A non-limiting exemplary N-oxide is pyridyl N-oxide.

[0143] As used herein, either by itself or as part of another group, the term “aminoheterocycline” refers to a heterocycline group substituted with an optionally substituted amino group. Non-limiting, illustrative examples of aminoheterocycline groups include: [ka]

[0144] The chemical terms used herein may be combined to describe larger substituents. In the case of monovalent substituents, the substituent listed last in the combined term is the substituent containing a bond point. For example, an "aryl C1-C4 alkyl" group contains a bond point on the alkyl group, while a "C1-C4 alkylaryl" group contains a bond point on the aryl group.

[0145] This disclosure encompasses any of the disclosed compounds in which one or more atoms are isotope-labeled (i.e., radioactively labeled) by replacing them with atoms having different atomic masses or mass numbers. Examples of isotopes that may be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example, respectively. 1 H, 2 H (i.e., Deuterium (D)), 3 H, 11 C, 12 C, 13 C, 14 C, 14 N 15 N, 18 O, 17 O, 16 O 31 P, 32 P, 32 S, 33 S, 34 S, 35 S, 36 S, 18 F, 19 F, 35 Cl, 37 Cl, and 36 Cl, for example, 3 H, 11 C, and 14C is an example. In some embodiments, a composition is provided in which substantially all of the atoms at a certain position in the disclosed compound are replaced by atoms having different atomic masses or mass numbers. In some embodiments, a composition is provided in which some of the atoms at a certain position in the disclosed compound are replaced, i.e., atoms having different atomic masses or mass numbers are concentrated at a certain position in the disclosed compound. The isotope-labeled disclosed compounds can be prepared by methods known in the art. The disclosure also encompasses any of the disclosed compounds in which a quaternary carbon atom is replaced by a silicon atom.

[0146] As described above, the compounds disclosed herein contain one or more chiral carbon atoms and therefore may give rise to enantiomers, diastereomers, and other stereoisomers. This disclosure encompasses the use of their racemic and divided forms, as well as mixtures thereof, as well as all such possible forms. Individual enantiomers can be separated according to methods known in the art in view of this disclosure. Where the compounds described herein contain olefin double bonds or other geometrically asymmetric centers, and unless otherwise specified, they are intended to include both E and Z geometric isomers. All tautomers are also encompassed by this disclosure.

[0147] As used herein, the term “stereoisomer” is a general term for all isomers of an individual molecule that differ only in the orientation of their atoms in space. It includes enantiomers and isomers (diastereomers) of compounds that have more than one chiral center and are not mirror images of each other.

[0148] The term "chiral center" or "chiral carbon atom" refers to a carbon atom to which four different groups are bonded.

[0149] The terms "enantiomer" and "of an enantiomer" refer to optically active molecules that cannot be superimposed on their mirror image, and therefore the enantiomer rotates its plane of polarization in one direction, while its mirror image rotates its plane of polarization in the opposite direction.

[0150] The term "racemic" refers to a mixture of equal enantiomers, such mixtures being optically inactive. In some embodiments, the disclosed compounds are racemic.

[0151] The term "absolute configuration" refers to the spatial arrangement of atoms in a chiral molecular entity (or group) and its stereochemical description (e.g., R or S).

[0152] The stereochemical terms and conventions used herein are intended to be consistent with those set forth in Pure & Appl. Chem 68:2193 (1996), unless otherwise specified.

[0153] The term "enantiomer excess" or "ee" refers to a measure of how much more of one enantiomer is present compared to the other. For a mixture of R and S enantiomers, the enantiomer excess percentage is defined as |RS| × 100, where R and S are the respective mole or weight fractions of the enantiomers in the mixture such that R + S = 1. With knowledge of the optical rotation of chiral substances, the enantiomer excess percentage is ([α] obs / [α] max Defined as ) × 100, in the formula, [α] obs [α] is the optical rotation of the enantiomer mixture. max This is the optical rotation of the pure enantiomer. The enantiomer excess can be determined using various analytical techniques, including NMR spectroscopy, chiral column chromatography, or optical rotation analysis.

[0154] The terms “administer,” “to administer,” and their grammatical variations refer to introducing compositions such as the PDGFRα inhibitors of this disclosure into a subject via a pharmaceutically acceptable route. Any preferred route of administration may be used when administering the PDGFRα inhibitors described herein to a subject. Non-limiting examples of such routes of administration are provided elsewhere in this disclosure.

[0155] The term "central nervous system" or "CNS" refers to the complex of nerve tissue that controls various bodily (e.g., voluntary and involuntary movements) and mental (e.g., thought, perception, and emotion) activities. The CNS generally consists of the brain and spinal cord.

[0156] As used herein, the term “desylenesis” refers to any disorder of the nervous system involving reduced myelination, including disorders in which myelin dysfunction or impairment occurs during development (e.g., hypomyelination), or disorders in which the myelin sheath of a neuron is damaged. “Myelin” and “myelin sheath” refer to the specialized membrane formed by oligodendrocytes that insulates the axon of a neuron. The insulation provided by the myelin sheath helps to increase the rate of nerve signal transmission along the axon, thereby promoting time- and energy-efficient nerve signaling. Myelin also provides metabolic support to the axon, maintaining its health and survival. In some embodiments, the nerve axons of an object suffering from or at risk of developing a demyelinesis are completely demyelinated. In some embodiments, nerve axons in subjects suffering from or at risk of developing a demyelinating disease are partially demyelinated (e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 95% compared to a fully myelinated reference nerve axon). Thus, unless otherwise indicated, the term “demyelination” includes any of the following: complete hypomyelination, complete demyelination, partial hypomyelination, partial demyelination, and any combination thereof. In some embodiments, due to partial and / or complete demyelination, the demyelinating diseases described herein are associated with impaired nerve signal conduction or impaired axonal or neuronal survival, which may then result in impairment of sensory, motor, cognitive, or other functions, depending on which neurons are affected. Non-limiting examples of demyelinating diseases are provided elsewhere in this disclosure. Generally, the term "hypermyelination" refers to cells, tissues, or subjects that lack a normal level of myelination (e.g., the level of myelination observed in corresponding cells, tissues, or subjects without demyelinating disease), regardless of the etiology.

[0157] As used herein, the term “myelin hypoplasia” refers to a deficiency of myelin for any reason (e.g., the body is unable to produce myelin at normal levels). Unless otherwise indicated, myelin hypoplasia includes demyelination (related to myelin destruction) and hypomyelination (related to abnormal myelin accumulation). Thus, myelin hypoplasia includes diseases of insufficient myelin that occur during development, as well as diseases associated with demyelination and / or hypomyelination. In some embodiments, myelin hypoplasia is present in all (i.e., “complete myelin hypoplasia”) or in part (i.e., “partial myelin hypoplasia”) of the nervous system, for example, the brain (e.g., myelin hypoplasia in white matter and / or gray matter), compared to a reference (e.g., the corresponding tissue of a subject without demyelinating disease, e.g., the brain), at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 95% of the nervous system (i.e., “complete myelin hypoplasia”) or in part (i.e., “partial myelin hypoplasia”), for example, the brain (e.g., myelin hypoplasia in white matter and / or gray matter).

[0158] As used herein, the term “neuron” includes electrically excitable cells that process and transmit information through electrical and chemical signals. Neurons are the main components of the brain and spinal cord of the central nervous system (CNS), as well as the ganglia of the peripheral nervous system (PNS), and can connect with each other to form neural networks. A typical neuron consists of a cell body (soma), dendrites, and axon. The “soma” of a neuron contains a nucleus. The “dendrites” of a neuron are cell extensions, generally with many branches, where most of the input to the neuron occurs. The “axon” (also referred to herein as “neural axon”) extends from the cell body and transports nerve signals far from the cell body and certain types of information back to the cell body.

[0159] As used herein, the terms “oligodendrocyte progenitor cell” or “OPC” (also known in the art as “oligodendrocyte precursor cell,” “polydendrocyte,” “NG2 cell,” and “O-2A cell”) refer to a subtype of glial cell in the central nervous system. They are progenitor cells of “oligodendrocytes” (also known as “oligodendroglia”), which are responsible for the development of the myelin sheath, which wraps around axons to provide insulation, assists in electrical conduction, and provides metabolic support. Nerve impulses can travel up to 200 times faster along myelinated axons compared to unmyelinated axons. OPCs and immature oligodendrocytes are generally positive for the following markers: A2B5, neuron-glia antigen 2 (NG2), and PDGFRα. Other suitable markers that may be used are known in the art.

[0160] As used herein, the term “remyelination” (or its derivatives) refers to the development of a new myelin sheath around a demyelinated (e.g., myelin hypoplasia) axon. The remyelination process involves the differentiation of OPCs into oligodendrocytes, which generate a functional myelin sheath around the demyelinated axon. Axonal remyelination can restore the conduction properties of action potentials to the axon, thereby promoting and / or improving neuronal function. Furthermore, remyelination can provide metabolic support to the axon, preventing its damage or loss. In the context of this application, unless otherwise specified, “remyelination” refers to any aspect of the process that may result in remyelination. For example, in some embodiments, “remyelination” includes the migration or colonization of OPCs to the site of the demyelinated axon. In some embodiments, “remyelination” includes the differentiation of OPCs into oligodendrocytes. In some embodiments, “remyelination” includes the development of a myelin sheath by oligodendrocytes around the demyelinated axon. In some embodiments, the term “remyelination” includes any combination of the following: (i) migration or colonization of OPCs to the site of the demyelinated axon, (ii) differentiation of OPCs into oligodendrocytes, and (iii) myelin sheath development by oligodendrocytes around the demyelinated (e.g., hypomyelin) axon.

[0161] As used herein, the term “restore” (and its derivatives) includes both complete and partial restoration. For example, in some embodiments, remyelination of a demyelinated axon restores the conduction properties of the axon to the same degree as those of the axon before demyelination (i.e., complete restoration). In some embodiments, remyelination of a demyelinated axon restores the conduction properties of the axon, where the conduction properties are improved, but not the same as those of the axon before demyelination (i.e., partial restoration). In some embodiments, remyelination provides metabolic support to the axon to prevent damage or loss. In some embodiments, remyelination provides partial metabolic support to the axon to delay damage or loss.

[0162] As used herein, the term “promote” refers to the ability of a drug (e.g., a PDGFRα inhibitor as described herein) to induce or increase a particular outcome (e.g., remyelination of demyelinated axons). In some embodiments, the term includes both inducing and increasing a particular outcome.

[0163] As used herein, the term “subject” refers to any animal subject, including humans, laboratory animals (e.g., non-human primates, rats, and mice), livestock (e.g., cattle, sheep, goats, pigs, turkeys, and chickens), and domestic pets (e.g., dogs, cats, and rodents).

[0164] As used herein, the terms “to treat,” “to treat,” and “treatment” refer to any type of intervention or process performed on a subject for the purpose of improving, alleviating, improving, inhibiting, or slowing a disease-related symptom, complication, condition, or one or more biochemical signs, or preventing its progression, onset, severity, or recurrence, or improving overall survival, or administering an active agent (e.g., a PDGFRα inhibitor as described herein) to a subject. As described herein, in some embodiments, the treatment may be for a subject that has the disease (e.g., exhibiting one or more disease-related symptoms). In some embodiments, the treatment may be for a subject that has some degree of demyelination but has not yet exhibited any disease-related symptoms. In relation to such subjects, administration of a PDGFRα inhibitor as described herein may help delay or prevent the onset of disease-related symptoms.

[0165] The term “effective dose” or “effective amount” is defined as the amount sufficient to achieve, or at least partially achieve, the desired effect (e.g., inducing remyelination of demyelinated nerve axons). The “therapeutably effective dose” or “therapeutably effective dosage” of a therapeutic agent (e.g., a PDGFRα inhibitor) is any amount of the therapeutic agent, when used alone or in combination with another therapeutic agent, that promotes disease regression, as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of asymptomatic periods of the disease, prevention of functional impairment or disability due to disease distress, or a reduction in disease progression. The therapeutically effective dose or dosage of a therapeutic agent also includes the “prophylactically effective dose” or “prophylactically effective dosage,” which is any amount of the therapeutic agent, when administered alone or in combination with another therapeutic agent, to subjects at risk of developing the disease or at risk of disease recurrence, that inhibits the onset or recurrence of the disease. The ability of a therapeutic agent to promote recovery from a disease or to inhibit the onset, progression, or recurrence of the disease can be evaluated using a variety of methods known to specialists, for example, by assaying the activity of the drug in human subjects during clinical trials, in animal model systems that predict efficacy in humans, or in in vitro assays.

[0166] As used herein, the term “dose interval” refers to the length of time elapsed between multiple (e.g., two or more) doses of the PDGFRα inhibitor described herein. While not bound by any single theory, in some embodiments, the PDGFRα inhibitors provided herein exert their therapeutic effect by inducing the differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes. Therefore, in some embodiments, for a PDGFRα inhibitor to have a therapeutic effect in a subject (e.g., by inducing OPC differentiation), the subject must have a pool of OPCs on which the PDGFRα inhibitor described herein can act. As demonstrated herein, administering PDGFRα inhibitors too frequently to a subject may be counterproductive in treating demyelinating diseases, as the OPC population may not have sufficient time to regrow in the brain. Accordingly, in some embodiments, a dosing interval preferred to the present disclosure is the length of time, or longer, required for the subject's OPC population to recover sufficiently after the administration of the initial dose of the PDGFRα inhibitor provided herein, so that a second (or additional) dose of the PDGFRα inhibitor has a therapeutic effect in the subject (e.g., an increase in the number of differentiated oligodendrocytes, as described herein). As will be apparent from the present disclosure, in some embodiments, the PDGFRα inhibitor described herein is administered to the subject at a dosing interval, where the dosing interval is the length of time required for the subject's OPC population (e.g., in size) to reach at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of that of the reference OPC population (e.g., the subject's OPC population before the initial administration of the PDGFRα inhibitor) after the administration of the initial dose of the PDGFRα inhibitor. In some embodiments, the OPC population may be measured by quantifying the density of NG2+ (neuronal glial antigen 2 / chondroitin sulfate proteoglycan-4) or PDGFRa+ cells in mice, rats, pigs, dogs, non-human primates, or other species using immunohistochemistry or fluorescence-activated cell sorting.Regeneration can be calculated as the OPC cell density at a given time after drug administration, compared to the OPC population in an untreated reference animal.

[0167] In some embodiments, a preferred dosing interval for the present disclosure includes the length of time required for the plasma level of the PDGFRα inhibitor to reach less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, or less than 5% of the reference level (e.g., the plasma level of the PDGFRα inhibitor in the corresponding subject immediately after administration of the initial dose of the PDGFRα inhibitor, e.g., about 4 hours later), and the preferred dosing interval is the time between the administration of the first dose of the PDGFRα inhibitor and the time when the plasma level of the PDGFRα inhibitor in the subject reaches the reduced level compared to the reference level. In some embodiments, an additional dose of the PDGFRα inhibitor is administered to the subject when the plasma level in the subject has decreased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or about 100% compared to the reference level. In some embodiments, the brain-to-plasma ratio may be evaluated in preclinical species using bioanalytical gases or liquid chromatography and mass spectrometry.

[0168] As used herein, the terms “binding,” “recognizing,” and “targeting” are synonymous and refer to molecules that can bind to and / or target specific regions of a protein (e.g., PDGFRα) (e.g., PDGFRα inhibitors as described herein), such binding and / or targeting will be understood by those skilled in the art. For example, as described herein, in some embodiments, a PDGFRα inhibitor is a small molecule that can target and inhibit the kinase portion of PDGFRα (e.g., by binding to the ATP binding site, by allosterically interfering with PDGFRα kinase activity, or both), thereby inhibiting its activity. Unless otherwise indicated, the above terms (i.e., binding, recognizing, and targeting) are used interchangeably and include any binding or targeting of PDGFRα such that its activity is reduced and / or inhibited (e.g., binding of a small molecule to the ATP binding site of PDGFRα, binding of a small molecule to the substrate binding site of PDGFRα, allosteric interference with PDGFRα kinase activity, or a combination thereof).

[0169] "Efficacy" is an expression of the activity of a therapeutic agent, expressed in units of the amount or concentration of the therapeutic agent that achieves the desired effect. Therefore, compounds useful for this disclosure can be identified using functional assays such as those described herein.

[0170] A molecule that "competes with another protein or compound for binding to its target" (e.g., PDGFRα inhibitors such as those described herein) refers to a molecule that (partially or completely) inhibits the binding of another protein (e.g., a naturally occurring PDGFRα ligand) to its target (e.g., PDGFRα). Whether two compounds compete with each other for binding to their target, i.e., whether and to what extent the PDGFRα inhibitors described herein inhibit the binding of a naturally occurring ligand to PDGFRα, can be determined using known competition experiments. In some embodiments, the PDGFRα inhibitors described herein compete with a naturally occurring ligand and inhibit the binding of the naturally occurring ligand to PDGFRα by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100%. Competitive assays may be performed as described herein, or, for example, as described in Ed Harlow and David Lane, Cold Spring Harbor Protoc.;2006, doi:10.1101 / pdb.prot4277, or as described in Chapter 11 of “Using Antibodies”, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999 by Ed Harlow and David Lane.

[0171] Additional non-limiting examples of other competitive binding assays that may be used in this disclosure include: solid-phase direct or indirect radioimmunoassay (RIA), solid-phase direct or indirect enzyme immunoassay (EIA), sandwich competitive assays (see Stahli et al., Methods in Enzymology 9:242 (1983)); solid-phase direct biotin-avidin EIA (see Kirkland et al., J.Immunol. 137:3614 (1986)); solid-phase direct labeling assays, solid-phase direct labeling sandwich assays (see Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Press, Second Edition (2014)); solid-phase direct labeling RIA using 1-125 labeling (see Morel et al., Mol.Immunol. 25(1):7 (1988)); solid-phase direct biotin-avidin EIA (see Cheung et al., Virology Examples include 176:546(1990)) and directly labeled RIA (Moldenhauer et al., Scand. J. Immunol. 32:77(1990)).

[0172] As further described elsewhere in this disclosure, in some embodiments, the antagonist activity of the PDGFRα inhibitors described herein does not depend on interference with the binding of another protein or compound to PDGFRα. Another non-limiting example is a small molecule that can bind to PDGFRα and inhibit (partially or completely) the binding of ATP or substrate, or otherwise inhibit PDGFRα activity. In some embodiments, the PDGFRα inhibitors described herein may inhibit or reduce PDGFRα activity via allosteric inhibition. In some embodiments, the PDGFRα inhibitors may reduce the protein level of PDGFRα by reducing synthesis or increasing degradation. Exemplary mechanisms of action are provided, but it will be apparent from this disclosure that the PDGFRα inhibitors described herein are not intended to be limited to such mechanisms.

[0173] As used herein, the terms "ug" and "uM" are interchangeable with "μg" and "μM," respectively.

[0174] The various embodiments described herein are described in further detail in the following subsections. The various embodiments and forms of this disclosure can be selected and combined as needed. For example, in some embodiments, any of the disclosed compounds can be used in any of the disclosed methods and uses.

[0175] II. PDGFRα inhibitors This specification discloses compounds that can inhibit PDGFRα activity and / or induce one or more activities related to the inhibition of the receptor. PDGFRα can bind with high affinity to several PDGF isoforms (i.e., PDGF-A, PDGF-B, PDGF-C, and PDGF-D). As used herein, a reference to inhibition of the binding of PDGFRα to its ligand refers to the inhibition of the binding of one or more PDGF isoforms that bind to PDGFRα. In some embodiments, the binding of all isoforms of PDGF that bind to PDGFRα is inhibited. The binding of platelet-derived growth factor (PDGF) isoforms to PDGFRα affects various cellular signaling pathways, including those involved in cell proliferation and differentiation. Therefore, PDGFRα activity is essential for both the proper development and long-term maintenance of certain tissues and organs. For example, animals that are completely lacking the functional PDGFRA gene die at birth or shortly after birth due to various developmental abnormalities (e.g., cardiac malformations). Bax et al., Dev.Dyn.239(8):2307-2317(Aug.2010). Conditional PDGFRα knockout in OPC during animal development results in severe myelin hypoplasia, and the animals die shortly after birth. Hamashima et al., Neuroscience 436:11-26(Jun.2020)

[0176] In humans, the gene encoding PDGFRα (i.e., PDGFRA) is located on chromosome 4 (e.g., nucleotides 54,229,127-54,298,245 of GenBank accession number NC_000004.12; plus-strand orientation). In addition to those provided above, other synonyms for PDGFRα are known, including “PDGFRa”, “PDGFR2”, “Platelet-derived growth factor receptor alpha”, “Platelet-derived growth factor receptor A”, “Platelet-derived growth factor receptor alpha”, “Platelet-derived growth factor receptor 2”, “CD140a”, and “CD140 antigen-like family member A”. As used herein, the term “PDGFRα” includes any variant or isoform of PDGFRα that is naturally expressed in cells.

[0177] Despite its importance in various biological processes, this disclosure describes the finding that inhibiting or reducing PDGFRα activity by, for example, one or more of the compounds described herein may have certain biological effects that may be useful in treating demyelinating diseases, such as those described herein. As demonstrated herein, in some embodiments, inhibiting or reducing PDGFRα activity by the PDGFRα inhibitors described herein may promote oligodendrocyte differentiation and induce myelin formation. In some embodiments, the PDGFRα inhibitors of this disclosure may promote (and, in some embodiments, be used for) remyelination of demyelinated nerve axons, which may be useful in treating various demyelinating diseases, such as multiple sclerosis. As used herein, the terms “PDGFRα inhibitor” and “PDGFRα antagonist” are interchangeable and refer to any compound that can reduce and / or inhibit (and, in some embodiments, be used for) PDGFRα activity (for example, by any of the exemplary mechanisms or methods described herein). In some embodiments, the PDGFRα inhibitors provided herein may inhibit the tyrosine kinase activity of PDGFRα. In some embodiments, the PDGFRα inhibitors provided herein may inhibit any other activity of PDGFRα. Where the term "may be" is used herein to describe a particular characteristic of a PDGFRα inhibitor, it means that the PDGFRα inhibitor may exhibit (i.e., have the ability to exhibit) such a characteristic, for example, under appropriate conditions. Unless otherwise indicated, it does not mean that the PDGFRα inhibitor will always exhibit such a characteristic when administered, for example, to a subject having a impaired oligodendrocyte progenitor cell population. The terms "may be" and "can be" are used interchangeably herein.

[0178] In some embodiments, PDGFRα inhibitors inhibit the kinase activity of PDGFRα. As used herein, unless otherwise indicated, the term “inhibit” (and its derivatives) includes both complete inhibition and partial inhibition (e.g., reduction of kinase activity). Thus, in some embodiments, when an OPC (expressing PDGFRα) interacts with a PDGFRα inhibitor described herein, the PDGFRα kinase activity is reduced by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or more (e.g., at least 50%, or more) compared to the PDGFRα kinase activity in the corresponding OPC that was not exposed to the PDGFRα inhibitor described herein (e.g., exposed to a vehicle control). In some embodiments, the kinase activity is completely inhibited. As will be apparent from this disclosure, (partial or complete) inhibition of PDGFRα kinase activity in OPCs may promote differentiation of OPCs into myelinating oligodendrocytes. Inhibition of PDGFRα kinase activity can be assayed using methods known in the art. In some embodiments, inhibition of PDGFRα kinase activity may be determined using an in vitro differentiation assay as described in Example 132 of this application. In some embodiments, inhibition of PDGFRα kinase activity may be evaluated using a cuprizone model for demyelination, such as that described in Torkildsen et al., Acta Neurol Scand Suppl 188:72-6 (2008). In some embodiments, inhibition of PDGFRα kinase activity may be determined using an enzymatic PDGFRα kinase assay, such as that described in Example 131 of this disclosure. In some embodiments, inhibition of PDGFRα kinase activity may be determined using an in vivo OPC differentiation assay, such as that described in Example 134.

[0179] As described herein, in some embodiments, PDGFRα inhibitors useful to this disclosure have the following properties: (1) they can promote (e.g., induce and / or enhance) the differentiation of OPCs into oligodendrocytes (in vitro, in vivo, or both) (and are used for this purpose in some embodiments); (2) they can promote (e.g., induce and / or enhance) the differentiation of oligodendrocytes into proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17 (GPR17), myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), and (1) can promote (e.g., induce and / or enhance) the expression of (combinations thereof) (and is used for that purpose in some embodiments), (2) can promote (e.g., induce and / or enhance) the myelination of nerve axons (and is used for that purpose in some embodiments), (3) can promote (e.g., induce and / or enhance) the remyelination of demyelinated nerve axons (and is used for that purpose in some embodiments), (4) can promote (e.g., induce and / or enhance) the remyelination of demyelinated nerve axons (and is used for that purpose in some embodiments), (5) can inhibit PDGFRα kinase activity (and is used for that purpose in some embodiments) (e.g., IC < 10,000 nM when measured using a Promega kinase assay (e.g., described in Example 131)). 50 (6) capable of penetrating the brain (and being used for that purpose in some embodiments) at a level sufficient to support activity in the CNS, and (7) comprising one or more of any combination thereof. Non-limiting examples of additional properties are provided elsewhere in this disclosure.

[0180] In addition to the characteristics described herein, the PDGFRα inhibitors described herein may include one or more additional features not present in other inhibitors known in the art. As will be apparent from this disclosure, such additional features may be useful in various clinical settings for treating demyelinating diseases, such as those described herein.

[0181] As described elsewhere in this disclosure, some exemplary PDGFRα inhibitors of this disclosure may have greater therapeutic efficacy than, for example, inhibitors known in the art and the exemplary inhibitors provided herein when assayed against a vehicle control. For example, in some embodiments, PDGFRα inhibitors may have higher potency than inhibitors known in the art. In some embodiments, PDGFRα inhibitors may be able to permeate the CNS better (e.g., cross the blood-brain barrier more effectively). Therefore, in some embodiments, the PDGFRα inhibitors described herein may be more effective in promoting OPC differentiation than inhibitors known in the art. For example, in some embodiments, the PDGFRα inhibitors of this disclosure may achieve more than a twofold increase in GPR17 expression compared to a vehicle control at doses of less than 50 mg / kg when measured using an in vivo GPR17 assay (such as that described in Example 134). In some embodiments, the PDGFRα inhibitors of this disclosure may achieve more than a twofold increase in GPR17 expression compared to a vehicle control at doses of less than 40 mg / kg, less than 30 mg / kg, less than 20 mg / kg, less than 10 mg / kg, less than 5 mg / kg, less than 4 mg / kg, less than 3 mg / kg, less than 2 mg / kg, or less than 1 mg / kg, as measured using an in vivo GPR17 assay (such as the one described in Example 134). As described elsewhere in this disclosure, GPR17 expression is a suitable marker for OPC differentiation. Additionally, in some embodiments, any of the other proteins associated with oligodendrocyte differentiation and / or myelination may be used instead of or in combination with GPR17. Non-limiting examples of such proteins include MBP, ASPA, GST-pi, CC1, MOG, oligodendrocyte-specific protein / claudin-11, CNPase, proteolipide protein 1 (PLP1), or combinations thereof.

[0182] In some embodiments, the PDGFRα inhibitor is of formula I: [ka] The compound,

[0183] or a pharmaceutically acceptable salt or solvate thereof, in the formula, [ka] However, it exhibits single or double bonds such that all valencies are satisfied. X 1 , X 2 , X 3 , and X 4 However, N and CR a Selected from, however, X 1 , X 2 , X 3 , and X 4 The condition is that two or fewer of them are N, Y 1 and Y 2 One of them is N, and Y 1 and Y 2 The other of these is C, Each R a These are independently selected from H, halo, C1-C4 alkyl, and C1-C4 alkoxy. R 1 However, all of these can be optionally substituted with 1, 2, 3, 4, 5, or 6 substituents selected from halo, hydroxy, oxo, C1-C4 alkyl, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxy C1-C4 alkyl, 3-8 membered heterocyclil, and 3-8 membered heterocyclil C1-C4 alkyl, provided that the number of substituents does not exceed the number of substituted positions. R 2However, these are selected from cycloalkyl, cycloalkenyl, alkyl, aminoalkylamino, amino, heterocyclyl, heteroaryl, aminoheterocyclyl, heterocyclylamino, and aminoalkylamino, all of which may be optionally substituted with one or more substituents selected from D, halo, oxo, and C1-C4 alkyl. R 2 However, 1, 2, or 3 R 3 Replaced by, R 3 However, aryl, heteroaryl, -C(O)R 31 , -C(O)OR 31 -C(O)NR 31 R 32 -S(O)2NR 31 R 32 -S(O)2R 31 -S(O)(NR 33 )R 31 -S(O)(NR 33 )NR 31 R 32 -C(S)NR 31 R 32 Selected from C3-C8 cycloalkyls, 3-8 membered heterocyclines, and C1-C4 alkyls, all of which are optional, with 1, 2, 3, 4, or 5 R groups. 30 It can be replaced by, Each R 30 These are independent of D, Halo, Ariel, -OR 300 , -NR 300 R 301 , -S(O) r R 300 , -C(O)R 300 -C(=CR) 34 R 35 )R 300 , and [ka] Selected from, r is selected from 0, 1, and 2. Each R 300These are independently selected from C1-C6 alkyl, C3-C7 cycloalkyl, aryl, heteroaryl, 3-8 membered heterocyclyl, and 3-8 membered heterocyclylaryl, all of which may be optionally substituted with 1, 2, 3, 4, or 5 substituents selected from D, halo, hydroxy, amino, alkylamino, cyano, C1-C4 alkyl, C1-C4 haloalkyl, and C1-C4 alkoxy. Each R 301 These are independently selected from H, halo, and C1-C4 alkyl groups. Each R 302 These are independently selected from H, F, hydroxy, amino, alkylamino, oxo, and C1-C4 alkoxy, Each R 303 These are independently selected from H and C1-C4 alkyl groups. n, o, and p are each independently selected from 0, 1, 2, 3, and 4. Each R 31 These are independently selected from C1-C8 alkyl, aryl C1-C4 alkyl, heteroaryl C1-C4 alkyl, heterocyclyl, heterocyclyl C1-C4 alkyl, cycloalkyl, and cycloalkyl C1-C4 alkyl, all of which are optional selections of D, halo, cyano, hydroxy, amino, -OCF3, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, hydroxy C1-C4 alkyl, and -S(O)2NR 304 R 305 , -C(O)OR 304 R 305 -C(O)NR 304 R 305 , and -NR 304 C(O)R 305 It may be substituted with 1, 2, 3, 4, or 5 substituents selected from the following: Each R 304 and R 305 These are independently selected from H and C1-C4 alkyl groups. Each R 32 These are independently selected from H and C1-C4 alkyl groups, or R 31 and R 32However, together with the atoms connected to them, D, halo, cyano, C1-C4 alkyl, and C1-C4 haloalkyl, and -C(O)NR 34 R 35 It forms a 5- to 8-membered heterocisyl which is optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the above, Each R 34 and R 35 These are independently selected from H, C1-C4 alkyl, and C1-C4 haloalkyl, Each R 33 These independently comprise H, C1-C4 alkyl, C1-C4 haloalkyl, and -C(O)R 34 Selected from, or R 31 and R 33 However, together with the atoms connected to them, they form 4- to 8-membered heterocisyl groups.

[0184] In some embodiments, Y 1 is N, and Y 2 C is C.

[0185] In some embodiments, Y 1 C is Y 2 It is N.

[0186] In some cases, X 1 is N, and X 2 CR a X 3 CR a X 4 CR a That is the case.

[0187] In some cases, X 1 CR a X 2 is N, and X 3 CR a X 4 CR a That is the case.

[0188] In some cases, X 1 CRa X 2 CR a X 3 is N, and X 4 CR a That is the case.

