LRRK2 inhibitors

Abstract

The present invention relates to imidazo [4, 5-c] quinoline compounds of formula (I) and pharmaceutically acceptable salts thereof. The invention also relates to pharmaceutical compositions comprising compounds of formula (I) and to the use of these compounds in the treatment of diseases associated with LRRK2, such as neurodegenerative diseases, including Parkinson's disease or Alzheimer's disease, or inflammatory bowel disease, such as Crohn's disease.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 341,065, filed on May 12, 2022, which is incorporated herein by reference in its entirety for all purposes. Background Art

[0003] LRRK2 is a 286 kDa protein with a complex multi-domain structure in the ROCO protein family. The identified protein motifs of LRRK2 include an armadillo-like (ARM) domain, ankyrin-like (ANK) domain, leucine-rich repeat (LRR) domain, Ras of complex (ROC) domain, C-terminal (COR) of the ROC domain, kinase domain, and C-terminal WD40 domain. The ROC domain binds to guanosine triphosphate (GTP), and the COR domain may be a regulator of the GTPase activity of the ROC domain. The kinase domain has structural homology with MAP kinase kinase kinase (MAPKKK) and has been shown to phosphorylate a variety of cellular proteins in vitro, but its endogenous substrates have not yet been determined. LRRK2 is present in different regions of the brain and in a variety of peripheral tissues, including the heart, lungs, spleen, and kidneys.

[0004] LRRK2 can play a complex role in multiple cellular processes because it has a multi-domain structure, each domain is associated with putative protein-protein interactions, guanosine triphosphatase (GTPase) activity and kinase activity. For example, LRRK2 is associated with NFAT inhibition in the immune system, and is associated with vesicle transport, presynaptic homeostasis, mammalian rapamycin target (mTOR) signaling, papillary renal carcinoma and thyroid cancer through receptor tyrosine kinase MET signaling, cytoskeleton dynamics, mitogen-activated protein kinase (MAPK) pathways, tumor necrosis factor-α (TNF-α) pathways, Wnt pathways and autophagy. Genome-wide association (GWA) genetic studies have shown that LRRK2 is associated with the pathogenesis of a variety of human diseases, such as Parkinson's disease and inflammatory bowel disease (such as Crohn's disease). (Lewis, PA and Manzoni, C. Science Signaling 2012, 5 (207), pe2).

[0005] Parkinson's disease (PD) is a relatively common age-related neurodegenerative disorder caused by the progressive loss of dopamine-producing neurons, with a prevalence of up to 4% in people over 80 years of age. Parkinson's disease is characterized by its motor symptoms, such as tremor, rigidity, dyskinesia and postural instability at rest, as well as non-motor symptoms such as impaired cognition, sleep and smell. GWA studies have linked LRRK2 to Parkinson's disease, and many patients with point mutations in LRRK2 present with symptoms indistinguishable from those with idiopathic Parkinson's disease. More than 20 LRRK2 mutations have been associated with autosomal dominant Parkinson's disease, of which the R1441C, R1441G, R1441H, Y1699C, G2019S, 12020T and N1437H missense mutations are considered pathogenic. Studies have shown that the LRRK2 R1441G mutation increases the release of proinflammatory cytokines in microglia from transgenic mice (increased levels of TNF-α, IL-10, and IL-12, and decreased levels of IL-10), and may therefore be directly toxic to neurons (Gillardon, F. et al. Neuroscience 2012, 208, 41-48). In a mouse model of neuroinflammation, LRRK2 was observed to be induced in microglia, and inhibition of LRRK2 kinase activity using small molecule LRRK2 inhibitors (LRRK2-IN-1 or sunitinib) or LRRK2 knockout resulted in attenuation of TNF-α secretion and induction of nitric oxide synthase (iNOS) (Moehle, M. et al. 2012, 32(5), 1602-1611). The most common LRRK2 mutation is the G2019S mutation, which is present in more than 85% of Parkinson's disease patients carrying LRRK2 mutations. This mutation is present in the LRRK2 kinase domain and results in increased LRRK2 kinase activity. In the human brain, LRRK2 is expressed at the highest levels in the same areas of the brain affected by Parkinson's disease, and LRRK2 has also been found in Lewy bodies (a hallmark of PD). Recent studies have shown that potent, selective, brain-penetrating LRRK2 kinase inhibitors can treat Parkinson's disease.

[0006] Dementia is caused by a wide variety of unique pathological processes. The most common pathological processes that lead to dementia are Alzheimer's disease (AD), cerebral amyloid angiopathy (CM), and prion-mediated diseases (see, e.g., Haan et al., Clin. Neurol. Neurosurg. 1990, 92(4): 305-310; Glenner et al., 1989, 94: 1-28). AD is a progressive neurodegenerative disease characterized by memory impairment and cognitive dysfunction. Nearly half of the population over 85 years old is affected by AD, which is also the fastest growing segment of the U.S. population. It is therefore expected that by 2050, the number of AD patients in the United States will increase from approximately 4 million to approximately 14 million. LRRK2 mutations are associated with AD-like lesions, suggesting that there may be some overlap in the neurodegenerative pathways of both AD and PD (Zimprach, A. et al., Neuron 2004, 44, 601-607). Furthermore, in certain populations, the LRRK2 R1628P variant (COR domain) is associated with an increased incidence of AD, which may be due to increased apoptosis and cell death (Zhao, Y. et al., Neurobiology of Aging 2011, 32, 1990-1993).

[0007] Inflammatory bowel disease (IBD), such as ulcerative colitis or Crohn's disease (CD), is considered a complex disease caused by an inappropriate immune response to the intestinal microbiota. Recent genome-wide association studies have found that LRRK2 is a major susceptibility gene for Crohn's disease, especially the M2397T polymorphism in the WD40 domain (Liu, Z. et al., Nat. Immunol. 2011, 12, 1063-1070). Compared with wild-type mice, LRRK2-deficient mice are more susceptible to dextransodium sulfate-induced colitis, suggesting that LRRK2 may play a role in the pathogenesis of IBD (Liu, Z. and Lenardo, M.; Cell Research 2012, 1-3).

[0008] Non-selective and selective small molecule compounds with LRRK2 inhibitory activity have been described, such as staurosporine, sunitinib, LRRK2-IN-1, CZC-25146, TAE684, and compounds in WO 2011 / 141756, WO 2012 / 028629, WO 2012 / 058193, WO 2017 / 046675, WO 2018 / 163030, WO 2018 / 163066, WO2021 / 080929, and US20210002260. It is desirable to provide compounds that are potent and selective inhibitors of LRRK2 with good pharmacokinetic characteristics and the ability to penetrate the blood-brain barrier. The present invention is intended to address these and other problems. Summary of the invention

[0009] In some embodiments, the present disclosure provides a compound of the present invention, which is a compound of formula (I):

[0010]

[0011] or a pharmaceutically acceptable salt thereof, wherein

[0012] Ring A is a 5- to 6-membered heterocycloalkyl group having 1 to 2 heteroatoms each independently being N, O or S, or a 5- to 6-membered heteroaryl group having 1 or 2 heteroatoms each independently being N, O or S;

[0013] Each R 1 Independently for C 1-6 Alkyl or =O;

[0014] Ring B is a 5- to 6-membered heteroaryl group having 1 to 4 heteroatoms each independently being N, O or S;

[0015] Each R 2 C 1-6 Alkyl, –OH, =O, C(O)R 2a 、C(O)OR 2b 、OC(O)R 2a 、S(O)2R 2a 、S(O)2OR 2b 、OS(O)2R 2a 、N(R 2b )S(O)2R 2a 、S(O)2N(R 2b )(R 2c ), C 3-6 Cycloalkyl, C 1-6 Alkyl-C 3-6Cycloalkyl or 3 to 6 membered heterocycloalkyl having 1 to 3 heteroatoms each independently being N, O or S, wherein each alkyl or cycloalkyl is surrounded by 1 to 3 R 2d and each heterocycloalkyl group is substituted by 0 to 3 R 2e group substitution;

[0016] Each R 2b and R 2c is hydrogen or C 1-6 alkyl;

[0017] Each R 2d independently C(O)R 2d1 or S(O)2R 2d1 ;

[0018] Each R 2e Independently for C 1-6 Alkyl, -OH, =O, C(O)R 2e1 or S(O)2R 2e1 ;

[0019] Each R 2a , R 2d1 and R 2e1 Independently for C 1-6 alkyl;

[0020] Each R 3 and R 4 For hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogen, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or –CN;

[0021] subscript n is 0, 1, or 2; and

[0022] The subscripts m and p are each independently an integer from 1 to 4.

[0023] In some embodiments, the present disclosure provides a pharmaceutical composition of the present invention, which comprises a compound of the present disclosure or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0024] In some embodiments, the present disclosure provides a method of inhibiting LRRK2 in a cell, comprising contacting the cell with an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0025] In some embodiments, the present disclosure provides methods of treating a LRRK2-related disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof. DETAILED DESCRIPTION

[0026] I. Overview

[0027] The compounds disclosed herein include compounds of formula (I), (Ia), (Ib) and (Ic), including the compounds in the Examples. These compounds can be used to inhibit LRRK2, and to treat LRRK2-mediated diseases, such as but not limited to Parkinson's disease, Lewy body dementia, frontotemporal dementia, corticobasal dementia, progressive supranuclear palsy, Alzheimer's disease, tauopathy or alpha-synucleinopathy.

[0028] II. Definitions

[0029] Unless otherwise specifically stated, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the invention belongs. In addition, any method or material similar or equivalent to the method or material described herein may be used in practicing the present invention. For purposes of the present invention, term definition is as follows.

[0030] "A", "an", or "the" refers to aspects that include not only one member, but also aspects that include more than one member. For example, unless the context clearly dictates otherwise, the singular forms "a", "an", and "the" include plural referents. Thus, for example, reference to "a cell" includes a plurality of such cells, reference to "the agent" includes one or more agents known to those of ordinary skill in the art, and so on.

[0031] When referring to a value, "about" includes + / - 10% of the value. For example, about 50% includes a range of 45% to 55%, and about 20 molar equivalents includes a range of 18 to 22 molar equivalents. Thus, when referring to a range, "about" refers to + / - 10% of the value at both ends of the range. For example, a ratio of about 1 to about 3 (weight / weight) includes a range of 0.9 to 3.3.

[0032] "Alkyl" refers to a straight or branched chain saturated monovalent alkane. An alkyl group may have 1 to 18 carbon atoms (i.e., C 1-18 Alkyl) or 1 to 8 carbon atoms (i.e. C 1-8 Alkyl) or 1 to 6 carbon atoms (i.e. C 1-6 Alkyl) or 1 to 4 carbon atoms (i.e. C 1-4Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2- Methyl-2-propyl (t-Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), 1-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH( 2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2) and 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3. Other alkyl groups include heptyl, octyl, nonyl, decyl, undecyl, dodecyl, pentadecyl, hexadecyl, heptadecyl and octadecyl. The alkyl group can be substituted or unsubstituted.

[0033] "Alkoxy" refers to an alkyl group with an oxygen atom attached to the alkyl group and a point of attachment: alkyl-O-. As with the alkyl group, the alkoxy group can have any suitable number of carbon atoms, such as C 1-6 Alkoxy groups include, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 2-butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like. The alkoxy group may be further substituted with various substituents as described. The alkoxy group may be substituted or unsubstituted.

[0034]

[0046] "Halo" or "halogen" as used herein refers to fluorine (-F), chlorine (-Cl), bromine (-Br), and iodine (-I).

[0035] An "oxo" substituent refers to a divalent substituent "=0" present on a single atom. For example, an oxo substituent combined with the carbon atom to which it is attached is a carbonyl (C=O).

[0036] As used herein, "haloalkyl" refers to an alkyl group as defined herein, wherein one or more hydrogen atoms of the alkyl group are independently replaced by a halo substituent, which halo substituents may be the same or different. 1-4 Haloalkyl is C 1-4 Alkyl, where C 1-4 One or more hydrogen atoms of the alkyl group have been replaced by a halo substituent. Examples of haloalkyl groups include, but are not limited to, fluoromethyl, fluorochloromethyl, difluoromethyl, difluorochloromethyl, trifluoromethyl, 1,1,1-trifluoroethyl, and pentafluoroethyl. Haloalkyl groups may be substituted or unsubstituted.

[0037] "Haloalkoxy" refers to an alkoxy group in which some or all of the hydrogen atoms are replaced by halogen atoms. As with alkyl, the haloalkoxy group may have any suitable number of carbon atoms, such as C 1-6 Alkoxy groups may be substituted with 1, 2, 3 or more halogens. When all hydrogens are replaced with halogens (e.g., fluorine), the compound is fully substituted, e.g., perfluorinated. Haloalkoxy groups include, but are not limited to, trifluoromethoxy, 2,2,2,-trifluoroethoxy, perfluoroethoxy, and the like. Haloalkoxy groups may be substituted or unsubstituted.

[0038] "Cycloalkyl" refers to a single saturated or partially unsaturated all-carbon ring (i.e., C 3-20Cycloalkyl) such as 3 to 12 annular atoms (annular atom), or 3 to 10 annular atoms, or 3 to 8 annular atoms, or 3 to 6 annular atoms, or 3 to 5 annular atoms, or 3 to 4 annular atoms.Term " cycloalkyl " also includes multiple condensed, saturated and partially unsaturated all-carbon ring systems (for example, the ring system comprising 2,3 or 4 carbocyclic rings).Therefore, cycloalkyl includes polycyclic carbocycles, such as bicyclic carbocycles (such as bicyclic carbocycles with 6 to 12 annular carbon atoms, such as bicyclic [3.1.0] hexane and bicyclic [2.1.1] hexane), and polycyclic carbocycles (such as tricyclic and tetracyclic carbocycles with up to 20 annular carbon atoms).When valence requirements allow, each ring of the polycondensed ring system can be connected by fusion, spiro connection and bridging. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl, and 1-cyclohex-3-enyl. The cycloalkyl may be substituted or unsubstituted.

[0039] "Alkyl-cycloalkyl" refers to a group having an alkyl component and a cycloalkyl component, wherein the alkyl component connects the cycloalkyl component to the point of attachment. The alkyl component is as defined above, but the alkyl component is at least divalent, i.e., alkylene, to connect to the cycloalkyl component and the point of attachment. The alkyl component may include any number of carbon atoms, such as C 1-6 , C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 The cycloalkyl component is as defined above. Exemplary alkyl-cycloalkyl groups include, but are not limited to, methyl-cyclopropyl, methyl-cyclobutyl, methyl-cyclopentyl, and methyl-cyclohexyl. The alkyl-cycloalkyl group may be substituted or unsubstituted.

[0040] As used herein, "heterocyclyl" or "heterocycle" or "heterocycloalkyl" refers to a single saturated or partially unsaturated non-aromatic ring or a polycyclic ring system having at least one heteroatom (i.e., at least one ring heteroatom selected from oxygen, nitrogen and sulfur) in the ring, wherein the polycyclic ring system includes at least a non-aromatic ring containing at least one heteroatom. The polycyclic ring system may also include other aromatic rings and non-aromatic rings. Unless otherwise specified, the heterocyclic group has 3 to 20 ring atoms, such as 3 to 12 ring atoms, such as 3 to 10 ring atoms, or 3 to 8 ring atoms, or 3 to 6 ring atoms, or 3 to 5 ring atoms, or 4 to 6 ring atoms, or 4 to 5 ring atoms. Thus, the term includes a single saturated ring or a partially unsaturated ring (such as a 3, 4, 5, 6 or 7-membered ring) having 1 to 6 ring carbon atoms and 1 to 3 ring heteroatoms selected from oxygen, nitrogen and sulfur. The heteroatoms may be optionally oxidized to form -N(-OH)-, =N(-OH)-, or -N(-OH)-. - )-, -S(=O)-, or -S(=O)2-. Where valence requirements permit, the rings of a polycondensed ring system (such as a bicyclic heterocyclic group) may be connected by fusion, spiro, and bridging. Heterocycles include, but are not limited to, azetidine, aziridine, imidazolidine, morpholine, oxirane (epoxide), oxetane, thiazole, piperazine, piperidine, pyrazolidine, piperidine, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, dihydropyridine, tetrahydropyridine, quinuclidine, 2-oxa-6-azaspiro[3.3]hept-6-yl, 6-oxa-1-azaspiro[3.3]hept-1-yl, 2-thia- [3.3]hept-6-yl, 2,6-diazaspiro[3.3]hept-2-yl, 2-azabicyclo[3.1.0]hex-2-yl, 3-azabicyclo[3.1.0]hexyl, 2-azabicyclo[2.1.1]hexyl, 2-azabicyclo[2.2.1]hept-2-yl, 4-azaspiro[2.4]heptyl, 5-azaspiro[2.4]heptyl and the like. The heterocycloalkyl group may be substituted or unsubstituted.