[0189] In some cases, X 1 CR a X 2 CR a X 3 CR a X 4 It is N.

[0190] In some cases, X 1 CR a X 2 CR a X 3 CR a X 4 CR a That is the case.

[0191] In some embodiments, R 1 These are 5-membered or 6-membered heteroaryl compounds.

[0192] In some embodiments, R 1 This is pyrazolyl, which is substituted by choice.

[0193] In some embodiments, R 1 teeth, [ka] Selected from, R 10 However, it is selected from H, C1-C4 alkyl, C1-C4 alkoxy, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, and C1-C4 alkylsulfonyl, with C1-C4 alkyl, C1-C4 alkoxy, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, and C1-C4 alkylsulfonyl being optional, and hydroxyl, C1-C4 alkoxy, NR 10a R 10bR may be substituted with one or more substituents selected from halo and deuterium, where R 10a and R 10b However, it is selected from hydrogen and C1-C4 alkyl, or R 10a and R 10b However, together with the nitrogen atoms bonded to them, they form 4- to 8-membered rings.

[0194] In some embodiments, R 1 teeth, [ka] That is the case.

[0195] In some embodiments, R 10 This is CH3.

[0196] In some embodiments, R 2 However, these are selected from cycloalkyl, cycloalkenyl, alkyl, oxoalkylamino, aminoalkylamino, amino, heterocyclyl, heteroaryl, aminoheterocyclyl, heterocyclylamino, and aminoalkylamino, all of which may be optionally substituted with six or more substituents selected from D, halo, hydroxy, oxo, and C1-C4 alkyl.

[0197] In some embodiments, R 2 It is a heterocycline that is optionally substituted with a C1-C4 alkyl or oxo molecule.

[0198] In some embodiments, R 2 is one R 3 It will be replaced by this.

[0199] In some embodiments, R 2 teeth, [ka] [ka] Selected from, [ka] However, it exhibits single or double bonds such that all valencies are satisfied. m is selected from 0, 1, 2, 3, 4, 5, and 6. Z 1 , Z 2 , and Z 3 However, N and CR a Selected from.

[0200] In some embodiments, R 2 teeth, [ka] Selected from.

[0201] In some embodiments, R 2 teeth, [ka] It is selected from the group consisting of the following.

[0202] In some embodiments, R 2 teeth, [ka] That is the case.

[0203] In some embodiments, R 3 These are aryl, heteroaryl, and -C(O)R 31 , -C(O)OR 31 -C(O)NR 31 R 32 -S(O)2NR 31 R 32 Selected from cycloalkyl and alkyl, all of which are optional, with 1, 2, 3, 4, or 5 R 30 It can be replaced with.

[0204] In some embodiments, R 3 is 1, 2, 3, 4, or 5 R 30Heteroaryls that are optionally substituted by -C(O)OR 31 Selected from.

[0205] In some embodiments, the PDGFRα inhibitor is expressed by formula Ia: [ka] The compound,

[0206] or a pharmaceutically acceptable salt or solvate thereof, where a and b are each independently selected from 1, 2, and 3, Q is selected from -CH- and -N-, provided that if Q is -N-, then a and b are not 1.

[0207] In some embodiments, the PDGFRα inhibitor is expressed as formula II: [ka] It is a compound of, or a pharmaceutically acceptable salt or solvate thereof.

[0208] In some embodiments, the PDGFRα inhibitor is expressed by formula IIa: [ka] It is a compound of, or a pharmaceutically acceptable salt or solvate thereof.

[0209] In some embodiments, R 3 teeth, [ka] [ka] Selected from, in the formula, A 1 However, O, S, and NR 37 Selected from, R 36 However, R is selected from hydrogen, optionally substituted C1-C6 alkyls, and optionally substituted C1-C6 alkylaryls. 37However, it is selected from hydrogen and C1-C6 alkyl groups.

[0210] In some embodiments, R 3 teeth, [ka] [ka] Selected from.

[0211] In some embodiments, R 3 teeth, [ka] Selected from.

[0212] In some embodiments, R 3 It is a heteroaryl compound.

[0213] In some embodiments, R 3 teeth, [ka] That is the case.

[0214] In some embodiments, R 30 teeth, [ka] That is the case.

[0215] In some embodiments, R 300 teeth, [ka] Selected from.

[0216] In some embodiments, R 300 teeth, [ka] That is the case.

[0217] In some embodiments, R 301 This is selected from H and CH3.

[0218] In some embodiments, R 301 H is H.

[0219] In some embodiments, R 302 H is H.

[0220] In some embodiments, R 301 and R 302 H is H.

[0221] In some embodiments, R 30 It is benzyl.

[0222] In some embodiments, R 3 teeth, [ka] Selected from.

[0223] In some embodiments, R 3 teeth, [ka] Selected from.

[0224] In some embodiments, R 32 H is H.

[0225] In some embodiments, the PDGFRα inhibitor is expressed as formula IV: [ka] A compound of, or a pharmaceutically acceptable salt or solvate thereof, in which L 1 However, it is either NH or O.

[0226] In some embodiments, the PDGFRα inhibitor is expressed as formula V: [ka] It is a compound of, or a pharmaceutically acceptable salt or solvate thereof.

[0227] In some embodiments, R 31 teeth, [ka] [ka] [ka] [ka] Selected from, In the formula, R 31a However, it is selected from H, D, alkylamino, optionally substituted C1-C4 alkyl, C1-C4 alkoxy, amino, and C1-C4 haloalkyl, Each R 31b The elements are independently selected from H, D, halo, hydroxy, amino, cyano, alkylamino, optionally substituted C1-C4 alkyl, C1-C4 haloalkyl, optionally substituted C1-C4 alkoxy, and C3-C6 cycloalkyl, where q is 1, 2, or 3.

[0228] Table 1 (below) provides structures of exemplary compounds useful for this disclosure (for example, those that can target and inhibit kinase activity related to PDGFRα expressed on OPCs). [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] Table 1-6 Table 1-7 Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12 Table 1-13 Table 1-14 Table 1-15 Table 1-16 Table 1-17 Table 1-18 Table 1-19 Table 1-20 Table 1-21 Table 1-22 [Table 1-23] [Table 1-24] [Table 1-25] [Table 1-26] [Table 1-27]

[0229] In some embodiments, the exemplary compounds of the present invention do not include compounds 156, 250, and 275. In some embodiments, the exemplary compounds of the present invention do not include the compounds listed in Table 1 on pp. 117-170 of International Application PCT / US2022 / 079480 (which is incorporated herein by reference in its entirety).

[0230] In some embodiments, the PDGFRa inhibitor is a compound listed in Table 1 on pp. 117-170 of International Application PCT / US2022 / 079480.

[0231] In some embodiments, the PDGFRa inhibitor is not one of the compounds listed in Table 1 on pp. 117-170 of International Application PCT / US2022 / 079480.

[0232] Mai. Pharmaceutical composition This specification provides compositions comprising a PDGFRα inhibitor of the present disclosure having a desired purity and a pharmaceutically acceptable carrier or excipient in a form suitable for administration to a subject. In some embodiments, the PDGFRα inhibitor is produced with a purity of about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or greater than about 99%. The pharmaceutically acceptable excipient or carrier may be determined in part by the particular composition to be administered and by the particular method used to administer the composition. Thus, a wide variety of suitable formulations of the pharmaceutical composition exist (e.g., Remington, 23). rd See Edition, The Science and Practice of Pharmacy, editor: A. Adejare, 2020, Adademic Press. Pharmaceutical compositions are generally sterilized and formulated in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.

[0233] In some embodiments, the pharmaceutical composition comprises a PDGFRα inhibitor as described herein and a pharmaceutically acceptable carrier. The acceptable carrier, additive, or stabilizer is nontoxic to the recipient at the dosage and concentration used and includes buffers, such as phosphoric acid, citrate, and other organic acids; antioxidants, preservatives (e.g., octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl alcohol or benzyl alcohol, alkylparabens, such as methylparaben or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, etc. For example, it includes serum albumin, gelatin, or immunoglobulin, hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine, monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin, chelating agents such as EDTA, sugars such as sucrose, mannitol, trehalose, or sorbitol, salt-forming counterions such as sodium, metal complexes (e.g., Zn-protein complexes), and / or nonionic surfactants such as TWEEN®, PLURONICS®, or polyethylene glycol (PEG).

[0234] In some embodiments, the pharmaceutical compositions disclosed herein include one or more additional components selected from bulking agents, stabilizers, surfactants, buffers, or combinations thereof.

[0235] Buffers useful in this disclosure may be weak acids or weak bases used to maintain the acidity (pH) of a solution near a selected value after the addition of another acid or base. A suitable buffer can maximize the stability of the pharmaceutical compositions disclosed herein by maintaining pH control of the composition. A suitable buffer can also ensure physiological compatibility or optimize solubility. Rheology, viscosity, and other properties may also depend on the pH of the composition. Common buffers include Tris buffer, Tris-Cl buffer, histidine buffer, TAE buffer, HEPES buffer, TBE buffer, sodium phosphate buffer, MES buffer, ammonium sulfate buffer, potassium phosphate buffer, potassium thiocyanate buffer, succinate buffer, tartaric acid buffer, DIPSO buffer, HEPPSO buffer, POPSO buffer, PIPES buffer, PBS buffer, MOPS buffer, acetate buffer, phosphate buffer, cacodylate buffer, glycine buffer, sulfate buffer, imidazole buffer, and glycine buffer. This includes, but is not limited to, anidine hydrochloride buffer, phosphate-citrate buffer, borate buffer, malonic acid buffer, 3-picoline buffer, 2-picoline buffer, 4-picoline buffer, 3,5-lutidine buffer, 3,4-lutidine buffer, 2,4-lutidine buffer, Aces, diethyl malonate buffer, N-methylimidazole buffer, 1,2-dimethylimidazole buffer, TAPS buffer, bis-Tris buffer, L-arginine buffer, lactate buffer, glycolic acid buffer, or combinations thereof.

[0236] In some embodiments, the pharmaceutical compositions disclosed herein further include bulking agents. Bulking agents may be added to pharmaceuticals to add volume and mass to the product, thereby facilitating its accurate measurement and handling. Bulking agents that may be used in this disclosure include, but are not limited to, sodium chloride (NaCl), mannitol, glycine, alanine, or combinations thereof.

[0237] In some embodiments, the pharmaceutical compositions disclosed herein may also include stabilizers. Non-limiting examples of stabilizers that may be used in this disclosure include sucrose, trehalose, raffinose, arginine, or combinations thereof.

[0238] In some embodiments, the pharmaceutical compositions disclosed herein include a surfactant. In some embodiments, the surfactant may be selected from the following: alkyl ethoxylates, nonylphenol ethoxylates, amine ethoxylates, polyethylene oxides, polypropylene oxides, aliphatic alcohols such as cetyl alcohol or oleyl alcohol, cocamide MEA, cocamide DEA, polysorbate, dodecyldimethylamine oxide, or a combination thereof. In some embodiments, the surfactant is polysorbate 20 or polysorbate 80.

[0239] In some embodiments, the pharmaceutical compositions disclosed herein (e.g., including PDGFRα inhibitors) further comprise amino acids. In some embodiments, the amino acids are selected from arginine, glutamate, glycine, histidine, or combinations thereof. In some embodiments, the composition further comprises sugar alcohols. Non-limiting examples of sugar alcohols include sorbitol, xylitol, maltitol, mannitol, or combinations thereof.

[0240] Pharmaceutical compositions disclosed herein (e.g., including PDGFRα inhibitors) may be formulated for any route of administration to a subject. Specific examples of routes of administration include intramuscular, cutaneous, subcutaneous, ocular, intravenous, intraperitoneal, intradermal, orbital, intracerebral, intracranial, intraspinal, intraventricular, intrathecal, intracapsular, oral, pulmonary, intranasal, intraarterial, intralymphatic, periorbital, topical, percutaneous, rectal, vaginal, or intratumoral, or intratympanic injection. Parenteral administration, characterized by, for example, cutaneous, subcutaneous, intramuscular, or intravenous injection, is also intended herein.

[0241] Injectable preparations can be prepared in the conventional form of either a liquid solution or suspension, or in a solid form suitable for a solution or suspension in a liquid before injection, or as an emulsion. Injectable preparations, solutions, and emulsions also contain one or more additives. Suitable additives include, for example, water, saline, dextrose, glycerol, or ethanol. If desired, the administered pharmaceutical composition may also contain small amounts of non-toxic adjuncts, such as osmotherapeutic or emulsifiers, pH buffers, stabilizers, solubility enhancers, and other such agents, such as sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrin.

[0242] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, non-aqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspensions and dispersants, emulsifiers, sequestering or chelating agents, and other pharmaceutically acceptable substances. Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose, and Ringer's lactate injection. Non-aqueous parenteral vehicles include plant-derived non-volatile oils, cottonseed oil, corn oil, sesame oil, and peanut oil. Antimicrobial agents in bacteriostatic or fungiostatic concentrates may be added to parenteral preparations packaged in multi-dose containers, for example, phenols or cresols, mercury, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoates, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. The buffer solution contains phosphates and citrates. The antioxidant contains sodium bisulfate. The local anesthetic contains procaine hydrochloride. The suspension and dispersant contains sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. The emulsifier contains polysorbate 80 (TWEEN® 80). The metal ion sequestering or chelating agent contains EDTA. The pharmaceutical carrier also contains ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible vehicles; as well as sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0243] Preparations for parenteral administration include sterile solutions ready for injection, sterile dried soluble products such as lyophilized powders ready to be combined with a solvent immediately before use, sterile suspensions ready for injection, sterile dried insoluble products ready to be combined with a vehicle immediately before use, and sterile emulsions. The solutions may be aqueous or non-aqueous.

[0244] When administered intravenously, suitable carriers include physiological saline or phosphate-buffered saline (PBS), as well as solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0245] Topical mixtures containing the compounds described herein are prepared as described for topical and systemic administration. The resulting mixtures may be solutions, suspensions, emulsions, etc., and may be formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, transdermal patches, or any other formulation suitable for topical administration.

[0246] The therapeutic agents described herein (e.g., PDGFRα inhibitors) may be formulated as aerosols for topical application, for example, by inhalation (see, for example, U.S. Patents 4,044,126, 4,414,209, and 4,364,923, which describe aerosols for the delivery of steroids useful in the treatment of inflammatory diseases, particularly asthma). These formulations for administration to the airways may be in the form of aerosols or solutions for sprayers, alone or in combination with an inert carrier such as lactose, or as ultrafine particle powders for blowing. In such cases, the particles of the formulation may have a diameter of less than about 50 microns, for example, less than about 10 microns. In some embodiments, the particle diameter may be less than about 50 microns, for example, less than 10 microns, as measured by dynamic light scattering (DLS) or static image analysis using a microscope.

[0247] The therapeutic agents disclosed herein (e.g., PDGFRα inhibitors) may be formulated in the form of gels, creams, and lotions for topical or local application, for example, for topical application to the skin and mucous membranes such as intraocularly, and for application to the eye, or for application to the cisterna magna or spinal cord. Topical administration is intended for transdermal delivery, and also for administration to the eye or mucous membranes, or for inhalation therapy.

[0248] Transdermal patches, including ionophoresis and electrophoresis apparatuses, are well known to those skilled in the art and can be used to administer therapeutic agents (e.g., those disclosed herein). For example, such patches are disclosed in U.S. Patents 6,267,983, 6,261,595, 6,256,533, 6,167,301, 6,024,975, 6,010,715, 5,985,317, 5,983,134, 5,948,433, and 5,860,957.

[0249] In some embodiments, the pharmaceutical composition comprising the therapeutic agent described herein (e.g., a PDGFRα inhibitor) is a lyophilized powder, which can be reconstituted as a solution, emulsion, and other mixture for administration. The pharmaceutical composition can also be reconstituted and formulated as a solid or gel. The lyophilized powder is prepared by dissolving the compound described herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. In some embodiments, the lyophilized powder is sterilized. The solvent may contain additives or other pharmacological components of the powder to improve stability, or a reconstituted solution prepared from the powder. Possible additives include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. The solvent may also contain a buffer, e.g., citrate, sodium phosphate, or potassium phosphate, or other such buffers known to those skilled in the art, at approximately a neutral pH in some embodiments. The desired formulation is obtained by subsequent sterile filtration of the solution under standard conditions known to those skilled in the art, followed by lyophilization. In some embodiments, the obtained solution can be distributed into vials for lyophilization. Each vial may contain a single dose or multiple doses of the compound. The lyophilized powder can be stored at room temperature or other suitable conditions at approximately 4°C.

[0250] Reconstitution of lyophilized powder with water for injection provides a formulation for parenteral administration. When reconstituting, the lyophilized powder is added to sterile water or another suitable carrier. The exact amount depends on the selected compound. Such amounts can be determined empirically.

[0251] The pharmaceutical compositions provided herein (including, for example, PDGFRα inhibitors) may also be formulated to target specific tissues, receptors, or other areas of the body being treated. Many such targeting methods are known to those skilled in the art. All such targeting methods are intended herein for use in the compositions. Non-limiting examples of targeting methods include, for example, U.S. Patents 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, and 6,0 See issues 60,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874.

[0252] Pharmaceutical compositions used for in vivo administration can be sterile. This can be achieved, for example, by filtration using a sterile filtration membrane.

[0253] IV. Kit This specification also provides kits comprising one or more PDGFRα inhibitors as described herein. In some embodiments, a pharmaceutical pack or kit is provided herein comprising one or more containers filled with one or more components of the pharmaceutical compositions described herein, such as one or more PDGFRα inhibitors provided herein, and optionally an instruction manual. In some embodiments, the kit contains the pharmaceutical compositions described herein (e.g., PDGFRα inhibitors) and any prophylactic or therapeutic agents, such as those described herein.

[0254] V. Method of Disclosure As demonstrated herein, the PDGFRα inhibitors of this disclosure are useful in reducing or inhibiting PDGFRα activity, which in some embodiments may promote axon myelination and / or remyelination of demyelinated nerve axons.

[0255] In some embodiments, the Disclosure relates to a method for inhibiting or reducing PDGFRα activity in cells or tissues of a subject requiring such inhibition, the method comprising contacting the cells or tissue with a PDGFRα inhibitor described herein. In some embodiments, after contact, the PDGFRα kinase activity in the cells or tissue is inhibited by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% compared to a reference (e.g., PDGFRα activity in cells before contact and / or PDGFRα activity in corresponding cells not contacted with the PDGFRα inhibitor). The PDGFRα activity state of the cells may be determined using any suitable method known in the art. In some embodiments, PDGFRα activity may be determined by measuring the expression level of the corresponding gene, for example, by qRT-PCR. In some embodiments, PDGFRα activity may be determined by measuring the PDGFRα protein level, for example, by immunoblotting. In some embodiments, PDGFRα activity can be determined by measuring receptor phosphorylation, for example, by Western blotting. An exemplary method for assaying such activity, a purified protein enzyme assay, is provided in Example 131 (also referred to herein as the “Promega assay”). In some embodiments, the enzymatic PDGFRα kinase assay comprises 20 ng of purified PDGFRα protein, 150 μM of ATP, and 1 μg of the substrate poly(Glu4Tyr1) in a volume of 15 μl. The potency of the compound can be determined using such an assay.Generally, the compounds most useful in the methods of this disclosure are those with an IC of less than 10,000 nM in such assays (e.g., less than 9,000 nM, less than 8,000 nM, less than 7,000 nM, less than 6,000 nM, less than 5,000 nM, less than 4,000 nM, less than 3,000 nM, less than 2,000 nM, less than 1,000 nM, less than 900 nM, less than 800 nM, less than 700 nM, less than 600 nM, less than 500 nM, less than 400 nM, less than 300 nM, less than 200 nM, less than 100 nM, less than 50 nM, less than 25 nM, less than 10 nM, less than 5 nM, less than 2.5 nM, less than 1 nM, less than 0.5 nM, or less than 0.2 nM, e.g., less than 500 nM). 50 It has. In some embodiments as described herein, the cells are oligodendrocyte progenitor cells (OPCs).

[0256] As described and demonstrated herein, inhibiting or reducing PDGFRα activity can promote myelination of neurons (e.g., neuronal axons), for example, by promoting differentiation from OPCs to oligodendrocytes, which then myelinate their axons. Similarly, inhibiting or reducing PDGFRα activity can also promote remyelination of demyelinated nerve axons. Furthermore, as will be apparent to those skilled in the art, in some embodiments, remyelination can repair damage to demyelinated (e.g., hypomyelinated) axons, thereby helping to restore axonal transmission or reduce axonal injury or loss.

[0257] Accordingly, in some embodiments, for example, in subjects requiring enhanced axonal myelination (e.g., subjects suffering from or at risk of developing demyelinating diseases), a method for enhancing axonal myelination is provided herein, the method comprising contacting OPC with an effective amount of a PDGFRα inhibitor described herein, the contact resulting in differentiation of OPC into oligodendrocytes, the oligodendrocytes being able to enhance axonal myelination (and, in some embodiments, used for this purpose). In some embodiments, after contact, the number of neurons with myelinated axons increases by at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 times compared to a reference (e.g., the number of myelinated axon neurons in a subject before administration and / or the number of myelinated axon neurons in a corresponding subject that did not receive the PDGFRα inhibitor). The increase in the number of myelinated axon neurons may be determined using any preferred method known in the art or described herein. In some embodiments, the increase in the number of myelinated axon neurons may be determined by visualizing and / or quantifying the expression of markers associated with myelinated neurons. For example, in some embodiments, markers associated with myelinated neurons include myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

[0258] In some embodiments, the Specified also provides a method for promoting the remyelination of neuronal axons (e.g., demyelinated nerve axons in subjects requiring remyelination of neuronal axons), the method comprising contacting OPC with an effective amount of any of the PDGFRα inhibitors described herein, the contact resulting in differentiation of OPC into oligodendrocytes, the oligodendrocytes being able to promote (and, in some embodiments, used for) the remyelination of demyelinated nerve axons. In some embodiments, after contact, the number of myelinated demyelinated axons increases by at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 times compared to a reference (e.g., the number of demyelinated nerve axons in a subject before administration and / or the amount of demyelinated nerve axons in a corresponding subject that did not receive the PDGFRα inhibitor). The increase in the number of myelinated demyelinated axons may be determined using any preferred method known in the art or described herein. In some embodiments, the increase in the number of myelinated demyelinated axons may be determined by visualizing and / or quantifying the expression of markers associated with myelinated neurons. For example, in some embodiments, markers associated with myelinated neurons include myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

[0259] In some embodiments, the Specified also provides a method for increasing remyelination in a subject (e.g., increasing remyelination of demyelinated nerve axons in a subject that requires increased remyelination of demyelinated nerve axons), the method comprising contacting OPC with an effective amount of any of the PDGFRα inhibitors described herein, the contact resulting in differentiation of OPC into oligodendrocytes, the oligodendrocytes being able to increase remyelination in the subject (and being used for this purpose in some embodiments). In some embodiments, after contact, remyelination is increased by at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50 times compared to a reference (e.g., the number of demyelinated nerve axons in the subject before administration and / or the amount of demyelinated nerve axons in the corresponding subject that did not receive the PDGFRα inhibitor). An increase in the number of myelinated demyelinated axons can be determined using any preferred method known in the art or described herein. In some embodiments, an increase in the number of myelinated demyelinated axons can be determined by visualizing and / or quantifying the expression of markers associated with myelinated neurons. For example, in some embodiments, markers associated with myelinated neurons include myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

[0260] In some embodiments, methods are provided herein for reducing myelinated axon demyelination in subjects requiring reduction of myelinated axon demyelination (e.g., reducing the rate at which myelinated axons become demyelinated), the methods comprising contacting OPCs with an effective amount of any of the PDGFRα inhibitors described herein, the contact resulting in differentiation of OPCs into oligodendrocytes, the oligodendrocytes being able to reduce myelinated axon demyelination (and, in some embodiments, used for this purpose). In some embodiments, after contact, myelinated axon demyelination in the subject is reduced by at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50 times compared to a reference (e.g., myelinated axon demyelination in a corresponding subject that did not receive the PDGFRα inhibitor). A decrease in the number of neurons with demyelinated axons can be determined using any preferred method known in the art or described herein. In some embodiments, a decrease in the number of neurons with demyelinated axons can be determined by visualizing and / or quantifying the expression of markers associated with myelinated neurons. For example, in some embodiments, markers associated with myelinated neurons include myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

[0261] In some embodiments, methods are provided herein for reducing the rate of demyelination in subjects requiring a reduction in the rate of demyelination (e.g., reducing the rate at which myelinated axons in a subject become demyelinated), the method comprising contacting OPCs with an effective amount of any of the PDGFRα inhibitors described herein, the contact resulting in differentiation of OPCs into oligodendrocytes, the oligodendrocytes being able to reduce the rate of demyelination in the subject (and, in some embodiments, used for this purpose). In some embodiments, after contact, the rate of demyelination in the subject is reduced by at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50 times compared to a reference (e.g., the rate of demyelination in a corresponding subject that did not receive the PDGFRα inhibitor). The reduction in the number of neurons having demyelinated axons may be determined using any preferred method known in the art or described herein. In some embodiments, a decrease in the number of neurons with demyelinated axons can be determined by visualizing and / or quantifying the expression of markers associated with myelinated neurons. For example, in some embodiments, markers associated with myelinated neurons include myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

[0262] In any of the methods provided herein, in some embodiments, contact may occur ex vivo (e.g., the OPC of interest may be isolated and brought into contact with a PDGFRα inhibitor in vitro to evaluate the therapeutic efficacy of the inhibitor). In some embodiments, contact may occur in vivo (e.g., in a subject requiring it). If contact occurs in vivo, the method may further include administering an effective amount of one of the PDGFRα inhibitors described herein to the subject.

[0263] While not bound by any single theory, in some embodiments, after administration to a subject, the PDGFRα inhibitors described herein can migrate into the subject's CNS (e.g., by crossing the blood-brain barrier) and interact with (and, in some embodiments, are used for) nervous system cells present in the CNS. For example, as described and demonstrated herein, the PDGFRα inhibitors of this disclosure can target (and, in some embodiments, are used for) PDGFRα expressed on OPCs, thereby inhibiting or reducing the PDGFRα activity of the OPCs. In some embodiments, this results in activation of OPCs and subsequent differentiation from OPCs into oligodendrocytes. Thus, in some embodiments, a method for activating OPCs in the CNS of a subject requiring OPC activation is provided herein, the method comprising administering the subject one of the PDGFRα inhibitors described herein.

[0264] As described herein, the myelin sheath surrounding nerve axons can affect various neuronal functions. For example, in some embodiments, by increasing myelination of nerve axons, the PDGFRα inhibitors described herein can increase the transmission rate of electrical impulses (i.e., nerve signals or action potentials) along the axons of neurons. Accordingly, in some embodiments, the present disclosure provides a method for increasing the transmission of electrical impulses along the axons of neurons, the method comprising contacting an OPC with an effective amount of one of the PDGFRα inhibitors described herein, the contact resulting in differentiation of the OPC into oligodendrocytes, the oligodendrocytes being able to myelinate (and, in some embodiments, used for this purpose) the axons of neurons, thereby increasing the transmission of electrical impulses. In some embodiments, after contact with the OPC, the transmission of electrical impulses along the neuronal axon increases by at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50 times compared to a reference (e.g., transmission of electrical impulses across the axon before or in the absence of contact with the PDGFRα inhibitor). Action potential conduction velocity can be measured as the action potential of the compound across the corpus callosum in vitro, or by recording auditory or visual evoked potentials in vivo. In some embodiments, remyelination may overcome conduction block and enable the transmission of axonal signals that could otherwise be terminated. Remyelination may improve axonal health and prevent axonal degradation in diseased states. Non-limiting examples of methods useful for measuring such nerve axon characteristics are described, for example, in Li et al., PLoS One 11(11):e0165637 (Nov. 2016), Maheras et al., Sci Rep 8(1):3798 (Feb. 2018), and Alqudah et al., Audiol Neurootol 23(1):20-31 (2018).

[0265] As will be apparent from this disclosure, the PDGFRα inhibitors described herein may be useful in the treatment of demyelinating diseases, for example, by inducing differentiation of OPCs into oligodendrocytes and thereby promoting remyelination of nerve axons. Accordingly, in certain embodiments, this disclosure relates to a method for treating a demyelinating disease in a subject requiring treatment of the demyelinating disease, the method comprising administering to the subject a therapeutically effective amount of the PDGFRα inhibitor described herein. References to a method for treating a demyelinating disease described herein also refer to the agents and compositions described herein for use in treating a demyelinating disease.

[0266] The PDGFRα inhibitors provided herein may also be used to treat a wide range of demyelinating diseases, including those associated with demyelination and / or myelin hypoplasia. In some embodiments, the demyelinating diseases that can be treated by this disclosure include those characterized by demyelination of one or more cells in the central nervous system (CNS).Non-limiting examples of demyelinating diseases that can be treated by this disclosure include acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis, acute optic neuritis, acute transverse myelitis, adrenoleukodystrophy, adrenal spinal neuropathy, Alexander disease, Alzheimer's disease, aminoaciduria, amyotrophic lateral sclerosis, anti-MAG peripheral neuropathy, anti-MOG related spectrum, Barlow concentric sclerosis, brain injury, CAMFAK syndrome, Canavan disease, carbon monoxide poisoning, central pontine myelinolysis, cerebral hypoxia, cerebral ischemia, and Charcot-Magnesia. Lee-Tooth disease, chronic inflammatory demyelinating polyneuropathy, chronic relapsing inflammatory optic neuropathy (CRION), chronic traumatic encephalopathy, syndrome consisting of the first episode (CIS), congenital cataract, copper deficiency-related conditions, delayed hypoxic leukoencephalopathy, Schilder's generalized encephalosclerosis, generalized myelin-destructive sclerosis, extrapontine myelin-disintegrating Gaucher disease, Guillain-Barré syndrome, hereditary neuropathy, hereditary pressure-fragility neuropathy, HTLV-1-associated myelopathy, Hurler syndrome, myelin hypoplasia, hypoxic brain injury, Krabbe disease, Leber gene Optic neuropathy and related mitochondrial disorders, leukodystrophy, Marburg multiple sclerosis, Marchia-Fava-Bignami disease, metachromatic leukodystrophy, multiple sclerosis, multiple system atrophy, myelin-destroying disorders, myelopathy, nerve injury, neuromyelitis optica (NMO), Niemann-Pick disease, optic neuropathy, neurospinal optic multiple sclerosis, osmotic demyelinating syndrome, Parkinson's disease, Pelizaeus-Merzbach disease, periventricular leukomalacia, peripheral neuropathy, phenylketonuria, primary progressive multiple sclerosis Examples of conditions treated include one or more of the following: pleural inflammatory neuropathy (PPMS), progressive inflammatory neuropathy, progressive multifocal leukoencephalopathy, progressive subcortical ischemic demyelination, progressive-onset multiple sclerosis, relapsing-onset multiple sclerosis, relapsing-remitting multiple sclerosis (RRMS), reperfusion injury, Schilder's disease, secondary progressive multiple sclerosis (SPMS), solitary sclerosis, spinal cord injury, subacute sclerosing panencephalitis, tabes dorsalis, Tay-Sachs disease, transverse myelitis, traumatic brain injury, tropical spastic paraplegia, tumor-like multiple sclerosis, vitamin B12 deficiency, and cerebral palsy. In some embodiments, the diseases that can be treated by this disclosure include tumors associated with abnormal (e.g., increased) PDGF or PDGFRα activity. Such tumors are referred to herein as “PDGF-associated tumors.”Accordingly, in certain embodiments, methods for treating PDGF-related tumors in subjects requiring treatment of such tumors are provided herein, the methods comprising administering to the subject one of the PDGFRα inhibitors described herein. Non-limiting examples of PDGF-related tumors include oligodendroglioma.

[0267] In some embodiments, demyelinating diseases that can be treated with the PDGFRα inhibitors described herein include multiple sclerosis. As used herein, the term “multiple sclerosis” (MS) refers to a chronic, often physically debilitating disease of the central nervous system characterized by progressive destruction of the myelin sheath. Multiple sclerosis is generally diagnosed as one of four internationally recognized subtypes or stages of MS: (1) primary progressive multiple sclerosis (PPMS), (2) relapsing-remitting multiple sclerosis (RRMS), (3) secondary progressive multiple sclerosis (SPMS), and (4) progressive-relapsing multiple sclerosis. Standards for diagnosis are known to those skilled in the art, and exemplary diagnostic criteria are described, for example, in the Merck Manual Professional Edition (www.merckmanuals.com / professional / neurologic-disorders / demyelinating-disorders / multiple-sclerosis-ms). Unless otherwise indicated, the term “multiple sclerosis” encompasses all different classifications of MS. Therefore, in certain embodiments, the PDGFRα inhibitors of this disclosure may be used to treat all types of MS.

[0268] In some embodiments, the PDGFRα inhibitors of this disclosure may be used to treat a first-episode syndrome (CIS). In some embodiments, the PDGFRα inhibitors of this disclosure may be used to treat a radiologically isolated syndrome (RIS).

[0269] In some embodiments, a method for treating relapsing-type multiple sclerosis in a subject requiring treatment for relapsing-type multiple sclerosis is provided herein, the method comprising administering to the subject a therapeutically effective amount of one of the PDGFRα inhibitors provided herein.

[0270] In some embodiments, the demyelinating disease that can be treated by this disclosure is optic neuritis. Accordingly, in some embodiments, PDGFRα inhibitors are used to improve optic neuritis, for example, optic neuritis resulting from multiple sclerosis.

[0271] As is evident from this disclosure, by promoting axon myelination and / or remyelination of demyelinated axons, the PDGFRα inhibitors of this disclosure may help restore neuronal cell function, thereby reducing and / or alleviating one or more symptoms associated with demyelinating diseases. Accordingly, in certain embodiments, treatment of demyelinating diseases includes reducing one or more symptoms associated with demyelinating diseases. Non-limiting examples of such symptoms include one or more of the following: fatigue, dizziness, malaise, high fever and high body temperature, extreme coldness of the hands and feet, weakness and stiffness of muscles and joints, weight changes, digestive or gastrointestinal disorders, hypotension, hypertension, irritability, anxiety, depression, blurred vision, double vision, ataxia, clonus, convulsions, dysarthria, weakness, clumsiness, paralysis of the hands, hemiplegia, loss of genital sensation, sexual dysfunction, incoordination, paresthesia, oculoparalysis, muscle coordination disorder, loss of sensation, tingling, anesthesia, pain, visual impairment, neurological symptoms, unsteady gait, balance problems, dizziness, spastic paraplegia, incontinence, hearing impairment, speech impairment, loss of smell, and anosmia.

[0272] Furthermore, the PDGFRα inhibitors described herein may also be used to prevent or delay the onset of one or more symptoms associated with demyelinating diseases. In many demyelinating diseases, subjects may not present any obvious symptoms of the disease, especially in the early stages. For example, in some embodiments, myelin sheath damage may be not very severe, allowing oligodendrocytes present in the subject's CNS to adequately repair any damage. However, as is the case with most demyelinating diseases (e.g., multiple sclerosis), myelin sheath damage can become more severe, leading to the onset of one or more symptoms of the disease. In some embodiments, administering the PDGFRα inhibitors described herein to a subject while myelin sheath damage is still minor may prevent or delay the onset of one or more symptoms of the disease. In some embodiments, the onset of one or more symptoms of the disease is delayed by at least 1.1 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, or at least 50 times compared to a reference subject (e.g., a corresponding subject that did not receive a PDGFRα inhibitor). In some embodiments, subjects treated with the PDGFRα inhibitors described herein exhibit fewer symptoms and / or reduced symptom severity compared to a reference subject.

[0273] Whether a subject exhibits one or more symptoms associated with demyelinating disease can be evaluated using any suitable method known in the art. In some embodiments, whether a subject presents with one or more symptoms associated with demyelinating disease may be determined by evaluating the subject's performance in one or more of the following tests: visual evoked potential (VEP) testing, multifocal visual evoked potential (mfVEP) testing, low-contrast visual acuity (LCVA) testing, magnetic resonance imaging (MRI) (e.g., magnetization transport rate (MTR), myelin water fraction (MWF), quantitative susceptibility mapping (QSM), and T2 imaging), electromyography (EMG), nerve conduction velocity (NCV) testing, expanded disability status scale (EDSS), gait time measurement (e.g., 25-foot walk time measurement), 9-hole peg test (9HPT), optical coherence tomography (OCT), quality of life measurement tests (e.g., quality of life in multiple sclerosis - 54 and visual quality of life), cognitive function assessments (e.g., code-digit modality tests or Montreal cognitive function assessment), or a combination thereof. In some embodiments, whether a subject exhibits one or more symptoms associated with demyelinating disease is determined by evaluating the subject's performance in the LC-VA test. In some embodiments, whether a subject exhibits one symptom associated with demyelinating disease is determined by evaluating the subject's performance in a cognitive function assessment.

[0274] In some embodiments, improved results compared to reference results (e.g., results of the corresponding subject in the examination when the reference subject was not treated with the PDGFRα inhibitor described herein and / or results of the subject before treatment with the PDGFRα inhibitor) indicate that one or more symptoms associated with demyelinating disease are reduced or alleviated in the subject. In some embodiments, equivalent or worse results compared to reference results (e.g., results of the corresponding subject in the examination when the subject was not treated with the PDGFRα inhibitor described herein and / or results of the subject before treatment with the PDGFRα inhibitor) indicate that one or more symptoms associated with demyelinating disease are maintained or worsened in the subject.

[0275] Accordingly, certain aspects of this disclosure relate to a method for treating a demyelinating disease in a subject requiring treatment for the demyelinating disease, the method comprising evaluating the subject's performance in a test for evaluating one or more symptoms associated with the demyelinating disease, the subject having received treatment comprising a PDGFRα inhibitor as described herein prior to the evaluation. In certain aspects, if the subject's performance in the test is improved compared to a reference performance (e.g., the performance of the corresponding subject in the test if the reference subject had not been treated with a PDGFRα inhibitor as described herein and / or the performance of the subject before treatment with a PDGFRα inhibitor), the treatment comprising a PDGFRα inhibitor may be maintained in the subject (e.g., the subject receives one or more additional doses of the PDGFRα inhibitor at the same dose and / or dosing interval). In certain aspects, if the subject shows improved performance, the treatment comprising a PDGFRα inhibitor may be reduced (e.g., the subject receives one or more additional doses of the PDGFRα inhibitor at a lower dose and / or a longer dosing interval) or discontinued. In some embodiments, if the subject's performance is equivalent to or worse than reference performance (e.g., the performance of the corresponding subject in the examination when the subject was not treated with the PDGFRα inhibitor described herein and / or the performance of the subject before treatment with the PDGFRα inhibitor), the treatment including the PDGFRα inhibitor is adjusted so that the subject receives one or more doses of the PDGFRα inhibitor at a higher dose and / or a shorter dosing interval.

[0276] Some aspects of this disclosure relate to a method for improving the performance of a subject in a test for evaluating one or more symptoms associated with a demyelinating disease, the method comprising administering to the subject one of the PDGFRα inhibitors described herein. In some aspects, after administration, the performance of the subject in the test is increased compared to a reference performance (e.g., the performance of the corresponding subject in the test if the reference subject had not been treated with a PDGFRα inhibitor described herein and / or the performance of the subject before treatment with a PDGFRα inhibitor). In some embodiments, the examination is one or more of the following: visual evoked potential (VEP) testing, multifocal visual evoked potential (mfVEP) testing, low-contrast visual acuity (LC-VA) testing, magnetic resonance imaging (MRI) (e.g., magnetization transport rate (MTR), myelin water fraction (MWF), quantitative susceptibility mapping (QSM), and T2 imaging), electromyography (EMG), nerve conduction velocity (NCV) testing, expanded disability status scale (EDSS), gait time measurement (e.g., 25-foot walk time measurement), 9-hole peg test (9HPT), optical coherence tomography (OCT), quality of life measurement tests (e.g., quality of life in multiple sclerosis - 54 and visual quality of life), cognitive function assessments (e.g., code-digit modality tests or Montreal cognitive function assessments), or a combination thereof. In some embodiments, the examination is an LCVA test. In some embodiments, the examination is a cognitive function assessment.

[0277] As will be apparent from this disclosure, PDGFRα inhibitors can be introduced into a target by any preferred route, including but not limited to intratumoral, oral, pulmonary, intranasal, parenteral (intravenous, intraarterial, intramuscular, intraperitoneal, or subcutaneous), rectal, vaginal, lymphatic, intrathecal, periophthalmos, skin, intradermal, intraorbital, intracerebral, intracranial, intraspinal, ventriculospheric, intravesicular, intrasacral, or topical administration. Administration may include self-administration and administration by another person. Preferred routes of administration allow the PDGFRα inhibitors described herein to perform their intended function (e.g., inhibiting PDGFRα activity and inducing OPC differentiation). In some embodiments, preferred routes of administration include those that would allow the PDGFRα inhibitor to reach the CNS or any other site where OPCs are present. Non-limited examples of such routes include intranasal delivery, intrathecal administration, intracranial administration, and combinations thereof. In some embodiments, the PDGFRα inhibitors described herein are administered orally. In some embodiments, the PDGFRα inhibitors of this disclosure are administered intravenously. In some embodiments, the PDGFRα inhibitors may be administered both orally and intravenously.

[0278] In any of the methods provided herein for administering a PDGFRα inhibitor to a subject, the PDGFRα inhibitor may be administered to the subject using any preferred dosing schedule. In some embodiments, the PDGFRα inhibitor is administered to the subject once. In some embodiments, multiple doses of the PDGFRα inhibitor are administered to the subject. In some embodiments, the PDGFRα inhibitor described herein is administered to the subject according to an intermittent dosing schedule. As used herein, the term “intermittent dosing schedule” (and any variation thereof) refers to a dosing schedule in which the therapeutic agent (e.g., a PDGFRα inhibitor) is administered non-continuously (i.e., there are intervening periods between doses). Intermittent dosing schedules useful to this disclosure may encompass any discontinuous dosing regimen that provides a therapeutically effective amount of the PDGFRα inhibitor to a subject in need. An intermittent dosing regimen may use doses of the PDGFRα inhibitor that are equivalent to, lower than, or higher than the doses used in a continuous dosing regimen. The advantages of intermittent dose administration include, but are not limited to, improved safety, reduced toxicity (e.g., reduced weight loss), acceptable levels of ADME criteria, acceptable levels of undesirable effects on organ systems such as the heart, lungs, liver, reproductive organs (e.g., ovaries or testes), or gastrointestinal tract, increased exposure, increased efficacy, and / or increased adherence to medication by the subject. These advantages may be realized when the PDGFRα inhibitor is administered as a single agent and / or in combination with one or more additional therapeutic agents, e.g., standard therapeutic agents. When administered in combination with one or more additional therapeutic agents, in some embodiments, the dosing regimens for the PDGFRα inhibitor (e.g., intermittent dosing) and the additional therapeutic agents are independent of each other. For example, in some embodiments, the PDGFRα inhibitor is administered using a first dosing regimen and the additional therapeutic agent is administered using a second dosing regimen, where the first and second dosing regimens are different.

[0279] In some embodiments, the intermittent dosing schedule includes administering the PDGFRα inhibitor to the subject every other day. In some embodiments, the PDGFRα inhibitor is administered to the subject once daily. In some embodiments, the PDGFRα inhibitor is administered to the subject twice daily. In some embodiments, the PDGFRα inhibitor is administered to the subject three times daily. In some embodiments, the PDGFRα inhibitor is administered to the subject four times daily. In some embodiments, the PDGFRα inhibitor is administered to the subject every three days. In some embodiments, the PDGFRα inhibitor is administered to the subject every four days. In some embodiments, the PDGFRα inhibitor is administered to the subject every five days. In some embodiments, the PDGFRα inhibitor is administered to the subject every six days. In some embodiments, the PDGFRα inhibitor is administered to the subject once weekly. In some embodiments, the PDGFRα inhibitor is administered to the subject once every eight days. In some embodiments, the PDGFRα inhibitor is administered to the subject once every nine days. In some embodiments, the PDGFRα inhibitor is administered to the subject every 10 days. In some embodiments, the PDGFRα inhibitor is administered to the subject every 11 days. In some embodiments, the PDGFRα inhibitor is administered to the subject every 12 days. In some embodiments, the PDGFRα inhibitor is administered to the subject every 13 days. In some embodiments, the PDGFRα inhibitor is administered to the subject once every two weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every three weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once a month. In some embodiments, the PDGFRα inhibitor is administered to the subject once every five weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every six weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every seven weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every two months. In some embodiments, the PDGFRα inhibitor is administered to the subject once every nine weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every 10 weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every 11 weeks. In some embodiments, the PDGFRα inhibitor is administered to the subject once every 3 months. In some embodiments, the PDGFRα inhibitor is administered to the subject once every 4 months. In some embodiments, the PDGFRα inhibitor is administered to the subject once every 5 months.In some embodiments, the PDGFRα inhibitor is administered to the subject once every six months. In some embodiments, the PDGFRα inhibitor is administered to the subject once every twelve months.

[0280] In some embodiments, the intermittent dosing schedule includes administering a first and second dose of a PDGFRα inhibitor to the subject, where the second dose is administered at least one day after the first dose. In some embodiments, the second dose is administered at least two days after the first dose. In some embodiments, the second dose is administered at least three days after the first dose. In some embodiments, the second dose is administered at least four days after the first dose. In some embodiments, the second dose is administered at least five days after the first dose. In some embodiments, the second dose is administered at least six days after the first dose. In some embodiments, the second dose is administered at least seven days after the first dose. In some embodiments, the second dose is administered at least eight days after the first dose. In some embodiments, the second dose is administered at least nine days after the first dose. In some embodiments, the second dose is administered at least 10 days after the first dose. In some embodiments, the second dose is administered at least 11 days after the first dose. In some embodiments, the second dose is administered at least 12 days after the first dose. In some embodiments, the second dose is administered at least 13 days after the first dose. In some embodiments, the second dose is administered at least 2 weeks after the first dose. In some embodiments, the second dose is administered at least 3 weeks after the first dose. In some embodiments, the second dose is administered at least 1 month after the first dose. In some embodiments, the second dose is administered at least 2 months after the first dose. In some embodiments, the second dose is administered at least 3 months after the first dose. In some embodiments, the second dose is administered at least 4 months after the first dose. In some embodiments, the second dose is administered at least 5 months after the first dose. In some embodiments, the second dose is administered at least six months after the first dose.In some embodiments, the second dose is administered at least 12 months after the first dose.

[0281] In some embodiments, the intermittent dosing schedule comprises administering a first dose set and a second dose set of a PDGFRα inhibitor to the subject, wherein the second dose set is administered at least one day after the first dose set. In some embodiments, the first dose set may be administered over, for example, one, two, three, or four days, for example, one, two, three, or four doses per day. In some embodiments, the second dose set may be administered over, for example, one, two, three, or four days, for example, one, two, three, or four doses per day.

[0282] In some embodiments, the second dose set is administered at least two days after the first dose set. In some embodiments, the second dose set is administered at least three days after the first dose set. In some embodiments, the second dose set is administered at least four days after the first dose set. In some embodiments, the second dose set is administered at least five days after the first dose set. In some embodiments, the second dose set is administered at least six days after the first dose set. In some embodiments, the second dose set is administered at least seven days after the first dose set. In some embodiments, the second dose set is administered at least eight days after the first dose set. In some embodiments, the second dose set is administered at least nine days after the first dose set. In some embodiments, the second dose set is administered at least ten days after the first dose set. In some embodiments, the second dose set is administered at least eleven days after the first dose set. In some embodiments, the second dose set is administered at least 12 days after the first dose set. In some embodiments, the second dose set is administered at least 13 days after the first dose set. In some embodiments, the second dose set is administered at least 2 weeks after the first dose set. In some embodiments, the second dose set is administered at least 3 weeks after the first dose set. In some embodiments, the second dose set is administered at least 1 month after the first dose set. In some embodiments, the second dose set is administered at least 2 months after the first dose set. In some embodiments, the second dose set is administered at least 3 months after the first dose set. In some embodiments, the second dose set is administered at least 4 months after the first dose set. In some embodiments, the second dose set is administered at least 5 months after the first dose set.In some embodiments, the second dose set is administered at least 6 months after the first dose set. In some embodiments, the second dose set is administered at least 12 months after the first dose set.

[0283] While not bound by any single theory, in some embodiments, the use of intermittent dosing schedules allows the OPC population of a treated subject (i.e., a subject previously administered a PDGFRα inhibitor) to recover sufficiently before the next dose of the PDGFRα inhibitor. As described herein, OPCs can self-replicate, and therefore, under steady state conditions (i.e., without inhibition of PDGFRα activity), the subject's OPC population is continuously replenished and maintained. As described and demonstrated herein, when the PDGFRα inhibitors described herein are administered to a subject, the PDGFRα inhibitors induce the differentiation of OPCs within the subject into oligodendrocytes. In some embodiments, this may result in a decrease in the OPC population within the subject, and therefore, immediate or rapid subsequent administration of the PDGFRα inhibitor may diminish its therapeutic activity (e.g., reduce further oligodendrocyte development). Accordingly, in some embodiments, an intermittent dosing schedule useful to the present disclosure comprises administering a first dose and a second dose of a PDGFRα inhibitor, wherein the second dose of the PDGFRα inhibitor is administered to the subject after the subject's OPC population has fully recovered from the effects of the first dose of the PDGFRα inhibitor. Where referring to an OPC population (e.g., a subject treated with a PDGFRα inhibitor), the expression “fully recovered” means that the number of OPCs in the subject has increased so that additional administration of the PDGFRα inhibitor may result in the development of additional oligodendrocytes. In some embodiments, a fully recovered OPC population is equivalent to that of a reference subject, wherein the reference subject includes (i) a subject prior to the first dose of the PDGFRα inhibitor, (ii) a corresponding normal healthy subject (i.e., not suffering from demyelinating disease) that has not received the PDGFRα inhibitor, or (iii) both (i) and (ii). In some embodiments, the number of OPCs in a fully recovered OPC population is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% of the number of OPCs in the reference OPC population.

[0284] Therefore, if the method provided herein involves administering a first and second dose of a PDGFRα inhibitor to a subject (for example, to treat a demyelinating disease), in some embodiments, the first dose of the PDGFRα inhibitor is administered to the subject, and the subject's OPC population is then evaluated, and if the subject's OPC population has recovered sufficiently compared to a reference subject (for example, as described above), the second dose of the PDGFRα inhibitor is administered to the subject. In some embodiments, the second dose of the PDGFRα inhibitor is administered to the subject if the size of the subject's OPC population (for example, the number of OPCs in the OPC population) is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% of the size of the reference subject's OPC population. In some embodiments, if an additional dose of the PDGFRα inhibitor is administered to the subject, the additional dose is administered to the subject at the same dosing interval as the first and second doses of the PDGFRα inhibitor were used. In some embodiments, when an additional dose of a PDGFRα inhibitor is administered to a subject, the method includes reassessing the subject's OPC population after a second dose, where the additional dose of the PDGFRα inhibitor is administered to the subject if the subject's OPC population has recovered sufficiently compared to a reference subject (e.g., as described above). In some embodiments, the additional dose of the PDGFRα inhibitor is administered to the subject if the size of the subject's OPC population (e.g., the number of OPCs in the OPC population) is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100% of the size of the reference subject's OPC population. As further described elsewhere in this disclosure, in some embodiments, the length of time required for the subject's OPC population to recover sufficiently after the initial dose of the PDGFRα inhibitor is the dosing interval of the PDGFRα inhibitor.

[0285] Whether the target OPC population has recovered sufficiently (and therefore the determination of the dosing interval for the PDGFRα inhibitor) can be determined using any suitable method known in the art. For example, in some embodiments, a cuprizon animal model can be used. As demonstrated herein, in some embodiments, a suitable dosing interval for the PDGFRα inhibitor described herein can be determined by administering a first and second dose of the PDGFRα inhibitor to a cuprizon animal model and then assessing the number of newly generated oligodendrocytes present in the brain of the animals (e.g., by quantifying the number of GPR17+ cells in the brain). If the number of newly generated oligodendrocytes in the brain is the same as or increases compared to a reference (e.g., the number of newly generated oligodendrocytes present in the brain of a corresponding animal that received a single dose of the PDGFRα inhibitor), the suitable dosing interval is the time between the administration of the first dose and the administration of the second dose of the PDGFRα inhibitor, or any longer time. If the number of newly generated oligodendrocytes in the brain is reduced compared to a reference (e.g., the number of newly generated oligodendrocytes present in the brain of a corresponding animal that received a single dose of a PDGFRα inhibitor), then the preferred dosing interval is longer than the time between the administration of the first and second doses of the PDGFRα inhibitor.

[0286] In some embodiments, the preferred dosing interval for a PDGFRα inhibitor is directly related to the plasma level of the PDGFRα inhibitor. Therefore, in some embodiments, a method for determining the preferred dosing interval for a PDGFRα inhibitor includes administering a first dose of the PDGFRα inhibitor to a subject and determining the plasma level of the PDGFRα inhibitor in the subject. In some embodiments, if the plasma level of the PDGFRα inhibitor is equivalent to that of a reference (e.g., a subject who has not previously received a dose of the PDGFRα inhibitor and / or a subject who has previously received a dose of the PDGFRα inhibitor but whose OPC population has recovered sufficiently as described herein), the preferred dosing interval is the time between administering the first dose of the PDGFRα inhibitor and when the plasma level of the PDGFRα inhibitor in the subject is equivalent to that of the reference. In some embodiments, if the plasma level of the PDGFRα inhibitor is reduced compared to a reference level (e.g., the plasma level of the PDGFRα inhibitor in the subject immediately after administration of the initial dose of the PDGFRα inhibitor, e.g., about 4 hours later), the preferred dosing interval is the time between the administration of the first dose of the PDGFRα inhibitor and the time when the plasma level of the PDGFRα inhibitor in the subject reaches the reduced level compared to the reference level. In some embodiments, an additional dose of the PDGFRα inhibitor is administered to the subject when the plasma level of the subject has decreased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or about 100% compared to the reference level. In some embodiments, the brain-to-plasma ratio may be assessed in preclinical species using bioanalytical gases or liquid chromatography and mass spectrometry.

[0287] In some embodiments, plasma levels of the PDGFRα inhibitor are related to the half-life of the PDGFRα inhibitor. In some embodiments, intermittent dosing schedules useful to the present disclosure include administering two or more doses of the PDGFRα inhibitor at dosing intervals longer than the half-life of the PDGFRα inhibitor. In some embodiments, the dosing interval is at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% longer than the half-life of the PDGFRα inhibitor. In some embodiments, the dosing interval is at least 1.5 times, at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least about 30 times, at least 40 times, or at least 50 times longer than the half-life of the PDGFRα inhibitor. In some embodiments, the dosing interval is at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, or at least 3 months longer than the half-life of the PDGFRα inhibitor.

[0288] In some embodiments, the methods described herein (e.g., for treating demyelinating diseases) may include the administration of additional therapeutic agents. For example, in some embodiments, the PDGFRα inhibitors described herein are used in combination with immunomodulatory agents. In some embodiments, the PDGFRα inhibitors are used alone in the methods provided herein.

[0289] As described herein, “immunomodulatory agent” refers to a therapeutic agent that acts by modulating (e.g., increasing and / or decreasing) one or more aspects of the immune response. In some embodiments, immunomodulatory agents useful to the present disclosure can reduce or mitigate the pro-inflammatory nature of demyelinating diseases (e.g., multiple sclerosis). For example, in some embodiments, immunomodulatory agents may block the production of pro-inflammatory mediators, promote the production of anti-inflammatory cytokines (e.g., IL-10 or TGF-β), or regulate T cells (T). reg This can involve promoting the production of ), influencing B cells, preventing immune cells from entering the brain, or any combination thereof. Non-limiting examples of immunomodulatory agents useful in this disclosure include interferon beta-1b (BETASERON®, EXTAVIA®), interferon beta-1a (AVONEX®, REBIF®), pegylated interferon beta-1a (PLEGRIDY®), alemtuzumab (LEMTRADA®), natalizumab (TYSABRI®), ocrelizumab (OCREVUS®), ofatumumab (KESIMPTA®), glatiramer acetate (COPAXONE®, GLATOPA®), teriflunomide (AUBAGIO®), dimethyl fumarate (TECFIDERA®), monomethyl fumarate (BAFIERTAM®), diroximel fumarate (Vumerity®), and combinations thereof.

[0290] In some embodiments, when an additional therapeutic agent (e.g., an immunomodulator) is administered to a subject, the additional therapeutic agent is administered to the subject before the administration of the PDGFRα inhibitor. Thus, in some embodiments, any of the methods provided herein (e.g., methods for treating demyelinating diseases) include administering a PDGFRα inhibitor to a subject, where the subject has previously received an additional therapeutic agent (e.g., an immunomodulator). In some embodiments, the additional therapeutic agent is administered after the administration of the PDGFRα inhibitor. In some embodiments, the additional therapeutic agent is administered concurrently with the administration of the PDGFRα inhibitor.

[0291] In some embodiments, to improve delivery to the CNS, the PDGFRα inhibitors described herein may be administered to a subject in combination with an agent that assists the delivery of the inhibitor to the CNS (e.g., enabling the inhibitor to cross the blood-brain barrier). In some embodiments, the PDGFRα inhibitors described herein are administered to a subject in combination with a peptide blood-brain barrier (BBB) ​​shuttle, where the peptide BBB shuttle enhances the ability of the PDGFRα inhibitor to cross the blood-brain barrier and reach the CNS. Non-limiting examples of such peptide BBB shuttles are provided in Table 2 (below). See, for example, Oller-Salvia et al., Chem Soc Rev 45:4690-4707 (2016) and Jafari et al., Expert Opinion on Drug Delivery 16:583-605 (2019). [Table 2]

[0292] VI. Enumerations This disclosure also provides the following enumeration clauses:

[0293] Clause 1. Compounds of Formula I: [ka] The compound,

[0294] or a pharmaceutically acceptable salt or solvate thereof,

[0295] [ka] However, it exhibits single or double bonds such that all valencies are satisfied.

[0296] X 1 , X 2 , X 3 , and X 4 However, N and CR a Selected from, however, X 1 , X 2 , X 3 , and X 4 The condition is that two or fewer of them are N,

[0297] Y 1 and Y 2 One of them is N, and Y 1 and Y 2 The other of these is C,

[0298] Each R a These are independently selected from H, halo, C1-C4 alkyl, and C1-C4 alkoxy.

[0299] R 1 However, all of these can be optionally substituted with 1, 2, 3, 4, 5, or 6 substituents selected from halo, hydroxy, oxo, C1-C4 alkyl, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkoxy C1-C4 alkyl, 3-8 membered heterocyclil, and 3-8 membered heterocyclil C1-C4 alkyl, provided that the number of substituents does not exceed the number of substituted positions.

[0300] R 2However, these are selected from cycloalkyl, cycloalkenyl, alkyl, oxoalkylamino, aminoalkylamino, amino, heterocyclyl, heteroaryl, aminoheterocyclyl, heterocyclylamino, and aminoalkylamino, all of which may be optionally substituted with one or more substituents selected from D, halo, hydroxy, oxo, and C1-C4 alkyl.

[0301] R 2 However, 1, 2, or 3 R 3 Replaced by,

[0302] R 3 However, aryl, heteroaryl, -C(O)R 31 , -C(O)OR 31 -C(O)NR 31 R 32 -S(O)2NR 31 R 32 -S(O)(NR 33 )R 31 -S(O)2R 31 -S(O)(NR 33 )NR 31 R 32 -C(S)NR 31 R 32 Selected from C3-C8 cycloalkyls, 3-8 membered heterocyclines, and C1-C4 alkyls, all of which are optional, with 1, 2, 3, 4, or 5 R groups. 30 It can be replaced by,

[0303] Each R 30 These are independent of D, Halo, Ariel, -OR 300 , -NR 300 R 303 , -S(O) r R 300 , -C(O)R 300 -C(=CR) 34 R 35 )R 300 , and [ka] Selected from,

[0304] r is selected from 0, 1, and 2.

[0305] Each R 300 These are independently selected from C1-C6 alkyl, C3-C7 cycloalkyl, aryl, heteroaryl, 3-8 membered heterocyclyl, and 3-8 membered heterocyclylaryl, all of which may be optionally substituted with 1, 2, 3, 4, or 5 substituents selected from D, halo, hydroxy, amino, alkylamino, cyano, C1-C4 alkyl, C1-C4 haloalkyl, and C1-C4 alkoxy.

[0306] Each R 301 These are independently selected from H, halo, and C1-C4 alkyl groups.

[0307] Each R 302 These are independently selected from H, F, hydroxyl, amino, alkylamino, oxo, and C1-C4 alkyloxy,

[0308] Each R 303 These are independently selected from H and C1-C4 alkyl groups.

[0309] n, o, and p are each independently selected from 0, 1, 2, 3, and 4.

[0310] Each R 31 These are independently selected from C1-C8 alkyl, aryl C1-C4 alkyl, heteroaryl C1-C4 alkyl, heterocyclyl, heterocyclyl C1-C4 alkyl, cycloalkyl, and cycloalkyl C1-C4 alkyl, all of which are optional selections of D, halo, cyano, hydroxy, amino, -OCF3, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 alkoxy, hydroxy C1-C4 alkyl, and -S(O)2NR 304 R 305 , -C(O)OR 304 R 305 -C(O)NR 304 R 305 , and -NR 304 C(O)R 305It may be substituted with 1, 2, 3, 4, or 5 substituents selected from the following:

[0311] Each R 304 and R 305 These are independently selected from H and C1-C4 alkyl groups.

[0312] Each R 32 These are independently selected from H and C1-C4 alkyl groups, or

[0313] R 31 and R 32 However, along with the atoms connected to them, D, halo, cyano, C1-C4 alkyl, C1-C4 haloalkyl, and -C(O)NR 34 R 35 It forms a 5- to 8-membered heterocisyl which is optionally substituted with 1, 2, 3, 4, or 5 substituents selected from the above,

[0314] Each R 34 and R 35 These are independently selected from H, C1-C4 alkyl, and C1-C4 haloalkyl,

[0315] Each R 33 These independently comprise H, C1-C4 alkyl, C1-C4 haloalkyl, and -C(O)R 34 Selected from, or

[0316] R 31 and R 33 The compound, or a pharmaceutically acceptable salt or solvate thereof, which forms a 4- to 8-membered heterocisyl group together with the atoms connected to them.

[0317] Clause 2.Y 1 N is Y 2 A compound as described in Clause 1, or a pharmaceutically acceptable salt or solvate thereof, wherein C is C.

[0318] Clause 3.Y 1 C is Y 2A compound as described in Clause 1, or a pharmaceutically acceptable salt or solvate thereof, wherein N is present.

[0319] Clause 4.X 1 N is X 2 CR a X 3 CR a X 4 CR a The compound described in any one of clauses 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

[0320] Clause 5.X 1 CR a X 2 N is X 3 CR a X 4 CR a The compound described in any one of clauses 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

[0321] Clause 6.X 1 CR a X 2 CR a X 3 N is X 4 CR a The compound described in any one of clauses 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

[0322] Clause 7.X 1 CR a X 2 CR a X 3 CR a X 4 A compound described in any one of clauses 1 to 3, wherein N is N, or a pharmaceutically acceptable salt or solvate thereof.