[0041] Heterocycloalkyl rings also include 9 to 15 membered fused ring heterocycloalkyls having 2, 3 or more rings, wherein at least one ring is aromatic and at least one ring is a non-aromatic ring containing at least one heteroatom. Representative fused bicyclic heterocycloalkyls include, but are not limited to, indoline (dihydroindoline), isoindoline (dihydroisoindoline), indazoline (dihydroindazole), benzo[d]imidazole, dihydroquinoline, dihydroisoquinoline, dihydrobenzofuran, dihydroisobenzofuran, benzo[d][1,3]dioxol, dihydrobenzo[b]dioxin, dihydrobenzo[d]oxazole, dihydrobenzo[b]thiophene, dihydroisobenzo[c]thiophene, dihydrobenzo[d]thiazole, dihydrobenzo[c]isothiazole and benzo[b][1,4]thiazine, as shown in the following structures:

[0042]

[0043] Fused bicyclic heterocycloalkyl groups can also be represented by the following structure:

[0044]

[0045] Where X 1 , X 2 , X 3 and X 4 are each independently absent, -CH2-, -NH-, -O- or -S-, X 1 , X 2 , X 3 and X 4 At least one of them is -NH-, -O- or -S-, and the dotted ring represents a saturated or partially unsaturated non-aromatic ring. The fused bicyclic heterocycloalkyl group may be substituted or unsubstituted.

[0046] "Alkyl-heterocycloalkyl" refers to a group having an alkyl component and a heterocycloalkyl component, wherein the alkyl component connects the heterocycloalkyl component to the point of attachment. The alkyl component is as defined above, but the alkyl component is at least divalent, i.e., alkylene, to connect the heterocycloalkyl component to the point of attachment. The alkyl component may include any number of carbon atoms, such as C 1-6 , C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 The heterocycloalkyl component is as defined above. The alkyl-heterocycloalkyl group may be substituted or unsubstituted. The alkyl-heterocycloalkyl group may be substituted or unsubstituted.

[0047] "Aryl" as used herein refers to a single all-carbon aromatic ring or a plurality of condensed all-carbon ring systems, wherein at least one ring is an aromatic ring. For example, in certain embodiments, the aryl group has 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms. Aryl includes phenyl. Aryl also includes a polycondensed ring system (such as a ring system comprising 2, 3 or 4 rings) having 9 to 20 carbon atoms (such as 9 to 16 carbon atoms), wherein at least one ring is an aromatic ring, and wherein the other rings can be aromatic rings or non-aromatic rings (i.e., carbocyclic rings). Such polycondensed ring systems may optionally be substituted by one or more (such as 1, 2 or 3) oxy groups in any carbocyclic ring portion of the polycondensed ring system. Where valence requirements permit, the rings of the polycondensed ring system may be connected by fusion, spiro connection and bridging. It should also be understood that when referring to a member aryl group (such as a 6-10 yuan aryl group) of a certain atomic range, the atomic range refers to the total ring atoms of the aryl group. For example, six-membered aryl groups include phenyl, and ten-membered aryl groups include naphthyl and 1,2,3,4-tetrahydronaphthyl. Non-limiting examples of aryl groups include, but are not limited to, phenyl, indenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, and the like. Aryl groups may be substituted or unsubstituted.

[0048] "Alkyl-aryl" refers to a group having an alkyl component and an aryl component, wherein the alkyl component connects the aryl component to the point of attachment. The alkyl component is as defined above, but the alkyl component is at least divalent (alkylene) to connect the aryl component to the point of attachment. The alkyl component may include any number of carbon atoms, such as C 1-6 , C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 In some cases, the aryl component may not be present. The aryl component is as defined above. Examples of alkyl-aryl groups include, but are not limited to, benzyl and ethylbenzene. The alkyl-aryl group may be substituted or unsubstituted.

[0049] As used herein, "heteroaryl" refers to a single aromatic ring having at least one atom other than a carbon atom in the ring, wherein the atom is selected from the group consisting of oxygen, nitrogen and sulfur; "heteroaryl" also includes multiple condensed ring systems having at least one such aromatic ring, which are further described below. Thus, "heteroaryl" includes a single aromatic ring having 1-6 carbon atoms and 1-4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur. Sulfur atoms and nitrogen atoms may also be present in oxidized form as long as they are aromatic rings. Exemplary heteroaryl ring systems include, but are not limited to, pyridyl, pyrimidinyl, oxazolyl or furanyl. "Heteroaryl" also includes polycondensed ring systems (e.g., ring systems containing 2, 3 or 4 rings), wherein the heteroaryl as defined above is condensed with one or more rings selected from the following: heteroaryl (forming, for example, 1,8-naphthyridinyl), heterocyclic ring (forming, for example, 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocyclic ring (forming, for example, 5,6,7,8-tetrahydroquinolinyl) and aryl (forming, for example, indazolyl) to form a polycondensed ring system. Thus, the heteroaryl (single aromatic ring or polycondensed ring system) has 1-20 carbon atoms and 1-6 heteroatoms in the heteroaryl ring. Such polycondensed ring systems may be optionally substituted with one or more (e.g., 1, 2, 3 or 4) oxy groups in the carbocyclic or heterocyclic ring portions of the condensed rings. Where valence requirements permit, the rings of the polycondensed ring system may be connected by fusion, spiro and bridging. It should be understood that the rings of the polycondensed ring system may be connected to each other in any order. It should be understood that the point of attachment of the heteroaryl or heteroaryl polycondensed ring system can be any suitable atom in the heteroaryl or heteroaryl polycondensed ring system, including carbon atoms and heteroatoms (such as nitrogen atoms). It should also be understood that when referring to a member heteroaryl of a certain atomic range (such as a 5- to 10-membered heteroaryl), the atomic range refers to the total ring atoms of the heteroaryl, including carbon atoms and heteroatoms. For example, a five-membered heteroaryl includes thiazolyl, and a ten-membered heteroaryl includes quinolinyl. Exemplary heteroaryl groups include, but are not limited to, pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furanyl, oxadiazolyl, thiadiazolyl, quinolyl, isoquinolyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalinyl (qunioxalyl), quinazolyl, 5,6,7,8-tetrahydroisoquinolyl, benzofuranyl, benzimidazolyl, thianthrandenyl, pyrrolo [2,3-b] pyridinyl, quinazolinyl-4 (3H) -one and triazolyl. The heteroaryl group can be substituted or unsubstituted.

[0050] "Alkyl-heteroaryl" refers to a group having an alkyl component and a heteroaryl component, wherein the alkyl component connects the heteroaryl component to the point of attachment. The alkyl component is as defined above, but the alkyl component is at least divalent (alkylene) to connect the heteroaryl component to the point of attachment. The alkyl component may include any number of carbon atoms, such as C 1-6 , C 1-2 , C1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 In some cases, the alkyl component may be absent. The heteroaryl component is as defined above. The alkyl-heteroaryl group may be substituted or unsubstituted.

[0051] "Compounds of the disclosure" include compounds disclosed herein, for example, compounds of the disclosure include compounds of Formula (I), (Ia), (Ib), and (Ic), including the compounds in the Examples.

[0052] Pharmaceutically acceptable salts, tautomers, and polymorphs of these compounds are also described herein. "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials that can be used to prepare pharmaceutical compositions suitable for veterinary or human use.

[0053] Examples of "pharmaceutically acceptable salts" of the compounds disclosed herein also include those derived from suitable bases, such as alkali metals (eg, sodium, potassium), alkaline earth metals (eg, magnesium), ammonium and NX4. + (Where X is C 1-4 Also included are base addition salts such as sodium or potassium salts.

[0054] When a compound is represented in chiral form, it is understood that embodiments of the present invention include, but are not limited to, specific diastereomers or enantiomerically enriched forms. Where chirality is not specified but is present, it is understood that embodiments of the present invention are directed to specific diastereomers or enantiomerically enriched forms; or racemic mixtures or scalemic mixtures of such compounds. As used herein, "scalemic mixture" refers to a mixture of stereoisomers in a ratio other than 1:1.

[0055] "Racemate" refers to a mixture of enantiomers. The mixture may contain equal or unequal amounts of each enantiomer.

[0056] "Stereoisomer" refers to a compound with different chirality at one or more stereocenters. Stereoisomers include enantiomers and diastereomers. If a compound has one or more asymmetric centers or has an asymmetrically substituted double bond, it can exist in the form of stereoisomers and can therefore be produced as a single stereoisomer or a mixture. Unless otherwise indicated, the present description includes both single stereoisomers and mixtures. Methods for determining stereochemistry and separation of stereoisomers are well known in the art (see, for example, Advanced Organic Chemistry 4th Edition Chapter 4, J. March, John Wiley and Sons, New York, 1992).

[0057] "Tautomer" refers to alternative forms of a compound that differ in the position of a proton, such as enol-keto and imine-enamine tautomers, or tautomeric forms of heteroaryl groups containing ring atoms attached to both the ring -NH- and the ring=N-, such as pyrazole, imidazole, benzimidazole, triazole, and tetrazole.

[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Dashes at the beginning or end of chemical groups are for convenience purposes; chemical groups may be described with or without one or more dashes without losing their ordinary meaning. Wavy lines passing through lines in structures indicate points of attachment for the groups. Dashed lines indicate optional bonds. Unless chemically or structurally required, no directionality is expressed or implied in the order in which chemical groups are written or at the points of attachment to the rest of the molecule. For example, the group "-SO2CH2-" is equivalent to "-CH2SO2-", and the two can be attached in either direction. Similarly, an "aralkyl" group, for example, can be attached to the rest of the molecule at either the aryl or alkyl portion of the group. Prefixes such as "C u-v " or (C u -C v ) indicates that the following groups have u to v carbon atoms. For example, "C 1-6 "C1-C6 alkyl" and "C1-C6 alkyl" both mean an alkyl group having 1 to 6 carbon atoms.

[0059] As used herein, "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product derived directly or indirectly from the combination of the specified ingredients in the specified amounts. By "pharmaceutically acceptable" is meant that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient.

[0060] "Pharmaceutically effective amount" refers to the amount of a compound of the present disclosure in a formulation or combination thereof that can provide the desired therapeutic or medicinal effect.

[0061] As used herein, "pharmaceutical composition" refers to a product comprising the specified ingredients in the specified amounts, as well as any product resulting directly or indirectly from the specified ingredients in the specified amounts. A pharmaceutical composition is generally safe for use in biological applications.

[0062] "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, vehicle, lubricant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that has been approved by the United States Food and Drug Administration for use in humans or livestock.

[0063] As used herein, "inhibiting" or "inhibiting" LRRK2 refers to reducing the activity and / or function of the LRRK2 enzyme. LRRK2 enzyme activity can be measured by any assay known in the art, including the assays described in WO 2011 / 141756, WO2012 / 028629, WO 2012 / 058193, WO 2017 / 046675, WO 2018 / 163030, WO 2018 / 163066, WO2021 / 080929 or US20210002260, or the assays described herein, such as the assays in the Examples.

[0064] As used herein, "treatment" or "therapy" or "treating" refers to a method for obtaining a beneficial or desired result. For purposes of this disclosure, a beneficial or desired result includes, but is not limited to, alleviating symptoms and / or alleviating the extent of symptoms and / or preventing worsening of symptoms associated with a disease or condition. In some embodiments, "treatment" or "treating" includes one or more of the following: a) inhibiting a disease or condition (e.g., alleviating one or more symptoms caused by a disease or condition, and / or alleviating the extent of a disease or condition); b) slowing or curbing the development of one or more symptoms associated with a disease or condition (e.g., stabilizing a disease or condition, delaying worsening or progression of a disease or condition); and c) alleviating a disease or condition, e.g., resolving clinical symptoms, improving the disease condition, slowing disease progression, improving quality of life, and / or prolonging survival.

[0065] As used herein, "therapeutically effective amount" or "effective amount" refers to an amount that effectively causes a desired biological or medical response, including an amount of a compound that is sufficient to produce a therapeutic effect on the disease when administered to a subject being treated for the disease. The effective amount may vary depending on the compound, the disease and its severity, and the age, weight, etc. of the subject to be treated. The effective amount may include a range of amounts. As understood in the art, an effective amount may be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired therapeutic endpoint. An effective amount may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to have administered an effective amount if, when used in conjunction with one or more other agents, the desired or beneficial results can or have been achieved. Due to the combined effects of the compounds (such as additive or synergistic effects), the appropriate dose of any co-administered compound may optionally be reduced.

[0066] "Administering" refers to oral administration, suppository administration, topical contact administration, parenteral administration, intravenous administration, intraperitoneal administration, intramuscular administration, intralesional administration, intranasal administration or subcutaneous administration, intrathecal administration, or implantation of a sustained-release device (such as a mini-osmotic pump) to a subject. Administration can be performed according to a schedule specifying the frequency of administration, the dosage of administration, and other factors.

[0067] As used herein, "combined administration" refers to the administration of a unit dose of a compound disclosed herein before or after the administration of a unit dose of one or more other therapeutic agents, for example, the administration of a compound disclosed herein is performed within a few seconds, minutes, or hours of the administration of one or more other therapeutic agents. For example, in certain embodiments, a unit dose of a compound disclosed herein is first administered, and then a unit dose of one or more other therapeutic agents is administered within a few seconds or minutes. Alternatively, in other embodiments, a unit dose of one or more other therapeutic agents is first administered, and then a unit dose of a compound disclosed herein is administered within a few seconds or minutes. In some embodiments, a unit dose of a compound disclosed herein is first administered, and then a unit dose of one or more other therapeutic agents is administered after a few hours (e.g., 1-12 hours). In other embodiments, a unit dose of one or more other therapeutic agents is first administered, and then a unit dose of a compound disclosed herein is administered after a few hours (e.g., 1-12 hours). The combined administration of a compound disclosed herein with one or more other therapeutic agents generally refers to the simultaneous or sequential administration of a compound disclosed herein and one or more other therapeutic agents, so that a therapeutically effective amount of each agent is present in the patient's body.

[0068] "Subject" refers to an animal, such as a mammal, including but not limited to primates (such as humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In certain embodiments, the subject is a human.

[0069] "Disease" or "condition" refers to a state of being or health in a patient or subject that can be treated with the compounds, pharmaceutical compositions or methods provided herein.

[0070] III. Compounds

[0071] Compounds of the present disclosure include compounds of Formula (I), (Ia), (Ib), and (Ic), including the compounds in the Examples.

[0072] In some embodiments, the compound of the present invention is a compound of formula (I):

[0073]

[0074] or a pharmaceutically acceptable salt thereof, wherein

[0075] Ring A is a 5- to 6-membered heterocycloalkyl group having 1 to 2 heteroatoms each independently being N, O or S, or a 5- to 6-membered heteroaryl group having 1 or 2 heteroatoms each independently being N, O or S;

[0076] Each R 1 Independently for C 1-6 Alkyl or =O;

[0077] Ring B is a 5- to 6-membered heteroaryl group having 1 to 4 heteroatoms each independently being N, O or S;

[0078] Each R 2 C 1-6 Alkyl, –OH, =O, C(O)R 2a 、C(O)OR 2b 、OC(O)R 2a 、S(O)2R 2a 、S(O)2OR 2b 、OS(O)2R 2a 、N(R 2b )S(O)2R 2a 、S(O)2N(R 2b )(R 2c ), C 3-6 Cycloalkyl, C 1-6 Alkyl-C 3-6 Cycloalkyl or 3 to 6 membered heterocycloalkyl having 1 to 3 heteroatoms each independently being N, O or S, wherein each alkyl or cycloalkyl is surrounded by 1 to 3 R 2d and each heterocycloalkyl group is substituted by 0 to 3 R 2e group substitution;

[0079] Each R 2b and R 2c is hydrogen or C 1-6 alkyl;

[0080] Each R 2d independently C(O)R 2d1 or S(O)2R 2d1 ;

[0081] Each R 2e Independently for C 1-6 Alkyl, -OH, =O, C(O)R 2e1 or S(O)2R 2e1 ;

[0082] Each R 2a , R 2d1 and R 2e1 Independently for C 1-6 alkyl;

[0083] Each R 3 and R 4 For hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogen, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or –CN;

[0084] subscript n is 0, 1, or 2; and

[0085] The subscripts m and p are each independently an integer from 1 to 4.

[0086] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript n is 1 or 2. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript n is 1.

[0087] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript m is 1, 2, 3, or 4. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript m is 1 or 2. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript m is 1.

[0088] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript p is 1, 2, 3, or 4. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript p is 1 or 2. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein subscript p is 1.

[0089] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein ring A is a 5- to 6-membered heterocycloalkyl group having 1 heteroatom of N, O or S or a 5- to 6-membered heteroaryl group having 1 or 2 heteroatoms each independently being N, O or S. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein ring A is a 5- to 6-membered heterocycloalkyl group having 1 heteroatom of N or O or a 5- to 6-membered heteroaryl group having 1 heteroatom of N. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein ring A is a pyrrolidinyl, piperidinyl, tetrahydropyranyl or pyridinyl group. In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein ring A is a tetrahydropyranyl group.

[0090] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 1 Independently for C 1-6 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 1 Independently for C 1-3 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 1 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein each R 1 A compound of Me.

[0091] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein R 3 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein R 3 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein R 3 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein R 3 A compound of hydrogen.

[0092] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 4 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 4 In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 4 A compound containing -CN.