[0323] Clause 8.X 1 CR a X 2 CR a X 3CR a X 4 CR a The compound described in any one of clauses 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

[0324] Clause 9.R 1 A compound described in any one of clauses 1 to 8, which is a five-membered or six-membered heteroaryl, or a pharmaceutically acceptable salt or solvate thereof.

[0325] Clause 10.R 1 However, the compound described in any one of clauses 1 to 9, or a pharmaceutically acceptable salt or solvate thereof, is a pyrazolyl that is optionally substituted.

[0326] Article 11.

[0327] R 1 but, [ka] Selected from,

[0328] R 10 However, it is selected from H, C1-C4 alkyl, C1-C4 alkoxy, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, and C1-C4 alkylsulfonyl, and the C1-C4 alkyl, C1-C4 alkoxy, amino C1-C4 alkyl, hydroxy C1-C4 alkyl, and C1-C4 alkylsulfonyl are optionally selected, and hydroxyl, C1-C4 alkoxy, NR 10a R 10b R may be substituted with one or more substituents selected from halo and deuterium, where R 10a and R 10b However, it is selected from hydrogen and C1-C4 alkyl, or R 10a and R 10b A compound described in any one of clauses 1 to 10, or a pharmaceutically acceptable salt or solvate thereof, which, together with the nitrogen atoms bonded to them, forms a 4-membered to 8-membered ring.

[0329] Clause 12.R 1 but, [ka] The compounds described in Clause 11, or their pharmaceutically acceptable salts or solvates.

[0330] Clause 13.R 10 However, the compounds described in Clause 11 or 12, or their pharmaceutically acceptable salts or solvates, are CH3.

[0331] Clause 14.R 2 The compounds described in any one of clauses 1 to 13, or pharmaceutically acceptable salts or solvates thereof, are heterocyclyls optionally substituted with C1-C4 alkyl or oxo groups.

[0332] Clause 15.R 2 However, one R 3 A compound described in any one of clauses 1 to 14, or a pharmaceutically acceptable salt or solvate thereof, which is substituted by [the compound].

[0333] Clause 16.R 2 but, [ka] [ka] Selected from,

[0334] [ka] However, it exhibits single or double bonds such that all valencies are satisfied.

[0335] m is selected from 0, 1, 2, 3, 4, 5, and 6.

[0336] Z 1 , Z 2 , and Z 3However, N and CR a A compound selected from any one of clauses 1 to 15, or a pharmaceutically acceptable salt or solvate thereof.

[0337] Clause 17.R 2 but, [ka] A compound selected from the compounds described in Clause 16, or a pharmaceutically acceptable salt or solvate thereof.

[0338] Article 18. Formula Ia: [ka] It has,

[0339] In the formula, a and b are each independently selected from 1, 2, and 3.

[0340] A compound described in any one of clauses 1 to 17, or a pharmaceutically acceptable salt or solvate thereof, wherein Q is selected from -CH- and -N-, provided that if Q is -N-, a and b are not 1.

[0341] Article 19. Formula II: [ka] A compound described in any one of clauses 1 to 18, or a pharmaceutically acceptable salt or solvate thereof, having the above characteristics.

[0342] Article 20. Formula IIa: [ka] A compound described in any one of clauses 1 to 19, or a pharmaceutically acceptable salt or solvate thereof, having the above characteristics.

[0343] Clause 21.R 3 However, 1, 2, 3, 4, or 5 R 30A compound described in any one of clauses 1 to 20, or a pharmaceutically acceptable salt or solvate thereof, which is a heteroaryl optionally substituted by [a specific compound].

[0344] Clause 22.R 3 but, [ka] [ka] Selected from,

[0345] In the formula, A 1 However, it is selected from O, S, and N, R 36 The compounds described in any one of Clauses 1 to 21, or pharmaceutically acceptable salts or solvates thereof, selected from hydrogen, optionally substituted C1-C6 alkyls, and optionally substituted C1-C6 alkylaryls.

[0346] Clause 23.R 3 but, [ka] A compound selected from any one of clauses 1 to 22, or a pharmaceutically acceptable salt or solvate thereof.

[0347] Clause 24.R 30 but, [ka] The compound described in any one of clauses 1 to 23, or a pharmaceutically acceptable salt or solvate thereof.

[0348] Clause 25.R 300 but, [ka] A compound selected from the compounds described in Clause 24, or a pharmaceutically acceptable salt or solvate thereof.

[0349] Clause 26.R 300 but, [ka] The compounds described in Clause 25, or their pharmaceutically acceptable salts or solvates.

[0350] Clause 27.R 301 H is R 302 A compound described in any one of clauses 1 to 26, or a pharmaceutically acceptable salt or solvate thereof, wherein the compound is H or CH3.

[0351] Clause 28.R 3 but, [ka] A compound selected from any one of clauses 1 to 22, or a pharmaceutically acceptable salt or solvate thereof.

[0352] Clause 29.R 3 but, [ka] A compound selected from the compounds described in Clause 28, or a pharmaceutically acceptable salt or solvate thereof.

[0353] Clause 30.R 32 A compound as described in Clause 29, or a pharmaceutically acceptable salt or solvate thereof, wherein the compound is H.

[0354] Clause 31.R 31 but, [ka] [ka] Selected from,

[0355] In the formula, R 31aHowever, it is selected from H, D, alkylamino, C1-C4 alkyl, and -CF3.

[0356] R 31b The compounds described in any one of clauses 28 to 30, selected from H, D, halo, hydroxy, amino, alkylamino, C1-C4 alkyl, -CF3, and -OCF3, or pharmaceutically acceptable salts or solvates thereof.

[0357] Clause 32. A compound described in Clause 1, or a pharmaceutically acceptable salt or solvate thereof, selected from any one of compounds 156, 250, 275, and 326-406 in Table 1.

[0358] Clause 33. A compound described in any one of Clauses 1 to 32 that exhibits any of the following properties: (i) promotes differentiation from OPCs to oligodendrocytes, (ii) promotes the expression of proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17 (GPR17), myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or a combination thereof), (iii) promotes axon myelination, (iv) promotes remyelination of demyelinated axons, (v) inhibits PDGFRα kinase activity, (vi) achieves a brain-to-plasma ratio greater than 0.1 when systemically administered to a subject, and (vii) exhibits one or more of any combination thereof.

[0359] Clause 34. A compound according to Clause 33 that can inhibit PDGFRα kinase activity.

[0360] Clause 35. IC of PDGFRα kinase activity less than 500 nM, less than 400 nM, less than 300 nM, less than 200 nM, less than 100 nM, less than 75 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, or less than 5 nM. 50 The compounds described in Clause 34 that can be inhibited by...

[0361] Clause 36. The IC of the PDGFRα inhibitor 50 The compound described in Clause 35, determined using an enzymatic PDGFRα kinase assay (e.g., the Promega kinase assay described in Example 131).

[0362] Clause 37. The compound according to Clause 36, wherein the enzymatic PDGFRα kinase assay comprises 20 ng of purified PDGFRα protein, 150 μM of ATP, and 1 μg of the substrate poly(Glu4Tyr1) in a volume of 15 μl.

[0363] Clause 38. A pharmaceutical composition comprising a compound described in any one of Clauses 1 to 37, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable additive.

[0364] Clause 39. A kit comprising a compound described in any one of Clauses 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a composition described in Clause 38, and instructions for use.

[0365] Clause 40. A compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38, for use in therapeutic purposes.

[0366] Clause 41. A method for treating a demyelinating disease in a subject requiring treatment for the demyelinating disease, comprising administering to the subject a therapeutically effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0367] Clause 42. A method for improving the performance of a subject in a test for evaluating one or more symptoms associated with a demyelinating disease, comprising administering to the subject a therapeutically effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38, wherein, after the administration, the performance of the subject in the test is improved compared to a reference subject (e.g., the subject before the administration).

[0368] Clause 43. The method according to Clause 42, wherein the examination is selected from visual evoked potential (VEP) testing, multifocal visual evoked potential (mfVEP) testing, low-contrast visual acuity (LC-VA) testing, magnetic resonance imaging (MRI) (e.g., magnetic transfer resonance, myelin water fraction (MWF), and quantitative susceptibility mapping (QSM)), electromyography (EMG), nerve conduction velocity (NCV) testing, expanded disability status scale (EDSS), gait time measurement (e.g., 25-foot gait time measurement), 9-hole peg test (9HPT), optical coherence tomography (OCT), quality of life measurement (e.g., quality of life in multiple sclerosis - 54 and visual quality of life), cognitive function assessment (e.g., Montreal cognitive function assessment), or a combination thereof.

[0369] Article 44. The demyelinating disease is defined as acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis, acute transverse myelitis, adrenoleukodystrophy, adrenal spinal neuropathy, Alexander disease, Alzheimer's disease, aminoaciduria, amyotrophic lateral sclerosis, anti-MAG peripheral neuropathy, anti-MOG related spectrum, Barlow concentric sclerosis, brain injury, CAMFAK syndrome, Canavan disease, carbon monoxide poisoning, central pontine myelinolysis, cerebral hypoxia, cerebral ischemia, Charcot-Marie-Tooth disease, chronic inflammatory demyelinating polyneuropathy Neuropathies, chronic traumatic encephalopathy, syndrome consisting of the first episode (CIS), congenital cataracts, copper deficiency-related conditions, delayed hypoxic leukoencephalopathy, Schilder's generalized encephalosclerosis, generalized myelin-destructive sclerosis, extrapontine myelin-disintegrating Gaucher disease, Guillain-Barré syndrome, hereditary neuropathy, hereditary pressure-fragility neuropathy, HTLV-1-associated myelopathy, Hurler syndrome, myelin hypoplasia, hypoxic brain injury, Krabbe disease, Leber's hereditary optic atrophy and associated mitochondrial disorders, leukodystrophy, Marky Afava-Bignami disease, metachromatic leukodystrophy, multiple sclerosis (e.g., primary progressive multiple sclerosis (PPMS), relapsing-remitting multiple sclerosis (RRMS), secondary progressive multiple sclerosis (SPMS), progressive-relapsing multiple sclerosis, Marburg multiple sclerosis, tumor-like multiple sclerosis, and neurospinal optic sclerosis), multiple system atrophy, myelin-destroying disorders, myelopathy, nerve injury, neuromyelitis optica (NMO), Niemann-Pick disease, optic neuropathy, optic neuritis (e.g., acute optic neuritis and chronic relapsing inflammatory optic neuritis) The methods described in Clauses 41-43, including, but not limited to, craniitis iontophoresis (CRION), osmotic demyelinating syndrome, Parkinson's disease, Pelizaeus-Merzbach disease, peripheral neuropathy, phenylketonuria, progressive inflammatory neuropathy, progressive multifocal leukoencephalopathy, progressive subcortical ischemic demyelination, reperfusion injury, Schilder's disease, isolated sclerosis, spinal cord injury, subacute sclerosing panencephalitis, tabes dorsalis, Tay-Sachs disease, transverse myelitis, traumatic brain injury, tropical spastic paraplegia, vitamin B12 deficiency, cerebral palsy, or any combination thereof.

[0370] Clause 45. The method according to any one of Clauses 41 to 44, wherein the demyelinating disease is characterized by the demyelinating of one or more cells in the CNS of the subject.

[0371] Clause 46. The method according to any one of Clauses 41 to 45, wherein the demyelinating disease is multiple sclerosis.

[0372] Clause 47. The method according to Clause 46, wherein the multiple sclerosis includes a syndrome consisting of a first episode ("CIS"), relapsing-remitting MS ("RRMS"), secondary progressive MS ("SPMS"), primary progressive MS ("PPMS"), optic neuritis, or transverse myelitis.

[0373] Clause 48. The method according to any one of Clauses 41 to 45, wherein the demyelinating disease is optic neuritis.

[0374] Clause 49. The method according to any one of Clauses 41 to 48, wherein the treatment of the demyelinating disease includes reducing one or more symptoms associated with the demyelinating disease.

[0375] Clause 50. The method according to Clause 49, wherein one or more of the symptoms include fatigue, dizziness, malaise, high fever and hyperthermia, extreme coldness of the hands and feet, weakness and stiffness of muscles and joints, weight changes, digestive or gastrointestinal disorders, hypotension, hypertension, irritability, anxiety, depression, visual impairment (e.g., blurred vision, diplopia, decreased low-contrast visual acuity (LC-VA)), ataxia, clonus, convulsions, dysarthria, weakness, clumsiness, paralysis of the hands, hemiplegia, genital loss of sensation, sexual dysfunction, incoordination, paresthesia, oculoparalysis, muscular incoordination, loss of sensation, tingling, anesthesia, pain, neurological symptoms, cognitive impairment, unsteady gait, balance problems, dizziness, spastic paraplegia, incontinence, hearing impairment, speech impairment, loss of smell, anosmia, or a combination thereof.

[0376] Clause 51. A method for promoting axonal myelination in a subject requiring promotion of axonal myelination, the method comprising administering to the subject an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0377] Clause 52. The method according to Clause 51, wherein the promotion of axon myelination results in increased expression of one or more of the following markers within the subject: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or any combination thereof.

[0378] Clause 53. The method according to Clause 51 or 52, wherein the myelination of the axon can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or any combination thereof.

[0379] Clause 54. A method for promoting the remyelination of demyelinated axons in a subject requiring promotion of said remyelination of said demyelinated axons, the method comprising administering to said subject an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0380] Clause 55. The method according to Clause 54, wherein the promotion of remyelination of the demyelinated axon results in increased expression of one or more of the following markers within the subject: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or any combination thereof.

[0381] Clause 56. The method according to Clause 54 or 55, wherein the remyelination of the demyelinated axon can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or any combination thereof.

[0382] Clause 57. A method for reducing demyelination of myelinated nerve axons in a subject requiring reduction of demyelination of said myelinated nerve axons, the method comprising administering to the subject an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0383] Clause 58. The method according to Clause 57, wherein the reduction in demyelination of the myelinated nerve axons results in an increase in the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or any combination thereof.

[0384] Clause 59. The method according to Clause 57 or 58, wherein the reduction of demyelination of the myelinated nerve axon can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or any combination thereof.

[0385] Clause 60. A method for activating oligodendrocyte progenitor cells (OPCs) in a subject requiring activation of such OPCs in the central nervous system (CNS), the method comprising administering to the subject an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0386] Clause 61. The method described in any one of Clauses 51 to 60, wherein the subject has or is at risk of developing a demyelinating disease, for example, one of the diseases described in any one of Clauses 44 to 50.

[0387] Clause 62. The method according to any one of Clauses 51 to 60, wherein the method is a method for treating or preventing a demyelinating disease, for example, a disease described in any one of Clauses 44 to 50.

[0388] Clause 63. The method according to any one of Clauses 41 to 62, wherein the compound or the pharmaceutical composition is administered once to the subject.

[0389] Clause 64. The method according to any one of Clauses 41 to 62, wherein the compound or the pharmaceutical composition is administered to the subject two or more times using intermittent dosing.

[0390] Clause 65. The method according to Clause 64, wherein the intermittent dosing comprises administering the compound or the pharmaceutical composition to the subject every other day, every three days, every four days, every five days, every six days, once a week, every eight days, every nine days, every ten days, every eleven days, every twelve days, every thirteen days, once every two weeks, once every three weeks, once a month, once every two months, once every three months, once every four months, once every five months, once every six months, or once every twelve months.

[0391] Clause 66. The method according to Clause 64 or 65, wherein the intermittent dosing comprises administering to the subject a first dose and a second dose of the compound or the pharmaceutical composition, the second dose being administered at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or at least 12 months after the administration of the first dose.

[0392] Clause 67. The method according to Clause 66, wherein the second dose is administered to the subject one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen days, two weeks, three weeks, one month, two months, three months, four months, five months, six months, or twelve months after the administration of the first dose.

[0393] Clause 68. The method according to any one of Clauses 41 to 67, wherein, after administration, the compound or the pharmaceutical composition can achieve a brain-to-plasma ratio of greater than 0.1, greater than 0.2, greater than 0.3, greater than 0.4, greater than 0.5, greater than 0.6, greater than 0.7, greater than 0.8, greater than 0.9, greater than 1.0, greater than 1.1, greater than 1.2, greater than 1.3, greater than 1.4, greater than 1.5, greater than 1.6, greater than 1.7, greater than 1.8, greater than 1.9, or greater than 2.0.

[0394] Clause 69. The method according to any one of Clauses 41 to 68, further comprising administering an additional therapeutic agent to the subject.

[0395] Clause 70. The method according to Clause 69, wherein the additional therapeutic agent includes a standard therapeutic agent.

[0396] Clause 71. The method according to Clause 69 or 70, wherein the additional therapeutic agent includes an immunomodulator.

[0397] Clause 72. The method according to Clause 71, wherein the additional therapeutic agent is selected from interferon beta-1b, interferon beta-1a, pegylated interferon beta-1a, alemtuzumab, natalizumab, ocrelizumab, ofatumumab, glatiramer acetate, teriflunomide, dimethyl fumarate, monomethyl fumarate, diloximel fumarate, fingolimod hydrochloride, siponimod fumarate, ozanimod hydrochloride, BTK inhibitors, or pharmaceutically acceptable salts thereof.

[0398] Clause 73. The method according to any one of Clauses 69 to 72, wherein the additional therapeutic agent is administered to the subject before, simultaneously with, or after the administration of the compound or the pharmaceutical composition.

[0399] Clause 74. A method for inducing differentiation from oligodendrocyte progenitor cells (OPCs) to oligodendrocytes, wherein the method comprises contacting the OPCs with an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0400] Clause 75. The method according to Clause 74, wherein the induction of differentiation from OPC to oligodendrocyte results in increased expression of the following markers in the subject: GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or a combination thereof.

[0401] Clause 76. The method according to Clause 74 or 75, wherein the differentiation from OPC to oligodendrocyte is measured by determining the expression of GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or a combination thereof.

[0402] Clause 77. A method for inhibiting PDGFRα activity in cells, the method comprising contacting the cells with an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0403] Clause 78. The method according to Clause 77, wherein the inhibition of PDGFRα activity is measured by one or more of the following: an in vitro OPC differentiation assay (e.g., as described in Example 132), a cuprizone model for demyelination (e.g., as described in Example 134), an in vivo OPC differentiation assay (e.g., as described in Example 134), an enzymatic PDGFRα kinase assay (e.g., as described in Example 131), or any combination thereof.

[0404] Clause 79. The method described in any one of Clauses 74-78, wherein the contact occurs ex vivo or in vivo.

[0405] Clause 80. The method described in any one of Clauses 74 to 79, wherein the method is a therapeutic treatment method.

[0406] Clause 81. A method for treating relapsing-type multiple sclerosis in a subject requiring treatment for said relapsing-type multiple sclerosis, the method comprising administering to the subject an effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38.

[0407] Clause 82. The method according to Clause 81, wherein the induction of differentiation from OPC to oligodendrocyte results in increased expression of the following markers in the subject: GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or a combination thereof.

[0408] Clause 83. The method according to Clause 81 or 82, wherein the differentiation from OPC to oligodendrocyte is measured by determining the expression of GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or a combination thereof.

[0409] Clause 84. The method according to any one of Clauses 81 to 83, wherein the relapsing-type multiple sclerosis includes a syndrome consisting of a first episode ("CIS"), relapsing-remitting MS ("RRMS"), secondary progressive MS ("SPMS"), primary progressive MS ("PPMS"), or transverse myelitis.

[0410] Clause 85. The following: A compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38 for use in one or more of the following ways: (i) promoting differentiation from OPCs to oligodendrocytes; (ii) promoting the expression of proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17, myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, or a combination thereof); (iii) promoting axon myelination; (iv) promoting remyelination of demyelinated axons; (v) inhibiting PDGFRα kinase activity; (vi) achieving a brain-to-plasma ratio greater than 0.1 when systemically administered to a subject; and (vii) any combination thereof.

[0411] Clause 86. A method for treating a PDGF-related tumor in a subject requiring treatment for the PDGF-related tumor, comprising administering to the subject a therapeutically effective amount of a compound described in any one of Clauses 1 to 37 or a pharmaceutical composition described in Clause 38, wherein, after the administration, PDGFRα activity is reduced in the subject.

[0412] Clause 87. The method according to Clause 86, wherein the PDGF-related tumor includes oligodendroglioma.

[0413] Clause 88. The method according to Clause 86 or 87, wherein the method is a therapeutic treatment method.

[0414] This specification shall be fully understood in light of the teachings of the references cited herein. The embodiments herein are for illustrative purposes only and should not be construed as limiting the scope. Those skilled in the art will readily recognize that many other embodiments are encompassed. All publications and patents cited herein are incorporated by reference in their entirety. In the event that any material incorporated by reference conflicts with or is inconsistent with this specification, this specification shall prevail over any such material. No reference herein constitutes prior art.

[0415] Unless otherwise indicated, all figures used herein, including in the claims, representing amounts of components, reaction conditions, etc., should be understood to be modified by the term "approximately" in all instances. Therefore, unless otherwise stated, numerical parameters are approximations and may vary depending on the desired properties to be obtained. At a minimum, and not as an attempt to limit the application of the doctrine of equivalents to the claims, each numerical parameter should be interpreted with regard to significant figures and customary rounding methods.

[0416] The following embodiments are provided as illustrative examples, not as limitations. All references cited throughout this application are expressly incorporated herein by reference. [Examples]

[0417] As further detailed below, this disclosure demonstrates that compounds inhibiting PDGFRα kinase activity in OPCs can induce oligodendrocyte differentiation (e.g., in vitro and in vivo), and that this effect is likely specific to the inhibition of PDGFRα kinase activity. Furthermore, this disclosure demonstrates that such compounds can induce remyelination in animal models of demyelination.

[0418] Table 3 below defines various abbreviations used in the following embodiments and elsewhere in this disclosure. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0419] Generally, for compounds having stereoisomers separated using, for example, chiral chromatography, the absolute stereochemistry of each compound was arbitrarily assigned.

[0420] Preparation of compounds for the synthesis of exemplary PDGFRα inhibitors To synthesize the exemplary compounds disclosed herein, intermediate compounds were first prepared as described below.

[0421] Synthesis of 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine (compound S3) [ka] Step 1. Preparation of 5-(1-methyl-1H-pyrazole-4-yl)pyridazine-3-ol [ka] To a solution of 5-chloropyridazine-3-ol (1.00 g, 7.66 mmol) and 1-methylpyrazole-4-boronic acid pinacol ester (1.59 g, 7.66 mmol) in dioxane (18 mL), 2 M Na2CO3 (15.3 mL, 30.6 mmol) was added. The two-phase mixture was sparged with argon for 10 minutes, and then Pd(dppf)Cl2 (0.28 g, 0.38 mmol) was added. The reaction mixture was sparged with argon for 10 minutes, and then heated under argon at 75°C for 6 hours. The reaction mixture was cooled to room temperature and stirred overnight. The mixture was concentrated under reduced pressure, and then water (30 mL) was added. The aqueous mixture was extracted with 10% EtOH in siRNA (4 × 50 mL) and 50% siRNA in THF (3 × 50 mL). Brine (30 mL) was added to the aqueous layer and then extracted with 10% EtOH in ethyl acetate (3 × 50 mL). The organic layers were combined, dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography while eluting with 0-10% MeOH in dimethylcellulose to obtain 0.93 g (69%) of 5-(1-methyl-1H-pyrazole-4-yl)pyridazin-3-ol as a pale yellow solid. MS(ESI)m / z[M+H] + Calculated value for C8H9N4O: 171.1; Measured value: 171.0.

[0422] Step 2. Preparation of 3-chloro-5-(1-methyl-1H-pyrazole-4-yl)pyridazine [ka] 5-(1-methyl-1H-pyrazole-4-yl)pyridazin-3-ol (6.0 g, 30 mmol, as prepared in the previous step) was treated with POCl3 (75 mL, 0.80 mol). The suspension was warmed at 80°C for 1.5 hours. The reaction mixture was cooled to room temperature, and excess POCl3 was removed under vacuum. Toluene was added to the residue, and then the solvent was removed under reduced pressure. This was repeated two more times. The residue was slowly treated with ice water (350 mL), and then solid NaHCO3 was slowly added to adjust the pH to 8. The reaction mixture was stirred for 30 minutes and then extracted with 10% MeOH in DCM (2 × 200 mL). The aqueous layer was diluted with water (100 mL) and extracted with 10% MeOH in DCM (2 × 200 mL) and 10% trifluoroethanol in DCM (3 × 200 mL). The organic layers were combined, dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography while eluting with 5% MeOH / DCM in 0-100% DCM solution to obtain 5.14 g (87%) of 3-chloro-5-(1-methyl-1H-pyrazole-4-yl)pyridazine as a white solid. MS(ESI)m / z[M+H] + Calculated value for C8H8ClN4: 195.0; measured value: 195.0.

[0423] Step 3. Preparation of N-(2,2-dimethoxyethyl)-5-(1-methyl-1H-pyrazole-4-yl)pyridazine-3-amine [ka] A suspension of 3-chloro-5-(1-methyl-1H-pyrazole-4-yl)pyridazine (5.10 g, 26.2 mmol, as prepared in the previous step) and aminoacetaldehyde dimethyl acetal (29 mL, 0.27 mol) was heated in an oil bath at 120°C for 20 hours, and then an additional aminoacetaldehyde dimethyl acetal (15 mL, 0.14 mol) was added. The reaction mixture was stirred at 120°C for 27 hours, then cooled to room temperature and concentrated under reduced pressure. The residue was partitioned into saturated aqueous NaHCO3 (100 mL) and 10% MeOH in DCM (100 mL). The aqueous layer was washed with 10% MeOH in DCM (4 × 100 mL). The organic layers were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography while eluting with 10% MeOH / DCM in a 0-100% DCM solution to obtain 6.56 g (95%) of N-(2,2-dimethoxyethyl)-5-(1-methyl-1H-pyrazole-4-yl)pyridazine-3-amine as a yellowish-brown solid. MS(ESI)m / z[M+H] + C 12 H 18 Calculated value for N5O2: 264.2; measured value: 264.0.

[0424] Step 4.7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine [ka] Concentrated H2SO4 (52 mL) was carefully added with stirring to N-(2,2-dimethoxyethyl)-5-(1-methyl-1H-pyrazole-4-yl)pyridazin-3-amine (15.7 g, 59.6 mmol, as prepared in the previous step). After 15 minutes, the reaction mixture was cooled in an ice bath, and 1 M NaOH (100 mL) was carefully added to the reaction mixture. 50% NaOH (approximately 210 mL) was slowly added to the reaction mixture, which was cooled to a basic pH (pH 12). The pH was adjusted to pH 8 by slowly adding 1 M H2SO4. The reaction mixture was partitioned into DCM (400 mL) and water (1200 mL). The aqueous layer was washed with DCM (2 × 300 mL). The aqueous layer containing the emulsion was filtered through a Celite pad. The filter pad was washed with DCM, and then used to wash the aqueous layer (2 × 300 mL). Next, the filter pad was washed with 10% MeOH in DCM, and then the aqueous layer (4 × 300 mL) was washed with this. The aqueous layer was washed again with 10% MeOH in DCM (4 × 100 mL), or by HPLC, until no product was evident in the aqueous layer. The organic layers were combined, dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure to obtain 10.5 g (89%) of 7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine as a yellowish-brown solid. MS(ESI)m / z[M+H] + C 10 H 10 Calculated value for N5: 200.1; Measured value: 200.0.

[0425] Step 5. Preparation of 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine (compound S3) [ka] A solution of 7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine (2.98 g, 15.0 mmol, as prepared in the previous step) in DMF (85.0 mL) was cooled in an ice bath, and then NBS (2.93 g, 16.4 mmol) was added. The reaction mixture was stirred at 0°C for 2.5 hours, and then poured into saturated Na2S2O3 aqueous solution (28.6 mL, 112 mmol) and saturated NaHCO3 aqueous solution (66.1 mL, 68.3 mmol). The reaction mixture was diluted with water (120 mL). The ice bath was removed, and the reaction mixture was allowed to rise to room temperature. After stirring for 1 hour, the reaction mixture was further diluted with water (270 mL). The solid was filtered, washed with water (2 × 45 mL), and dried under nitrogen press to obtain 3.80 g (91%) of 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine (compound S3) as a yellow solid. (400 MHz, DMSO-d6) δ(ppm) 9.00 (d, J=1.96 Hz, 1 H) 8.45 (s, 1 H) 8.33 (d, J=2.08 Hz, 1 H) 8.17 (s, 1 H) 7.85 (s, 1 H) 3.90 (s, 3 H); MS(ESI) m / z [M+H] + C 10 H 9Br Calculated value for N5: 278.0; Measured value: 277.9; HPLC purity: 210nm: 97.9%; 254nm: 97.5%.

[0426] Synthesis of O-(mesitylsulfonyl)hydroxylamine (compound S4) [ka] To a solution of ethyl N-hydroxyacetoimidate (40.0 g, 329.2 mmol) and 2,4,6-trimethylbenzenesulfonyl chloride (40.0 g, 182.8 mmol) in DMF (300 mL), TEA (27.7 g, 274.3 mmol) was added dropwise at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into ice water (900 mL), and the mixture was filtered. The filter cake was washed with H2O (3 × 200 mL) and dried under reduced pressure. The solid was dissolved in dioxane (26 mL), and the solution was cooled to 0°C. Then HClO4 (16 mL) was added dropwise at 0°C. After the addition was complete, the mixture was stirred at 0°C for 30 minutes, and then poured into ice water (400 mL). The mixture was filtered, and the filter cake was washed with H2O (3 × 200 mL). The solid was dissolved in DCM (200 mL) to obtain a DCM solution of O-(mesitylsulfonyl)hydroxylamine (compound S4).

[0427] Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine (compound S5) [ka] Step 1. Synthesis of 1-amino-3-bromopyrazine-1-ium-2,4,6-trimethylbenzenesulfonate [ka] To a solution of 2-bromopyrazine (22.0 g, 138.3 mmol) in DCM (154 mL), a solution of O-(mesitylsulfonyl)hydroxylamine (compound S4) (38.6 g, 179.8 mmol) in DCM (308 mL) was added dropwise at room temperature, and the mixture was stirred overnight at room temperature. The reaction product was concentrated under reduced pressure, and the residue was triturated with MTBE (100 mL) and dried under reduced pressure to obtain 34.0 g (66%) of 1-amino-3-bromopyrazine-1-ium-2,4,6-trimethylbenzenesulfonate as a black solid.

[0428] Step 2.6 Synthesis of ethyl bromopyrazolo[1,5-a]pyrazine-3-carboxylate [ka] To a solution of K2CO3 (12.4 g, 97.4 mmol) and ethyl propioate (27.0 g, 108.2 mmol) in DMF (270 mL), 1-amino-3-bromopyrazine-1-ium 2,4,6-trimethylbenzenesulfonate (27.0 g, 72.1 mmol, as prepared in the previous step) was added in small amounts at room temperature, and the reaction mixture was stirred overnight at room temperature. The mixture was diluted with H2O and extracted with MTBE (3 × 200 mL). The combined organic layer was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain 2.0 g (10%) ethyl 6-bromopyrazolo[1,5-a]pyrazine-3-carboxylate as a yellow solid. 1 H NMR(400MHz, CDCl3)δ(ppm)9.41(s,1H),8.66(d,J=0.8Hz,1H),8.46(s,1H),4.45(q,J=7.2Hz,2H),1.45(t,J=7.2Hz,3H).