[0093] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound having the structure of Formula Ia:

[0094]

[0095] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound having the structure of Formula Ib:

[0096]

[0097] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound having the structure of Formula Ic:

[0098]

[0099] In some embodiments, the compound or its pharmaceutically acceptable salt is a compound wherein ring B is a 5- to 6-membered heteroaryl group having 1 to 3 heteroatoms each independently being N, O or S. In some embodiments, the compound or its pharmaceutically acceptable salt is a compound wherein ring B is a 5- to 6-membered heteroaryl group having 2 to 3 heteroatoms each independently being N, O or S. In some embodiments, the compound or its pharmaceutically acceptable salt is a compound wherein ring B is pyrrolyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl. In some embodiments, the compound or its pharmaceutically acceptable salt is a compound wherein ring B is pyrazolyl, isoxazolyl, 1,2,3-triazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl or pyridinyl.

[0100] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein the group for of compounds.

[0101] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound wherein

[0102] Each R 2 C 1-3 Alkyl, =O, C(O)OR 2b 、OC(O)R 2a 、S(O)2R 2a 、N(R 2b )S(O)2R 2a 、S(O)2N(R 2b )(R 2c ), C 3-6 Cycloalkyl or 3 to 6 membered heterocycloalkyl having 1 to 3 heteroatoms each independently being N, O or S, wherein each alkyl or cycloalkyl is surrounded by 1 to 2 R 2d and each heterocycloalkyl group is optionally substituted by 1 to 2 R2e group substitution;

[0103] Each R 2b and R 2c is hydrogen or C 1-3 alkyl;

[0104] Each R 2d independently C(O)R 2d1 or S(O)2R 2d1 ;

[0105] Each R 2e Independently for C 1-3 Alkyl, -OH, =O, C(O)R 2e1 or S(O)2R 2e1 ;as well as

[0106] Each R 2a , R 2d1 and R 2e1 Independently for C 1-3 alkyl.

[0107] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein each R 2 are independently =O, -COOH, -C(O)OMe, -SO2Me, -NHSO2Me, -CH2CH2SO2Me, of compounds.

[0108] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is wherein the group for of compounds.

[0109] In some embodiments, the compound or a pharmaceutically acceptable salt thereof is a compound having the structure of the compound in Table 1.

[0110] Table 1. Compounds

[0111]

[0112]

[0113]

[0114]

[0115]

[0116]

[0117] The compounds of the present disclosure described herein can be prepared and / or formulated into pharmaceutically acceptable salts, or formulated into free bases when appropriate. Pharmaceutically acceptable salts are non-toxic salts of the free base form of the compound, having the desired free base pharmacological activity. These salts can be from inorganic or organic acids or bases. Examples of pharmaceutically acceptable salts of the compounds of formula (I) disclosed herein include inorganic acid salts, such as hydrochlorides, sulfates, carbonates and phosphates, and organic acid salts, such as fumarate, maleate, mesylate and p-toluenesulfonate, etc. In addition, salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as magnesium or calcium, salts with organic amines such as lower alkylamines or lower alcoholamines, and salts or ammonium salts of basic amino acids such as lysine, arginine, ornithine. For example, compounds containing basic nitrogen can be prepared into pharmaceutically acceptable salts by contacting with inorganic or organic acids. Non-limiting examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, hexanoates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyrates, The pharmaceutically acceptable salts of the present invention include benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, benzenesulfonate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate and mandelate. A list of other suitable pharmaceutically acceptable salts is found in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott Wiliams and Wilkins, Philadelphia, Pa., 2006.

[0118] In certain embodiments, the compounds of the present disclosure described herein or pharmaceutically acceptable salts, isomers or mixtures thereof are compounds in which 1 to n hydrogen atoms attached to a carbon atom may be replaced by deuterium atoms or D, where n is the number of hydrogen atoms in the molecule. As is known in the art, deuterium atoms are non-radioactive isotopes of hydrogen atoms. Such compounds can increase resistance to metabolism and can therefore be used to increase the half-life of the compounds described herein or their pharmaceutically acceptable salts, isomers or mixtures after administration to mammals. See, for example, Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism", Trends Pharmacol. Sci., 5(12): 524-527 (1984). Such compounds are synthesized by methods well known in the art, such as using starting materials in which one or more hydrogen atoms are replaced by deuterium.

[0119] Examples of isotopes that may be incorporated into the disclosed compounds also include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. Using positron emitting isotopes (such as 11 C. 18 F. 15 O and 13 N) substitution can be used for positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotope-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the examples below, using appropriate isotope-labeled reagents instead of the non-labeled reagents previously used.

[0120] The compounds of the embodiments disclosed herein, or pharmaceutically acceptable salts thereof, may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms, which may be defined, in terms of absolute stereochemistry, as (R)- or (S)-, or (D)- or (L)- for amino acids. The present disclosure is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques such as chromatography and fractional crystallization. Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers. Likewise, all tautomeric forms are also included.

[0121] Activity against LRRK2 can be measured by any biochemical assay known in the art for evaluating LRRK2, such as commercially available assays such as the LRRK2 ELISA kit (Aviva Systems, San Diego, CA USA) and the LRRK2 kinase system (Promega Corp.), assays described in U.S. Pat. Nos. 10039753 and 11161844, and assays described herein. In some embodiments, the compounds of the present disclosure comprise activity against LRRK2, wherein the IC in the biochemical assay is 50 Less than about 30 μM, such as less than about 20 μM, less than about 10 μM, less than about 1 μM, less than about 0.1 μM, less than about 0.01 μM, less than about 0.001 μM, or less than about 0.0001 μM.

[0122] Activity against LRRK2 can also be measured by any cell-based assay known in the art for evaluating LRRK2, such as the Phospho-LRRK2 (Ser935) Cellular Kit (Cisbio Bioassays, France), as described in Hermanson, SB et al., PLOS ONE 7(8): e43580, and the assays described herein. In some embodiments, the compounds of the present disclosure comprise activity against LRRK2, wherein the IC 50Less than about 30 μM, such as less than about 20 μM, less than about 10 μM, less than about 1 μM, less than about 0.1 μM, less than about 0.01 μM, less than about 0.001 μM, or less than about 0.0001 μM in a cellular assay.

[0123] In some embodiments, the compounds of the present disclosure are selective for LRRK2 over one or more other kinases, such as LRRK1, LIMK1, LIMK2, RIPK1, RIPK2, RIPK3, ANKRD3, SgK288, IRAK1, IRAK2, IRAK3, IRAK4, JAK1, JAK2, JAK3, TESK1, and / or TESK2. Selectivity can be measured by relative values ​​in corresponding biochemical assays, for example, inhibiting the activity of LRRK2 is higher than inhibiting the activity of LRRK1, LIMK1, LIMK2, RIPK1, RIPK2, RIPK3, ANKRD3, SgK288, IRAK1, IRAK2, IRAK3, IRAK4, JAK1, JAK2, JAK3, TESK1, and / or TESK2.

[0124] In some embodiments, the compounds of the present disclosure are selective for LRRK2 over one or more (e.g., 2, 3, 4, 5, 6, 7, 8, or 9, or more) other kinases (including LRRK1, LIMK1, LIMK2, RIPK1, RIPK2, RIPK3, ANKRD3, SgK288, IRAK1, IRAK2, IRAK3, IRAK4, JAK1, JAK2, JAK3, T The ESK1 and / or TESK2) are at least about 1.2, about 1.5, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 200, about 300, about 400, about 500, about 1000, about 2000, about 3000, about 4000, about 5000, about 10000 times or more.

[0125] IV. Pharmaceutical Compositions

[0126] In some embodiments, the pharmaceutical composition comprises a pharmaceutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises a pharmaceutically effective amount of a compound of formula (I), (Ia), (Ib) or (Ic) or a pharmaceutically acceptable salt and / or solvate thereof, and a pharmaceutically acceptable carrier or excipient.

[0127] In some embodiments, the pharmaceutical composition further comprises one or more additional therapeutic agents. Any suitable additional therapeutic agent or combination therapy may be used with a compound of Formula (I), (Ia), (Ib) or (Ic) or a pharmaceutically acceptable salt thereof (such as the agents and therapies described herein).

[0128] In some embodiments, the pharmaceutical composition comprises a compound of Formula (I), (Ia), (Ib) or (Ic) and an additional therapeutic agent, wherein the additional therapeutic agent is an anti-Parkinson's disease agent.

[0129] In some embodiments, the pharmaceutical composition comprises a compound of Formula (I), (Ia), (Ib) or (Ic) and an additional therapeutic agent, wherein the additional therapeutic agent is an anti-inflammatory bowel disease agent.

[0130] The compounds herein are formulated with conventional carriers and excipients. Tablets contain excipients, lubricants, fillers, binders and the like. Water-based formulations are prepared in a sterile form and are generally isotonic formulations when used for delivery by non-oral administration. All formulations optionally contain excipients, such as those listed in the "Handbook of Pharmaceutical Excipients" (1986). Excipients include ascorbic acid and other antioxidants, chelating agents (such as EDTA), carbohydrates (such as polydextrose), hydroxyalkyl cellulose, hydroxyalkyl methyl cellulose, stearic acid, etc. The pH range of the formulation is from about 3 to about 11, for example, from about 7 to 10.

[0131] Although the active ingredient can be administered alone, it is preferably presented in the form of a pharmaceutical formulation. Both veterinary and human formulations include at least one active ingredient as defined above, together with one or more acceptable carriers and optionally other therapeutic ingredients, particularly those discussed herein. The carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not physiologically harmful to the recipient.

[0132] Preparations include preparations suitable for the above-mentioned routes of administration. Preparations can be conveniently provided in unit dosage form and can be prepared by any appropriate method. Preparation techniques and preparations can generally be found in Remington's Pharmaceutical Sciences (Mack Publishing Co.). These methods include the step of associating the active ingredient with a carrier constituting one or more auxiliary ingredients. In general, the method for preparing the preparation is to uniformly and closely associate the active ingredient with a liquid carrier or a finely powdered solid carrier or both, and then, if necessary, the product is molded.

[0133] Formulations suitable for oral administration may be in the form of discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; in the form of a powder or granules; in the form of a solution or suspension in water or a non-aqueous liquid; or in the form of an oil-in-water emulsion or a water-in-oil emulsion. The active ingredient may also be administered in the form of a bolus, electuary or paste.

[0134] Tablets are made by compression or molding, optionally with the addition of one or more auxiliary ingredients. Compressed tablets are prepared by compressing the active ingredient in a free-flowing form such as a powder or granules in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersant. Molded tablets can be made by molding a mixture of powdered active ingredients moistened with an inert liquid diluent in a suitable machine. Tablets may optionally be coated or scored, or optionally formulated to provide sustained or controlled release of the active ingredient.

[0135] The pharmaceutical preparations herein include a combination of one or more pharmaceutically acceptable carriers or excipients and optionally other therapeutic agents. The pharmaceutical preparation containing the active ingredient can be in any form suitable for the intended method of administration. For example, when used for oral administration, tablets, lozenges, pastilles, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, solutions, syrups or elixirs can be prepared. Compositions intended for oral administration can be prepared according to the production method of any pharmaceutical composition, and such compositions can contain one or more medicaments, including sweeteners, flavoring agents, colorants and preservatives to provide a palatable preparation. It is acceptable to mix tablets containing active ingredients with non-toxic, pharmaceutically acceptable excipients suitable for making tablets. These excipients can be, for example, inert diluents such as calcium carbonate or sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrants such as corn starch or alginic acid; binders such as starch, gelatin or gum arabic (acacia); and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or coated by known techniques (including microencapsulation coating) to delay disintegration and absorption in the gastrointestinal tract, thereby providing a longer sustained effect. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used alone or with a wax.

[0136] Oral preparations can also be hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (such as calcium phosphate or kaolin); or soft gelatin capsules in which the active ingredient is mixed with water or an oil medium (such as peanut oil, liquid paraffin or olive oil).

[0137] Aqueous suspensions contain active substances and are mixed with excipients suitable for making aqueous suspensions. These excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth and gum arabic, and dispersants or wetting agents, such as naturally occurring phosphatides (such as lecithin), condensation products of alkylene oxide and fatty acids (such as polyoxyethylene stearate), condensation products of ethylene oxide and long-chain fatty alcohols (such as heptadecaethyleneoxycetanol), condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol anhydrides (such as polyoxyethylene sorbitol monooleate). Aqueous suspensions may also contain one or more preservatives, such as ethyl p-hydroxybenzoate or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweeteners, such as sucrose or saccharin.

[0138] Oily suspensions can be prepared by suspending the active ingredient in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (e.g., liquid paraffin). Oral suspensions may contain thickeners, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners (as described above) and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant (e.g., ascorbic acid).

[0139] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide the active ingredient, which is mixed with a dispersant or wetting agent, a suspending agent and one or more preservatives. Examples of suitable dispersants or wetting agents and suspending agents are disclosed above. In addition, other excipients, such as sweeteners, flavoring agents and coloring agents, may also be provided.

[0140] The pharmaceutical composition can also be an oil-in-water emulsion. The oil phase can be a vegetable oil (such as olive oil or peanut oil), a mineral oil (such as liquid paraffin) or a mixture thereof. Suitable emulsifiers include naturally occurring gums such as gum arabic and tragacanth; naturally occurring phospholipids such as soybean lecithin; esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate; and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavorings. Syrups and elixirs may be formulated with sweeteners such as glycerol, sorbitol or sucrose. Such preparations may also contain demulcents, preservatives, flavorings or coloring agents.

[0141] The pharmaceutical composition can be in the form of a sterile injection or intravenous preparation, such as a sterile injection aqueous suspension or oily suspension. Such a suspension can be prepared according to known techniques using the appropriate dispersants or wetting agents and suspending agents mentioned above. The sterile injection or intravenous preparation can also be a sterile injection solution or suspension in a non-toxic enterally acceptable diluent or solvent, such as a solution in 1,3-butanediol or prepared as a lyophilized powder. Acceptable carriers and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils can generally be used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can also be used to prepare injections.

[0142] The amount of active ingredient that can be combined with a carrier material to form a single dosage form will vary depending on the subject being treated and the specific mode of administration. For example, a sustained-release formulation for oral administration to humans may contain about 1 to 1000 mg of the active ingredient and be compounded with an appropriate and convenient amount of carrier material, which may account for about 5% to about 95% (by weight) of the total ingredients. The pharmaceutical composition can be formulated into an amount that is easy to measure. For example, an aqueous solution for intravenous infusion may contain about 3 to 500 μg of the active ingredient per milliliter of solution so that an appropriate amount can be infused at a rate of about 30 mL / hr.

[0143] Formulations suitable for topical administration to the eye also include eye drops in which the active ingredient is dissolved or suspended in a suitable carrier, particularly an aqueous solvent for the active ingredient. The concentration of the active ingredient in such formulations is preferably 0.5% to 20%, advantageously 0.5% to 10%, particularly about 1.5% w / w.

[0144] Formulations suitable for topical administration in the mouth include pastilles comprising the active ingredient in a flavored base (usually sucrose and acacia or tragacanth); lozenges comprising the active ingredient in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0145] Formulations for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

[0146] The particle size range of the formulation suitable for intrapulmonary or intranasal administration is 0.1 to 500 microns, such as 0.5, 1, 30, 35, etc., which is administered by rapid inhalation through the nasal cavity or by inhalation through the mouth to reach the alveolar sacs. Suitable formulations include aqueous or oily solutions of the active ingredient.

[0147] Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

[0148] Formulations suitable for parenteral administration include aqueous and nonaqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and nonaqueous sterile suspensions which may contain suspending agents and thickening agents.

[0149] The formulations are provided in unit dose or multi-dose containers (such as sealed ampoules and vials) and can be stored in a freeze-dried (lyophilized) state, requiring only the addition of a sterile liquid carrier (such as water for injection) immediately before use. Ready-to-use injection solutions and suspensions are prepared from sterile powders, granules and tablets of the type described above. Preferred unit dose formulations are formulations containing a daily dose or daily unit sub-dose of the above-mentioned active ingredient or an appropriate portion thereof.

[0150] It should be understood that in addition to the ingredients particularly mentioned above, the formulations may include other conventional agents in the art having regard to the type of formulation concerned. For example, formulations suitable for oral administration may include flavoring agents.

[0151] In some embodiments, the veterinary composition comprises at least one active ingredient as defined above and a veterinary carrier therefor.

[0152] Veterinary carriers are materials used to administer the composition and may be solid, liquid or gaseous materials which are inert or acceptable in veterinary technology and are compatible with the active ingredient. These veterinary compositions may be administered orally, parenterally or by any other desired route.

[0153] The compounds herein can be used to provide controlled release pharmaceutical formulations containing one or more compounds as active ingredients ("controlled release formulations") in which the release of the active ingredient is controlled and regulated to allow less frequent administration or to improve the pharmacokinetic or toxicity profile of a given active ingredient.