[0429] Step 3.6 Synthesis of ethyl (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine-3-carboxylate [ka] To a mixture of 6-bromopyrazolo[1,5-a]pyrazine-3-carboxylate ethyl (5.0 g, 18.5 mmol, as prepared in the previous step) in dioxane (80 mL) and water (20 mL), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (5.7 g, 27.7 mmol), Na2CO3 (5.7 g, 55.4 mmol), and Pd(dppf)Cl2 (1.0 g) were added under nitrogen. The mixture was heated under reflux and stirred under nitrogen for 2.5 hours. The mixture was cooled to room temperature and filtered. The filtrate was poured into cold water and extracted 10 times with DCM / MeOH(10 / 1). The organic phase was washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain 3.5 g (70%) of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine-3-carboxylate ethyl as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)9.56(d,J=1.6Hz,1H),8.59(d,J=1.2Hz,1H),8.45(s,1H) ),7.96(d,J=7.2Hz,2H),4.45(q,J=7.2Hz,2H),4.01(s,3H),1.46(t,J=7.2Hz,3H).

[0430] Step 4.6 Synthesis of (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine [ka] A mixture of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine-3-carboxylate ethyl (3.5 g, 12.9 mmol, as prepared in the previous step) in a 40% aqueous H2SO4 solution (35 mL) was stirred overnight at 100°C, then the mixture was poured into water and the pH was adjusted to 8 with aqueous NaOH solution. The precipitate was isolated by filtration, the filtration cake was washed with water and dried to obtain 1.7 g (66%) of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)9.19(d,J=1.6Hz,1H),9.14(d,J=0.8Hz,1H),8.26(s, 1H), 8.12(d,J=2.4Hz,1H),8.05(s,1H),6.956(dd,J=0.8,2.4Hz,1H),3.89(s,3H).

[0431] Step 5. Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine (compound S5) [ka] To a mixture of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine (1.0 g, 5.0 mmol, as prepared in the previous step) in DMF (30 mL), NBS (1.76 g, 5.0 mmol) was added in small amounts. The mixture was stirred at room temperature for 1 hour, poured into water, and extracted three times with ethyl acetate. The combined organic phase was washed with water and brine, dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography while eluting with DCM / MeOH in a 1:0 to 100:1 ratio to obtain 600 mg (43%) of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine (compound S5) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 9.19 (d, J = 1.2 Hz, 1H), 9.09 (d, J = 1.2 Hz, 1H), 8.28 (s, 2H), 8.07 (s, 1H), 3.90 (s, 3H).

[0432] Synthesis of 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-a]pyridine (compound S7) [ka] Step 1.7 Synthesis of (1-methyl-1H-pyrazole-4-yl)imidazo[1,2-a]pyridine [ka] To a mixture of 7-bromoimidazo[1,2-a]pyridine (4.5 g, 22.8 mmol) in dioxane (100 mL) and H2O (20 mL), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (7.1 g, 34.3 mmol), Na2CO3 (5.3 g, 50.3 mmol), and Pd(dppf)Cl2 (0.9 g) were added under nitrogen. The mixture was heated under reflux and stirred under nitrogen for 6 hours. The mixture was cooled to room temperature and filtered. The filtrate was poured into cold water and extracted 10 times with DCM / MeOH (10 / 1). The combined organic phase was then washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography to obtain 3.4 g (75%) of 7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-a]pyridine as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.50(d,J=7.2Hz,1H),8.29(s,1H),8.03(s,1 H),7.86(s,1H),7.76(s,1H),7.52(s,1H),7.14-7.16(m,1H),3.88(s,3H).

[0433] Step 2.3 Synthesis of 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-a]pyridine (compound S7) [ka] To a mixture of 7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-a]pyridine (4.1 g, 20.7 mmol, as prepared in the previous step) in ACN (120 mL), NBS (3.7 g, 20.7 mmol) was added in small amounts. The mixture was stirred at room temperature for 2 hours, poured into water, and extracted 10 times with DCM / MeOH (10 / 1). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain 2.2 g (39%) of 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-a]pyridine (compound S7) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.36(s,1H),8.29(d,J=7.2Hz,1H),8.08(s,1H),7.85(s,1H),7.67(s,1H),7.32-7.35(m,1H),3.88(s,3H).

[0434] Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine (compound S8) [ka] Step 1. Synthesis of 1-amino-3-bromopyridazine-1-ium-2,4,6-trimethylbenzenesulfonate [ka] To a solution of 3-bromopyridazine (20.0 g, 125.7 mmol) in DCM (300 mL), a solution of O-(mesitylsulfonyl)hydroxylamine (compound S4) (35.1 g, 125.7 mmol) in DCM (300 mL) was added dropwise at room temperature, and the reaction mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure, and the residue was triturated with MTBE (80 mL). The mixture was filtered to obtain 24.0 g (51%) of 1-amino-3-bromopyridazine-1-ium-2,4,6-trimethylbenzenesulfonate as a black solid.

[0435] Step 2.6 - Synthesis of ethyl bromopyrazolo[1,5-b]pyridazine-3-carboxylate [ka] To a solution of K2CO3 (12.4 g, 89.8 mmol) and ethyl propioate (7.5 g, 77.0 mmol) in DMF (230 mL), 1-amino-3-bromopyridazine-1-ium 2,4,6-trimethylbenzenesulfonate (24.0 g, 64.1 mmol, as prepared in the previous step) was added in small amounts at room temperature, and the reaction mixture was stirred overnight at room temperature. The mixture was diluted with H2O and extracted with MTBE (3 × 200 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by chromatography to obtain 2.2 g (13%) of ethyl 6-bromopyrazolo[1,5-b]pyridazine-3-carboxylate as a yellow solid. 1 H NMR(400MHz, CDCl3)δ(ppm)8.40-8.44(m,2H),7.40(d,J=9.6Hz,1H),4.43(q,J=7.2Hz,2H),1.44(t,J=7.2Hz,3H).

[0436] Step 3.6 Synthesis of ethyl (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine-3-carboxylate [ka] To a mixture of ethyl 6-bromopyrazolo[1,5-b]pyridazine-3-carboxylate (9.0 g, 33.3 mmol, as prepared in the previous step) in dioxane (144 mL) and water (36 mL), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (10.4 g, 49.9 mmol), Na2CO3 (10.6 g, 99.9 mmol), and Pd(dppf)Cl2 (1.0 g) were added under nitrogen. The mixture was heated under reflux and stirred under nitrogen for 2 hours. The mixture was cooled to room temperature and filtered. The filtrate was poured into cold water and extracted 10 times with DCM / MeOH(10 / 1). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography to obtain 9 g (99%) of ethyl 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine-3-carboxylate as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.48(d,J=9.2Hz,1H),8.42(s,1H),8.06(d,J=4.4Hz,1H),7.4 5(d,J=9.2Hz,1H),7.86(s,1H),4.43(q,J=7.2Hz,2H),4.03(s,3H),1.45(t,J=7.2Hz,3H).

[0437] Step 4.6 - Synthesis of (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine [ka] A mixture of ethyl 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine-3-carboxylate (4.5 g, 16.6 mmol, as prepared in the previous step) in a 40% aqueous H2SO4 solution (45 mL) was stirred overnight at 100°C, then the mixture was poured into water and extracted 10 times with DCM / MeOH(10 / 1). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was washed with MTBE to obtain 2.5 g (76%) of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine as a yellow solid. 1 H NMR(400MHz, CDCl3)δ(ppm)8.04(d,J=7.2Hz,2H),7.98(d,J=2.4Hz,1H),7.93(d,J=9.6Hz,1H),7.21(d,J=9.2Hz,1H),6.61(d,J=2.4Hz,1H),4.00(s,3H).

[0438] Step 5. Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine (compound S8) [ka] To a mixture of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine (2.5 g, 12.5 mmol, as prepared in the previous step) in ACN (50 mL), NBS (2.2 g, 12.5 mmol) was added in small amounts. The mixture was stirred at room temperature for 1 hour, then poured into water and extracted 10 times with DCM / MeOH (10 / 1). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 2.5 g (71%) of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-b]pyridazine (compound S8) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 8.51 (s, 1H), 8.14-8.19 (m, 3H), 7.65 (d, J = 9.6Hz, 1H), 3.93 (s, 3H).

[0439] Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S9) [ka] Step 1.6 Synthesis of (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine [ka] To a mixture of 6-bromopyrazolo[1,5-a]pyridine (4.5 g, 22.8 mmol) in dioxane (90 mL) and water (18 mL), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (7.1 g, 34.2 mmol), Na2CO3 (7.2 g, 68.8 mmol), and Pd(dppf)Cl2 (0.45 g) were added under nitrogen. The mixture was heated under reflux and stirred under nitrogen for 3 hours. The mixture was cooled to room temperature and filtered. The filtrate was poured into cold water and extracted 10 times with DCM / MeOH(10 / 1). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography to obtain 3.5 g (78%) of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.61(s,1H),7.95(d,J=1.6Hz,1H),7.77(s,1H),7.63(s ,1H),7.57(d,J=9.2Hz,1H),7.23-7.26(m,1H),6.52(d,J=1.6Hz,1H),3.99(s,3H).

[0440] Step 2.3 Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S9) [ka] To a mixture of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (4.0 g, 20.2 mmol, as prepared in the previous step) in ACN (80 mL), NBS (4.3 g, 24.2 mmol) was added in small amounts. The mixture was stirred at room temperature for 1 hour, then poured into water and extracted 10 times with DCM / MeOH (10 / 1). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain 2.5 g (45%) of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S9) as a yellow solid. 1 H NMR(400MHz, CDCl3)δ(ppm)8.55(s,1H),7.92(s,1H),7.77(s,1H),7.65(s,1H),7.53(d,J=8.8Hz,1H),7.53(dd,J=0.8,9.2Hz,1H),4.00(s,3H).

[0441] Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine (compound S10) [ka] Step 1.6 Synthesis of (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine [ka] To a mixture of 6-bromopyrazolo[1,5-a]pyrimidine (4.5 g, 22.7 mmol) in dioxane (90 mL) and water (18 mL), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (7.0 g, 34.1 mmol), Na2CO3 (7.2 g, 68.1 mmol), and Pd(dppf)Cl2 (0.45 g) were added under nitrogen. The mixture was heated under reflux and stirred under nitrogen for 3 hours. The mixture was cooled to room temperature and filtered. The filtrate was poured into cold water and extracted 10 times with DCM / MeOH(10 / 1). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography to obtain 3.5 g (78%) of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine as a yellow solid. 1 H NMR(400MHz, CDCl3)δ(ppm)8.75-8.76(m,1H),8.64(d,J=2Hz,1H),8.12(d,J=2.4Hz,1H),7.80(s,1H),7.70(s,1H),6.72-6.73(m,1H),4.02(s,3H).

[0442] Step 2.3 Synthesis of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine (compound S10) [ka] To a mixture of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine (7.0 g, 35.1 mmol, as prepared in the previous step) in ACN (140 mL), NBS (7.5 g, 42.1 mmol) was added in small amounts. The mixture was stirred at room temperature for 1 hour, poured into water, and extracted 10 times with DCM / MeOH (10 / 1). The combined organic phase was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain 5 g (51%) of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine (compound S10) as a yellow solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 8.72-8.69 (m, 2H), 8.10 (s, 1H), 7.79 (s, 1H), 7.72 (s, 1H), 4.02 (s, 3H).

[0443] Synthesis of tert-butyl 4-[6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl]piperazine-1-carboxylate (compound 11) [ka] tBuOH (202 mL) and dioxane (124 mL) were added to a round-bottom flask containing 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S9) (4.00 g, 14.4 mmol) and tert-butyl-1-piperazine carboxylate (3.23 g, 17.3 mmol). The solution was sparged with nitrogen at 25°C for 20 minutes. tBuXPhos Pd G1 (248 mg, 5.41 mmol) and NaOtBu (520 mg, 0.54 mmol) were added to the solution, and the reaction mixture was further sparged with nitrogen at 25°C for 10 minutes. The flask was heated in a 55°C bath for 1.5 hours, cooled to room temperature, and diluted with ethyl acetate. The mixture was washed with H2O and brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography while eluting with 10-90% siRNA / DCM. The resulting solid was dried under reduced pressure to obtain 1.62 g (29%) of tert-butyl 4-[6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl]piperazine-1-carboxylate (compound 11) as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.46(s,1H),7.74(s,1H),7.70(s,1H),7.63(s,1H),7.50(d,J=7.3Hz,1H),7.14 (d,J=1.47Hz,1H),4.03-4.01(m,4H),3.98(s,3H),3.64(m,4H),3.02(m,4H),1.50(s,9H);MS(ESI)m / z[M+H] + C 20 H 27 Calculated value for N6O2: 383.2; Measured value: 383.3; HPLC purity: 210nm: 99.4%; 254nm: 99.2%.

[0444] Synthesis of 6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine (compound 12) [ka] In a round-bottom flask containing tert-butyl 4-[6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl]piperazine-1-carboxylate (compound 11) (0.340 g, 0.89 mmol), 6.80 mL of dimethylcellulose (DCM) was added. The solution was then cooled in an ice bath, and 1.70 mL of tallow-containing phosphate (TFA) (22.1 mmol) was slowly added. After 1 hour, the solvent was removed under reduced pressure, and then MTBE was added to the residue. The solution was concentrated under reduced pressure to remove excess TFA. The residue was dissolved in DCM, saturated aqueous NaHCO3 solution was added and stirred, diluted with H2O, and then diluted with MTBE. The aqueous layer was washed with MTBE and extracted with DCM. The DCM layer was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain 105 mg (42%) of 6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine (compound 12) as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.45(s,1H),7.74(s,1H),7.70(s,1H),7.60(s,1H),7.52(d,J=9. 17Hz,1H),7.11(d,J=9.05Hz,1H),3.97(s,3H),3.08(bs,5H),3.06(bs,4H);MS(ESI+,m / z):C 15 H 19 Calculated value for N6: 283.2. Measured value: 283.1; HPLC purity: 210nm: 100%; 254nm: 100%.

[0445] Synthesis of (6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine dihydrochloride) (compound S19) [ka] A round-bottom flask containing tert-butyl 4-[6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl]piperazine-1-carboxylate (compound 11) (0.250 g, 0.65 mmol) was mixed with MeOH (5.00 mL) and 4 M HCl in dioxane (2 mL). The reaction mixture was stirred at 27°C for 24 hours, then at 24°C for a further 5 hours. The mixture was filtered through frit, and the solid was rinsed with MeOH (5 mL). The solid was dried under reduced pressure to obtain 227 mg (98%) of 6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine dihydrochloride (compound S19) as a pale gray solid. MS(ESI)m / z[M+H] + C 15 H 21 Calculated value for Cl2N6: 283.2; measured value: 283.2.

[0446] Synthesis of di-tert-butyl 4,4'-(1,2-bis(1H-benzo[d][1,2,3]triazole-1-yl)ethane-1,2-diyl)bis(piperazine-1-carboxylate) (compound S47) [ka] A solution of 1H-1,2,3-benzotriazole (19.2 g, 161.1 mmol) and tert-butylpiperazine-1-carboxylate (30.0 g, 161.1 mmol) in EtOH (300 mL) was stirred under nitrogen at 25°C for 20 minutes. Then, glyoxal (40 wt% in H2O, 4.7 g, 80.5 mmol) was added to the mixture at 25°C. The reaction mixture was stirred at 30°C for 16 hours. The reaction mixture was concentrated under reduced pressure, and then PE was added. The mixture was stirred for 30 minutes, filtered, and the filtered cake was dried to obtain 42.0 g (32%) of di-tert-butyl 4,4'-(1,2-bis(1H-benzo[d][1,2,3]triazole-1-yl)ethane-1,2-diyl)bis(piperazine-1-carboxylate (compound S47)) as a white solid.

[0447] Using the procedure described for compound S47, as well as reagents, starting materials, and conditions known to those skilled in the art, the following compounds representative of this disclosure were prepared. [Table 4]

[0448] Synthesis of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-amine (compound S56) [ka] Step 1. Synthesis of 6-(1-methyl-1H-pyrazole-4-yl)-3-nitropyrazolo[1,5-a]pyridine [ka] To a solution of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (4.0 g, 5.05 mmol, compound S9, as prepared in step 1) dissolved in concentrated H2SO4 (10 mL), KNO3 (2.24 g, 5.55 mmol) was added. The reaction mixture was stirred under nitrogen at room temperature for 1 hour. The mixture was cooled to 0°C and diluted with water (300 mL). The mixture was neutralized to pH 8 with saturated Na2CO3 aqueous solution. The precipitated solid was collected by filtration and washed with water (3 × 300 mL). The filtration cake was dried under reduced pressure to obtain 3.5 g (71%) of 6-(1-methyl-1H-pyrazole-4-yl)-3-nitropyrazolo[1,5-a]pyridine as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)9.34(s,1H),8.90(s,1H),8.38(s,1H),8.24(d,J =9.1Hz,1H),8.15(d,J=9.3Hz,1H),8.11(s,1H),3.90(s,3H);MS(ESI)m / z[M+H] + C 11 Calculated value for H9N5O2: 244.1; Measured value: 244.1.

[0449] Step 2.6 Synthesis of (1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-amine (compound S56) [ka] To a solution of 6-(1-methyl-1H-pyrazole-4-yl)-3-nitropyrazolo[1,5-a]pyridine (3.0 g, 12.33 mmol, as prepared in the previous step) dissolved in HBr aqueous solution (50 mL), SnCl2·2H2O (8.42 g, 37.00 mmol) was added. The resulting solution was stirred under nitrogen at 90°C for 4 hours. The mixture was cooled to room temperature, the pH was adjusted to pH 9 with NaOH aqueous solution, and extracted with ELISA (3 × 500 mL). The combined organic extract was washed with brine (3 × 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 2.5 g (90%) of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-amine (compound S56) as a yellowish-brown solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.63(s,1H),8.15(s,1H),7.92(s,1H),7.59(d,J=9.2Hz ,1H),7.44(s,1H),7.11(d,J=9.2Hz,1H),4.33(s,2H),3.86(s,3H);MS(ESI)m / z[M+H] + C 11 H 11 Calculated value for N5: 214.1; Measured value: 214.1.

[0450] Synthesis of 6-(1-methyl-1H-pyrazole-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (compound S57) [ka] Step 1. Preparation of 3-iodo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine [ka] To a solution of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (7.0 g, 35.3 mmol, compound S9, as prepared in step 1) in DMF (70 mL), NIS (9.53 g, 42.4 mmol) was added. The resulting mixture was stirred at room temperature for 1 hour. The mixture was poured into a stirred mixture of 2.8 M Na2S2O3 aqueous solution (64 mL) and 1.14 M NaHCO3 aqueous solution (144 mL). The mixture was vigorously stirred for 1 hour, resulting in the formation of an off-white precipitate. The solid was collected by filtration, washed with H2O (2 × 10 mL), and dried under high vacuum to obtain 10 g (79%) of 3-iodo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)9.06(s,1H),8.28(s,1H),8.08(s,1H),8.02(d,J=0.8H z,1H),7.60(d,J=9.2Hz,1H),7.52(d,J=9.2Hz,1H),3.88(s,3H).;MS(ESI)m / z[M+H] + C 11 Calculated value for H9IN4: 325.0; Measured value: 325.1.

[0451] Step 2.6 Preparation of (1-methyl-1H-pyrazole-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (compound S57) [ka] To a solution of 3-iodo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (9.0 g, 27.8 mmol, as prepared in the previous step) in THF (180 mL), 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (17.1 g, 91.7 mmol) was added, the mixture was cooled to 0°C under nitrogen, and 1.3 M iPrMgCl·LiCl solution in THF (31.9 mL, 41.7 mmol) was added dropwise. The reaction mixture was stirred under nitrogen at 0°C for 3 hours, diluted with DCM (160 mL), and filtered through a silica gel pad. The pad was washed with ELISA (100 mL), and the combined filtrate was concentrated under reduced pressure to obtain 5.0 g (42%) of 6-(1-methyl-1H-pyrazole-4-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (compound S57) as a grayish-white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)9.08(s,1H),8.26(s,1H),8.12(s,1H),8.01(d,J=0.8Hz,1H), 7.82(d,J=9.1Hz,1H),7.65(d,J=9.1Hz,1H),3.89(s,3H),1.32(s,12H).;MS(ESI)m / z[M+H] + C 17 H 21 Calculated value for BN4O2: 325.2; Measured value: 325.2.

[0452] Synthesis of 2-chloro-5-(pyridine-2-ylmethyl)pyrimidine (compound S80, steps 1-2) [ka] Step 1. Preparation of potassium pyridine-2-ylacetate [ka] In a dried glass container, a solution of methyl 2-pyridylacetate (0.500 g, 3.31 mmol), anhydrous ethanol (6.6 mL), and water (0.63 mL, 3.47 mmol) was heated at 60°C. In a dried glass container, a solution of potassium tert-butoxide (0.390 g, 3.47 mmol) was prepared in anhydrous ethanol (3.3 mL) and then added dropwise to the first solution over 30 minutes at 60°C. After complete conversion, the solvent was removed under vacuum, and the residue was stirred in anhydrous Et2O (6.6 mL) for 30 minutes. The solid was filtered through frit and quickly washed sequentially with ethanol / ether (1:1, 0.6 mL × 2) and Et2O (3 mL × 3). The white solid was transferred to a dried flask and left in a 30°C bath under vacuum for 2 hours to obtain 468 mg (81%) of potassium pyridine-2-ylacetate as a white solid. 1 H NMR(400MHz,DMSO-d6)d(ppm)8.33(d,J=4.3Hz,1H)7.57(td,J=7.6,1.8,1H)7.27(d,J=7.8Hz,1H)7.06(dd,J=6.6,5.4Hz1H)3.32(s,2H); 13 C NMR(100MHz,DMSO-d6)d(ppm)49.5,119.9,123.8,135.1,147.8,160.5,171.5;MS(ESI)m / z[M+H] + Calculated value for C7H8NO2: 138.1; measured value: 138.0.

[0453] Step 2. 2-Chloro-5-(pyridine-2-ylmethyl)pyrimidine (compound S80) [ka] 5-bromo-2-chloropyrimidine (110 mg, 0.590 mmol), xanthophos (22 mg, 0.04 mmol), Pd2(dba)3 (10 mg, 0.01 mmol), potassium pyridine-2-ylacetate (125 mg, 0.713 mmol, as prepared in the previous step), and mesitylene (1.2 mL) were added to a pressure tube and pre-flashed with argon. It was then sparged with further argon, sealed, and heated at 150°C for 24 hours. The mixture was cooled to room temperature and purified by silica gel flash chromatography (acetone / DCM containing 0.1% Et3N) to obtain 72 mg (59%) of 2-chloro-5-(pyridine-2-ylmethyl)pyrimidine (compound S80) as oil. 1 H NMR(400MHz,CDCl3)d(ppm)8.59(s,2H)8.57(m,1H)7.68(td,J=7.6,1.8Hz1H)7.21(m,2H)4.13(s,2H);MS(ESI)m / z[M+H] + C 10 Calculated value for H9ClN3: 206.5; Measured value: 206.0; HPLC purity: 210nm: 93.3%; 254nm: 100.0%.

[0454] Synthesis of 7-(1-methyl-1H-pyrazole-4-yl)-3-(piperazin-1-yl)imidazo[1,2-b]pyridazine hydrochloride (compound S83) [ka] Step 1. Synthesis of tert-butyl 4-(7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazin-3-yl)piperazine-1-carboxylate [ka] To a 20 mL vial containing 3-bromo-7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazine (compound S3) (300.4 mg, 1.080 mmol) and tert-butyl 1-piperazine carboxylate (1.00 g, 5.40 mmol), tBuOH (3.0 mL) and dioxane (1.5 mL) were added, and the solution was sparged with argon for 10 minutes. KOtBu (182 mg, 1.62 mmol) was added, and the mixture was again sparged with argon for 10 minutes. tBuXPhos Pd G1 (111 mg, 0.162 mmol) was added, and the mixture was sparged with argon for 10 minutes. The mixture was stirred under argon at 55°C for 1 hour. The reaction mixture was cooled to room temperature, diluted with DCM (40 mL), and washed with 10% citric acid solution (3 × 10 mL), H₂O (3 × 10 mL), and saturated NaHCO₃ aqueous solution (2 × 10 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel flash chromatography eluted with 0 to 100% (10% MeOH in DCM) / DCM to obtain 280.8 mg (68%) of tert-butyl 4-(7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazin-3-yl)piperazine-1-carboxylate as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.51(d,J=2.0Hz,1H),7.91(d,J=2.0Hz,1H),7.84(s,1H),7.75(s,1H) ),7.25(s,1H),4.00(s,3H),3.75-3.64(m,4H),3.29-3.20(m,4H),1.51(s,9H);MS(ESI)m / z[M+H] + C 19 H 25 Calculated value for N7O2: 384.2, measured value: 384.2; HPLC purity: 210nm: 97.8%; 254nm: 100.0%.

[0455] Step 2. Synthesis of 7-(1-methyl-1H-pyrazole-4-yl)-3-(piperazin-1-yl)imidazo[1,2-b]pyridazine hydrochloride (compound S83) [ka] A solution of tert-butyl 4-[7-(1-methyl-1H-pyrazole-4-yl)imidazo[1,2-b]pyridazin-3-yl]piperazine-1-carboxylate (274.5 mg, 0.7159 mmol, as prepared in the previous step) in DCM (30 mL) was cooled to 0°C, and then a 4 M HCl solution in dioxane (1.8 mL, 7.3 mmol) was added dropwise. After the addition was complete, the mixture was warmed to room temperature and stirred for 7 hours. The reaction was concentrated under reduced pressure, and the residue was triturated with ether and filtered. The solid was dried under reduced pressure to obtain 214.6 mg (94%) of 7-(1-methyl-1H-pyrazole-4-yl)-3-(piperazine-1-yl)imidazo[1,2-b]pyridazine hydrochloride (compound S83) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)9.69(br s,2H),9.34(d,J=2.0Hz,1H),8.69(s,1H),8.44(d,J=2.0Hz,1H),8.34(s,1H),7 .98(s,1H),3.94(s,3H),3.54-3.49(m,4H),3.37-3.25(m,4H);MS(ESI)m / z[M+H] + C 14 H 17 Calculated value for N7: 284.2, measured value: 284.2; HPLC purity: 210nm: 95.9%; 254nm: 100.0%.

[0456] Synthesis of 5-benzyl-2-chloropyrimidine (compound S88) [ka] A mixture of (2-chloropyrimidine-5-yl)boronic acid (20 g, 0.126 mol) and Na2CO3 (40.43 g, 0.381 mol) in dioxane (200 mL) and H2O (80 mL), stirred under nitrogen at 25°C, was mixed with (Ph3P2PdCl2 (4.43 g, 0.0063 mol)) and after 20 minutes, benzyl bromide (23.76 g, 0.138 mol) was added. The reaction mixture was stirred at 100°C for 1 hour. The mixture was diluted with water and extracted with HCl (3 × 50 mL). The combined organic layers were concentrated under vacuum to obtain the crude product, which was purified by silica gel column with elution using DCM / MeOH (30:1 to 15:1) to obtain 18.18 g (69%) of 5-benzyl-2-chloropyrimidine (compound S88) as a white solid. 1 H NMR (400MHz, CDCl3) δ (ppm) 8.48 (s, 2H), 7.34~7.37 (m, 2H), 7.27~7.31 (m, 1H), 7.19 (d, J = 7.2Hz, 2H), 3.98 (s, 2H).

[0457] Synthesis of (R)-1-(4-chlorophenyl)ethyl(2,5-dioxopyrrolidine-1-yl) carbonate (compound S95) [ka] To a solution of (1R)-1-(4-chlorophenyl)ethanol (20.00 g, 127.7 mmol) in ACN (240 mL), di(N-succinimidyl) carbonate (49.07 g, 191.6 mmol) was added, and the mixture was then cooled to 0°C. TEA (53 mL, 380 mmol) was added dropwise over 5 minutes, and the mixture was stirred at 0°C for 30 minutes. The solution was warmed to room temperature, stirred overnight, and the mixture was then concentrated under reduced pressure. The residue was diluted with siRNA (250 mL) and washed with saturated aqueous NaHCO3 solution. An additional siRNA (100 mL) was added, and the organic layer was washed with water and brine, then dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. MTBE (150 mL) was added to the residue, and the mixture was heated to 45°C. Hexane (100 mL) was added at high temperature, and the mixture was then cooled while being efficiently stirred. The solid was isolated by filtration, washed with hexane (100 mL), and dried under reduced pressure to obtain 31.85 g (84%) of (R)-1-(4-chlorophenyl)ethyl(2,5-dioxopyrrolidine-1-yl) carbonate (compound S95) as a pale yellowish-brown solid. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 7.49 (s, 4H), 5.89 (q, J = 6.8Hz, 1H), 2.80 (s, 4H), 1.62 (d, J = 6.8Hz, 3H).

[0458] Synthesis of 6-(1-methyl-1H-pyrazole-4-yl)-3-(piperazin-1-yl)pyrazolo[1,5-a]pyrimidine (compound S98) [ka] Step 1. Preparation of tert-butyl 4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine-3-yl)piperazine-1-carboxylate [ka] 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine (compound S10) (0.150 g, 0.539 mmol) and tert-butyl 1-piperazine carboxylate (0.120 g, 0.647 mmol), followed by tBuOH (1.5 mL) and dioxane (3 mL), and the mixture was sparged with argon for 5 minutes. NaOtBu (0.0778 g, 0.809 mmol) was added, and the mixture was sparged with argon for 5 minutes. tBuXPhos Pd G1 (37.0 mg, 0.0539 mmol) was added, and the mixture was sparged with argon for 5 minutes. The reaction mixture was heated to 80°C for 2.5 hours, then an additional tBuXPhos Pd G1 (15 mg) was added as a slurry in dioxane (0.5 mL), and the mixture was stirred overnight at 80°C. The reaction mixture was cooled to room temperature, quenched with saturated NaHCO3 aqueous solution, and extracted with DCM (3×). The combined organic extracts were washed with water, then dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography to obtain 95 mg (46%) of tert-butyl 4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine-3-yl)piperazine-1-carboxylate as a yellow to orange solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.58(d,J=2.0Hz,1H),8.45(d,J=2.4Hz,1H),7.78(s,1H),7. 77(s,1H),7.68(s,1H),4.01(s,3H),3.71-3.65(m,4H),3.30-3.24(m,4H),1.51(s,9H). MS(ESI)m / z[M+H] + C 19 H 25 Calculated value for N7O2: 384.2; measured value: 384.3.

[0459] Step 2. Preparation of 6-(1-methyl-1H-pyrazole-4-yl)-3-(piperazine-1-yl)pyrazolo[1,5-a]pyrimidine (compound S98) [ka] To a solution of tert-butyl 4-[6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrimidine-3-yl]piperazine-1-carboxylate (0.09 g, 0.2 mmol, as prepared in the previous step) in DCM (2 mL), TFA (0.2 mL) was added, and the reaction mixture was stirred overnight at room temperature. The mixture was diluted with DCM (20 mL) and washed with saturated NaHCO3 aqueous solution (25 mL). The aqueous layer was extracted with DCM (2 × 20 mL), and the combined organic extracts were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to obtain 65 mg (93%) of 6-(1-methyl-1H-pyrazole-4-yl)-3-(piperazine-1-yl)pyrazolo[1,5-a]pyrimidine (compound S98) as an orange solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.58(d,J=2.0Hz,1H),8.45(d,J=2.4Hz,1H),7.79(s, 1H),7.77(s,1H),7.68(s,1H),4.01(s,3H),3.37-3.31(m,4H),3.20-3.14(m,4H). MS(ESI)m / z[M+H] + C 14 H 17 Calculated value for N7: 284.2; measured value: 284.1.