[0154] The effective dose of the active ingredient depends at least on the nature of the disease being treated, toxicity, whether the compound is used for prevention (lower dose) or for the treatment of active viral infection, the method of delivery and the pharmaceutical dosage form, and will be determined by the clinician through routine dose escalation studies. The expected dose is about 0.0001 to about 100 mg / kg body weight per day; usually about 0.01 to about 10 mg / kg body weight per day; more typically about 0.01 to about 5 mg / kg body weight per day; most typically about 0.05 to about 0.5 mg / kg body weight per day. For example, the candidate dose range for an adult weighing about 70 kg per day is 1 mg to 1000 mg, preferably between 5 mg and 500 mg, and can be in the form of a single dose or multiple doses.

[0155] V. Route of Administration

[0156] One or more compounds of Formula (I), (Ia), (Ib) or (Ic) may be administered by any route appropriate to the condition to be treated. Suitable routes include oral, rectal, nasal, pulmonary, topical (including oral and sublingual), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural), etc. It should be understood that the preferred route may vary depending on the condition of the recipient, etc. One advantage of the compounds described herein is that they have oral bioavailability and can be administered orally.

[0157] The compounds of the present disclosure can be administered by any route suitable for the condition to be treated. Suitable routes include oral, rectal, nasal, topical (including oral and sublingual), transdermal, vaginal administration and parenteral administration (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural) and the like. It should be understood that the preferred route may vary depending on the condition of the recipient, etc. The advantage of some compounds disclosed herein is that they have oral bioavailability and can be administered orally.

[0158] The compounds of the present disclosure can be administered to the patient in a desired period of time or duration according to an effective administration regimen, such as at least about one month, at least about two months, at least about three months, at least about six months, or at least about twelve months or longer. In a variation, the compound is administered daily or intermittently over the patient's life cycle.

[0159] During the course of treatment, the dosage or frequency of administration of the compounds of the present disclosure may be adjusted according to the judgment of the physician administering the drug.

[0160] The compounds of the present disclosure can be administered to a patient (eg, a human) in an effective amount. In some embodiments, the compound is administered once a day.

[0161] The disclosed compounds can be administered by any useful route and method, such as oral or parenteral (such as intravenous injection). The therapeutically effective amount of the compound may include about 0.00001 mg / kg body weight to about 10 mg / kg body weight per day, such as about 0.0001 mg / kg body weight to about 10 mg / kg body weight per day, or such as about 0.001 mg / kg body weight to about 1 mg / kg body weight per day, or such as about 0.01 mg / kg body weight to about 1 mg / kg body weight per day, or such as about 0.05 mg / kg body weight / day to about 0.5 mg / kg body weight / day, or such as from about 0.3 mg to about 30 mg / day, or such as from about 30 mg to about 300 mg / day.

[0162] The compounds of the present disclosure may be combined with one or more additional therapeutic agents in any dose of the compounds of the present disclosure (e.g., 1 mg to 1000 mg of the compounds of the present disclosure). The therapeutically effective amount may include about 1 mg to about 1000 mg per dose, such as about 50 mg to about 500 mg per dose, or such as about 100 mg to about 400 mg per dose, or such as about 150 mg to about 350 mg per dose, or such as about 200 mg to about 300 mg per dose. Other therapeutically effective amounts of the compounds of the present disclosure are about 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475 or about 500 mg per dose. Other therapeutically effective amounts of the disclosed compounds are about 100 mg per dose, or about 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450 or about 500 mg per dose. A single dose can be administered hourly, daily or weekly. For example, a single dose can be administered once every 1 hour, 2, 3, 4, 6, 8, 12, 16 hours or even every 24 hours. A single dose can also be administered once every 1 day, 2 days, 3 days, 4 days, 5 days, 6 days or every 7 days. A single dose can also be administered once every 1 week, 2 weeks, 3 weeks or even every 4 weeks. In certain embodiments, a single dose can be administered once a week. A single dose can also be administered once a month.

[0163] Other therapeutically effective amounts of the compounds of the present disclosure are about 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or about 100 mg per dose.

[0164] The frequency of administration of the compounds of the present disclosure will be determined according to the needs of each patient, for example, it can be once a day, or it can be twice a day or more. As long as it is required for the treatment of viral infection, it can be continuously administered. For example, the compounds of the present disclosure can be administered to people infected with viruses for a period of 20 to 180 days, or for example 20 to 90 days, or for example 30 to 60 days.

[0165] Administration can be intermittent, with the patient receiving a compound of the present disclosure every day for a few days or more, and then not receiving a compound of the present disclosure every day for a few days or more. For example, a patient can receive a compound of the present disclosure every other day or three times a week. For another example, a patient can receive a dose of a compound of the present disclosure every day for 1 to 14 days; followed by a period of 7 to 21 days during which the patient does not receive a dose of a compound of the present disclosure; followed by a subsequent period of time (e.g., 1 to 14 days) during which the patient again receives a dose of a compound of the present disclosure every day. Depending on the clinical needs of the patient's treatment, the cycle of alternating administration of the compound and subsequent non-administration of the compound can be repeated.

[0166] In certain embodiments, a pharmaceutical composition comprises a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, in combination with one or more (e.g., one, two, three, four, one or two, one to three, or one to four) additional therapeutic agents and a pharmaceutically acceptable excipient.

[0167] VI. Method or purpose

[0168] In some embodiments, the method or use of inhibiting LRRK2 in a cell in need thereof comprises administering to the cell an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the present disclosure. In some embodiments, the method or use of inhibiting LRRK2 in a cell comprises contacting the cell with an effective amount of a compound of Formula (I), (Ia), (Ib) or (Ic), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or a salt thereof.

[0169] In some embodiments, the present disclosure provides a method of inhibiting LRRK2 in a cell, the method comprising contacting the cell with an effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0170] Inhibition of LRRK2 enzyme activity can be measured by any assay method known in the art, such as the in vitro assay methods described in WO 2011 / 141756, WO 2012 / 028629, WO 2012 / 058193, WO2017 / 046675, WO 2018 / 163030, WO 2018 / 163066, WO 2021 / 080929 or US20210002260. Other exemplary in vitro assay methods can be found in the examples herein. In some embodiments, the in vitro assay includes an enzyme assay or a cell assay.

[0171] In certain embodiments, inhibition of LRRK2 enzymatic activity is determined in an in vivo model. Illustrative in vivo models of LRRK2-related diseases are described in Xiong, Y. et al., Adv Neurobiol. 2017; 14: 163-191.

[0172] In some embodiments, the method of inhibiting LRRK2 comprises administering an effective amount of a compound of the present disclosure, thereby reducing the activity of LRRK2 in the assay described herein compared to a control to which the compound of the present disclosure is not administered. In some embodiments, the activity of LRRK2 is reduced by about 5% to about 100%, for example, from about 10% to about 97%, from about 20% to about 95%, from about 20% to about 90%, from about 20% to about 80%, or from about 20% to about 70%. In certain embodiments, the activity of LRRK2 is reduced by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%.

[0173] In some embodiments, the method or use of inhibiting LRRK2 in a subject in need thereof comprises administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the present disclosure. In some embodiments, the method or use of inhibiting LRRK2 in a subject in need thereof comprises administering to the subject a therapeutically effective amount of a compound of Formula (I), (Ia), (Ib) or (Ic), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the compound or a salt thereof.

[0174] In some embodiments, the present disclosure provides a method or use for treating LRRK2-related diseases or conditions, such as nervous system diseases (e.g., Parkinson's disease) and certain immune system diseases (e.g., inflammatory bowel diseases, such as ulcerative colitis or Crohn's disease), comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0175] In some embodiments, the present disclosure provides a method of treating an LRRK2-related disease or condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof.

[0176] In some embodiments, the present disclosure provides a method or use for treating a LRRK2-related disease or condition, comprising administering a compound of the present disclosure or a pharmaceutically acceptable salt thereof. In some embodiments, LRRK2-related diseases or conditions include Parkinson's disease; brain injury; stroke; cerebrovascular disease (including cerebral arteriosclerosis, cerebral amyloid angiopathy, hereditary cerebral hemorrhage and cerebral hypoxia-ischemia); cognitive disorders (including amnesia, senile dementia, HIV-related dementia, Alzheimer's disease, Huntington's disease, Lewy body dementia, vascular dementia, drug-related dementia, tardive dyskinesia, myoclonus, dystonia, delirium, Pick's disease, Creutzfeldt-Jacob disease, AIDS, Tourette's syndrome, epilepsy, muscle spasms and diseases associated with muscle spasms or weakness (including tremors and mild cognitive impairment); intellectual disability (including spasticity, Down syndrome and fragile X syndrome). Xsyndrome); sleep disorders (including hypersomnia, circadian rhythm sleep disorders, insomnia, parasomnias, and sleep deprivation) and psychiatric disorders, such as anxiety disorders (including acute stress disorder, generalized anxiety disorder, social anxiety disorder, panic disorder, post-traumatic stress disorder, agoraphobia, and obsessive-compulsive disorder); factitious disorder (including acute hallucinatory mania); impulse control disorders (including compulsive gambling and intermittent explosive disorder); mood disorders (including bipolar I mood disorder, bipolar II mood disorder, mania, mixed affective states, major depressive disorder, chronic depression, seasonal depression, psychotic depression, premenstrual syndrome (PMS), premenstrual dysphoric disorder (PMS), and The following are some of the most common psychiatric disorders: psychotic disorders (including schizophrenia, schizoaffective disorder, schizoaffective disorder, and delusional disorder); substance dependence (including narcotic dependence, alcohol abuse, amphetamine dependence, cocaine addiction, nicotine dependence, and drug withdrawal syndrome); eating disorders (including anorexia, bulimia, binge eating disorder, overeating, obesity, compulsive eating disorder, and pagophagia); sexual dysfunction; urinary incontinence; neuronal damage (including eye damage, retinal disease or macular degeneration, tinnitus, hearing impairment and loss, and cerebral edema) and pediatric psychiatric disorders (including attention deficit disorder, attention-deficit / hyperactivity disorder, conduct disorder, and autism).

[0177] In certain embodiments, the LRRK2-related disease or condition is Parkinson's disease, Lewy body dementia, frontotemporal dementia, corticobasal dementia, progressive supranuclear palsy, Alzheimer's disease, tauopathy, or alpha-synucleinopathy. In some embodiments, the LRRK2-related disease or condition is Parkinson's disease. In some embodiments, the LRRK2-related disease or condition is frontotemporal dementia. In certain embodiments, the LRRK2-related disease or condition is corticobasal dementia. In some embodiments, the LRRK2-related disease or condition is progressive supranuclear palsy. In some embodiments, the LRRK2-related disease or condition is Alzheimer's disease. In some embodiments, the LRRK2-related disease or condition is tauopathy. In certain embodiments, the LRRK2-related disease or condition is alpha-synucleinopathy.

[0178] In certain embodiments, the LRRK2-related disease or condition is inflammatory bowel disease. In some embodiments, the inflammatory bowel disease is ulcerative colitis or Crohn's disease. In some embodiments, the inflammatory bowel disease is ulcerative colitis. In certain embodiments, the inflammatory bowel disease is Crohn's disease.

[0179] In certain cell types of patients with Parkinson's disease, increases in LRRK2 levels and / or activity are associated with abnormalities in autophagy levels. For example, LRRK2 G2019S and LRRK2 R1441C mutations are associated with increased kinase activity and reduced autophagic flux due to blocked autophagosome clearance. See Madureira, M. et al., Frontiers in Neuroscience 2020, 14, Chapter 498, pp. 1-19. Inhibition of LRRK2 G2019S kinase activity in cell models can enhance the formation of autolysosomes. See, Obergasteiger et al., Cell Death Discovery 2020, 6(45), pp. 1-13.

[0180] Many diseases are associated with abnormal levels of autophagy, especially decreased levels of autophagy compared to healthy people. See, Ichimiya et al., Intl. J. Mol. Sci. 2020, 21, 8974, pp. 1-21. By administering the compounds of the present disclosure or pharmaceutically acceptable salts thereof, any autophagy-related disease or condition may benefit from the inhibition of LRRK2.

[0181] Thus, in some embodiments, the LRRK2-related disease or condition is an autophagy-related disease or condition. In some embodiments, the autophagy-related disease or condition involves a decrease in the level of one or more of mitochondrial autophagy, homophagy, ER autophagy, lysosomal autophagy, nuclear autophagy, peroxisomal autophagy, lipophagy, xenophagy, aggresome autophagy, ribosomal autophagy, NPC autophagy, and RN / RN-autophagy compared to the level in a control subject. In some embodiments, the disease or condition associated with autophagy is a liver disease (e.g., non-alcoholic fatty liver disease (NAFLD), alpha1-antitrypsin deficiency (alpha1-antitrypsin deficiency; AATD) or hereditary hypofibrinogenemia with hepatic storage (hereditary hypofibrinogenemia with hepatic storage; HHHS)), a kidney disease (e.g., type 1 diabetes, type 2 diabetes, acute kidney injury, and chronic kidney disease caused by diabetes, hypertension or chronic nephritis), a heart disease (e.g., heart failure), an inflammatory bowel disease (e.g., Crohn's disease), or a neurodegenerative disease (e.g., Parkinson's disease). In certain embodiments, the autophagy-related disease or condition is alpha-1-antitrypsin deficiency (AATD).

[0182] In some embodiments, the use of the invention for the preparation of a medicament for treating a LRRK2-related disease or condition comprises a compound or pharmaceutical composition described herein.

[0183] In some embodiments, a compound or composition of the invention for treating a LRRK2-related disease or condition comprises a compound or pharmaceutical composition described herein.

[0184] In some embodiments, the kit is suitable for carrying out the above methods or uses. In some embodiments, the kit of the present invention comprises one or more compounds of the present invention. In some embodiments, the kit comprises a first dosage form comprising one or more compounds of the present invention and a container for holding the dosage, the amount of which is sufficient to implement the methods or uses of the present invention.

[0185] VII. Examples

[0186] Many general references provide commonly known chemical synthesis schemes and conditions useful for synthesizing the disclosed compounds (eg, see Smith, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th ed., Wiley-Interscience, 2013).

[0187] The compounds described herein can be purified by any method known in the art, including chromatography, such as high performance liquid chromatography (HPLC), preparative thin layer chromatography, flash column chromatography, and ion exchange chromatography. Any suitable stationary phase can be used, including normal and reverse phase and ionic resins. For example, the disclosed compounds can be purified by silica gel chromatography. For example, see "Introduction to Modern Liquid Chromatography", 2nd Edition, LRSnyder and JJKirkland, John Wiley and Sons, 1979; and Thin Layer Chromatography, E.Stahl (ed.), Springer-Verlag, New York, 1969.

[0188] The compounds were characterized using standard instrumental methods. Compounds were identified by H NMR spectroscopy ( 1 H-NMR) and mass spectrometry (MS). 1 The measurement frequency of H-NMR is 400 MHz, unless otherwise specified. In some cases, depending on the compound and the measurement conditions, exchangeable hydrogen cannot be clearly observed. Br. or broad used in this article refers to a broad signal. Unless otherwise specified, HPLC preparative chromatography uses a commercially available ODS column, water / methanol (containing formic acid) as the eluent, and is performed in gradient mode.

[0189] Certain abbreviations and acronyms are used in describing the experimental details. Although most of them are understood by those skilled in the art, many of the abbreviations and acronyms are listed in the following table.

[0190] Table 2. List of abbreviations and acronyms

[0191]

[0192]

[0193] The examples provided herein describe the synthesis of the compounds disclosed herein and the intermediates used to prepare these compounds. It should be understood that the various steps described herein can be combined. It should also be understood that individual batches of compounds can also be combined and then proceed to the next synthesis step.

[0194] In the following description of the embodiments, specific embodiments will be introduced. The description of these embodiments is detailed enough to enable those skilled in the art to implement certain embodiments of the present disclosure. Without departing from the scope of the present disclosure, other embodiments may also be used, and logical and other changes may be made. Therefore, the following description is not intended to limit the scope of the present disclosure.

[0195] Representative syntheses of compounds of the present disclosure are described in the Schemes below and in the specific Examples that follow.

[0196] Intermediate 1

[0197]

[0198] (R)-3-((tert-butyldimethylsilyl)oxy)butyric acid methyl ester: TBSCl (41.5 mL, 339 mmol, 2.0 eq) and imidazole (46 g, 677 mmol, 4.0 eq) were added to a solution of (R)-3-hydroxybutyric acid methyl ester (19.4 mL, 169 mmol, 1.0 eq) in DCM (400 mL, 0.4 M) at 0°C. The mixture was stirred at 20°C under nitrogen atmosphere for 12 hours. The mixture was diluted with water (100 mL), extracted with DCM (3 x 100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (petroleum ether: ethyl acetate = 1 / 0 to 9 / 1) to give the title compound as a colorless oil. 1 H NMR (400MHz, CDCl3) δ4.36-4.23(m,1H),3.67(s,3H),2.53-2.34(m,2H),1.20(d,J=6.0Hz,3H),0.87(s,9H),0.06(d,J=9.2Hz,6H).

[0199] (R)-1-(2-((tert-butyldimethylsilyl)oxy)propyl)cyclopropan-1-ol: To a solution of (R)-3-((tert-butyldimethylsilyl)oxy)butanoic acid methyl ester (39 g, 151 mmol, 1.0 eq) in THF (350 mL, 0.4 M) at 0°C was added tetraisopropoxytitanium (44 mL, 151 mmol, 1.0 eq) and EtMgBr (3M in THF, 151 mL, 453 mmol, 3.0 eq). The mixture was stirred at 20°C for 2 hours. The mixture was quenched at 0°C with saturated NH4Cl (200 mL) and 20% citric acid (200 mL). MTBE (200 mL) was added. The mixture was stirred at 0°C for 20 minutes and filtered. The filtrate was extracted with MTBE (3 x 200 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to afford the title compound as a brown oil, which was used in the next step without further purification. 1 H NMR (400MHz, CDCl3) δ4.31-4.18(m,1H),1.85-1.76(m,1H),1.65-1.58(m,1H),1.26(d,J=6.4Hz,3H),0.93-0.8 9(m,9H),0.82-0.76(m,1H),0.72-0.65(m,1H),0.49-0.43(m,1H),0.40-0.34(m,1H),0.13(s,3H),0.12(s,3H).

[0200] (R)-5-((tert-Butyldimethylsilyl)oxy)hex-1-en-3-one: To a solution of (R)-1-(2-((tert-Butyldimethylsilyl)oxy)propyl)cyclopropan-1-ol (35 g, 152 mmol, 1.0 eq) in DCM (400 mL, 0.4 M) was added NBS (27 g, 152 mmol, 1.0 eq). The mixture was stirred at 0°C for 1 hour. TEA (42.3 mL, 304 mmol, 2.0 eq) was added and the mixture was stirred at 0°C for 2 hours. The mixture was diluted with 20% citric acid (200 mL) and the layers were separated. The organic layer was washed with saturated NaHCO3 (3 x 200 mL), dried over Na2SO4 and filtered, and hydroquinone (200 mg) was added to the organic layer. The organic layer was concentrated under reduced pressure. The residue was filtered through a silica gel pad (300 g) and the filter cake was washed with DCM (800 mL). The filtrate and the washings were combined and concentrated under atmospheric pressure to give the title compound as a brown oil, which was used in the next step without further purification. 1H NMR (400MHz, CDCl3) δ6.46-6.30(m,1H),6.29-6.17(m,1H),5.96-5.79(m,1H),4.39-4.28(m,1H),2.85(dd,J=7. 2,14.8Hz,1H),2.54(dd,J=5.2,14.8Hz,1H),1.19(d,J=6.0Hz,3H),0.86-0.82(m,9H),0.05(m,3H),0.01(s,3H).

[0201] (R)-2-Methyl-2,3-dihydro-4H-pyran-4-one: A mixture of (R)-5-((tert-butyldimethylsilyl)oxy)hex-1-en-3-one (10 g, 39 mmol, 1.0 eq), PdCl2(MeCN)2 (102 mg, 394 μmol, 0.01 eq), benzoquinone (8.9 mL, 39 mmol, 1.0 eq) and H2O (3.4 mL, 189.14 mmol, 4.8 eq) in acetone (50 mL, 0.8 M) was stirred at 40 °C under nitrogen atmosphere for 6 hours. The solvent was removed under reduced pressure and the residue was dissolved in DCM (30 mL). After filtering the solution, the filtrate was concentrated under reduced pressure to obtain a residue. The residue was distilled under vacuum (50 °C, oil pump, 10 mmHg) to obtain the title compound as a colorless oil. 1 H NMR (400MHz, CDCl3) δ7.33(d,J=6.0Hz,1H),5.39(dd,J=1.2,6.0Hz,1H),4.61-4.48(m,1H),2.62-2.34(m,2H),1.45(d,J=6.4Hz,3H).[α] D 25 = +167.009 (c = 0.109, CHCl3).

[0202] (R)-2-Methyltetrahydro-4H-pyran-4-one: A mixture of (R)-2-methyl-2,3-dihydro-4H-pyran-4-one (1.0 g, 8.03 mmol, 1.0 eq) and Pd / C (500 mg, 10 wt%) in THF (20 mL, 0.4 M) was stirred at 20°C under H2 (15 psi) for 1 hour. The mixture was filtered and concentrated under reduced pressure to give the title compound as a colorless oil, which was used in the next step without further purification. 1 H NMR (400MHz, CDCl3) δ4.31-4.23(m,1H),3.79-3.63(m,2H),2.64-2.51(m,1H),2.44-2.21(m,3H),1.32(d,J=6.4Hz,3H).

[0203] Intermediate 1: A mixture of (R)-2-methyltetrahydro-4H-pyran-4-one (200 mg, 1.75 mmol, 1.0 eq) and (2,4-dimethoxyphenyl)methanamine (396 μL, 2.63 mmol, 1.5 eq) in MeOH (5 mL, 0.4 M) was stirred at 20 °C for 1 hour. The mixture was cooled to -78 °C and LiBH4 (38.2 mg, 1.75 mmol, 1.0 eq) was added. The mixture was stirred at 20 °C for 12 hours. The mixture was then quenched with saturated Na2CO3 (20 mL) and diluted with water (20 mL). The mixture was extracted with DCM (3 x 20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude material obtained was purified by flash silica gel chromatography (DCM / MeOH=1 / 0 to 9 / 1) to give the title compound as a brown oil. LCMS [M+H] + =266.2. 1 H NMR (400MHz, CDCl3) δ7.12 (d, J = 8.0Hz, 1H), 6.51-6.34 (m, 2H), 4.06-3.92 (m, 1H), 3.80 (s, 3H), 3.79 (s, 3H), 3.75 (s, 2H), 3.44-3 .34(m,2H),2.73-2.61(m,1H),1.92-1.85(m,1H),1.84-1.77(m,1H),1.46-1.30(m,1H),1.18(d,J=6.0Hz,3H),1.15-1.03(m,1H).

[0204] Intermediate 2

[0205]

[0206] 4-Hydroxy-3-nitroquinoline-6-carbonitrile: DMF (60 mL, 0.3 M) was added to a flask containing 6-bromo-3-nitroquinolin-4-ol (5 g, 18.6 mmol, 1 eq), tetrapotassium hexacyanoferrate (II) trihydrate (4.71 g, 11.15 mmol, 0.6 eq), dppf (2.06 g, 3.72 mmol, 0.2 eq), Pd(OAc)2 (417.22 mg, 1.86 mmol, 0.1 eq) and K2CO3 (3.08 g, 22.3 mmol, 1.2 eq) under nitrogen. The mixture was stirred at 130 °C for 12 h, then the reaction mixture was cooled to room temperature and filtered through a pad of celite. The filter cake was slowly washed with DMF (50 mL) and MTBE (600 mL) while stirring the filtrate. A dark solid precipitate formed during stirring. The resulting mixture was stirred at 20°C for 15 minutes and then filtered. The second filtrate was concentrated in vacuo to a volume of approximately 10 mL, which was diluted with MTBE (30 mL). The resulting dark precipitate was collected by filtration and triturated with ethyl acetate (30 mL) to afford the title compound as a dark green solid, which was used directly in the next step without further purification. LCMS [M+H] + =215.9.

[0207] Intermediate 2: POCl3 (2.18 mL, 23.4 mmol, 2.8 eq) was added to a DMF (60 mL, 0.1 M) solution of 4-hydroxy-3-nitroquinoline-6-carbonitrile (1.8 g, 8.37 mmol, 1.0 eq) under nitrogen atmosphere. Stirred at 20 ° C for 12 h, the mixture was quenched by adding water (30 mL), and the solution was extracted with EtOAc (3 x 30 mL). The organic phases were combined and dried over Na2SO4, and the filtered filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether / EtOAc=1 / 0 to 85 / 15) to give the title compound as a white solid. 1 H NMR (400MHz, CDCl3) δ9.39 (s, 1H), 8.84 (d, J = 1.2Hz, 1H), 8.36 (d, J = 8.8Hz, 1H), 8.10 (dd, J = 1.6, 8.4Hz, 1H).

[0208] Intermediate 3

[0209]

[0210] 4-((3,4-Dimethylbenzyl)((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)-3-nitroquinoline-6-carbonitrile: DIPEA (64.3 μL, 369 μmol, 1.15 eq) was added to a solution of 4-chloro-3-nitroquinoline-6-carbonitrile (79.0 mg, 321 μmol, 1.0 eq) and (2R,4R)-N-(3,4-dimethylbenzyl)-2-methyltetrahydro-2H-pyran-4-amine (94.7 mg, 321 μmol, 1.0 eq) in MeCN (5 mL, 0.06 M) under nitrogen atmosphere. The mixture was stirred at 20 °C for 2 h. After removing the solvent in vacuo, the residue was dissolved with EtOAc (30 mL). The solution was washed with brine (3 x 10 mL), dried over Na2SO4, filtered and the filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether / EtOAc = 1 / 0 to 4 / 1) to give the title compound as an orange oil. LCMS [M+H] + =463.2. 1 H NMR (400MHz, DMSO-d6) δ9.19(s,1H),8.55(s,1H),8.16(d,J=0.8Hz,2H),6.88(d,J=8.0Hz,1H),6.31-6.24(m,2H),4.39-4.27(m,2H),3.92-3 .77(m,2H),3.63(s,3H),3.42(s,3H),3.41-3.35(m,2H),1.97-1.85(m ,2H),1.83-1.71(m,1H),1.48(q,J=11.6Hz,1H),1.09(d,J=6.2Hz,3H).

[0211] 4-(((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)-3-nitroquinoline-6-carbonitrile: TFA (43.2 μL, 583 μmol, 3.0 eq) was added to a solution of 4-((3,4-dimethylbenzyl)((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)-3-nitroquinoline-6-carbonitrile (94.7 mg, 195 μmol, 1.0 eq) in DCM (2 mL, 0.1 M) under nitrogen atmosphere. The mixture was stirred at 20 °C for 2 h. The mixture was concentrated to a volume of 5 mL and washed with saturated aqueous sodium bicarbonate solution (20 mL). The aqueous layer was extracted with DCM (3 x 20 mL), and the combined organic layers were dried over Na2SO4 and filtered and concentrated in vacuo to give the title compound as a yellow solid which was used in the next step without further purification. LCMS [M+H] + =313.2. 1H NMR (400MHz, DMSO-d6) δ9.08(s,2H),8.29(d,J=8.8Hz,1H),8.15(d,J=8.8Hz,1H),8.01(d,J=8.8Hz,1H),3.91(dd,J=4.0,11.2 Hz,1H),3.80-3.75(m,1H),3.44-3.37(m,2H),2.04-1.97(m,2H),1.95-1.88(m,1H),1.73-1.64(m,1H),1.12(d,J=6.0Hz,3H).

[0212] Intermediate 3: Water (0.5 mL) and EtOH (2 mL) were added to a mixture of 4-(((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)-3-nitroquinoline-6-carbonitrile (70 mg, 224 μmol, 1.0 eq), NH4Cl (120 mg, 2.24 mmol, 10 eq) and Fe (125 mg, 2.24 mmol, 10 eq). The reaction mixture was heated to 80 ° C for 1 hour. The mixture was then diluted with ethanol (20 mL) and filtered. The filtrate was concentrated in vacuo and the solid formed was partitioned between saturated aqueous sodium bicarbonate solution (20 mL) and DCM (30 mL). The organic layer was dried over Na2SO4 and filtered, and concentrated under reduced pressure to give the title compound as a brown solid, which was used directly in the next step without further purification. LCMS [M+H] + =283.2. 1 H NMR (400MHz, MeOD-d4) δ8.55(d,J=1.6Hz,1H),8.51(s,1H),7.90(d,J=8.4Hz,1H),7.60(dd,J=1.6,8.8Hz,1H),3.99-3.94(m ,1H),3.65-3.58(m,1H),3.48-3.40(m,2H),1.93-1.79(m,2H),1.67-1.58(m,1H),1.30-1.28(m,1H),1.17(d,J=6.4Hz,3H). Intermediate 4

[0213]

[0214] Intermediate 4 was prepared by the same method as Intermediate 3 using 4,6-dichloro-3-nitroquinoline as a starting material.

[0215] 6-Chloro-N-(3,4-dimethylbenzyl)-N-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-3-nitroquinolin-4-amine: LCMS [M+H] + =472.2.1 H NMR (400MHz, DMSO-d6) δ9.06 (s, 1H), 8.15 (d, J = 2.4Hz, 1H), 8.03 (d, J = 8.8Hz, 1H ),7.88(dd,J=2.4,8.9Hz,1H),6.82(d,J=8.4Hz,1H),6.30-6.23(m,2H),4.27(br s,2H),3.93-3.83(m,1H),3.80-3.70(m,1H),3.62(s,3H),3.46(s,3H),3.41-3.34(m,2H),1.94( d,J=13.6Hz,1H),1.89-1.81(m,1H),1.79-1.64(m,1H),1.50-1.38(m,1H),1.08(d,J=6.0Hz,3H).

[0216] 6-Chloro-N-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-3-nitroquinolin-4-amine: LCMS [M+H] + =322.1. 1 H NMR (400MHz, CDCl3) δ9.38(s,1H),9.13(d,J=8.4Hz,1H),8.14(d,J=2.4Hz,1H),7.99(d,J=8.8Hz,1H),7.75(dd,J=2.4,8.8Hz,1H),4. 38-4.22(m,1H),4.17-4.09(m,1H),3.63-3.51(m,2H),2.25-2.11(m,2H),1.86-1.73(m,1H),1.55-1.45(m,1H),1.29(d,J=6.4Hz,3H).

[0217] Intermediate 4: LCMS [M+H] + =292.1. 1 H NMR (400MHz, CDCl3) δ8.48(s,1H),7.90(d,J=8.8Hz,1H),7.74(d,J=2.0Hz,1H),7.40(dd,J=2.0,8.8Hz,1H),4.02(dd,J=3.6,11.6Hz,1H),3.89(br s,2H),3.55-3.35(m,4H),1.97-1.78(m,2H),1.62-1.50(m,1H),1.21(d,J=6.0Hz,3H).

[0218] Intermediate 5

[0219]

[0220] 2-Chloro-N-(6-cyano-4-(((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)quinolin-3-yl)acetamide: NEt3 (739 μL, 5.31 mmol, 3.0 eq) was added to a solution of compound intermediate 3 (500 mg, 1.77 mmol, 1.0 eq) in DCM (10 mL). Chloroacetyl chloride (141 μL, 1.77 mmol, 1.0 eq) in DCM (5 mL) was added dropwise to the reaction mixture at -10°C. The resulting mixture was stirred at -10°C under nitrogen atmosphere for one hour. The mixture was slowly warmed to 10°C and stirred for another hour under nitrogen atmosphere. The reaction mixture was poured into water (20 mL) and then extracted with DCM (3 x 20 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4 and filtered. The filtrate was concentrated in vacuo. The crude material was purified by silica gel chromatography (90% EtOAc / petroleum ether) to afford the title compound as a yellow solid. LCMS [M+H] + =359.1. 1 H NMR (400MHz, DMSO-d6) δ10.06(s,1H),9.03(s,1H),8.34(s,1H),7.94-7.91(m,2H),6.50(d,J=8.8Hz,1H),4.35(s,2H),4.12-4.04(m,1H) ,3.92-3.85(m,1H),3.42-3.39(m,2H),1.90-1.85(m,1H),1.82-1.76(m,1H),1.61-1.51(m,1H),1.20-1.14(m,1H),1.10(d,J=6.4Hz,3H).

[0221] Intermediate 5: AcOH (1.0 mL, 17.5 mmol, 20 eq) was added to a solution of 2-chloro-N-(6-cyano-4-(((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)quinolin-3-yl)acetamide (337 mg, 845 μmol, 1.0 eq) in dioxane (6 mL, 0.1 M) under nitrogen atmosphere. The mixture was stirred at 100 °C for 12 h. The reaction was poured into water (20 mL) and then extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and filtered. The filtrate was concentrated in vacuo and the resulting crude material was chromatographed on silica gel (100% EtOAc) to give Intermediate 5 as a yellow solid. LCMS [M+H] + =341.1. 1H NMR (400MHz, DMSO-d6) δ9.39(s,1H),9.01(br s,1H),8.35(d,J=8.4Hz,1H),8.09(dd,J=1.6,8.4Hz,1H),5.33(br s,2H),4.30-4.13(m,1H),3.89-3.66(m,2H),3.40-3.37(m,1H),2.55-2.51(m,1H),2.29-2.01(m,3H),1.26(d,J=6.4Hz,3H).

[0222] Intermediate 6

[0223]

[0224] Ethyl 5-(2-hydroxyethyl)isoxazole-3-carboxylate: To a solution of homopropargyl alcohol (5.40 mL, 71.34 mmol, 1 eq.) and ethyl (chlorohydroxyimino)acetate (32.43 g, 214 mmol, 3 eq.) in EtOAc (500 mL, 0.1 M) was added NaHCO (17.98 g, 214 mmol, 3 eq.). The reaction mixture was stirred at 100 °C for 2 hours. The mixture was filtered and concentrated in vacuo. The resulting residue was purified via silica gel chromatography (15% EtOAc in petroleum ether) to give the title compound as a colorless oil. 1 H NMR (400MHz, CDCl3) δppm6.52 (s, 1H), 4.39 (q, J = 7.2Hz, 2H), 3.94 (t, J = 6.4Hz, 2H), 3.04 (t, J = 6.4Hz, 2H), 2.64 (br s, 1H), 1.37 (t, J = 7.2Hz, 3H).