[0460] Synthesis of 5-benzyl-2-pyrrolidine-3-ylpyrimidine (compound S100) [ka] Step 1. Preparation of tert-butyl 3-(5-benzylpyrimidine-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate [ka] A mixture of tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (500.0 mg, 1.69 mmol), 5-benzyl-2-chloropyrimidine (compound S88) (289 mg, 1.41 mmol), and K3PO4 (599 mg, 2.82 mmol) in dioxane (14.3 mL) and H2O (5.1 mL) was sparged with argon for 20 minutes. (tBu3P)2Pd (108 mg, 0.21 mmol) was added, and the mixture was then heated to 80°C for 4 hours. The reaction mixture was cooled to room temperature, diluted with H2O (40 mL), and extracted with siRNA (2 × 25 mL). The combined organic extract was washed with H2O (25 mL), dried over anhydrous MgSO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography while eluting with 20-30% siRNA / hexane to obtain 420 mg (88%) of tert-butyl 3-(5-benzylpyrimidine-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate as a white solid. 1 H NMR(400MHz,CDCl3)δ(ppm)8.56(s,2H),7.31-7.39(m,2H),7.24-7.31(m,1H),7.20(d,J=6.97Hz,2H),6. 83-6.95(m,1H),4.53-4.69(m,2H),4.33-4.49(m,2H),3.89-4.03(m,2H),1.53(s,9H);MS(ESI)m / z[M+H] + C 20 H 24 Calculated value for N3O2: 338.2; Measured value: 338.3; HPLC purity: 210nm: 98.3%; 254nm: 100.0%.

[0461] Step 2. Preparation of tert-butyl 3-(5-benzylpyrimidine-2-yl)pyrrolidine-1-carboxylate [ka] To a solution of tert-butyl 3-(5-benzylpyrimidine-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (437 mg, 1.30 mmol, as prepared in the previous step) in siRNA (20 mL), 10% Pd / C (100 mg, 50% H2O) was added. The reaction flask was evacuated three times and filled with hydrogen gas, and the mixture was stirred under a hydrogen atmosphere. After 19 hours, the reaction mixture was filtered through a Celite pad, and the pad was washed with an additional 40 mL of siRNA. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography with elution using 25-40% siRNA / hexane to obtain 377 mg (86%) of tert-butyl 3-(5-benzylpyrimidine-2-yl)pyrrolidine-1-carboxylate as a colorless, viscous oil. 1 H NMR(400MHz,CDCl3)δ(ppm)8.52(s,2H),7.30-7.39(m,2H),7.22-7.30(m,1H),7.19(d,J=7.09Hz,2H),3. 95(s,2H),3.83(m,1H),3.53-3.73(m,3H),3.43(m,1H),2.18-2.43(m,2H),1.47(s,9H);MS(ESI)m / z[M+H] + C 20 H 25 Calculated value for N3O2: 340.2; Measured value: 340.3; HPLC purity: 210nm: 100.0%; 254nm: 100.0%.

[0462] Step 3. Preparation of 5-benzyl-2-pyrrolidin-3-ylpyrimidine (compound S100) [ka] A solution of tert-butyl 3-(5-benzylpyrimidine-2-yl)pyrrolidine-1-carboxylate (377 mg, 1.11 mmol, as prepared in the previous step) in DCM (9.0 mL) was cooled to 0°C, and TFA (1.0 mL) was added dropwise. The mixture was stirred at 0°C for 30 minutes, then warmed to room temperature for 3 hours. The reaction product was concentrated under reduced pressure, and the residue was dissolved in DCM (10 mL) and concentrated again. The residue was dissolved in DCM (20 mL) and washed with 20 mL of saturated NaHCO3 aqueous solution. The organic phase was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to obtain 211 mg (79%) of 5-benzyl-2-pyrrolidine-3-ylpyrimidine (compound S100) as a pale pink, viscous oil. 1 H NMR(400MHz,CD3OD)δ(ppm)8.58(s,2H),7.27-7.36(m,2H),7.19-7.27(m,3H),3.99(s,2H),3.56-3.69(m,1H),3.32-3.38(m,1H) ),3.23-3.30(m,1H),3.13-3.23(m,1H),3.08(dt,J=11.19,7.37Hz,1H),2.24-2.38(m,1H),2.07-2.24(m,1H);MS(ESI)m / z[M+H] + C 15 H 17 Calculated value for N3: 240.2; Measured value: 240.1; HPLC purity: 210nm: 100.0%; 254nm: 100.0%.

[0463] Synthesis of 1-benzyl-3-(pyrrolidine-3-yl)-1H-1,2,4-triazole (compound S101) [ka] Step 1. Preparation of 1-benzyl-3-bromo-1H-1,2,4-triazole [ka] A mixture of 3-bromo-1H-1,2,4-triazole (5.0 g, 33.8 mmol), benzyl bromide (5.8 g, 33.8 mmol), and NaOMe (1.8 g, 33.8 mmol) in DMF (50 mL) was stirred under nitrogen at 35°C for 6 hours, then diluted with water and extracted with toluene (5 × 100 mL). The combined organic extract was concentrated under reduced pressure, and the residue was purified by silica gel chromatography while eluting with toluene / PE at a ratio of 1:50 to 1:20 to obtain 2.5 g (31%) of 1-benzyl-3-bromo-1H-1,2,4-triazole as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 8.72 (s, 1H), 7.30~7.40 (m, 5H), 5.40 (s, 2H).

[0464] Step 2.3 - Preparation of (1-benzyl-1H-1,2,4-triazol-3-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate [ka] To a solution of 1-benzyl-3-bromo-1H-1,2,4-triazole (3.0 g, 12.6 mmol, as prepared in the previous step) in a mixture of dioxane (10 mL) and H2O (30 mL), tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (4.5 g, 15.1 mmol), K3PO4 (5.4 g, 25.2 mmol), and Pd(PPh3)2Cl2 (0.4 g, 0.6 mmol) were added under nitrogen. The reaction mixture was stirred under nitrogen at 80°C for 4 hours, then poured into water (100 mL) and extracted with ELISA (3 × 50 mL). The combined organic extract was washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography while eluting with 1:10 siRNA / PE to 1:1 siRNA / PE to obtain 3.3 g (80%) of tert-butyl 3-(1-benzyl-1H-1,2,4-triazole-3-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.67(d,J=4.4Hz,1H),7.26~7.35(m,5H),6.42( d,J=12.8Hz,1H),5.41(s,1H),4.32(s,2H),4.18(s,2H),1.41~1.43(m,12H).

[0465] Step 3. Preparation of tert-butyl 3-(1-benzyl-1H-1,2,4-triazole-3-yl)pyrrolidine-1-carboxylate [ka] A mixture of tert-butyl 3-(1-benzyl-1H-1,2,4-triazole-3-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (3.0 g, 9.2 mmol, as prepared in the previous step) and 10% Pd / C (50% water, 1.6 g) in Â10% (30 mL) was stirred at 25°C for 5 hours under a hydrogen (20 psi) atmosphere. The mixture was filtered, and the filtrate was concentrated to obtain 3.0 g (99%) of tert-butyl 3-(1-benzyl-1H-1,2,4-triazole-3-yl)pyrrolidine-1-carboxylate as a yellow solid. 1 H NMR(400MHz,CDCl3)δ(ppm)7.94(s,1H),7.37(d,J=7.2Hz,3H),7.25-7.29(m,2H),5.28(d,J=7.2 Hz, 2H), 3.79 (s, 1H), 3.42~3.52 (m, 3H), 3.26~3.28 (m, 1H), 2.20~2.23 (m, 2H), 1.37~1.45 (m, 9H).

[0466] Step 4. Preparation of 1-benzyl-3-(pyrroridine-3-yl)-1H-1,2,4-triazole (compound S101) [ka] To a solution of tert-butyl 3-(1-benzyl-1H-1,2,4-triazole-3-yl)pyrrolidine-1-carboxylate (2.9 g, 8.8 mmol, as prepared in the previous step) in DCM (30 mL), TFA (10 mL) was added at 0°C. The reaction mixture was stirred under nitrogen at 25°C for 3 hours. The pH of the mixture was adjusted to pH 9 with saturated NaHCO3 aqueous solution, and then extracted with ELISA (3 × 50 mL). The combined organic extracts were washed with brine, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 1.1 g (55%) of 1-benzyl-3-(pyrrolidine-3-yl)-1H-1,2,4-triazole (compound S101) as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.53(s,1H),7.31~7.34(m,3H),7.26~7.30(m ,2H),5.33(s,2H),3.21~3.38(m,2H),2.86~2.96(m,3H),1.87~2.05(m,2H).

[0467] Synthesis of 3-(4-(1H-1,2,4-triazol-3-yl)piperazine-1-yl)-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S102) [ka] Step 1. Preparation of a mixture of 3-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole, 3-bromo-4-(tetrahydro-2H-pyran-2-yl)-4H-1,2,4-triazole, and 5-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole. [ka] To a solution of 3-bromo-1H-1,2,4-triazole (500 mg, 3.37 mmol) and 3,4-dihydro-2H-pyran (426.3 mg, 5.06 mmol) in toluene (20 mL), TsOH·H2O (64.2 mg, 0.33 mmol) was added, and the mixture was heated to 105°C and stirred overnight under nitrogen. The reaction product was cooled to room temperature and then concentrated under reduced pressure. The residue was dissolved in water (100 mL) and extracted with DCM (3 × 100 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, and then the filtrate was concentrated under reduced pressure to obtain a mixture of 700 mg (89%) of 3-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole, 3-bromo-4-(tetrahydro-2H-pyran-2-yl)-4H-1,2,4-triazole, and 5-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole as a black oil. MS(ESI)m / z[M+H] + C7H 10Calculated value for BrN3O: 232.0; Measured value: 232.0. LCMS purity: 254nm: 85%.

[0468] Step 2. Preparation of a mixture of 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole-3-yl)piperazin-1-yl)pyrazolo[1,5-a]pyridine, 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(4-(tetrahydro)-2H-pyran-2-yl)-4H-1,2,4-triazole-3-yl)piperazin-1-yl)pyrazolo[1,5-a]pyridine, and 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole-5-yl)piperazin-1-yl)pyrazolo[1,5-a]pyridine [ka] A mixture of 3-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole, 3-bromo-4-(tetrahydro-2H-pyran-2-yl)-4H-1,2,4-triazole, and 5-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole (600 mg, 2.58 mmol, as prepared in the previous step) and a solution of 6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine (compound 12) (364.9 mg, 1.29 mmol) in dioxane (20 mL) was prepared by adding Cs2CO3 (1.68 g, 5.17 mmol) and Pd-PEPPSI-IPent. Cl2-methylpyridine (326.1 mg, 0.38 mmol) was added, and the reaction mixture was heated to 100°C and stirred overnight under nitrogen. The mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was dissolved in water (100 mL) and extracted with DCM (3 × 100 mL). The combined organic extracts were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 10% to 50% ACN / water (10 mM NH4HCO3) to obtain 450 mg (40%) of 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole-3-yl)piperazine-1-yl)pyrazolo[1,5-a]pyridine, 6-(1-methyl A mixture of ru-1H-pyrazole-4-yl)-3-(4-(4-(tetrahydro-2H-pyran-2-yl)-4H-1,2,4-triazole-3-yl)piperazin-1-yl)pyrazolo[1,5-a]pyridine and 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole-5-yl)piperazin-1-yl)pyrazolo[1,5-a]pyridine was obtained as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.83-8.76(m,1H),8.36(s,1H),8.22(s,1H),7.9 7(s,1H),7.78(s,1H),7.67(dd,J=9.3,0.9Hz,1H),7.32(dd,J=9.2,1.5Hz,1H), 5.33(dd,J=9.9,2.3Hz,1H),3.97-3.90(m,1H),3.87(s,3H),3.67-3.57(m,1H), 3.49-3.47(m,4H),3.07(t,J=5.0Hz,4H),2.05-1.90(m,3H),1.67-1.52(m,3H). MS(ESI)m / z[M+H] + C 22 H 27 Calculated value for N9O: 434.2; Measured value: 434.2. LCMS purity: 254nm: 90%.

[0469] Step 3.3-(4-(1H-1,2,4-triazol-3-yl)piperazine-1-yl)-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S102) [ka] 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole-3-yl)piperazine-1-yl)pyrazolo[1,5-a]pyridine in DCM (20 mL), 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(4-(tetrahydro)-2H-pyran-2-yl)-4H-1,2,4-triazole-3-yl)piperazine-1-yl)pyrazolo[1,5 To a solution of a mixture of -a]pyridine and 6-(1-methyl-1H-pyrazole-4-yl)-3-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-1,2,4-triazole-5-yl)piperazin-1-yl)pyrazolo[1,5-a]pyridine (400 mg, 0.92 mmol, as prepared in the previous step), 4 M HCl in dioxane (336.4 mg, 9.23 mmol) was added, and the reaction was stirred under nitrogen at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the pH of the residue was adjusted to pH 7 with aqueous NH3 solution. The residue was purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 10% to 50% ACN / water (10 mM NH4HCO3) to obtain 270 mg (84%) of 3-(4-(1H-1,2,4-triazole-3-yl)piperazin-1-yl)-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S102) as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δ(ppm)13.13-12.70(m,1H),8.82(s,1H),8.57-8.20(m,2H),7.97(s,1H),7.78(s, 1H),7.68(d,J=9.2Hz,1H),7.34-7.30(m,1H),3.87(s,3H),3.50(t,J=4.8Hz,4H),3.08(t,J=4.8Hz,4H). MS(ESI)m / z[M+H]+ C 17 H 19 Calculated value for N9: 350.2; Measured value: 350.2. LCMS purity: 254nm: 95%.

[0470] Synthesis of 1-[6-(1-methylpyrazole-4-yl)pyrazolo[1,5-a]pyrazine-3-yl]piperazine (compound S103) [ka] A solution of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyrazine (compound S5) (900 mg, 3.25 mmol) in dioxane (20 mL) is prepared by adding piperazine (1.39 g, 16.18 mmol) and Pd-PEPPSI-IPent. Cl 2-methylpyridine (272.2 mg, 0.32 mmol) and Cs2CO3 (2.11 g, 6.47 mmol) were added, and the mixture was heated to 100°C and stirred under nitrogen for 16 hours. The mixture was cooled to room temperature, diluted with water (50 mL), and extracted with siRNA (3 × 50 mL). The combined organic extract was washed with brine (3 × 10 mL), dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 10% to 100% ACN / water (0.1% FA) to obtain 500 mg (42%) of 1-[6-(1-methylpyrazole-4-yl)pyrazolo[1,5-a]pyrazine-3-yl]piperazine as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)9.17(s,1H),8.89(s,1H),8.28(d,J=5.6Hz,1H),8.21(s,1H),8 .00(s,1H),7.81(s,1H),3.88(s,3H),3.21-3.11(m,4H),3.08-2.94(m,4H);MS(ESI)m / z[M+H] + C 14 H 17 Calculated value for N7: 284.1; Measured value: 284.1; LCMS purity: 254nm: 98.6%.

[0471] Synthesis of 1-(2-methylbenzo[d]thiazole-6-yl)ethane-1-one (compound S104) [ka] To a solution of 6-bromo-2-methylbenzo[d]thiazole (5 g, 21.92 mmol) and tributyl(1-ethoxyvinyl) stannane (11.87 g, 32.88 mmol) in dioxane (500 mL), Pd(PPh3)4 (2.53 g, 2.19 mmol) was added, and the mixture was then heated to 90°C and stirred under nitrogen for 16 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. Water (150 mL) was added to the residue, and the mixture was then extracted with SiO2 (3 × 100 mL). The organic extracts were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 3.5 g (83%) of 1-(2-methylbenzo[d]thiazole-6-yl)ethane-1-one (compound S104) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.72(d,J=1.8Hz,1H),8.06-7.95(m,2H),2.85(s,3H),2.65(s,3H);MS(ESI)m / z[M+H] + C 10 Calculated value for H9NOS: 192.0; Measured value: 192.1.

[0472] Synthesis of (2-methylbenzo[d]thiazole-6-yl)methanol (compound S105) [ka] To a stirred solution of 2-methylbenzo[d]thiazole-6-carboxylic acid (1 g, 5.17 mmol) in THF (20 mL) cooled to 0°C, a solution of 2 M LiAlH4 in THF (5.2 mL) was added, and the mixture was warmed to room temperature and stirred for 3 hours. The reaction mixture was cooled to 0°C and quenched by adding saturated NaHCO3 aqueous solution. The mixture was filtered, and the filter cake was washed with DCM (3 × 30 mL). The filtrate was extracted with DCM / MeOH (10:1) (3 × 80 mL), then the organic extracts were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography and eluted with PE / siRNA (1:1) to obtain 400 mg (43%) of (2-methylbenzo[d]thiazole-6-yl)methanol (compound S105) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.93(d,J=1.5Hz,1H),7.84(d,J=8.3Hz,1H),7.41(dd,J=8. 3,1.5Hz,1H),5.31(t,J=5.7Hz,1H),4.61(d,J=5.7Hz,2H),2.78(s,3H);MS(ESI)m / z[M+H] + Calculated value for C9H9NOS: 180.0; Measured value: 180.1.

[0473] Synthesis of 2-(hydroxymethyl)-5-methylbenzonitrile (compound S106) [ka] To a stirred solution of methyl 2-cyano-4-methylbenzoate (500 mg, 2.85 mmol) and NaBH4 (323.91 mg, 8.56 mmol) in MeOH (10 mL) cooled to 0°C, LiCl (241.97 mg, 5.70 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 1 hour. The reaction product was diluted with water (30 mL) and extracted with  (3 × 50 mL). The organic extracts were combined, washed with brine (2 × 2 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 400 mg (38%) of 2-(hydroxymethyl)-5-methylbenzonitrile (compound S106) as a yellow solid. MS(ESI)m / z[M+H] + Calculated value for C9H9NO: 148.2; Measured value: 148.0.

[0474] Synthesis of 2-(hydroxymethyl)-5-(trifluoromethyl)benzonitrile (compound S107) [ka] Step 1. Preparation of methyl 2-cyano-4-(trifluoromethyl)benzoate [ka] To a solution of 2-bromo-5-(trifluoromethyl)benzonitrile (5 g, 19.99 mmol) in MeOH (30 mL), Pd(dppf)Cl2 (1.46 g, 2 mmol) was added in a pressure vessel, and the mixture was then pressurized to 15 atm with carbon monoxide at 80°C overnight. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with elution using PE / siRNA (1:1) to obtain 3 g (65%) of methyl 2-cyano-4-(trifluoromethyl)benzoate as a brown solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.55-8.50(m,1H),8.30(d,J=8.2Hz,1H),8.24(dd,J=8.3,1.9Hz,1H),3.96(s,3H);MS(ESI)m / z[M+H] + C10 Calculated value for H6F3NO2: 229.0; measured value: 229.2.

[0475] Step 2. Preparation of 2-(hydroxymethyl)-5-(trifluoromethyl)benzonitrile (compound S107) [ka] To a stirred solution of methyl 2-cyano-4-(trifluoromethyl)benzoate (2.9 g, 12.65 mmol, as prepared in the previous step) in MeOH (29 mL) cooled to 0°C, NaBH4 (0.96 g, 25.3 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction product was concentrated under reduced pressure, and the residue was dissolved in water (300 mL) and extracted with siRNA (2 × 200 mL). The organic extracts were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 1.5 g (59%) of 2-(hydroxymethyl)-5-(trifluoromethyl)benzonitrile (compound S107) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.05-7.94(m,2H),7.82(d,J=3.8Hz,1H),5.43(d,J=8.7Hz,2H),4.55-4.53(m,1H);MS(ESI)m / z[M+H] + Calculated value for C9H6F3NO: 201.0; Measured value: 201.1.

[0476] Synthesis of 1-(4-(fluoromethyl)phenyl)ethane-1-ol (compound S108) [ka] Step 1. Preparation of 1-(4-(fluoromethyl)phenyl)ethane-1-one [ka] To a stirred solution of 1-(p-tolyl)ethane-1-one (3 g, 22.3 mmol) and K2S2O8 (18.13 g, 67 mmol) in ACN (30 mL) and H2O (30 mL), F-TEDA (23.7 g, 67 mmol) was added in small amounts at room temperature, and the mixture was then heated to 80 °C and stirred overnight. The reaction product was cooled to room temperature and extracted with siRNA (3 × 200 mL). The organic extracts were combined, washed with brine (3 × 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / siRNA (1:1) to obtain 2 g (59%) of 1-(4-(fluoromethyl)phenyl)ethane-1-one as a colorless liquid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.00(dd,J=8.3,1.3Hz,2H),7.62-7.49(m,2H),5.53(d,J=48Hz,2H),2.60(d,J=0.8Hz,3H);MS(ESI)m / z[M+H] + Calculated value for C9H9FO: 153.0; Measured value: 153.0.

[0477] Step 2. Preparation of 1-(4-(fluoromethyl)phenyl)ethane-1-ol (compound S108) [ka] To a solution of 1-(4-(fluoromethyl)phenyl)ethane-1-one (800 mg, 5.2 mmol, as prepared in the previous step) in MeOH (10 mL) cooled to 0°C, NaBH4 (238 mg, 6.3 mmol) was added, and the mixture was stirred for 1 hour. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 600 mg (74%) of 1-(4-(fluoromethyl)phenyl)ethane-1-ol (compound S108) as a colorless liquid. 1H NMR(400MHz,DMSO-d6)δ(ppm)7.43-7.29(m,4H),5.38(d,J=48Hz,2H),5.20(d,J=4.2Hz,1H),4.87-4.68(m,1H),1.33(d,J=6.4Hz,3H).

[0478] Synthesis of 1-(4-(difluoromethyl)phenyl)ethane-1-ol (compound S109) [ka] To a solution of 1-(4-(difluoromethyl)phenyl)ethane-1-one (600 mg, 3.5 mmol) in MeOH (8 mL) cooled to 0°C, NaBH4 (267 mg, 7 mmol) was added, and the mixture was stirred for 2 hours. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 550 mg (91%) of 1-(4-(difluoromethyl)phenyl)ethane-1-ol (compound S109) as a colorless oil. 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.76-7.41(m,4H),7.01(t,J=56.0Hz,1H),5.29(d,J=4.3Hz,1H),4.95-4.61(m,1H),1.34(d,J=6.5Hz,3H).

[0479] Synthesis of 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethane-1-ol (compound S110) and 2-((tert-butyldimethylsilyl)oxy)-2-(p-tolyl)ethane-1-ol (compound S111) [ka] Step 1. Preparation of 2-hydroxy-1-(p-tolyl)ethane-1-one [ka] To a solution of 2-bromo-1-(4-methylphenyl)ethanone (3 g, 14.08 mmol) in MeOH (20 mL), NaOAc (9.24 g, 112.64 mmol) was added, and the mixture was then heated to 80°C and stirred under nitrogen for 12 hours. The reaction product was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / SiO(5:1) to obtain 1.47 g (70%) of 2-hydroxy-1-(4-methylphenyl)ethanone as a white solid. 1 H NMR (400MHz, CDCl3) δ(ppm)7.82(d,J=8.2Hz,2H),7.30(d,J=8.0Hz,2H),4.85(s,2H),3.50(d,J=7.4Hz,1H),2.43(s,3H).

[0480] Step 2. Preparation of 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethane-1-one [ka] To a solution of 2-hydroxy-1-(4-methylphenyl)ethanone (1.27 g, 8.45 mmol, as prepared in the previous step) in THF (20 mL), TBSCl (1.91 g, 12.68 mmol) and imidazole (0.87 g, 12.68 mmol) were added, and the mixture was stirred at room temperature for 2 hours. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with elution using PE / SiO(5:1) to obtain 2.17 g (97%) of 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethan-1-one as a colorless liquid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.83(d,J=8.2Hz,2H),7.32(d,J=8.0Hz,2H),4.99(s,2H),2.37(s,3H),0.90(s,9H),0.07(s,6H).

[0481] Step 3. Preparation of 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethane-1-ol (compound S110) and 2-((tert-butyldimethylsilyl)oxy)-2-(p-tolyl)ethane-1-ol (compound S111) [ka] To a stirred solution of 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethane-1-one (1 g, 3.78 mmol, as prepared in the previous step) in MeOH (10 mL) cooled to 0°C, NaBH4 (0.22 g, 5.673 mmol) was added in small amounts, and the mixture was then warmed to room temperature and stirred under nitrogen at room temperature for 2 hours. The reaction product was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a Prep Phenyl column while eluting with 35-65% MeOH / water (10 mM NH4HCO3) to obtain 250 mg (25%) of 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethane-1-ol (compound S110) as a colorless liquid, and 200 mg (20%) of 2-((tert-butyldimethylsilyl)oxy)-2-(p-tolyl)ethane-1-ol (compound S111) as a colorless oil.

[0482] 2-((tert-butyldimethylsilyl)oxy)-1-(p-tolyl)ethane-1-ol (compound S110). 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.21(d,J=8.0Hz,2H),7.10(d,J=7.9Hz,2H),5.16(d,J=4.3Hz,1H),4.49(q, J=5.7Hz,1H),3.65-3.58(m,1H),3.55-3.48(m,1H),2.27(s,3H),0.81(s,9H),-0.04(s,3H),-0.07(s,3H).

[0483] 2-((tert-butyldimethylsilyl)oxy)-2-(p-tolyl)ethane-1-ol (compound S111). 1H NMR(400MHz,DMSO-d6)δ(ppm)7.20(d,J=8.0Hz,2H),7.12(d,J=7.9Hz,2H),4.70(t,J=5.7Hz,1 H),4.66-4.61(m,1H),3.44-3.31(m,2H),2.28(s,3H),0.85(s,9H),0.05(s,3H),-0.05(s,3H).

[0484] Synthesis of 1-(1,5-dimethyl-1H-pyrazole-3-yl)ethane-1-ol (compound S112) [ka] Step 1. Preparation of 1-(1,5-dimethyl-1H-pyrazole-3-yl)ethane-1-one [ka] To a solution of 1-(5-methyl-1H-pyrazole-3-yl)ethane-1-one (1 g, 8.05 mmol) in DCM (20 mL), MeI (1.72 g, 12.08 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction product was filtered, the filter cake was washed with DCM (2 × 20 mL), and the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 20-30% ACN / water (10 mM NH4HCO3) to obtain 240 mg (22%) of 1-(1,5-dimethyl-1H-pyrazole-3-yl)ethane-1-one as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)6.47(s,1H),3.81(s,3H),2.41(s,3H),2.27(s,3H);MS(ESI)m / z[M+H] + C7H 10 Calculated value for N2O: 139.1; measured value: 139.2.

[0485] Step 2. Preparation of 1-(1,5-dimethyl-1H-pyrazole-3-yl)ethane-1-ol (compound S112) [ka] To a stirred mixture of 1-(1,5-dimethyl-1H-pyrazole-3-yl)ethane-1-one (220 mg, 1.59 mmol, as prepared in the previous step) in MeOH (5 mL) cooled to 0°C, NaBH4 (90.3 mg, 2.38 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction mixture was cooled to 0°C, quenched with water, and extracted with ELISA (3 × 20 mL). The organic extracts were combined, washed with water (3 × 20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 5-10% ACN / water (10 mM NH4HCO3) to obtain 140 mg (63%) of 1-(1,5-dimethyl-1H-pyrazole-3-yl)ethane-1-ol (compound S112) as a yellow liquid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)5.92(s,1H),4.83(d,J=4.7Hz,1H),4.63-4.52(m,1H),3.63(s,3H),2.19(s,3H),1.29(d,J=6.5Hz,3H).

[0486] Synthesis of 3-(4-cyclopropylphenyl)propanoic acid (compound S113) [ka] To a stirred mixture of 3-(4-bromophenyl)propanoic acid (1 g, 4.36 mmol) and cyclopropylboronic acid (449.9 mg, 5.23 mmol) in dioxane (10 mL), Pd(dppf)Cl2 (319.4 mg, 0.43 mmol) and Cs2CO3 (2.8 g, 8.73 mmol) were added in small amounts under nitrogen at room temperature. The mixture was heated to 100 °C and stirred for 2 hours. The reaction product was cooled to room temperature, diluted with water (200 mL), and extracted with DCM (2 × 150 mL). The organic extracts were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 5-100% ACN / water (10 mM NH4HCO3) to obtain 500 mg (60%) of 3-(4-cyclopropylphenyl)propanoic acid (compound S113) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.08(d,J=7.9Hz,2H),6.97(d,J=7.9Hz,2H),2.75(t,J=7. 7Hz, 2H), 2.44 (t, J = 7.7Hz, 2H), 1.92-1.78 (m, 1H), 0.96-0.82 (m, 2H), 0.67-0.55 (m, 2H).

[0487] Synthesis of 1-(4-(2,2,2-trifluoroethyl)phenyl)ethane-1-ol (compound S114) [ka] Step 1. Preparation of 1-(4-(2,2,2-trifluoroethyl)phenyl)ethane-1-one [ka] To a stirred mixture of 4-acetylphenylboronic acid (1 g, 6.1 mmol) and 1,1,1-trifluoro-2-iodoethane (5.1 g, 24.3 mmol) in dioxane (20 mL), Cs2CO3 (5.96 g, 18.3 mmol), Pd2(dba)3 (1.1 g, 1.2 mmol), and XantPhos (705 mg, 1.2 mmol) were added in small amounts at room temperature under nitrogen. The mixture was then heated to 100 °C and stirred overnight under nitrogen. The reaction product was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with elution using PE / siRNA (5:1) to obtain 800 mg (65%) of 1-(4-(2,2,2-trifluoroethyl)phenyl)ethane-1-one as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.02-7.95(m,2H),7.52(d,J=8.0Hz,2H),3.77(q,J=11.5Hz,2H),2.59(s,3H);MS(ESI)m / z[M+H] + C 10 Calculated value for H9F3O: 203.1; Measured value: 203.1.

[0488] Step 2. Preparation of 1-(4-(2,2,2-trifluoroethyl)phenyl)ethane-1-ol (compound S114) [ka] To a stirred solution of 1-(4-(2,2,2-trifluoroethyl)phenyl)ethane-1-one (400 mg, 2 mmol) in MeOH (10 mL) cooled to 0°C, NaBH4 (89.8 mg, 2.4 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 1 hour. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 150 mg (37%) of 1-(4-(2,2,2-trifluoroethyl)phenyl)ethane-1-ol (compound S114) as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)7.38-7.32(m,2H),7.32-7.23(m,2H),5.16(d,J =4.2Hz,1H),4.85-4.53(m,1H),3.61(q,J=11.6Hz,2H),1.32(d,J=6.4Hz,3H).

[0489] Synthesis of 1-(4-(1,1-difluoroethyl)phenyl)ethane-1-ol (compound S115) [ka] Step 1. Preparation of 1-bromo-4-(1,1-difluoroethyl)benzene [ka] To a solution of 1-(4-bromophenyl)ethane-1-one (10 g, 50.23 mmol) in DCM (60 mL), DAST (64.7 g, 401.9 mmol) was added dropwise at room temperature, and the mixture was then heated to 50°C and stirred overnight. The reaction mixture was cooled to 0°C, quenched with water, and extracted with DCM (2 × 100 mL). The organic extracts were combined, washed with water (2 × 100 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with DCM / PE (50:1) to obtain 3.4 g (31%) of 1-bromo-4-(1,1-difluoroethyl)benzene as a colorless liquid. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 7.70 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 8.7 Hz, 2H), 1.97 (t, J = 18.9 Hz, 3H).

[0490] Step 2. Preparation of 1-(4-(1,1-difluoroethyl)phenyl)ethane-1-ol (compound S115) [ka] To a stirred solution of 1-bromo-4-(1,1-difluoroethyl)benzene (500 mg, 2.26 mmol, as prepared in the previous step) in THF (10 mL) cooled to -78°C, 2.5 M nBuLi (1.4 mL, 3.39 mmol) in hexane was added dropwise under nitrogen, and the mixture was stirred under nitrogen at -78°C for 30 minutes. A solution of 5 M acetaldehyde in THF (0.6 mL, 2.71 mmol) was added dropwise over 5 minutes at -78°C, and the reaction mixture was then warmed to room temperature and stirred for 1 hour. The reaction mixture was cooled to 0°C, quenched with saturated NH4Cl aqueous solution, and extracted with RINKAN (2 × 30 mL). The organic extracts were combined, washed with water (2 × 30 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 30-40% ACN / water (10 mM NH4HCO3) to obtain 180 mg (43%) of 1-(4-(1,1-difluoroethyl)phenyl)ethane-1-ol (compound S115) as a yellow liquid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.50(d,J=8.3Hz,2H),7.44(d,J=8.2Hz,2H),5.25( d,J=4.3Hz,1H),4.81-4.71(m,1H),1.95(t,J=18.8Hz,3H),1.32(d,J=6.5Hz,3H).

[0491] Synthesis of 2-(4-(trifluoromethyl)phenyl)propanoic acid (compound S116) [ka] Step 1. Preparation of methyl 2-(4-(trifluoromethyl)phenyl) acetate [ka] To a stirred solution of 2-(4-(trifluoromethyl)phenyl)acetic acid (1.2 g, 5.878 mmol) in MeOH (20 mL), concentrated H2SO4 (1.15 g, 11.756 mmol) was added dropwise at room temperature. The mixture was then heated to 80°C and stirred overnight. The reaction mixture was cooled to room temperature, diluted with water (50 mL), and the pH was adjusted to 7 with saturated NaHCO3 aqueous solution. The resulting mixture was extracted with Et2O (3 × 100 mL), the organic extracts were combined, washed with brine (3 × 10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with elution using PE / siRNA (1:1) to obtain 800 mg (62%) of methyl 2-(4-(trifluoromethyl)phenyl)acetate as a yellow oil. The product was used without further purification.

[0492] Step 2. Preparation of methyl 2-(4-(trifluoromethyl)phenyl)propanoate [ka] To a stirred solution of methyl 2-(4-(trifluoromethyl)phenyl)acetate (500 mg, 2.292 mmol, as prepared in the previous step) in THF (20 mL) cooled to -78°C, NaH (60 wt%, 183.2 mg, 4.584 mmol) was added in small amounts under nitrogen, and the mixture was then stirred under nitrogen at -78°C for 30 minutes. MeI (650.5 mg, 4.584 mmol) was added in small amounts to the stirred reaction at -78°C, and the mixture was then warmed to room temperature and stirred overnight. The reaction was poured into a water / ice mixture and then extracted with RINKAN (3 × 100 mL). The organic extracts were combined, washed with brine (3 × 50 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 130 mg (24%) of methyl 2-(4-(trifluoromethyl)phenyl)propanoate as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)7.70(d,J=8.0Hz,2H),7.55(d,J=8.0Hz,2H),3.97(q,J=7.2Hz,1H),3.61(s,3H),1.43(d,J=7.2Hz,3H);MS(ESI)m / z[M+H] + C 11 H 11 Calculated value for F3O: 231.0; Measured value: 231.2.

[0493] Step 3. Preparation of 2-(4-(trifluoromethyl)phenyl)propanoic acid (compound S116) [ka] To a stirred solution of methyl 2-(4-(trifluoromethyl)phenyl)propanoate (110 mg, 0.474 mmol, as prepared in the previous step) in MeOH (10 mL), LiOH (22.6 mg, 0.948 mmol) was added in small amounts at room temperature, and the mixture was then heated to 50 °C and stirred overnight. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and the pH was adjusted to pH 4 with 1 M HCl solution. The mixture was extracted with ethyl acetate (3 × 50 mL), the organic extracts were combined, washed with brine (3 × 10 mL), dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with elution using PE / ethyl acetate (1:1) to obtain 150 mg (145%) of 2-(4-(trifluoromethyl)phenyl)propanoic acid (compound S116) as a yellow-green solid. The solid was used without further purification. MS(ESI)m / z[MH] - C 10 Calculated value for H9F3O2: 217.2; Measured value: 217.0.

[0494] Synthesis of 1-(2-methylquinoline-6-yl)ethane-1-ol (compound S117) [ka] Step 1. Preparation of N-methoxy-N,2-dimethylquinoline-6-carboxamide [ka] To a stirred mixture of 2-methylquinoline-6-carboxylic acid (2 g, 10.7 mmol) and N,O-dimethylhydroxylamine hydrochloride (1.5 g, 16.11 mmol) in DCM (30 mL), HATU (6.1 g, 16 mmol) and DIEA (5.5 g, 42.71 mmol) were added, and the mixture was stirred at room temperature for 3 hours. The reaction product was concentrated under reduced pressure, and the residue was then purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 1.6 g (65%) of N-methoxy-N,2-dimethylquinoline-6-carboxamide as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.35(d,J=8.4Hz,1H),8.22(d,J=1.8Hz,1H),8.00-7.93(m,1H),7.88(d d,J=8.7,1.9Hz,1H),7.48(d,J=8.4Hz,1H),3.57(s,3H),3.32(s,3H),2.68(s,3H);MS(ESI)m / z[M+H] + C 13 H 14 Calculated value for N2O2: 231.1; measured value: 231.1.

[0495] Step 2.1-(2-methylquinoline-6-yl)ethane-1-one [ka] To a stirred solution of N-methoxy-N,2-dimethylquinoline-6-carboxamide (1.5 g, 6.5 mmol, as prepared in the previous step) in THF (20 mL) cooled to -78°C, 3 M MeMgBr (5 mL, 19.5 mmol) in THF was added dropwise under nitrogen, and the mixture was stirred under nitrogen at -78°C for 1 hour. The reaction mixture was warmed to 0°C, quenched with saturated NH4Cl aqueous solution, and extracted with DCM (3 × 300 mL). The organic extracts were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with elution using PE / siRNA (1:1) to obtain 800 mg (66%) of 1-(2-methylquinoline-6-yl)ethane-1-one as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.67(d,J=2.1Hz,1H),8.49-8.39(m,1H),8.17(dd,J=8.8,2.0Hz ,1H),7.99(d,J=8.8Hz,1H),7.53(d,J=8.4Hz,1H),2.71(s,3H),2.70(s,3H);MS(ESI)m / z[M+H] + C 12 H 11 Calculated value for NO: 186.1; Measured value: 186.1.

[0496] Step 3. 1-(2-methylquinoline-6-yl)ethane-1-ol (compound S117) [ka] To a mixture of 1-(2-methylquinoline-6-yl)ethane-1-one (500 mg, 2.7 mmol, as prepared in the previous step) in MeOH (10 mL) cooled to 0°C, NaBH4 (204 mg, 5.4 mmol) was added, and the mixture was stirred for 1 hour. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / siRNA (1:1) to obtain 320 mg (63%) of 1-(2-methylquinoline-6-yl)ethane-1-ol (compound S117) as a colorless oil. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.21(dd,J=8.5,0.8Hz,1H),7.88(d,J=8.7Hz,1H),7.84(d,J=2.0Hz,1H),7.71(dd,J=8.7,2.0H z,1H),7.38(d,J=8.4Hz,1H),5.37(d,J=4.2Hz,1H),5.07-4.86(m,1H),2.64(s,3H),1.42(d,J=6.4Hz,3H);MS(ESI)m / z[M+H] + C 12 H 13 Calculated value for NO: 188.1; Measured value: 188.1.

[0497] Synthesis of 1-(2-methylquinoline-7-yl)ethane-1-ol (compound S118) [ka] To a solution of 7-bromo-2-methylquinoline (500 mg, 2.25 mmol) in THF (10 mL) cooled to -78°C, 2.5 M nBuLi (1.4 mL, 3.37 mmol) in hexane was added dropwise over 5 minutes under nitrogen. Subsequently, 5 M acetaldehyde solution in THF (0.54 mL, 2.70 mmol) was added dropwise at -78°C. The mixture was then warmed to room temperature and stirred for 1 hour. The reaction mixture was cooled to 0°C, quenched with saturated NH4Cl aqueous solution, and extracted with DCM (3 × 20 mL). The organic extracts were combined, washed with water (3 × 20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 5-10% ACN / water (10 mM H4HCO3) to obtain 210 mg (50%) of 1-(2-methylquinoline-7-yl)ethanol (compound S118) as a yellow solid. MS(ESI)m / z[M+H] + C 12 H 13 Calculated value for NO: 188.1; Measured value: 188.1.

[0498] Synthesis of 1-(4-(oxetan-3-yl)phenyl)ethane-1-ol (compound S119) [ka] To a stirred solution of 3-(4-bromophenyl)oxetane (500 mg, 2.34 mmol) in THF (6 mL) cooled to -78°C, 2.5 M nBuLi (1.4 mL, 3.52 mmol) in hexane was added dropwise under nitrogen, and the mixture was stirred for 30 minutes. To the stirred reaction, 5 M acetaldehyde solution in THF (0.56 mL, 2.81 mmol) was added dropwise at -78°C, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction was quenched with water (100 mL) and extracted with RINKAN (2 × 150 mL). The organic extracts were combined, dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 5-100% ACN / water (10 mM H4HCO3) to obtain 200 mg (48%) of 1-(4-(oxetan-3-yl)phenyl)ethane-1-ol (compound S119) as a yellow solid. MS(ESI)m / z[M+H] + C 11 H 14 Calculated value for O2: 178.1; measured value: 178.2.

[0499] Synthesis of 1-(4-isopropylphenyl)ethane-1-ol (compound S120) [ka] To a stirred solution of 1-(4-isopropylphenyl)ethane-1-one (500 mg, 3.1 mmol) in MeOH (10 mL) cooled to 0°C, NaBH4 (140 mg, 3.7 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / siRNA (1:1) to obtain 500 mg (99%) of 1-(4-isopropylphenyl)ethane-1-ol (compound S120) as a colorless oil. 1H NMR(400MHz,DMSO-d6)δ(ppm)7.28-7.22(m,2H),7.18-7.16(m,2H),5.04(d,J=4.2Hz,1 H),4.73-4.62(m,1H),2.92-2.79(m,1H),1.30(d,J=6.5Hz,3H),1.19(d,J=6.9Hz,6H).

[0500] Synthesis of 1-(quinoline-7-yl)ethane-1-ol (compound S121) [ka] To a stirred solution of 1-(quinoline-7-yl)ethane-1-one (400 mg, 2.33 mmol) in MeOH (10 mL) cooled to 0°C, NaBH4 (132.5 mg, 3.50 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction product was quenched with water (50 mL) and extracted with DCM (2 × 150 mL). The organic extracts were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 350 mg (86%) of 1-(quinoline-7-yl)ethane-1-ol (compound S121) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.89(dd,J=4.2,1.8Hz,1H),8.33(dd,J=8.6,2.2Hz,1H),8.00-7.90(m,2H),7.62( dd,J=8.5,1.7Hz,1H),7.49(dd,J=8.3,4.2Hz,1H),5.45-5.38(m,1H),5.01-4.91(m,1H),1.44(d,J=6.4Hz,3H).

[0501] Synthesis of 1-(4-(oxetan-2-yl)phenyl)ethane-1-ol (compound S122) [ka] To a stirred solution of 2-(4-bromophenyl)oxetane (500 mg, 2.34 mmol) in THF (10 mL) cooled to -78°C, 2.5 M nBuLi (1.4 mL, 3.52 mmol) in hexane was added dropwise under nitrogen. Subsequently, 5 M acetaldehyde solution in THF (0.56 mL, 2.81 mmol) was added dropwise at -78°C. The mixture was then warmed to room temperature and stirred for 1 hour. The reaction mixture was cooled to 0°C, quenched with saturated NH4Cl aqueous solution, and extracted with DCM (3 × 20 mL). The organic extracts were combined, washed with water (3 × 20 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 15-20% ACN / water (10 mM NH4HCO3) to obtain 170 mg (41%) of 1-(4-(oxetan-2-yl)phenyl)ethane-1-ol (compound S122) as a yellow liquid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.40-7.33(m,4H),5.69(t,J=7.5Hz,1H),5.13(d,J=4.2Hz,1H),4. 77-4.62(m,2H),4.56-4.48(m,1H),3.00-2.90(m,1H),2.60-2.52(m,1H),1.31(d,J=6.4Hz,3H).

[0502] Synthesis of 1-(quinoline-6-yl)ethane-1-ol (compound S123) [ka] To a stirred solution of 1-(quinoline-7-yl)ethane-1-one (300 mg, 1.75 mmol) in MeOH (5 mL) cooled to 0°C, NaBH4 (132.5 mg, 3.50 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction product was quenched with water (20 mL) and extracted with siRNA (3 × 20 mL). The organic extracts were combined, washed with brine (2 × 3 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 300 mg (99%) of 1-(quinoline-7-yl)ethane-1-ol (compound S123) as a colorless oil.1 H NMR(400MHz,CDCl3)δ(ppm)8.89(dd,J=4.2,1.6Hz,1H),8.17(d,J=8.2Hz,1H),8.11(d,J=8.7Hz,1H),7.82(d,J=1.5Hz, 1H),7.74(dd,J=8.7,1.9Hz,1H),7.41(dd,J=8.3,4.3Hz,1H),5.12(q,J=6.5Hz,1H),3.49(s,1H),1.60(d,J=6.5Hz,3H).

[0503] Synthesis of 1-(2-chloroquinoline-6-yl)ethane-1-ol (compound S124) [ka] Step 1. Preparation of 1-(2-chloroquinoline-6-yl)ethane-1-one [ka] To a stirred solution of 6-bromo-2-chloroquinoline (1 g, 4.1 mmol) and tributyl(1-ethoxyethenyl) stannane (1.8 g, 4.9 mmol) in dioxane (15 mL), Pd(dppf)Cl2 (0.6 g, 0.8 mmol) was added in small amounts under nitrogen. The mixture was then heated to 100 °C and stirred overnight. The reaction mixture was cooled to room temperature, and a 0.1 M HCl solution (15 mL, 2 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at room temperature for 1 hour, and then extracted with DCM (3 × 300 mL). The organic extracts were combined, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel column chromatography while eluting with DCM / MeOH (10:1) to obtain 200 mg (24%) of 1-(2-chloroquinoline-6-yl)ethane-1-one as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.80(d,J=2.0Hz,1H),8.65(d,J=8.6Hz,1H),8.28( dd,J=8.8,2.0Hz,1H),8.05(d,J=8.9Hz,1H),7.73(d,J=8.6Hz,1H),2.73(s,3H).

[0504] Step 2. Preparation of 1-(2-chloroquinoline-6-yl)ethane-1-ol (compound S124) [ka] To a solution of 1-(2-chloroquinoline-6-yl)ethane-1-one (190 mg, 0.9 mmol, as prepared in the previous step) in MeOH (10 mL) cooled to 0°C, NaBH4 (41.9 mg, 1.1 mmol) was added in small increments, and the mixture was stirred at 0°C for 1 hour. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / siRNA (1:1) to obtain 170 mg (89%) of 1-(2-chloroquinoline-6-yl)ethane-1-ol (compound S124) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.45(dd,J=8.7,0.8Hz,1H),7.98(d,J=1.9Hz,1H),7.92(d,J=8.7Hz,1H),7.83( dd,J=8.8,2.0Hz,1H),7.57(d,J=8.6Hz,1H),5.43(d,J=4.2Hz,1H),4.98-4.88(m,1H),1.42(d,J=6.5Hz,3H).

[0505] Synthesis of 2-((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-ol (compound S125) and 2-((tert-butyldimethylsilyl)oxy)-2-(4-fluorophenyl)ethane-1-ol (compound S126) [ka] Step 1.2 Preparation of ((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-one [ka] To a stirred mixture of 1-(4-fluorophenyl)-2-hydroxyethane-1-one (1 g, 6.49 mmol) and imidazole (0.88 g, 12.9 mmol) in THF (20 mL) cooled to 0°C, TBSCl (1.6 g, 9.73 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred overnight. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 600 mg (34%) of 2-((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-one as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 8.06-7.98 (m, 2H), 7.40-7.32 (m, 2H), 5.03 (s, 2H), 0.90 (s, 9H), 0.08 (s, 6H).

[0506] Step 2. Preparation of 2-((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-ol (compound S125) and 2-((tert-butyldimethylsilyl)oxy)-2-(4-fluorophenyl)ethane-1-ol (compound S126) [ka] To a stirred solution of 2-((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-one (1 g, 3.72 mmol, as prepared in the previous step) in MeOH (20 mL) cooled to 0°C, NaBH4 (84.5 mg, 2.23 mmol) was added in small increments, and the mixture was then warmed to room temperature and stirred overnight. The reaction product was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography while eluting with PE / SiO(1:1) to obtain 700 mg (69%) of 2-((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-ol (compound S125) as a yellow solid, and 350 mg (35%) of 2-((tert-butyldimethylsilyl)oxy)-2-(4-fluorophenyl)ethane-1-ol (compound S126) as a yellow solid.

[0507] 2-((tert-butyldimethylsilyl)oxy)-1-(4-fluorophenyl)ethane-1-ol (compound S125). 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.42-7.32(m,2H),7.18-7.08(m,2H),5.33(d,J=4.4Hz,1H),4.60-4.51(m, 1H),3.66(dd,J=10.1,6.0Hz,1H),3.53(dd,J=10.1,5.9Hz,1H),0.81(s,9H),-0.05(s,3H),-0.08(s,3H).

[0508] 2-((tert-butyldimethylsilyl)oxy)-2-(4-fluorophenyl)ethane-1-ol (compound S126). 1 H NMR(400MHz,DMSO-d6)δ(ppm)7.40-7.30(m,2H),7.18-7.09(m,2H),4.77(t,J=5.7 Hz, 1H), 4.68 (dd, J=6.8, 5.2Hz, 1H), 3.47-3.38 (m, 2H), 0.84 (s, 9H), 0.05 (s, 6H).

[0509] Synthesis of 2-(4-(1-hydroxyethyl)phenyl)propan-2-ol (compound S127) [ka] Step 1. Preparation of 1-(4-(2-hydroxypropan-2-yl)phenyl)ethane-1-one [ka] To a stirred mixture of N,O-dimethylhydroxylamine hydrochloride (5.47 g, 56.12 mmol) in THF (30 mL) cooled to 0°C, 1.5 M DIBAL-H (38 mL, 56.12 mmol) in toluene was added dropwise under nitrogen, and the mixture was then warmed to room temperature and stirred for 2 hours. The reaction mixture was cooled to 0°C, methyl 4-acetylbenzoate (5 g, 28.06 mmol) was added, and the mixture was stirred at 0°C for 1 hour. To the stirred mixture at 0°C, 1 M MeMgBr (33.7 mL, 33.672 mmol) in THF was added dropwise over 30 minutes, and the reaction mixture was then stirred at 0°C for 2 hours. The reaction mixture was cooled to -78°C, 3 M MeLi (11.2 mL, 33.672 mmol) in Et2O was added dropwise over 30 minutes, and the mixture was then stirred at -78°C for 1 hour. The reaction mixture was poured into a water / ice mixture and extracted with ethyl acetate (3 × 100 mL). The organic extracts were combined, washed with brine (3 × 10 mL), dried over anhydrous sodium 2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel column chromatography while eluting with PE / ethyl acetate (1:1) to obtain 1 g (20%) of 1-(4-(2-hydroxypropan-2-yl)phenyl)ethane-1-one as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ (ppm) 7.92-7.90 (m, 2H), 7.59-7.57 (m, 2H), 2.58 (s, 3H), 1.59 (s, 6H).

[0510] Step 2. Preparation of 2-(4-(1-hydroxyethyl)phenyl)propan-2-ol (compound S127) [ka] To a stirred solution of 1-(4-(2-hydroxypropan-2-yl)phenyl)ethane-1-one (1.8 g, 10.09 mmol, as prepared in the previous step) in MeOH (10 mL) cooled to 0°C, NaBH4 (460 mg, 12.11 mmol) was added in small amounts under nitrogen, and the mixture was then warmed to room temperature and stirred overnight. The reaction product was concentrated under reduced pressure to obtain 300 mg of 2-(4-(1-hydroxyethyl)phenyl)propan-2-ol (compound S127), which was used directly without further purification.

[0511] Example 1: Synthesis of Exemplary Compound 156 3-(4-(1-benzyl-1H-1,2,4-triazol-3-yl)piperazine-1-yl)-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound 156) [ka] Step 1. Preparation of 1-benzyl-3-chloro-1H-1,2,4-triazole [ka] To a solution of 3-chloro-1H-1,2,4-triazole (5 g, 48.3 mmol) and benzyl bromide (9.09 g, 53.14 mmol) in DMF (200 mL), K2CO3 (13.35 g, 96.61 mmol) was added, and the reaction mixture was stirred under nitrogen at 80°C for 16 hours. The mixture was cooled to room temperature and then concentrated under reduced pressure. The residue was filtered, and the filter cake was washed with DCM (3 × 100 mL). The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 10% to 100% ACN / water (10 mM NH4HCO3) to obtain 3.9 g (42%) of 1-benzyl-3-chloro-1H-1,2,4-triazole as a white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.76(s,1H),7.46-7.26(m,5H),5.40(s,2H).;MS(ESI)m / z[M+H] +Calculated value for C9H8ClN3: 194.1; Measured value: 194.2.

[0512] Step 2.3-(4-(1-benzyl-1H-1,2,4-triazol-3-yl)piperazine-1-yl)-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound 156) [ka] A solution of 1-benzyl-3-chloro-1H-1,2,4-triazole (5 g, 25.82 mmol, as prepared in the previous step) and 6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine (compound 12) (7.29 g, 25.82 mmol) in dioxane (200 mL) is prepared by adding Cs2CO3 (16.83 g, 51.64 mmol) and Pd-PEPPSI-IPent. Cl 2-methylpyridine (2.17 g, 2.58 mmol) was added. The reaction mixture was stirred under nitrogen at 90°C for 16 hours, and the resulting mixture was concentrated under reduced pressure. The residue was diluted with water and extracted with DCM (6 × 100 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography with elution using MeOH / DCM (1:15), and then recrystallized from ACN / THF (5:1) to obtain 5.14 g (45%) of 3-(4-(1-benzyl-1H-1,2,4-triazole-3-yl)piperazine-1-yl)-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound 156) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.82(s,1H),8.32(s,1H),8.23(s,1H),7.97(s,1H),7.77(s,1H),7.66(d,J=9.2Hz, 1H),7.43-7.25(m,6H),5.24(s,2H),3.87(s,3H),3.46(t,J=4.8Hz,4H),3.06(t,J=4.8Hz,4H);MS(ESI)m / z[M+H] + C24 H 25 Calculated value for N9: 440.2; Measured value: 440.05; LCMS purity: 254nm: 99.7%.

[0513] Example 2: Synthesis of Exemplary Compound 275 1-(5-benzylpyrimidine-2-yl)-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)-1,4-diazepan-2-one (compound 275). [ka] 6-(1-methyl-1H-pyrazole-4-yl)-3-piperazine-1-ylpyrazolo[1,5-a]pyridine hydrochloride (compound S19) (5.25 g, 16.5 mmol) was added to a solution of 1-chloro-4-(isocyanatomethyl)benzene (2.76 g, 16.5 mmol) in ACN (110 mL) and DIEA (14.0 mL, 80.4 mmol). The reaction mixture was stirred at room temperature for 1 hour, and then the mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography while eluting with acetone (with 0.1% TEA additive) / siRNA to obtain 5.90 g (80%) of N-(4-chlorobenzyl)-4-[6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl]piperazine-1-carboxamide (compound 275) as a solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.81(s,1H),8.22(s,1H)7.96(s,1H)7.77(s,1H),7.66(d,J=8.7Hz,1H),7.37(m,2H), 7.31(m,3H)7.21(t,J=5.9Hz,1H),4.25(d,J=5.7Hz,2H),3.87(s,3H),3.52(m,4H),2.95(m,4H);MS(ESI)m / z[M+Na] + C 23 H 24 Calculated value for ClN7NaO: 472.2. Measured value: 472.2; HPLC purity: 210nm: 98.7%; 254nm: 98.3%.

[0514] Example 3: Synthesis of exemplary compound 326 1-(5-benzylpyrimidine-2-yl)-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)-1,4-diazepan-2-one (compound 326) [ka] Step 1. Preparation of tert-butyl(3-((6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)amino)propyl)carbamate [ka] To a solution of 6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-amine (compound S56) (1.0 g, 4.69 mmol) in DMF (30 mL), tert-butyl(3-bromopropyl)carbamate (1.34 g, 5.63 mmol) and K2CO3 (1.94 g, 14.07 mmol) were added. The reaction mixture was stirred under nitrogen at 90°C for 16 hours, cooled to room temperature, and diluted with water (50 mL). The mixture was extracted with RINKAN (3 × 200 mL), and the combined organic extract was then washed with brine (3 × 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by preparative TLC (MeOH / DCM 1:20) to obtain 500 mg (19%) of tert-butyl(3-((6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)aminopropyl)carbamate as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.67(s,1H),8.17(s,1H),7.93(s,1H),7.60(dd,J=9.2,1.2Hz,1H),7.49(s,1H),7.14(dd,J=9.2,1.6Hz ,1H),6.88(t,J=5.6Hz,1H),4.70(t,J=6.4Hz,1H),3.86(s,3H),3.09-2.98(m,4H),1.73-1.62(m,2H),1.38(s,9H);MS(ESI)m / z[M+H]+ C 19 H 26 Calculated value for N6O2: 371.2; measured value: 371.2.

[0515] Step 2. Preparation of ethyl N-(3-((tert-butoxycarbonyl)amino)propyl)-N-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)glycinate [ka] To a solution of tert-butyl(3-((6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)amino)propyl) carbamate (500 mg, 1.35 mmol, as prepared in the previous step) and ethyl 2-iodoethyl (346.6 mg, 1.62 mmol) in DMF (10 mL), K2CO3 (373.1 mg, 2.70 mmol) was added, and the reaction was stirred under nitrogen at room temperature for 16 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in water (30 mL). The mixture was extracted with RINKAN (3 × 50 mL), and the combined organic extract was washed with brine (10 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 10% to 100% ACN / water (0.1% FA) to obtain 300 mg (49%) of ethyl N-(3-((tert-butoxycarbonyl)amino)propyl)-N-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)glycinate as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.77(s,1H),8.20(s,1H),7.95(s,1H),7.76(s,1 H),7.57(d,J=9.3Hz,1H),7.29(dd,J=9.3,1.2Hz,1H),6.79(t,J=5.3Hz,1H),4 .04(q,J=7.1Hz,2H),3.93-3.82(m,5H),3.22-3.13(m,2H),2.96(q,J=6.3Hz,2 H),1.52(q,J=6.3Hz,2H),1.35(s,9H),1.14(t,J=7.1Hz,3H);MS(ESI)m / z[M+H] + C 23 H 32 Calculated value for N6O4: 457.2; measured value: 457.3.

[0516] Step 3. Preparation of ethyl N-(3-aminopropyl)-N-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)glycinate hydrochloride [ka] To a solution of ethyl N-(3-((tert-butoxycarbonyl)amino)propyl)-N-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)glycinate (300 mg, 0.68 mmol, as prepared in the previous step) in DCM (10 mL), 4 M HCl in dioxane (3 mL) was added, and the mixture was stirred under nitrogen at room temperature for 2 hours. The reaction product was concentrated under reduced pressure to obtain 200 mg (77%) of ethyl N-(3-aminopropyl)-N-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)glycinate hydrochloride as a white solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.77(s,1H),8.19(s,1H),7.95(s,1H),7.77( s,1H),7.63-7.54(m,1H),7.29(d,J=9.3Hz,1H),4.04(q,J=7.1Hz,2H),3.94 -3.88(m,2H),3.86(s,3H),3.26-3.18(m,2H),2.98(q,J=5.8Hz,1H),2.58( t,J=6.7Hz,1H),1.58-1.43(m,2H),1.14(t,J=7.1Hz,3H);MS(ESI)m / z[M+H] + C 19 H 25 Calculated value for N2O2: 356.2; Measured value: 356.2.

[0517] Step 4. Preparation of 4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)-1,4-diazepan-2-one [ka] To a solution of ethyl N-(3-aminopropyl)-N-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)glycinate hydrochloride (200 mg, 0.55 mmol, as prepared in the previous step) in MeOH (5 mL), 1,5,7-triazabicyclo[4.4.0]deca-5-ene (152.6 mg, 1.10 mmol) was added, and the mixture was stirred under nitrogen at 90°C for 16 hours. The reaction product was concentrated under reduced pressure, and the residue was dissolved in water (30 mL). The mixture was extracted with Âx (3 × 50 mL), and the combined organic extract was washed with brine (10 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by preparative TLC (MeOH / DCM 1:3) to obtain 160 mg (84%) of 4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)-1,4-diazepan-2-one as a white solid. 1H NMR(400MHz,CD3OD)δ(ppm)8.56(s,1H),8.03(s,1H),7.87(s,1H),7.80(s,1H),7.66(d,J=9.3Hz,1H),7.34(dd,J= 9.3,1.4Hz,1H),3.95(d,J=5.5Hz,1H),3.59-3.52(m,2H),3.45-3.39(m,2H),1.94-1.85(m,2H);MS(ESI)m / z[M+H] + C 16 H 18 Calculated value for N6O: 311.2; Measured value: 311.3.

[0518] Step 5. 1-(5-benzylpyrimidine-2-yl)-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)-1,4-diazepan-2-one (compound 326) [ka] To a solution of 4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridin-3-yl)-1,4-diazepan-2-one (150.0 mg, 0.48 mmol, as prepared in the previous step) and 5-benzyl-2-chloropyrimidine (compound S88) (118.7 mg, 0.58 mmol) in dioxane (10 mL), Pd2(dba)3 (88.5 mg, 0.01 mmol), XantPhos (55.9 mg, 0.01 mmol), and K3PO4 (205.2 mg, 0.97 mmol) were added, and the mixture was then stirred under nitrogen at 90°C for 16 hours. The reaction product was concentrated under reduced pressure, and the residue was then purified by preparative TLC (MeOH / DCM 1:10) and preparative HPLC to obtain 42.3 mg (18%) of 1-(5-benzylpyrimidine-2-yl)-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)-1,4-diazepan-2-one (compound 326) as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.87(s,1H),8.70(m,3H),8.21(s,1H),7.90(s,1H),7.65(dd,J=9.2,1.0Hz,1H),7.35-7.27(m, 5H),7.26-7.18(m,1H),4.15(d,J=7.2Hz,4H),4.00-3.80(m,5H),3.52(t,J=5.4Hz,2H),1.94-1.81(m,2H);MS(ESI)m / z[M+H] + C 27 H 26 Calculated value for N8O: 479.2; Measured value: 479.1; LCMS purity: 254nm: 99.0%

[0519] Example 4: Synthesis of exemplary compound 327 (R)-1-phenylethyl(R)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate (compound 327) [ka] Step 1. Preparation of tert-butyl(R)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate [ka] To a solution of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S9) (2.0 g, 7.22 mmol) dissolved in dioxane (30 mL) and tBuOH (30 mL), tert-butyl(R)-2-methylpiperazine-1-carboxylate (7.21 g, 36.08 mmol), tBuXPhos Pd G1 (0.99 g, 1.44 mmol), and KOtBu (1.62 g, 14.43 mmol) were added, and the reaction mixture was stirred under nitrogen at 90°C for 16 hours. The reaction mixture was cooled to room temperature and diluted with water (200 mL). The mixture was extracted with ELISA (3 × 200 mL), and the combined organic extract was washed with brine (3 × 200 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by silica gel chromatography while eluting with siRNA / PE (3:2) to obtain 1.2 g (42%) of tert-butyl(R)-2-methyl-4-[6-(1-methylpyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl]piperazine-1-carboxylate as a pale blue solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.82(s,1H),8.22(s,1H),7.96(s,1H),7.75(s,1H),7.57( d,J=9.2Hz,1H),7.33(dd,J=9.2,1.6Hz,1H),4.22(dd,J=7.6,4.0Hz,1H),3.87(s,3H),3 .84-3.76(m,1H),3.22-3.19(m,1H),3.14-3.07(m,1H),2.77(dd,J=11.6,3.6Hz,1H),2. 68-2.58(m,1H),2.08-1.93(m,1H),1.43(s,9H),1.34(d,J=6.8Hz,3H);MS(ESI)m / z[M+H] + C 21 H 28 Calculated value for N6O2: 397.2; Measured value: 397.2.