[0225] Ethyl 5-(2-((methylsulfonyl)oxy)ethyl)isoxazole-3-carboxylate: A mixture of ethyl 5-(2-hydroxyethyl)isoxazole-3-carboxylate (4 g, 20.5 mmol, 1 eq.) and MsCl (2.05 mL, 26.5 mmol, 1.29 eq.) in DCM (80 mL, 0.25 M) was cooled to 0 ° C. Et3N (8.57 mL, 61.5 mmol, 3 eq.) was added dropwise, and the mixture was warmed to room temperature for 2.5 hours. The reaction mixture was then slowly poured into ice water (100 mL), and the mixture was extracted with DCM (2×100 mL). The combined organic layers were washed with brine (2×100 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as a yellow oil, which was used directly in the next step without further purification. LCMS[M+H] + =264.0. 1H NMR (400MHz, CDCl3) δppm 6.61-6.57(m,1H),4.56-4.49(m,2H),4.47-4.38(m,2H),3.33-3.25(m,2H),3.02-2.98(m,3H),1.44-1.38(m,3H).

[0226] Ethyl 5-(2-(Methylthio)ethyl)isoxazole-3-carboxylate: A mixture (60 mL) of sodium methanethiolate (2.64 mL, 41.5 mmol, 2 eq.) in DMF was sealed in a round-bottom flask with a rubber septum and placed under a N2 atmosphere. A solution of ethyl 5-(2-((methylsulfonyl)oxy)ethyl)isoxazole-3-carboxylate (5.46 g, 20.7 mmol, 1 eq.) in DMF (100 mL) was added to the reaction mixture, and the mixture was stirred at 80 °C for 6 hours. The reaction mixture was poured into water (200 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (3×200 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as a yellow oil, which was used directly in the next step without further purification. LCMS[M+H] + =216.1. 1 H NMR (400MHz, CDCl3) δppm6.50 (s, 1H), 4.42 (q, J = 7.2Hz, 2H), 3.10 (t, J = 7.2Hz, 2H), 2.83 (t, J = 7.2Hz, 2H), 2.12 (s, 3H), 1.40 (t, J = 7.2Hz, 3H).

[0227] Ethyl 5-(2-(methylsulfonyl)ethyl)isoxazole-3-carboxylate: To a solution of ethyl 5-(2-(methylthio)ethyl)isoxazole-3-carboxylate (1.6 g, 6.47 mmol, 1 eq.) in DCE (50 mL, 0.1 M) was added m-CPBA (4.18 g, 19.4 mmol, 3 eq.) in portions at 0 °C. The reaction mixture was stirred at 85 °C for 3 hours. The reaction mixture was poured into a 1:1 mixture of saturated NaHCO3 (100 mL) and saturated Na2SO3 (100 mL). The solution was extracted with DCM (3×100 mL), and the combined organic layers were washed with brine (3×200 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as a yellow solid, which was used in the next step without further purification. LCMS [M+H] + =248.0. 1H NMR (400MHz, CDCl3) δppm 6.59 (s, 1H), 4.44 (q, J = 7.2Hz, 2H), 3.45-3.40 (m, 4H), 2.95 (s, 3H), 1.41 (t, J = 7.2Hz, 3H).

[0228] (5-(2-(methylsulfonyl)ethyl)isoxazol-3-yl)methanol: To a solution of ethyl 5-(2-(methylsulfonyl)ethyl)isoxazole-3-carboxylate (900 mg, 3.64 mmol, 1 eq) in MeOH (10 mL, 0.4 M) was slowly added NaBH4 (1.38 g, 36.4 mmol, 10 eq) at 0 °C. Under N2 atmosphere, the reaction mixture was stirred at 20 °C for 12 hours. The mixture was quenched with H2O (5 mL) and saturated NH4Cl (5 mL), and the reaction mixture was concentrated in vacuo. The resulting crude material was purified via silica gel chromatography (3% MeOH in DCM) to give the title compound as a white solid. 1 H NMR (400MHz, DMSO-d6) δppm6.37(s,1H),5.47(t,J=6.0Hz,1H),4.46(d,J=6.0Hz,2H),3.57-3.50(m,2H),3.23-3.16(m,2H),3.03(s,3H).

[0229] Methyl (5-(2-(methylsulfonyl)ethyl)isoxazol-3-yl)methanesulfonate: To a solution of (5-(2-(methylsulfonyl)ethyl)isoxazol-3-yl)methanol (930 mg, 4.08 mmol, 1 eq.) in DCM (20 mL, 0.2 M) was slowly added TEA (1.70 mL, 12.2 mmol, 3 eq.) and MsCl (574 μL, 7.42 mmol, 1.82 eq.) at 0 °C. Under N2 atmosphere, the reaction mixture was stirred at 20 °C for 1 hour. The reaction mixture was slowly poured into ice water (50 mL) and extracted with DCM (2×50 mL). The combined organic layers were washed with brine (2×50 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as a white solid, which was used directly in the next step without further purification. LCMS [M+H] + =284.0. 1 H NMR (400MHz, DMSO-d6) δppm 6.57(s,1H),5.33(s,2H),3.59-3.54(m,2H),3.28(s,3H),3.28-3.23(m,2H),3.03(s,3H).

[0230] 2-(5-(2-(methylsulfonyl)ethyl)isoxazol-3-yl)acetonitrile: To a solution of methyl (5-(2-(methylsulfonyl)ethyl)isoxazol-3-yl)methanesulfonate (1.07 g, 3.78 mmol, 1 eq) in DMF (15 mL, 0.25 M) at 0 °C was added NaCN (277 mg, 5.65 mmol, 1.5 eq). The reaction mixture was stirred at 20 °C for 2 hours under N2 atmosphere. The reaction mixture was slowly poured into ice water (50 mL), and the mixture was extracted with DCM (2×50 mL). The combined organic layers were washed with brine (2×50 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified via silica gel chromatography (5% MeOH in DCM) to give the title compound as a yellow oil. Note: 2M NaOH aqueous solution was added to the separated aqueous phase until pH = 14, and the mixture was quenched with saturated NaClO aqueous solution (100 mL). LCMS [M+H] + =215.0. 1 H NMR (400MHz, DMSO-d6) δppm 6.48 (s, 1H), 4.19 (s, 2H), 3.58-3.52 (m, 2H), 3.27-3.21 (m, 2H), 3.03 (s, 3H).

[0231] Intermediate 6: To a solution of 2-(5-(2-(methylsulfonyl)ethyl)isoxazol-3-yl)acetonitrile (150 mg, 595 μmol, 1 eq) in H2O (0.5 mL) was added concentrated HCl solution (2 mL, 20.1 mmol, 33 eq). The reaction mixture was stirred at 90 °C under N2 atmosphere for 7 hours. The reaction mixture was directly lyophilized to give Example 6 as a white solid. The crude product was used directly in the next step without further purification. 1 H NMR (400MHz, DMSO-d6) δppm12.67(s,1H),6.38(s,1H),3.66(s,2H),3.53(t,J=6.4Hz,2H),3.20(t,J=6.4Hz,2H),3.02(s,3H).

[0232] Intermediate 7

[0233]

[0234] Methyl 2-(5-(methylsulfonyl)pyridin-2-yl)acetate: To a solution of methyl 2-(5-bromopyridin-2-yl)acetate (500 mg, 2.17 mmol, 1 eq.) in DMSO (4 mL, 0.5 M) was added L-proline (300.26 mg, 2.61 mmol, 1.2 eq.), Cs2CO3 (495.69 mg, 1.52 mmol, 0.7 eq.), CuI (290 mg, 1.52 mmol, 0.7 eq.) and sodium methanesulfinate (333 mg, 3.26 mmol, 1.5 eq.). The mixture was bubbled with N2 and stirred at 100 °C under microwave (2 bar) for 6 h. The mixture was quenched with water (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (2×50 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified via silica gel chromatography (0-100% EtOAc in petroleum ether) to give the title compound as a yellow solid. LCMS [M+H] + =230.0. 1 H NMR (400MHz, CDCl3) δppm: 9.09 (s, 1H), 8.21 (dd, J = 2.0, 8.0Hz, 1H), 7.54 (d, J = 8.0Hz, 1H), 3.99 (s, 2H), 3.76 (s, 3H), 3.12 (s, 3H).

[0235] Intermediate 7: To a solution of methyl 2-(5-(methylsulfonyl)pyridin-2-yl)acetate (180 mg, 785 μmol, 1 eq.) in THF (3 mL) and H2O (3 mL) was added LiOH HO (65.90 mg, 1.57 mmol, 2 eq.) at 0°C. The mixture was then stirred at 20°C for 2 hours. The reaction mixture was slowly poured into water (5 mL) and extracted with EtOAc (1×5 mL). The aqueous phase was purified directly via reverse phase chromatography (0% MeCN in water) to give Intermediate 7 as a white solid. 1 H NMR (400MHz, DMSO-d6) δppm 8.88-8.82(m,1H),8.16-8.09(m,1H),7.59-7.51(m,1H),3.55-3.49(m,2H),3.27(s,3H).

[0236] Intermediate 8

[0237]

[0238] (Z)-tert-Butyl 3-amino-3-(hydroxyimino)propanoate: Water (6 mL) was added dropwise to a vigorously stirred mixture of tert-butyl cyanoacetate (1.0 mL, 7.08 mmol, 1 eq.), NH2OH·HCl (0.5 mL, 10.6 mmol, 1.5 eq.) and Na2CO3 (578 mg, 5.45 mmol, 0.77 eq.) in EtOH (10 mL) at 25 °C. The resulting solution was stirred at room temperature for 10 hours. The mixture was concentrated in vacuo, diluted with water (10 mL), and extracted with EtOAc (2×10 mL). The combined organic layers were washed with brine (2×10 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as an off-white solid, which was used in the next step without further purification. LCMS [M+H] + =175.1. 1 HNMR(400MHz,DMSO-d6)δppm:8.98(s,1H),5.41(s,2H),2.90(s,2H),1.40(s,9H).

[0239] (Z)-tert-Butyl 3-amino-3-(((1-(methylsulfonyl)cyclopropane-1-carbonyl)oxy)imino)propanoate: To a solution of 1-(methylsulfonyl)cyclopropanecarboxylic acid (500 mg, 3.05 mmol, 1 eq) and (Z)-tert-butyl 3-amino-3-(hydroxyimino)propanoate (589 mg, 3.05 mmol, 1 eq) in DCM (15 mL, 0.2 M) at 25 °C was added PyBOP (1.58 g, 3.05 mmol, 1 eq) and DIPEA (2.12 mL, 12.2 mmol, 4 eq). The mixture was then stirred at 25 °C for 12 h. The mixture was quenched with water (40 mL) and extracted with DCM (2×40 mL). The combined organic layers were washed with brine (2×40 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified via silica gel chromatography (10-50% EtOAc in petroleum ether) to give the title compound as a brown oil. LCMS [M+H] + =321.0. 1 H NMR (400MHz, DMSO-d6) δppm: 6.91-6.17(m,2H),3.25(s,3H),3.08(s,2H),1.80-1.73(m,2H),1.66-1.60(m,2H),1.42(s,9H).

[0240] Tert-butyl 2-(5-(1-(methylsulfonyl)cyclopropyl)-1,2,4-oxadiazol-3-yl)acetate: A solution of tert-butyl (Z)-3-amino-3-(((1-(methylsulfonyl)cyclopropane-1-carbonyl)oxy)imino)propanoate (650 mg, 1.83 mmol, 1 eq) in pyridine (7 mL, 87 mmol, 48 eq) was stirred at 90 °C under N2 atmosphere for 12 h. The mixture was concentrated in vacuo and the resulting crude material was purified by preparative HPLC (NH4HCO3 conditions) to give the title compound as a brown oil. LCMS [M-56+H] + =247.0. 1 H NMR (400MHz, CDCl3) δppm: 3.76 (s, 2H), 3.32 (s, 3H), 2.11-2.05 (m, 2H), 1.86-1.81 (m, 2H), 1.48 (s, 9H).

[0241] Intermediate 8: To a solution of tert-butyl 2-(5-(1-(methylsulfonyl)cyclopropyl)-1,2,4-oxadiazol-3-yl)acetate (90 mg, 267 μmol, 1 eq) in DCM (2 mL) was added TFA (2 mL, 27 mmol, 100 eq). The mixture was stirred at 25 °C for 6 hours. The solution was concentrated in vacuo to give Intermediate 8 as a brown oil, which was used directly in the next step without further purification. 1 H NMR (400MHz, CDCl3) δppm: 3.92 (s, 2H), 3.32 (s, 3H), 2.13-2.05 (m, 2H), 1.89-1.82 (m, 2H).

[0242] Intermediate 9

[0243]

[0244] 3-Azido-1-(methylsulfonyl)azetidine: To a 50 mL round-bottom flask containing DMF (0.76 mL) and aqueous KHCO3 (3 M, 887 μL, 4 eq) was added 1-methylsulfonylazetidine-3-amine (100 mg, 665 μmol, 1 eq) and fluorosulfonyl azide (0.26 M in MTBE, 2.56 mL, 1 eq). The reaction mixture was stirred at room temperature for 12 h. The reaction mixture was then diluted with EtOAc (20 mL) and washed with brine (3×10 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to obtain the title compound, which was used in the next step without further purification. 1H NMR (400MHz, DMSO-d6) δppm: 4.48 (tt, J=5.2, 7.2Hz, 1H), 4.17-4.08 (m, 2H), 3.79 (dd, J=5.2, 9.6Hz, 2H).

[0245] Intermediate 9: CuSO4 (1M in water, 178.78 μL, 0.3 eq) and sodium L-ascorbate (1M in water, 178 μL, 0.3 eq) were added to a solution of 3-azido-1-(methylsulfonyl)azetidine (105 mg, 595 μmol, 1 eq), citric acid (125 mg, 595 μmol, 1 eq) and 3-butynoic acid (50.1 mg, 595 μmol, 1 eq) in t-BuOH (2 mL), DMSO (2 mL) and H2O (2 mL) under N2 atmosphere. The reaction mixture was stirred at 20 °C for 12 h. The solution was then extracted with DCM (3×10 mL) and EtOAc (1×10 mL). The aqueous layer was lyophilized under reduced pressure to give a pink solid, which was dissolved with DCM (100 mL). The solution was filtered, and the filtrate was concentrated in vacuo to give the title compound, which was used in the next step without further purification. 1 H NMR (500MHz, CDCl3) δppm: 8.18 (br s, 1H), 5.62-5.47 (m, 1H), 4.45-4.35 (m, 2H), 4.33-4.24 (m, 2H), 3.71 (s, 2H), 3.14 (s, 3H).

[0246] Example 1. Procedure A: Synthesis of Compound 1

[0247]

[0248] 1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-2-((5-(trichloromethyl)-1,2,4-oxadiazol-3-yl)methyl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile: DIPEA (416 μL, 2.39 mmol, 3 eq) was added to a microwave vessel containing intermediate 4 (225 mg, 796 μmol, 1 eq), 2-(5-(trichloromethyl)-1,2,4-oxadiazol-3-yl)acetic acid (254 mg, 1.04 mmol, 1.3 eq), T3P (50% in EtOAc, 1.14 mL, 1.91 mmol, 2.4 eq) and EtOAc (2 mL, 0.4 M) under N2 atmosphere at 0°C. The reaction mixture was irradiated using a microwave reactor for 13 h (85°C, 2 bar). After cooling to 20°C, the mixture was diluted with DCM (70 mL) and washed with brine (3×15 mL). The organic layer was dried over Na 2 SO 4 , filtered and concentrated in vacuo. The resulting residue was purified via silica gel chromatography (50% EtOAc in petroleum ether) to give the title compound as an orange solid. LCMS [M+H] + =491.1. 1 HNMR(400MHz,DMSO-d6)δppm 9.35(s,1H),9.03(br s,1H),8.34(d,J=8.4Hz,1H),8.08(d,J=7.6Hz,1H),5.48-5.22(m,1H),5.01(s,2H),4.19(br s,1H),3.71(br s,2H),2.21(br s,2H),2.13(br s,2H),1.24(d,J=6.0Hz,3H).

[0249] 2-{[5-(3-hydroxyazetidin-1-yl)-1,2,4-oxadiazol-3-yl]methyl}-1-[(2R,4R)-2-methyloxan-4-yl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile (Compound 1): Under N2 atmosphere, Cs2CO3 (53.7 mg, 164 μmol, 3 equivalents) was added to 1-((2R,4R)-2 -methyltetrahydro-2H-pyran-4-yl)-2-((5-(trichloromethyl)-1,2,4-oxadiazol-3-yl)methyl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile (30 mg, 54.9 μmol, 1 eq.) and azetidin-3-ol hydrochloride (18.1 mg, 164 μmol, 3 eq.) in DMF (1 mL, 0.05 M). The reaction mixture was stirred at 20 °C for 12 hours. The mixture was diluted with water (20 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified by preparative HPLC (NH4OH conditions) to give Compound 1 as a white solid. LCMS [M+H] + =446.2. 1 H NMR (400MHz, DMSO-d6) δppm 9.35 (s, 1H), 9.02 (br s, 1H), 8.34 (d, J = 8.8Hz, 1H), 8.06 (d, J = 8.4Hz, 1H), 5.90 (d, J = 6.8Hz, 1H), 5.32 (br s,1H),4.63-4.58(m,1H),4.56(br s,2H),4.36-4.30(m,2H),4.26-4.14(m,1H),3.90(dd,J=4.4,8.8Hz,2H),3.83-3.58(m,2H),2.42-2.38(m,1H),2.17(br s,2H),2.09-1.99(m,1H),1.24(d,J=6.0Hz,3H).