[0520] Step 2. Preparation of (R)-6-(1-methyl-1H-pyrazole-4-yl)-3-(3-methylpiperazine-1-yl)pyrazolo[1,5-a]pyridine [ka] To a solution of tert-butyl(R)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate (800 mg, 2.02 mmol, as prepared in the previous step) in DCM (10 mL) cooled to 0°C, 4 M HCl in dioxane (3 mL) was added, and the reaction mixture was stirred under nitrogen at 0°C for 1 hour. The mixture was neutralized to pH 8 with saturated Na2CO3 aqueous solution, and then extracted with MeOH / DCM (1:10) (3 × 100 mL). The combined organic extracts were washed with brine (3 × 100 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain 500 mg (83%) of (R)-6-(1-methyl-1H-pyrazole-4-yl)-3-(3-methylpiperazine-1-yl)pyrazolo[1,5-a]pyridine as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.79(s,1H),8.21(s,1H),7.96(d,J=0.8Hz,1H ),7.70(s,1H),7.60(dd,J=9.2,1.0Hz,1H),7.27(dd,J=9.2,1.2Hz,1H),3.8 6(s,3H),3.21-3.11(m,2H),2.91(dd,J=8.4,2.8Hz,3H),2.67-2.54(m,1H), 2.28(d,J=10.4Hz,1H),1.23(s,1H),1.00(d,J=6.4Hz,3H);MS(ESI)m / z[M+H] + C 16 H 20 Calculated value for N6: 297.2; Measured value: 297.2.

[0521] Step 3. Preparation of (R)-1-phenylethyl(R)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate (compound 327) [ka] To a solution of (R)-6-(1-methyl-1H-pyrazole-4-yl)-3-(3-methylpiperazine-1-yl)pyrazolo[1,5-a]pyridine (400 mg, 1.35 mmol, as prepared in the previous step) and (R)-1-phenylethane-1-ol (494.6 mg, 4.05 mmol) in pyridine (15 mL) cooled to 0°C, triphosgene (400.5 mg, 1.35 mmol) was added in small amounts over 1 minute. The reaction mixture was warmed to room temperature, stirred for 1 hour, then diluted with water (100 mL) and extracted with ethyl acetate (6 × 100 mL). The combined organic extract was washed with brine (6 × 100 mL), dried over anhydrous sodium 2 SO4, and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by preparative HPLC to obtain 158 mg (26%) of (R)-1-phenylethyl(R)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate (compound 327) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ(ppm)8.84(s,1H),8.23(s,1H),7.98(d,J=0.8Hz,1H),7.76(s,1H),7.59(dd, J=9.2,1.0Hz,1H),7.43-7.31(m,5H),7.31-7.26(m,1H),5.76(q,J=6.4Hz,1H),4.33(s,1H),3.92(d, J=13.2Hz,1H),3.88(s,3H),3.38-3.28(m,1H),3.26(d,J=12.4Hz,1H),3.18-3.09(m,1H),2.81(dd,J =11.4,4.0Hz,1H),2.73-2.63(m,1H),1.49(d,J=6.8Hz,3H),1.38(d,J=6.0Hz,3H);MS(ESI)m / z[M+H] + C 25 H 28 Calculated value for N6O2: 445.2; Measured value: 445.1; LCMS purity: 254nm: 98.5%; Chiral purity: 100%.

[0522] Example 5: Synthesis of Exemplary Compound 329 (R)-1-phenylethyl(S)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate (compound 329) [ka] Step 1. Preparation of tert-butyl(S)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate [ka] To a solution of 3-bromo-6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine (compound S9) (2 g, 7.21 mmol) and tert-butyl(S)-2-methylpiperazine-1-carboxylate (7.23 g, 36.08 mmol) in dioxane (50 mL) and tBuOH (50 mL), KOtBu (1.62 g, 14.43 mmol) and tBuXPhos Pd G1 (0.99 g, 1.44 mmol) were added, and the mixture was stirred overnight under nitrogen at 90°C. The reaction product was cooled to room temperature and concentrated under reduced pressure. The mixture was diluted with water (100 mL) and extracted with DCM (3 × 100 mL). The combined organic extract was dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure, and the residue was then purified by reverse-phase flash chromatography using a C18 silica gel column while eluting with 10% to 50% ACN / water (0.1% FA) to obtain 370 mg (13%) of tert-butyl(S)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate as a yellow solid. 1H NMR(400MHz,DMSO-d6)δ(ppm)8.84(t,J=1.3Hz,1H),8.23(s,1H),7.97(d,J=0. 8Hz,1H),7.76(s,1H),7.59-7.57(m,1H),7.35-7.33(m,1H),4.22(s,1H),3.87 (s,3H),3.85-3.77(m,1H),3.25-3.20(m,2H),3.13-3.09(m,1H),2.79-2.75(m ,1H),2.69-2.58(m,1H),1.43(s,9H),1.34(d,J=6.7Hz,3H);MS(ESI)m / z[M+H] + C 21 H 28 Calculated value for N6O2: 397.2; Measured value: 397.2.

[0523] Step 2. Preparation of (S)-6-(1-methyl-1H-pyrazole-4-yl)-3-(3-methylpiperazine-1-yl)pyrazolo[1,5-a]pyridine hydrochloride [ka] To a solution of (S)-2-methyl-4-(6-(1-methyl-1H-pyrazole-4-yl)pyrazolo[1,5-a]pyridine-3-yl)piperazine-1-carboxylate (360 mg, 0.90 mmol, as prepared in the previous st...

Claims

1. Formula I: 【Chemistry 1】 A compound thereof, or a pharmaceutically acceptable salt or solvate thereof, 【Chemistry 2】 However, it exhibits single or double bonds such that all valencies are satisfied. X １ , X ２ , X ３ , and X ４ are selected from N and CR ａ , provided that X １ , X ２ , X ３ , and X ４ are two or less than two of N, on the condition that Y １ and Y ２ One of them is N, and Y １ and Y ２ The other of these is C, Each R ａ They became independent as H, Halo, and C １ ~C ４ Alkyl and C １ ~C ４ Selected from alkoxy, R １ However, C １ ~C ４ Alkyl, C ３ ~C ８ Cycloalkyl, 3-8 membered heterocyclyl, heteroaryl, aryl, and C １ ~C ８ Selected from alkoxys, all of which are optional, including halo, hydroxy, oxo, and C. １ ~C ４ Alkyl, amino C １ ~C ４ Alkyl, hydroxy C １ ~C ４ Alkyl, C １ ~C ４ Alkoxy, C １ ~C ４ Alkoxy C １ ~C ４ Alkyl, 3-8 membered heterocyclyl, and 3-8 membered heterocyclyl C １ ~C ４ It can be substituted with 1, 2, 3, 4, 5, or 6 substituents selected from alkyl groups, provided that the number of substituents does not exceed the number of substituted positions. R ２ However, these are selected from cycloalkyl, cycloalkenyl, alkyl, oxoalkylamino, aminoalkylamino, amino, heterocyclyl, heteroaryl, aminoheterocyclyl, heterocyclylamino, and aminoalkylamino, all of which are optional, and include D, halo, hydroxy, oxo, and C. １ ~C ４ It may be substituted with one or more substituents selected from alkyl groups. R ２ However, 1, 2, or 3 R ３ Replaced by, R ３ However, aryl, heteroaryl, -C(O)R ３１ , -C(O)OR ３１ , -C(O)NR ３１ R ３２ , -S(O) ２ NR ３１ R ３２ , -S(O)(NR ３３ ) R ３１ , -S(O) ２ R ３１ , -S(O)(NR ３３ ) NR ３１ R ３２ , -C(S)NR ３１ R ３２ , C ３ ~C ８ Cycloalkyl, 3-8 membered heterocyclyl, and C １ ~C ４ Selected from alkyl groups, all of which are optional, with 1, 2, 3, 4, or 5 Rs. ３０ It can be replaced by, Each R ３０ These are independent of D, Halo, Arial, -OR ３００ , -NR ３００ R ３０３ , -S(O) ｒ R ３００ , -C(O)R ３００ -C(=CR ３４ R ３５ ) R ３００ , and 【Transformation 3】 Selected from, r is selected from 0, 1, and 2. Each R ３００ C １ ~C ６ Alkyl, C ３ ~C ７ Selected from cycloalkyl, aryl, heteroaryl, 3- to 8-membered heterocyclyl, and 3- to 8-membered heterocyclylaryl, all of which are optional, and include D, halo, hydroxy, amino, alkylamino, cyano, and C. １ ~C ４ Alkyl, C １ ~C ４ Haloalkyl and C １ ~C ４ It may be substituted with one, two, three, four, or five substituents selected from alkoxys. Each R ３０１ is independently selected from H, halo, and C １ -C ４ to C alkyl Each R ３０２ These independently comprise H, F, hydroxyl, amino, alkylamino, oxo, and C １ ~C ４ Selected from alkyloxy, Each R ３０３ H and C are independent of each other. １ ~C ４ Selected from alkyl groups, n, o, and p are each independently selected from 0, 1, 2, 3, and 4. Each R ３１ is independently C １ to C ８ alkyl, aryl C １ to C ４ alkyl, heteroaryl C １ to C ４ alkyl, heterocyclyl, heterocyclyl C １ to C ４ alkyl, cycloalkyl, and cycloalkyl C １ to C ４ / / 这里似乎有重复的格式,推测可能是文档排版问题,按照原样翻译 alkyl selected, and all of these are optionally D, halo, cyano, hydroxy, amino, -OCF ３ C １ to C ４ alkyl, C １ to C ４ haloalkyl, C １ to C ４ alkoxy, hydroxy C １ to C ４ alkyl, -S(O) ２ NR ３０４ R ３０５ -C(O)OR ３０４ R ３０５ -C(O)NR ３０４ R ３０５ and -NR ３０４ C(O)R ３０５ and may be substituted with 1, 2, 3, 4, or 5 substituents selected from Each R ３０４ and R ３０５ H and C are independent of each other. １ ~C ４ Selected from alkyl groups, Each R ３２ H and C are independent of each other. １ ~C ４ Selected from alkyl groups, or R ３１ and R ３２ However, along with the atoms connected to them, D, halo, cyano, C １ ~C ４ Alkyl, C １ ~C ４ Haloalkyl and -C(O)NR ３４ R ３５ A 5- to 8-membered heterocycloyl is formed by optionally substituting with 1, 2, 3, 4, or 5 substituents selected from the above. Each R ３４ and R ３５ H and C became independent. １ ~C ４ Alkyl and C １ ~C ４ Selected from haloalkyl groups, Each R ３３ H and C became independent. １ ~C ４ Alkyl, C １ ~C ４ Haloalkyl and -C(O)R ３４ Selected from, or R ３１ and R ３３ The compound, or a pharmaceutically acceptable salt or solvate thereof, which forms a 4- to 8-membered heterocycloyl group together with the atoms connected thereto.

2. Y １ N is Y ２ The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein C is C.

3. Y １ C is Y ２ The compound according to claim 1, or a pharmaceutically acceptable salt or solvate thereof, wherein is N.

4. X １ N is X ２ CR ａ X ３ CR ａ X ４ CR ａ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

5. X １ CR ａ X ２ N is X ３ CR ａ X ４ CR ａ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

6. X １ CR ａ X ２ CR ａ X ３ N is X ４ CR ａ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

7. X １ CR ａ X ２ CR ａ X ３ CR ａ X ４ A compound according to any one of claims 1 to 3, wherein is N, or a pharmaceutically acceptable salt or solvate thereof.

8. X １ CR ａ X ２ CR ａ X ３ CR ａ X ４ CR ａ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or solvate thereof.

9. R １ The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt or solvate thereof, wherein the compound is a five-membered or six-membered heteroaryl compound.

10. R １ The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt or solvate thereof, wherein the pyrazolyl is optionally substituted.

11. R １ but, 【Chemistry 4】 Selected from, R １０ However, H, C １ ~C ４ Alkyl, C １ ~C ４ Alkoxy, Amino C １ ~C ４ Alkyl, hydroxy C １ ~C ４ Alkyl and C １ ~C ４ Selected from alkylsulfonyl, the C １ ~C ４ alkyl, the C １ ~C ４ Alkoxy, the amino C １ ~C ４ Alkyl, the hydroxy C １ ~C ４ Alkyl, and the C １ ~C ４ Alkyl sulfonyl is optional, hydroxyl, C １ ~C ４ Alkoxy, NR １０ａ R １０ｂ R may be substituted with one or more substituents selected from halo and deuterium, where R １０ａ and R １０ｂ However, hydrogen and C １ ~C ４ Selected from alkyl groups, or R １０ａ and R １０ｂ The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt or solvate thereof, wherein the compounds, together with the nitrogen atoms bonded thereto, form a 4-membered to 8-membered ring.

12. R １ but, 【Transformation 5】 The compound according to claim 11, or a pharmaceutically acceptable salt or solvate thereof.

13. R １０ However, CH ３ The compound according to claim 11 or 12, or a pharmaceutically acceptable salt or solvate thereof.

14. R ２ However, C １ ~C ４ A compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt or solvate thereof, which is a heterocyclyl optionally substituted with an alkyl or oxo group.

15. R ２ However, one R ３ A compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt or solvate thereof, substituted with.

16. R ２ but, 【Chemistry 6-1】 【Chemistry 6-2】 Selected from, 【Transformation 7】 However, it exhibits single or double bonds such that all valencies are satisfied. m is selected from 0, 1, 2, 3, 4, 5, and 6. Z １ Z ２ , and Z ３ However, N and CR ａ A compound according to any one of claims 1 to 15, selected from, or a pharmaceutically acceptable salt or solvate thereof.

17. R ２ but, 【Transformation 8】 A compound according to claim 16, or a pharmaceutically acceptable salt or solvate thereof, selected from the above.

18. Equation Ia: 【Chemistry 9】 It has, In the formula, a and b are each independently selected from 1, 2, and 3. A compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt or solvate thereof, wherein Q is selected from -CH- and -N-, provided that if Q is -N-, a and b are not 1.

19. Formula II: 【Chemistry 10】 A compound according to any one of claims 1 to 18, or a pharmaceutically acceptable salt or solvate thereof, having the above.

20. Formula IIa: 【Chemistry 11】 A compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt or solvate thereof, having the above.

21. R ３ However, 1, 2, 3, 4, or 5 R ３０ A compound according to any one of claims 1 to 20, or a pharmaceutically acceptable salt or solvate thereof, wherein the heteroaryl is optionally substituted.

22. R ３ but, 【Chemistry 12-1】 【Chemistry 12-2】 Selected from, In the formula, A １ However, O, S, and NR ３７ Selected from, R ３６ However, hydrogen is optionally substituted with C. １ ~C ６ Alkyl and optionally substituted C １ ~C ６ Selected from alkylaryls, R ３７ However, hydrogen and C １ ~C ６ A compound selected from alkyl groups, according to any one of claims 1 to 21, or a pharmaceutically acceptable salt or solvate thereof.

23. R ３ but, 【Chemistry 13-1】 【Chemistry 13-2】 A compound according to any one of claims 1 to 22, selected from, or a pharmaceutically acceptable salt or solvate thereof.

24. R ３０ but, 【Chemistry 14】 The compound according to any one of claims 1 to 23, or a pharmaceutically acceptable salt or solvate thereof.

25. R ３００ but, 【Chemistry 15】 A compound according to claim 24, or a pharmaceutically acceptable salt or solvate thereof, selected from the above.

26. R ３００ but, 【Chemistry 16】 The compound according to claim 25, or a pharmaceutically acceptable salt or solvate thereof.

27. R ３０１ H is R ３０２ However, H or CH ３ The compound according to any one of claims 1 to 26, or a pharmaceutically acceptable salt or solvate thereof.

28. R ３ but, 【Chemistry 17】 A compound according to any one of claims 1 to 22, selected from, or a pharmaceutically acceptable salt or solvate thereof.

29. R ３ but, [Chemistry 18] A compound according to claim 28, or a pharmaceutically acceptable salt or solvate thereof, selected from the above.

30. R ３２ The compound according to claim 29, or a pharmaceutically acceptable salt or solvate thereof, wherein is H.

31. R ３１ but, 【Chemistry 19-1】 【Chemistry 19-2】 【Chemistry 19-3】 Selected from, In the formula, R ３１ａ However, C is substituted with H, D, alkylamino, or of any choice. １ ~C ４ Alkyl, C １ ~C ４ Alkoxy, amino, and C １ ~C ４ Selected from haloalkyl groups, Each R ３１ｂ C is independently substituted with H, D, halo, hydroxy, amino, cyano, alkylamino, or any other element. １ ~C ４ Alkyl, C １ ~C ４ Haloalkyl, optionally substituted C １ ~C ４ Alkoxy, and C ３ ~C ６ A compound according to any one of claims 28 to 30, selected from cycloalkyls, wherein q is 1, 2, or 3, or a pharmaceutically acceptable salt or solvate thereof.

32. A compound according to claim 1, selected from any one of the compounds in Table 1, or a pharmaceutically acceptable salt or solvate thereof.

33. A compound according to any one of claims 1 to 32, or a pharmaceutically acceptable salt or solvate thereof, that can exhibit one or more of the following properties: (i) promote differentiation from OPCs to oligodendrocytes; (ii) promote the expression of proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17 (GPR17), myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof); (iii) promote axon myelination; (iv) promote remyelination of demyelinated axons; (v) inhibit PDGFRα kinase activity; (vi) achieve a brain-to-plasma ratio greater than 0.1 when systemically administered to a subject; and (vii) exhibit one or more of any combination thereof.

34. A compound according to claim 33, or a pharmaceutically acceptable salt or solvate thereof, that can inhibit PDGFRα kinase activity.

35. PDGFRα kinase activity is reduced to less than 500 nM, less than 400 nM, less than 300 nM, less than 200 nM, less than 100 nM, less than 75 nM, less than 50 nM, less than 40 nM, less than 30 nM, less than 20 nM, less than 10 nM, less than 5 nM, less than 2.5 nM, less than 1 nM, less than 0.5 nM, or less than 0.2 nM IC ５０ A compound according to claim 34, or a pharmaceutically acceptable salt or solvate thereof, which can be inhibited by [unclear].

36. The IC of the PDGFRα inhibitor ５０ The compound according to claim 35, or a pharmaceutically acceptable salt or solvate thereof, determined using an enzymatic PDGFRα kinase assay (for example, the Promega kinase assay described in Example 131).

37. The compound according to claim 36, or a pharmaceutically acceptable salt or solvate thereof, wherein the enzymatic PDGFRα kinase assay comprises 20 ng of purified PDGFRα protein, 150 μM of ATP, and 1 μg of the substrate poly(Glu4Tyr1) in a volume of 15 μl.

38. A pharmaceutical composition comprising a compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable additive.

39. A kit comprising a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a composition according to claim 38, and instructions for use.

40. A compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38, for use in therapeutic purposes.

41. A method for treating a demyelinating disease in a subject requiring treatment for the said demyelinating disease, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

42. A method for improving the performance of a subject in a test for evaluating one or more symptoms associated with a demyelinating disease, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38, wherein, after the administration, the performance of the subject in the test is improved compared to a reference subject (e.g., the subject before the administration).

43. The method according to claim 42, wherein the examination is one or more of the following: visual evoked potential (VEP) testing, multifocal visual evoked potential (mfVEP) testing, low-contrast visual acuity (LCVA) testing, magnetic resonance imaging (MRI) (e.g., magnetization transport (MTR), myelin water fraction (MWF), quantitative susceptibility mapping (QSM), and T2 imaging), electromyography (EMG), nerve conduction velocity (NCV) testing, expanded disability status scale (EDSS), gait time measurement (e.g., 25-foot walk time measurement), 9-hole peg test (9HPT), optical coherence tomography (OCT), quality of life measurement tests (e.g., quality of life in multiple sclerosis - 54 and visual-related quality of life), cognitive function assessment (e.g., code-digit modality test or Montreal cognitive function assessment), or a combination thereof.

44. The aforementioned demyelinating diseases include acute disseminated encephalomyelitis (ADEM), acute hemorrhagic leukoencephalitis, acute transverse myelitis, adrenoleukodystrophy, adrenal spinal neuropathy, Alexander disease, Alzheimer's disease, aminoaciduria, amyotrophic lateral sclerosis, anti-MAG peripheral neuropathy, anti-MOG-related spectrum, Barlow concentric sclerosis, brain injury, CAMFAK syndrome, Canavan disease, carbon monoxide poisoning, central pontine myelin breakdown, cerebral hypoxia, and brain. Ischemia, Charcot-Marie-Tooth disease, chronic inflammatory demyelinating polyneuropathy, chronic traumatic encephalopathy, syndrome of the first episode (CIS), congenital cataract, copper deficiency-related conditions, delayed hypoxic leukoencephalopathy, Schilder's generalized encephalosclerosis, generalized myelin-destructive sclerosis, extrapontine myelin-disintegrating Gaucher disease, Guillain-Barré syndrome, hereditary neuropathy, hereditary pressure-fragility neuropathy, HTLV-1-associated myelopathy, Hurler syndrome Myelin hypoplasia, hypoxic brain injury, Krabbe disease, Leber hereditary optic nerve atrophy and associated mitochondrial disorders, leukodystrophy, Marquiafava-Bignami disease, metachromatic leukodystrophy, multiple sclerosis (e.g., primary progressive multiple sclerosis (PPMS), relapsing-remitting multiple sclerosis (RRMS), secondary progressive multiple sclerosis (SPMS), progressive relapsing multiple sclerosis, Marburg multiple sclerosis, tumor-like multiple sclerosis) Multiple sclerosis and neuromyelitis optica), multiple system atrophy, myelin-destroying disorders, myelopathy, nerve injury, neuromyelitis optica (NMO), Niemann-Pick disease, optic neuropathy, optic neuritis (e.g., acute optic neuritis and chronic relapsing inflammatory optic neuritis (CRION)), osmotic demyelinating syndrome, Parkinson's disease, Pelizaeus-Merzbach disease, peripheral neuropathy, phenylketonuria, progressive inflammatory neuropathy, progressive multifocal leukemia The method according to claims 41 to 43, comprising one or more of the following: encephalopathy, progressive subcortical ischemic demyelination, reperfusion injury, Schilder's disease, isolated sclerosis, spinal cord injury, subacute sclerosing panencephalitis, tabes dorsalis, Tay-Sachs disease, transverse myelitis, traumatic brain injury, tropical spastic paraplegia, vitamin B12 deficiency, and cerebral palsy.

45. The method according to any one of claims 41 to 44, wherein the demyelinating disease is characterized by the demyelination of one or more cells in the target CNS.

46. The method according to any one of claims 41 to 45, wherein the demyelinating disease is multiple sclerosis.

47. The method according to claim 46, wherein the multiple sclerosis comprises a syndrome consisting of a first episode ("CIS"), relapsing-remitting MS ("RRMS"), secondary progressive MS ("SPMS"), primary progressive MS ("PPMS"), optic neuritis, or transverse myelitis.

48. The method according to any one of claims 41 to 45, wherein the demyelinating disease is optic neuritis.

49. The method according to any one of claims 41 to 48, wherein the treatment of the demyelinating disease includes reducing one or more symptoms associated with the demyelinating disease.

50. The method according to claim 49, wherein one or more of the above symptoms include one or more of the following: fatigue, dizziness, malaise, high fever and high body temperature, extreme coldness of the hands and feet, weakness and stiffness of muscles and joints, weight changes, digestive or gastrointestinal disorders, hypotension, hypertension, irritability, anxiety, depression, visual impairment (e.g., blurred vision, diplopia, decreased low contrast visual acuity (LCVA)), ataxia, clonus, convulsions, dysarthria, weakness, clumsiness, paralysis of the hands, hemiplegia, loss of genital sensation, sexual dysfunction, incoordination, paresthesia, oculoparalysis, muscle coordination disorder, loss of sensation, tingling, anesthesia, pain, neurological symptoms, cognitive impairment, unsteady gait, balance problems, dizziness, spastic paraplegia, incontinence, hearing impairment, speech disorders, loss of smell, and anosmia.

51. A method for promoting axon myelination in a subject requiring promotion of axon myelination, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

52. The method according to claim 51, wherein the promotion of axon myelination results in an increase in the expression of one or more of the following markers within the subject: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

53. The method according to claim 51 or 52, wherein the myelination of the axon can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

54. A method for promoting the remyelination of demyelinated axons in a subject requiring promotion of the remyelination of said demyelinated axons, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

55. The method according to claim 54, wherein the promotion of remyelination of the demyelinated axons results in an increase in the expression of one or more of the following markers within the subject: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

56. The method according to claim 54 or 55, wherein the remyelination of the demyelinated axon can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

57. A method for reducing demyelination of myelinated nerve axons in subjects requiring reduction of demyelination of myelinated nerve axons, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

58. The method according to claim 57, wherein the reduction in demyelination of the myelinated nerve axons results in an increase in the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

59. The method according to claim 57 or 58, wherein the reduction in demyelination of the myelinated nerve axon can be determined by visualizing and / or quantifying the expression of one or more of the following markers: myelin basic protein (MBP), myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or any combination thereof.

60. A method for activating oligodendrocyte progenitor cells (OPCs) in a subject requiring activation of such OPCs in the central nervous system (CNS), the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

61. The method according to any one of claims 51 to 60, wherein the subject has a demyelinating disease, for example, the disease described in any one of claims 44 to 50, or is at risk of developing such a disease.

62. The method according to any one of claims 51 to 60, wherein the method is a method for treating or preventing a demyelinating disease, for example, a disease described in any one of claims 44 to 50.

63. The method according to any one of claims 41 to 62, wherein the compound or the pharmaceutical composition is administered to the subject once.

64. The method according to any one of claims 41 to 62, wherein the compound or the pharmaceutical composition is administered to the subject two or more times using intermittent dosing.

65. The method according to claim 64, wherein the intermittent administration includes administering the compound or the pharmaceutical composition to the subject every other day, every three days, every four days, every five days, every six days, once a week, every eight days, every nine days, every ten days, every eleven days, every twelve days, every thirteen days, once every two weeks, once every three weeks, once a month, once every two months, once every three months, once every four months, once every five months, once every six months, or once every twelve months.

66. The method according to claim 64 or 65, wherein the intermittent administration comprises administering a first dose and a second dose of the compound or the pharmaceutical composition to the subject, the second dose being administered at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, or at least 12 months after the administration of the first dose.

67. The method according to claim 66, wherein the second dose is administered to the subject one day, two days, three days, four days, five days, six days, seven days, eight days, nine days, ten days, eleven days, twelve days, thirteen days, two weeks, three weeks, one month, two months, three months, four months, five months, six months, or twelve months after the administration of the first dose.

68. The method according to any one of claims 41 to 67, wherein, after administration, the compound or the pharmaceutical composition can achieve a brain-to-plasma ratio of greater than 0.1, greater than 0.2, greater than 0.3, greater than 0.4, greater than 0.5, greater than 0.6, greater than 0.7, greater than 0.8, greater than 0.9, greater than 1.0, greater than 1.1, greater than 1.2, greater than 1.3, greater than 1.4, greater than 1.5, greater than 1.6, greater than 1.7, greater than 1.8, greater than 1.9, or greater than 2.

0.

69. The method according to any one of claims 41 to 68, further comprising administering an additional therapeutic agent to the subject.

70. The method according to claim 69, wherein the additional therapeutic agent includes a standard therapeutic agent.

71. The method according to claim 69 or 70, wherein the additional therapeutic agent comprises an immunomodulator.

72. The method according to claim 71, wherein the additional therapeutic agent is selected from interferon beta-1b, interferon beta-1a, pegylated interferon beta-1a, alemtuzumab, natalizumab, ocrelizumab, ofatumumab, ubrituximab-xiiy, glatiramer acetate, teriflunomide, dimethyl fumarate, monomethyl fumarate, diloximel fumarate, fingolimod hydrochloride, siponimod fumarate, ozanimod hydrochloride, ponesimod, cladribine, mitoxantrone, BTK inhibitors, statins, or pharmaceutically acceptable salts thereof.

73. The method according to any one of claims 69 to 72, wherein the additional therapeutic agent is administered to the subject before, simultaneously with, or after the administration of the compound or the pharmaceutical composition.

74. A method for inducing differentiation from oligodendrocyte progenitor cells (OPCs) to oligodendrocytes, the method comprising contacting the OPCs with an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition according to claim 38.

75. The method according to claim 74, wherein the induction of differentiation from OPC to oligodendrocyte results in increased expression of the following markers in the subject: GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

76. The method according to claim 74 or 75, wherein the differentiation from OPC to oligodendrocyte is measured by determining the expression of GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

77. A method for inhibiting PDGFRα activity in cells, the method comprising contacting the cells with an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

78. The method according to claim 77, wherein the inhibition of PDGFRα activity is measured by one or more of the following: an in vitro OPC differentiation assay (e.g., as described in Example 132), a cuprizone model for demyelination, an in vivo OPC differentiation assay (e.g., as described in Example 134), an enzymatic PDGFRα kinase assay (e.g., as described in Example 131), or any combination thereof.

79. The method according to any one of claims 74 to 78, wherein the contact occurs ex vivo or in vivo.

80. The method according to any one of claims 74 to 79, wherein the method is a treatment method.

81. A method for treating relapsing-type multiple sclerosis in a subject requiring treatment for the relapsing-type multiple sclerosis, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38.

82. The method according to claim 81, wherein the induction of differentiation from OPC to oligodendrocyte results in increased expression of the following markers in the subject: GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

83. The method according to claim 81 or 82, wherein the differentiation from OPC to oligodendrocyte is measured by determining the expression of GPR17, MBP, ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof.

84. The method according to any one of claims 81 to 83, wherein the relapsing multiple sclerosis includes a syndrome consisting of a first episode ("CIS"), relapsing-remitting MS ("RRMS"), secondary progressive MS ("SPMS"), primary progressive MS ("PPMS"), or transverse myelitis.

85. A compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38, for use in one or more of the following ways: (i) promoting differentiation from OPCs to oligodendrocytes; (ii) promoting the expression of proteins associated with oligodendrocyte differentiation and / or myelination (e.g., G protein-coupled receptor 17, myelin basic protein (MBP), ASPA, GST-pi, CC1, myelin oligodendrocyte glycoprotein (MOG), oligodendrocyte-specific protein / claudin-11, CNPase, proteolipidoprotein 1 (PLP1), or a combination thereof); (iii) promoting axon myelination; (iv) promoting remyelination of demyelinated axons; (v) inhibiting PDGFRα kinase activity; (vi) achieving a brain-to-plasma ratio greater than 0.1 when systemically administered to a subject; and (vii) any combination thereof.

86. A method for treating a PDGF-related tumor in a subject requiring treatment for the PDGF-related tumor, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 37 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to claim 38, wherein the PDGF-α activity in the subject is reduced after the administration.

87. The method according to claim 86, wherein the PDGF-related tumor includes oligodendrocyte.

88. The method according to claim 86 or 87, wherein the method is a treatment method.

Images