[0250] Example 2. Procedure B: Synthesis of Compound 2

[0251]

[0252] 1-((8-cyano-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)methyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid methyl ester: A mixture of intermediate 5 (10 mg, 29.3 μmol, 1 eq), methyl 2-hydroxynicotinate (8.99 mg, 58.7 μmol, 2 eq) and Cs2CO3 (28.7 mg, 88.0 μmol, 3 eq) in DMF (2 mL, 0.01 M) was stirred at 35 °C for 12 h. The mixture was filtered and the filtrate was purified by preparative HPLC (NH4HCO3 conditions) to give the title compound as a white solid. LCMS [M+H] + =458.2. 1 H NMR (400MHz, DMSO-d6) δppm: 9.28 (s, 1H), 9.01 (s, 1H), 8.31 (d, J = 8.4Hz, 1H), 8.22 (dd,J=2.0,6.8Hz,1H),8.15(dd,J=2.0,7.2Hz,1H),8.05(dd,J=1.6,8.4Hz,1H),6 .49(t,J=6.8Hz,1H),5.69(s,2H),5.60-5.41(m,1H),4.23-4.10(m,1H),3.94-3.7 5(m,2H),3.71(s,3H),2.47-2.36(m,1H),2.30-2.05(m,3H),1.27(d,J=6.4Hz,3H).

[0253] 1-({8-Cyano-1-[(2R,4R)-2-methyloxan-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)-2-oxo-1,2-dihydropyridine-3-carboxylic acid (Compound 2): A mixture of methyl 1-((8-cyano-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)methyl)-2-oxo-1,2-dihydropyridine-3-carboxylate (67 mg, 146 μmol, 1 eq) and LiOH·H2O (30.7 mg, 732 μmol, 5 eq) in THF (5 mL, 0.03 M) was stirred at 25° C. for 12 h. The mixture was adjusted to pH=3 using 1N HCl and concentrated in vacuo. The resulting residue was purified by preparative HPLC (NH4HCO3 conditions) to give Example 2 as a white solid. LCMS [M+H] + =444.2. 1H NMR(400MHz,DMSO-d6)δppm:14.03(s,1H),9.28(s,1H),9.02(s,1H),8.53-8.47(m,1H),8.45-8.40(m ,1H),8.31(d,J=8.8Hz,1H),8.07(dd,J=1.6,8.8Hz,1H),6.87(t,J=6.8Hz,1H),5.91(s,2H),5.53(br s,1H),4.17(br s,1H),3.97-3.76(m,2H),2.46-2.39(m,1H),2.31-2.05(m,3H),1.27(d,J=6.0Hz,3H).

[0254] Example 3. Procedure C: Synthesis of Compound 3

[0255]

[0256] The compound of Example 3 was prepared by the same method as Example 1 using Intermediate 4 as a starting material.

[0257] 3-((8-Chloro-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)methyl)-5-(trichloromethyl)-1,2,4-oxadiazole: LCMS [M+H] + =502.0.

[0258] 1-[3-({8-chloro-1-[(2R,4R)-2-methyloxan-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)-1,2,4-oxadiazol-5-yl]azetidin-3-ol (Compound 3): LCMS [M+H] + =455.2. 1 H NMR (400MHz, DMSO-d6) δppm 9.20 (s, 1H), 8.68 (br s, 1H), 8.20 (d, J = 8.8Hz, 1H), 7.76 (dd, J = 2.0, 8.8Hz, 1H), 5.90 (d, J = 6.4Hz, 1H), 5.24 (br s,1H),4.64-4.57(m,1H),4.53(s,2H),4.32(t,J=7.8Hz,2H),4.22-4.13(m,1H),3.90(dd,J=4.4,8.8Hz ,2H),3.76-3.58(m,2H),2.47-2.38(m,1H),2.27-2.11(m,2H),2.06-1.97(m,1H),1.23(d,J=6.0Hz,3H).

[0259] Example 4. Procedure D: Synthesis of Compound 4

[0260]

[0261] 2-((3-Bromo-1H-pyrazol-1-yl)methyl)-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile: To a solution of 3-bromo-1H-pyrazole (63.1 mg, 429 μmol, 1.5 eq) in THF (5 mL) was added NaH (60% in mineral oil, 22.9 mg, 572 μmol, 2 eq) at 0°C. The reaction mixture was stirred at 0°C for 30 min. Thereafter, a solution of intermediate 5 (100 mg, 286 μmol, 1 eq) in THF (3 mL) was added dropwise. The reaction mixture was then stirred at 0-5°C for another 2 h. Thereafter, the mixture was slowly poured into ice water (10 mL), and the mixture was extracted with DCM (2×10 mL). The combined organic layers were washed with brine (2×20 mL), dried over Na 2 SO 4 , filtered and concentrated in vacuo. The resulting crude mixture of isomers was purified via silica gel chromatography (80-100% EtOAc in petroleum ether) to give the title compound as a yellow solid. LCMS [M+H] + =451.0 / 453.0. 1 H NMR (500MHz, DMSO-d6) δppm 9.42-9.34(m,1H),8.97(br s,1H),8.34(d,J=9.0Hz,1H),8.07(d,J=8.5Hz,1H),8.00(br s,1H),6.50(d,J=2.5Hz,1H),5.99(s,2H),5.50-5.28(m,1H),4.24-4.10(m,1H),3.81-3. 59(m,2H),2.48-2.39(m,1H),2.22-2.05(m,1H),2.02-1.69(m,2H),1.23(d,J=6.0Hz,3H).

[0262] N-(1-{[8-cyano-1-(2-methyloxan-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl]methyl}-1H-pyrazol-3-yl)methanesulfonamide (Compound 4): To a solution of 2-((3-bromo-1H-pyrazol-1-yl)methyl)-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile (40 mg, 82.7 μmol, 1 eq) and methanesulfonamide (78.7 mg, 826 μmol, 10 eq) in dioxane (2 mL, 0.04 M) were added t-BuBrettPhos Pd G3 (7.1 mg, 8.27 μmol, 0.1 eq.), tBuONa (23.8 mg, 248 μmol, 3 eq.). The reaction mixture was heated to 120 °C and stirred under N2 atmosphere for 16 hours. The reaction mixture was concentrated in vacuo. The resulting residue was diluted with DCM (10 mL) and ice water (10 mL). The mixture was extracted with DCM (2×20 mL). The combined organic layers were washed with brine (2×20 mL), dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified by preparative HPLC (NH4HCO3 conditions) to give Example 4 compound as a white solid. LCMS [M+H] + =466.1. 1 H NMR (400MHz, DMSO-d6) δppm9.79(br s,1H),9.38(s,1H),8.98(br s,1H),8.34(d,J=8.8Hz,1H),8.07(dd,J=1.2,8.8Hz,1H),7.89(br s,1H),6.09(d,J=2.4Hz,1H),5.88(s,2H),5.46-5.24(m,1H),4.25-4.04(m,1H),3.79-3.56(m,2 H),2.96(s,3H),2.46-2.35(m,1H),2.19-2.03(m,1H),2.02-1.68(m,2H),1.22(d,J=6.0Hz,3H).

[0263] Example 5. Procedure E: Synthesis of Compound 5

[0264]

[0265] 2-{[1-(1-methylsulfonylazetidin-3-yl)-1H-1,2,3-triazol-4-yl]methyl}-1-[(2R,4R)-2-methyloxan-4-yl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile (Compound 5): T3P (50% in EtOAc, 205 μL, 346 μmol, 3 eq) was added to a solution of Intermediate 9 (30 mg, 115 μmol, 1 eq), Intermediate 3 (32.5 mg, 115 μmol, 1 eq) and DIPEA (200 μL, 1.15 mmol, 10 eq) in DMF (2 mL) under N2 atmosphere. The mixture was stirred at 20°C for 1 hour and then at 130°C for 3 hours. The mixture was poured into water (10 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified by preparative HPLC (NH4HCO3 conditions) to give Example 5 as a white solid. LCMS [M+H] + =507.2. 1 H NMR(500MHz,DMSO-d6)δppm:9.37(s,1H),9.02(br s,1H),8.34(d,J=8.5Hz,1H),8.32(s,1H),8.07(dd,J=1.5,8.5Hz,1H),5.60-5.51(m,1H),5.48-5.32(m,1H),4.71(s,2H),4.40(t,J=8.5Hz ,2H),4.29(dd,J=6.0,9.0Hz,2H),4.17(s,1H),3.81-3.61(m,2H),3.13(s,3H),2.49-2.39(m,1H),2.25-1.84(m,3H),1.23(d,J=6.0Hz,3H).

[0266] Example 6. Procedure F: Synthesis of Compound 6

[0267]

[0268] 2-((6-Chloropyridin-3-yl)methyl)-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile: T3P (50% in EtOAc, 1.01 mL, 1.70 mmol, 2.4 eq) was added to a microwave vessel containing Intermediate 3 (200 mg, 708 μmol, 1 eq), 2-(6-chloropyridin-3-yl)acetic acid (145.85 mg, 850 μmol, 1.2 eq), DIPEA (370 μL, 2.13 mmol, 3 eq) and EtOAc (7 mL, 0.1 M) under N2 atmosphere at 0°C. The reaction mixture was irradiated using a microwave reactor for 10 h (85°C, 2 bar). The mixture was diluted with DCM (50 mL) and washed with brine (3×15 mL). The organic phase was dried over Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified by preparative HPLC (HCl conditions) to give the title compound as a yellow solid. LCMS [M+H] + =418.1.

[0269] N-(5-{[8-cyano-1-(2-methyloxan-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl]methyl}pyridin-2-yl)methanesulfonamide (Compound 6): Under N2 atmosphere, to a solution of 2-((6-chloropyridin-3-yl)methyl)-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile (50 mg, 87.34 μmol, 1 eq) and methanesulfonamide (83.1 mg, 873 μmol, 10 eq) in dioxane (2 mL, 0.04 M) were added tBuBrettPhos PdG3 (7.46 mg, 8.73 μmol, 0.1 eq) and t-BuONa (25.2 mg, 262 μmol, 3 eq) in sequence. The reaction mixture was then heated to 120 °C and stirred under N2 atmosphere for 16 hours. The reaction mixture was diluted with EtOAc (50 mL) and washed with brine (3×10 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting residue was purified using preparative HPLC (NH4OH conditions) to give Example 6 as a white solid. LCMS [M+H] + =477.2. 1H NMR(400MHz,DMSO-d6)δppm 10.56(br s,1H),9.31(s,1H),8.97(s,1H),8.35-8.21(m,2H),8.03(dd,J=1.2,8.4Hz,1H),7.69(dd,J=2. 0,8.4Hz,1H),6.97(d,J=7.6Hz,1H),5.43-5.15(m,1H),4.53(s,2H),4.19-4.10(m,1H),3.25(br s,3H),2.22-2.02(m,2H),1.89(br s,2H),1.21(d,J=6.1Hz,3H).

[0270] Example 7. Procedure G: Synthesis of Compound 7

[0271]

[0272] Ethyl 3-((6-cyano-4-(((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)quinolin-3-yl)amino)-3-oxopropanoate: To a solution of compound intermediate 3 (400 mg, 1.42 mmol, 1 eq.), monoethyl malonate (251 mg, 2.13 mmol, 1.5 eq.) in pyridine (10 mL, 0.14 M) was added EDCI (543 mg, 2.83 mmol, 2.0 eq.). The reaction mixture was stirred at 25 °C for 2 hours. The mixture was concentrated in vacuo and the resulting crude residue was purified via silica gel chromatography (0-100% EtOAc in petroleum ether followed by 0-10% MeOH in DCM) to give the title compound as a brown solid. LCMS [M+H] + =383.1. 1 H NMR (400MHz, CDCl3) δppm: 10.88 (br s, 1H), 8.51-8.38 (m, 2H), 7.99 (d, J = 8.8Hz, 1H), 7.72-7.53 (m, 1H), 4.38 (br s,1H),4.19-4.10(m,1H),3.84(d,J=8.8Hz,3H),3.81-3.75(m,2H),3.64-3.49 (m,2H),3.47(s,1H),2.20-1.99(m,3H),1.32-1.28(m,3H),1.27-1.25(m,1H).

[0273]

[0136] Ethyl 2-(8-cyano-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)acetate: To a solution of ethyl 3-((6-cyano-4-(((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)amino)quinolin-3-yl)amino)-3-oxopropanoate (500 mg, 941 μmol, 1 eq) in DMF (5 mL, 0.2 M) was added AcOH (0.5 mL, 8.74 mmol, 0.5 mL, 9.3 eq). The mixture was stirred at 130 °C under microwave (2 bar) for 9 h. The reaction mixture was concentrated in vacuo, and the resulting crude material was purified via silica gel chromatography (50-100% EtOAc in petroleum ether) to give the title compound as a brown solid. LCMS [M+H] + =365.1. 1 H NMR (400MHz, CDCl3) δppm: 9.43 (s, 1H), 9.10 (d, J = 18.8Hz, 1H), 8.66 (br s, 1H), 7.96 (d, J = 8.8Hz, 1H), 4.93 (br s, 1H), 4.37 (br s,1H),4.30(s,2H),3.83(s,3H),3.76(br s,2H),2.41(br s,1H),2.17(br s,1H),2.02(s,2H),1.40(d,J=6.0Hz,3H).

[0274] 2-(8-Cyano-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)acetohydrazide: A solution of ethyl 2-(8-cyano-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)acetyl hydrazide (50 mg, 112 μmol, 1 eq.), hydrazine hydrate (11.2 μL, 225 μmol, 2.0 eq.) in EtOH (2 mL, 0.05 M) was stirred at 0 °C for 2 h. The reaction mixture was filtered, purified by preparative HPLC (NH4HCO3 conditions) and lyophilized to give the title compound as a white solid. LCMS [M+H] + =365.1. 1H NMR (400MHz, DMSO-d6) δppm: 9.48 (br s, 1H), 9.33 (s, 1H), 9.04 (br s, 1H), 8.33 (d, J = 8.4Hz, 1H), 8.05 (dd, J = 1.6, 8.4Hz, 1H), 5.29 (br s,1H),4.35(d,J=4.0Hz,2H),4.20(br s,1H),4.12(br s,2H),3.74(br s,2H),2.28-2.04(m,4H),1.25(d,J=6.0Hz,3H).

[0275] 1-[(2R,4R)-2-methyloxan-4-yl]-2-[(5-oxo-4,5-dihydro-1,3,4-oxadiazol-2-yl)methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile (Compound 7): To a solution of 2-(8-cyano-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinolin-2-yl)acetohydrazide (17 mg, 45.1 μmol, 1 eq) in DMSO (0.5 mL, 0.1 M) was added CDI (14.6 mg, 90.2 μmol, 2.0 eq). The mixture was stirred at 70 °C under microwave (2 bar) for 1 hour. The reaction mixture was filtered, purified by preparative HPLC (formic acid conditions) and lyophilized to give Example 7 as a white solid. LCMS [M+H] + =391.2. 1 H NMR (400MHz, DMSO-d6) δppm: 12.37 (s, 1H), 9.40 (s, 1H), 9.02 (br s, 1H), 8.34 (d, J = 8.4Hz, 1H), 8.08 (dd, J = 1.2, 8.8Hz, 1H), 5.21 (br s,1H),4.71(s,2H),4.18(br s,1H),3.92-3.60(m,2H),2.48-2.40(m,1H),2.23-1.92(m,3H),1.24(d,J=6.0Hz,3H).

[0276] Example 8. Procedure H: Synthesis of Compound 8

[0277]

[0278] 2-(1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)ethan-1-ol: To a suspension of 2-(1H-pyrazol-4-yl)ethanol (5 g, 44.6 mmol, 1 eq.) and Cs2CO3 (21.8 g, 66.9 mmol, 1.5 eq.) in DMF (50 mL, 0.9 M) at 25°C was added SEM-Cl (11.8 mL, 66.9 mmol, 1.5 eq.). The reaction mixture was stirred at 25°C for 2 h. The mixture was quenched with water (250 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were washed with water (3×100 mL) and brine (100 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified via silica gel chromatography (30% EtOAc in petroleum ether) to give the title compound as a colorless oil. LCMS [M+H] + =243.1. 1 H NMR (500MHz, CDCl3) δppm: 7.44 (s, 1H), 7.43 (s, 1H), 5.38 (s, 2H), 3.79 (t, J = 6.5H z,2H),3.58-3.53(m,2H),2.74(t,J=6.5Hz,2H),0.92-0.88(m,2H),-0.03(s,9H).

[0279] 2-(1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)acetaldehyde: A suspension of 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)ethan-1-ol (4 g, 16.5 mmol, 1 eq.), Dess-Martin periodinane (7.30 mL, 23.6 mmol, 1.43 eq.) in DCM (50 mL) was stirred at 25 °C for 4 hours. The reaction mixture was diluted with DCM (250 mL) and washed with saturated aqueous NaHCO3 (250 mL), water (250 mL) and brine (100 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified by silica gel chromatography (20% EtOAc in petroleum ether) to give the title compound as a yellow oil. LCMS [M+H] + =241.1. 1H NMR(400MHz, CDCl3)9.74(t,J=1.6Hz,1H),7.55(s,1H),7.47(s,1H),5.42-5.41(m,2H),3.63 -3.60(m,1H),3.61(d,J=1.5Hz,1H),3.56(d,J=8.0Hz,2H),0.93-0.89(m,2H),-0.02(s,9H).

[0280] 2-(1-((2-(Trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)acetic acid: To a solution of 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)acetaldehyde (1 g, 4.16 mmol, 1 eq), 2-methylbut-2-ene (2.64 mL, 25.0 mmol, 6 eq) in t-BuOH (20 mL), THF (10 mL) and water (5 mL) at 25°C was added NaH2PO4 (1.5 g, 12.5 mmol, 3 eq) and NaClO2 (933 μL, 12.5 mmol, 3 eq). The reaction mixture was stirred at 25°C for 1 hour and then concentrated in vacuo. The residue was diluted with water (100 mL) and extracted with EtOAc (2×100 mL). The combined organic layers were washed with water (100 mL) and brine (100 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound as a colorless oil, which was used in the next step without further purification. LCMS [M+H] + =257.1.

[0281] 1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-2-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)methyl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile: A solution of 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)acetic acid (600 mg, 2.34 mmol, 1.32 equiv), Intermediate 3 (500 mg, 1.77 mmol, 1 equiv), DIPEA (1.8 g, 13.93 mmol, 2.43 mL, 7.86 equiv) and T3P (50% in EtOAc, 1.40 mL, 4.71 mmol, 2.7 equiv) in EtOAc (10 mL) was stirred at 80 °C under microwave irradiation for 6 h. The reaction mixture was diluted with EtOAc (50 mL) and washed with water (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4, filtered and concentrated in vacuo. The resulting crude material was purified via silica gel chromatography (100% EtOAc) to give the title compound as a yellow oil. LCMS [M+H]+ =503.2.

[0282] 2-((1H-pyrazol-4-yl)methyl)-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile: To a solution of 1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-2-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)methyl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile (280 mg, 557 μmol, 1 eq) in DCM (10 mL) was added TFA (3 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 3 hours. The reaction mixture was then concentrated in vacuo to afford the title compound as a yellow oil which was used in the next step without further purification. LCMS [M+H] + =373.1.

[0283] 1-[(2R,4R)-2-methyloxan-4-yl]-2-{[1-(oxetan-3-yl)-1H-pyrazol-4-yl]methyl}-1H-imidazo[4,5-c]quinoline-8-carbonitrile (Compound 8): In a glove box, 3-iodooxetane (160 mg, 869 μmol, 4.1 eq.) was added to a suspension of 2-((1H-pyrazol-4-yl)methyl)-1-((2R,4R)-2-methyltetrahydro-2H-pyran-4-yl)-1H-imidazo[4,5-c]quinoline-8-carbonitrile (80 mg, 214 μmol, 1 eq.) and Cs2CO3 (160 mg, 491 μmol, 2.3 eq.) in DMF (1 mL) at 25° C. The reaction mixture was stirred at 70° C. for 4 h in the glove box. The reaction mixture was then cooled to 25°C, diluted with MeOH (2 mL) and filtered. The filtrate was purified by preparative HPLC (NH4HCO3 conditions) to give Example 8 as a yellow solid. LCMS [M+H] + =429.1. 1H NMR (400MHz, DMSO-d6) δppm: 9.35 (s, 1H), 8.99-8.95 (m, 1H), 8.33 (d, J = 8.4Hz, 1H), 8.06-8.02 (m, 1H), 7.83 (s, 1H), 7 .57(s,1H),5.54(quin,J=7.2Hz,1H),5.47-5.09(m,1H),4.92-4.86(m,2H),4.85-4.81(m,2H),4.42(s,2H),4.13(br s,1H),3.73(br s,2H),2.16-1.63(m,4H),1.20(d,J=6.0Hz,3H).

[0284] Example 9. Synthesis of other compounds of formula (I)

[0285] Other compounds of the present invention as described in Table 3 below were synthesized according to the above procedures.

[0286] Table 3. Synthesis

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293] Table 4 provides characterizations of exemplary compounds of the present disclosure.

[0294] Table 4. LCMS and NMR data

[0295]

[0296]

[0297]

[0298] Example 10. Biological Activity Biochemical assays

[0299]

[0300] Prepare 10 mM compound solution in DMSO. Perform 11-point, 3-fold dilutions with the highest concentration being 10 μM. Add 10 mM compound DMSO solution to Labcyte LDV plate, and the source plate has a compound concentration of 10 mM. Transfer 1.5 μL of 10 mM compound from the source plate to 28.9 μL DMSO to prepare the first compound concentration of 4.938 × 10 -1 mM. Transfer 60 nL of 10 mM compound from the source plate into 32.7 μL DMSO to prepare the second compound concentration for the Inter plate at 1.829 × 10 -2 mM. Transfer 2.5 nL of 10 mM compound from the source plate to 36.9 μL DMSO to prepare the third compound concentration of the Inter plate at 6.774 × 10 -4 Dispense 100 nL of reference compound in column 1 of the low control wells and 100 nL of DMSO in column 24 of the high control wells. Dispense compounds into columns 2 to 23 of the assay plate and top up with DMSO to a total volume of 100 nL.

[0301] Prepare 2X LRRK2 enzyme solution (final concentration 3nM) in assay buffer (Tris-HCl pH8.0: 50mM, MgCl2: 5mM, EDTA: 1mM, Brij-35: 0.01%, 2mM DTT). Prepare 2X substrate solution: LRRK2tide substrate (final concentration 400nM) and ATP (final concentration 25μM) in assay buffer. Dispense 5μL of 2X LRRK2 enzyme solution into each well of the assay plate using Multidrop. The assay plate was centrifuged at 1000rpm for 1 minute and then incubated at 23°C for 15 minutes. Dispense 5μL of 2X ATP / LRRKtide solution into each well of the assay plate using Multidrop. The assay plate was centrifuged at 1000rpm for 1 minute and then incubated at 23°C for 120 minutes.

[0302] Prepare 2X detection solution: Tb-pERM (pLRRKtide) antibody (final concentration 0.25 nM) and EDTA (final concentration 10 mM) were prepared in TR-FRET dilution buffer. 10 μL of 2X detection solution was dispensed into each well of the assay plate using Multidrop to stop the kinase reaction. The assay plate was centrifuged at 1000 rpm for 1 minute and then incubated at 23°C for 30 minutes. Assay plates were read on Envision in the TR-FRET configuration.

[0303] pS935 LRRK2 cell assay

[0304] The following protocol describes an in vitro method for measuring phosphorylation at Ser935 of wild-type LRRK2 overexpressed in recombinant HEK-293T cells. The method is based on the HTRF technology, which combines fluorescence resonance energy transfer (FRET) and time-resolved measurement (TR). Phosphorylated-LRRK2 (Ser935) is detected in a sandwich method using two different specific antibodies, one labeled with Eu3+-Cryptate (donor) and the other labeled with d2 (acceptor). When the fluorophores are in close proximity, the donor is excited with a light source (flashlight), stimulating the acceptor to undergo FRET, thereby emitting fluorescence at a specific wavelength (665nm). The fluorescence emission of the donor at a wavelength of 615nm is also measured to perform a ratiometric reduction on the data. The specific signal is proportional to phosphorylated-LRRK2 (Ser935).

[0305] Table 5. Materials used for cell-based assays

[0306]

[0307]

[0308] plan:

[0309] Day 0:

[0310] Plasmid transient transfection: Warm DMEM medium, FBS, DPBS, Trans-IT and OPTI-MEM reagents to room temperature. Before transfection, place HEK293T cells in a T150 flask and culture them in DMEM + 10% FBS complete medium to about 80% confluency. Then rinse the cells with 10mL PBS and detach the cells with 3mL 0.25% trypsin. Seed 30x 10E6 HEK293T cells in DMEM + 10% FBS complete medium into a 15cm culture dish.

[0311] Preparation of DNA, TransIT-LT1, OPTI-MEM complex: Add 2000 μL OPTI-MEM to a 15 mL conical tube, then add 20 μg of plasmid to OPTI-MEM and mix, then add 60 μL TransIT-LT1 to the plasmid OPTI-MEM mixture and mix. Incubate the mixture for 15 minutes.

[0312] Add the above plasmid, DNA and OPTI-MEM mixture dropwise to a 15 cm culture dish, ensuring that the droplets are evenly distributed. Gently shake the culture dish back and forth and left and right to evenly distribute the complex. Incubate the transfection culture dish at 37°C, 5% CO2 for 24 hours.

[0313] Day 1:

[0314] Collect transfected HEK293T in a 15 cm dish. Aspirate the medium from the tissue culture dish and dispense 10 mL of 1X DPBS into the 15 cm dish for washing. Aspirate the 1X DPBS and dispense 3 mL of trypsin into the 15 cm dish. Incubate the dish with trypsin at room temperature for 3 minutes until the cells are detached. Add 10 mL of DMEM + 10% FBS medium into the 15 cm dish and grind to ensure a uniform cell suspension.

[0315] Transfer the homogenized cell suspension to a 50 mL tube and centrifuge at 1000 rpm for 5 minutes. Aspirate the supernatant and resuspend in 20 mL of complete medium. Transfer 1 mL of the cell suspension for cell counting. Dilute the cell suspension to 2x10E5 cells / ml. Add 50 μL of the cell suspension to a 384-well plate. Quickly centrifuge at 800 rpm for 1 minute and then incubate overnight at 37°C, 5% CO2.

[0316] Day 2:

[0317] Compound distribution: Compounds were diluted (10 mM DMSO stock solution) and added to the assay plate in duplicate using a Tecan liquid handler (highest concentration: 10 μM, 3-fold serial dilutions, 9 doses). The DMSO concentration in each well was normalized to 0.2%. The plate was quickly centrifuged at 1000 rpm for 1 minute. The plate was incubated at 37°C, 5% CO2 for 2 hours.

[0318] Prepare a 1X lysis buffer solution with added blocking reagent (e.g. 1 mL lysis buffer 4X + 3 mL water + 40 μL of stock blocking reagent 100X). Prepare antibody working solution by diluting d2 and Cryptate antibodies 40 times with detection buffer (e.g. 1520 μL detection buffer + 40 μL d2 antibody stock solution + 40 μL Cryptate antibody stock solution).

[0319] After 2 hours of incubation, remove the cell plate from the incubator. Remove the culture medium with a plate washer, then add 16 μL of lysis buffer 1X to each well and incubate at room temperature with shaking (800 rpm / min) for 30 minutes. Add 4 μL of antibody working solution to each well, cover with the top seal, and incubate overnight in a 23°C incubator.

[0320] Day 3:

[0321] In Wallac 2104 HTRF signals (665 nm and 615 nm) were read on a multi-label reader and data were analyzed by XL fit software.

[0322] The biochemical data of the exemplary compounds of the present disclosure are shown in Table 8.

[0323] Table 6. LRRK2 inhibition

[0324]

[0325]

[0326] Although the above invention has been described in some detail by way of examples and embodiments for the purpose of clear understanding, it will be appreciated by those skilled in the art that certain changes and modifications may be implemented within the scope of the appended claims. In addition, each reference provided herein is incorporated in its entirety by reference to the same extent as each reference is incorporated individually. In the event of a conflict between the present application and the reference provided herein, the present application shall prevail.

Claims

1. Compounds of formula I: or a pharmaceutically acceptable salt thereof, wherein Ring A is a 5- to 6-membered heterocycloalkyl group having 1 to 2 heteroatoms each independently being N, O or S, or a 5- to 6-membered heteroaryl group having 1 or 2 heteroatoms each independently being N, O or S; Each R 1 Independently for C 1-6 Alkyl or =O; Ring B is a 5- to 6-membered heteroaryl group having 1 to 4 heteroatoms each independently being N, O or S; Each R 2 C 1-6 Alkyl, –OH, =O, C(O)R 2a 、C(O)OR 2b 、OC(O)R 2a 、S(O)2R 2a 、S(O)2OR 2b 、OS(O)2R 2a 、N(R 2b )S(O)2R 2a 、S(O)2N(R 2b )(R 2c ), C 3-6 Cycloalkyl, C 1-6 Alkyl-C 3-6 Cycloalkyl or 3 to 6 membered heterocycloalkyl having 1 to 3 heteroatoms each independently being N, O or S, wherein each alkyl or cycloalkyl is surrounded by 1 to 3 R 2d and each heterocycloalkyl group is substituted by 0 to 3 R 2e Group substitution; Each R 2b and R 2c is hydrogen or C 1-6 alkyl; Each R 2d independently C(O)R 2d1 or S(O)2R 2d1 ; Each R 2e Independently for C 1-6 Alkyl, -OH, =O, C(O)R 2e1 or S(O)2R 2e1 ; Each R 2a , R 2d1 and R 2e1 Independently for C 1-6 alkyl; Each R 3 and R 4 For hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogen, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or –CN; subscript n is 0, 1, or 2; and The subscripts m and p are each independently an integer from 1 to 4.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein subscript n is 1 or 2.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the subscript m is 1. 4 . The compound or pharmaceutically acceptable salt thereof according to claim 1 , wherein subscript p is 1.

5. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein Ring A is a 5- to 6-membered heterocycloalkyl group having 1 heteroatom which is N, O or S, or a 5- to 6-membered heteroaryl group having 1 or 2 heteroatoms which are each independently N, O or S. 6 . The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 , wherein Ring A is tetrahydropyranyl.

7. A compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein each R 1 Independently for C 1-6 alkyl.

8. A compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, wherein each R 1 For me.

9. A compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof, wherein R 3 is hydrogen or halogen.

10. The compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, wherein R 3 For hydrogen.

11. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, wherein each R 4 are independently halogen or -CN.

12. A compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, wherein each R 4 are independently Cl or -CN.

13. A compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, which has a structure of Formula Ia:

14. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, which has the structure of Formula Ib:

15. A compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, which has the structure of Formula Ic: 16 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein Ring B is a 5- to 6-membered heteroaryl group having 1 to 3 heteroatoms each independently being N, O or S. 17 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein Ring B is pyrazolyl, isoxazolyl, 1,2,3-triazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl or pyridinyl.

18. A compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, wherein Group for 19. A compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, wherein Each R 2 C 1-3 Alkyl, =O, C(O)OR 2b 、OC(O)R 2a 、S(O)2R 2a 、N(R 2b )S(O)2R 2a 、S(O)2N(R 2b )(R 2c ), C 3-6 Cycloalkyl or 3 to 6 membered heterocycloalkyl having 1 to 3 heteroatoms each independently being N, O or S, wherein each alkyl or cycloalkyl is surrounded by 1 to 2 R 2d and each heterocycloalkyl group is optionally substituted by 1 to 2 R 2e Group substitution; Each R 2b and R 2c is hydrogen or C 1-3 alkyl; Each R 2d independently C(O)R 2d1 or S(O)2R 2d1 ; Each R 2e Independently for C 1-3 Alkyl, -OH, =O, C(O)R 2e1 or S(O)2R 2e1 ;as well as Each R 2a , R 2d1 and R 2e1 Independently for C 1-3 alkyl.

20. A compound according to any one of claims 1 to 19 or a pharmaceutically acceptable salt thereof, wherein each R 2 are independently =O, -COOH, -C(O)OMe, -SO2Me, -NHSO2Me, -CH2CH2SO2Me, 21. A compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof, wherein Group for 22. A compound according to any one of claims 1 to 21 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of the compound in Table 1.

23. A pharmaceutical composition comprising a compound according to any one of claims 1 to 22 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

24. A method of inhibiting LRRK2 in a cell, the method comprising contacting the cell with an effective amount of a compound according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof.

25. A method of treating a LRRK2-related disease or condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof.

26. The method of claim 25, wherein the LRRK2-associated disease or condition is Parkinson's disease, dementia with Lewy bodies, frontotemporal dementia, corticobasal dementia, progressive supranuclear palsy, Alzheimer's disease, tauopathy, or alpha-synucleinopathy.

27. The method of claim 25, wherein the LRRK2-associated disease or condition is inflammatory bowel disease.

28. The method of claim 25, wherein the LRRK2-associated disease or condition is an autophagy-associated disease or condition.

29. The method of claim 28, wherein the autophagy-related disease or condition is alpha 1 -antitrypsin deficiency (AATD).

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