WO2025136898A1

WO2025136898A1 - Agonists of trem2 activity

Info

Publication number: WO2025136898A1
Application number: PCT/US2024/060448
Authority: WO
Inventors: Manasi ANANTPUR; Yaroslav BOYKO; Joanna L. CHEN; Erin F. Dimauro; Timothy J. Henderson; Ping Liu; Vladimir SIMOV; Brandon A. VARA; Xiao Mei Zheng
Original assignee: Merck Sharp and Dohme LLC
Current assignee: Merck Sharp and Dohme LLC
Priority date: 2023-12-18
Filing date: 2024-12-17
Publication date: 2025-06-26
Anticipated expiration: 2026-06-18

Abstract

The present disclosure is directed to compounds of Formula (I) and their use as TREM2 agonists for treatment and prevention of a neurodegenerative disorder associated with a loss of function of human TREM2. The disclosed TREM2 agonists may be useful for the treatment of Alzheimer's Disease and associated neurological conditions.

Description

AGONISTS OF TREM2 ACTIVITY

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The application claims the benefit of priority to U.S. Provisional Application Nos. 63/692.975, filed September 10, 2024. and 63/611,363, filed December 18, 2023, the contents of each of which are incorporated herein by reference in their entireties.

FIELD

[0002] The present disclosure provides certain compounds that are TREM2 agonists. The compounds are useful for treatment and prevention of a neurodegenerative disorder associated with a loss of function of human TREM2. The disclosed TREM2 agonists may be useful for the treatment of Alzheimer’s Disease and associated neurological conditions.

BACKGROUND

[0003] Alzheimer’s disease (AD) is a neurodegenerative disease that is the most common cause of dementia in the United States. Worldwide, over 50 million people are living with dementia, and this prevalence is expected to triple by 2050. Of the top 10 causes of death in the United States, AD is the only major cause of morbidity and mortality without suitable treatments for prevention, slowing, or cure (2017 Alzheimer’s Association Report). Current therapies for AD such as acetylcholinesterase inhibitors (e.g.. donepezil) and N-methyl-D-aspartate receptor antagonists (e.g., memantine) show modest and transient benefits to cognition and behavior parameters in AD patients, but do not slow or halt the progression of the disease (Cummings (2004) NEngl J Med, 351 :56-67).

[0004] Triggering Receptor Expressed on Myeloid cells-2 (TREM2). an immunoglobulin-like transmembrane receptor, appears to play a key role in AD pathology. Heterozygous mutations in the TREM2 gene have been found to increase the risk of AD by up to 3-fold (Guerreiro et al. (2013), NEngl J Med, 368: 117-127; Jonsson et al. (2013) NEngl J Med, 368: 107-116), and increase the rate at which brain volume shrinks (Rajagopalan et al. (2013) N Engl J Med, 369: 1565-1567). Even individuals without AD who carry a heterozygous TREM2 mutation show impaired cognition compared to individuals with tw o normal TREM2 alleles. In the context of AD pathology, TREM2 expression impacts amyloid pathology⁷, modulates neuritic dystrophy, tau hyperphosphorylation and aggregation, and affects synaptic and neuronal loss (Jay et al. (2017) Mol Neurodegener, 12(1 ): 56). In addition, it has been shown that TREM2 plays a key role in limiting the development of peri-plaque tau pathologies (Leyns et al. (2019) Nat Neurosci). Recent mouse genetic model studies also strongly support a key role for TREM2 in AD, with loss or deficiency of TREM2 being associated with increased pathology (Cheng-Hathaway et al. (2018) Mol Neurodegcner. 13(1):29; Wang et al. (2015) Cell, 160: 1061-1071; Wang et al. (2016) J Exp Med, 213:667-675; Yuan et al. (2016) Neuron, 90:724-739).

[0005] As reflected in its name, TREM2 is expressed primarily on myeloid lineage cells, including microglia (Colonna & Wang (2016) Nat Rev Neurosci, 17:201-207). Microglia are resident macrophages of the central nervous system (CNS) that, when activated appropriately, are thought to sen e an important protective role in Alzheimer’s disease through their housekeeping functions such as facilitating clearance of cellular debris through phagocytosis, as well as secretion of growth factors. In the CNS, TREM2 is exclusively expressed on microglia. It has been shown that TREM2 expression regulates microglial chemotaxis and phagocytosis, and enhances microglial cell survival, proliferation, and differentiation. In addition, it is well known that TREM2 is required to sustain microglial trophic function in the aging brain, and studies showed that an overlap exists betw een aged microglia phenoty pe and microglial molecular signatures found in animal models of AD, which include TREM2 pathways (Krasemann et al. (2017) Immunity, 47(3):566-581).

[0006] Certain reports have suggested that the role of TREM2 in experimental amyloid-based AD models is that the protein has an important role in plaque entrainment by microglia, where it appears to be overexpressed and drive transit of these innate immune cells to a disease-associated microglial (DAM) state. This transition is marked by microglial priming that is evident by a set of overexpressed genes and an improved phagocytic capacity. George, Neural Regeneration Research, 18(12): 2680-81 (2023).

[0007] These findings suggest that activation of TREM2 may ameliorate AD symptoms and result in improvements in cognitive function through activation of the innate immune system, and as such agonists of TREM2 may be useful in the treatment of Alzheimer’s disease and other dementias and related neurodegenerative disorders. There is a need in the art for novel agonists of TREM2 and methods of treating neurodegenerative disorders with such agonists.

SUMMARY

[0008] The present invention is directed to certain carbonyl -substituted fused heteroaryl derivative compounds. These compounds are shown to exhibit agonism of the TREM2 receptor, surprisingly and advantageously. The present invention is further directed to the use of these compounds in the treatment or prevention of a neurodegenerative disorder, in a subject in need thereof. The present invention provides compounds that may be adapted for pharmaceutical compositions that may be administered to a subject suffering from a neurodegenerative disorder. The compounds of the disclosure contain a core having two fused nitrogen-containing aryl groups, and exhibit excellent potency in activating the TREM2 receptor, such as the human TREM2 receptor. In some embodiments, the compounds of the invention exhibit superior potency as agonists of TREM2, as evidenced by the data reported herein. The compounds of the invention may be useful in the treatment or prevention of neurodegenerative disorders (or one or more symptoms associated with such disorders) in which TREM2 is involved, including Alzheimer's disease and other indications, diseases and disorders as described herein. The invention is also directed to pharmaceutical compositions comprising a compound of the invention and to methods for the use of such compounds and compositions for the treatments described herein.

[0009] In some aspects, provided herein are compounds having the Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

X¹ and X² are independently N, C(F) or C(H);

X4 is N or C(R⁸); wherein when Xl is N then X4 is C(R ), and when X4 is N then Xl is C(F) or C(H);

R¹ is H, -Ci-4 alkyl, -N(CH₃)₂. -(CH₂)z-OH, or -O(CH₂)_ZCH₃;

R² is H, -Ci-4 alkyl; or, alternatively, R¹ and R² are optionally taken together with one or more intervening atoms to form a ring CZ; wherein ring CZ is:

(i) non-aromatic, partially unsaturated 5- to 6-membered mono- or a 6- to 8- membered bicyclic cycloalkyl; or

(ii) 5- to 6-membered heterocycloalkyl, wherein said 5- to 6-membered heterocycloalkyl is non-aromatic and partially unsaturated and contains one heteroatom selected from the group consisting of N, O and S; wherein ring

is unsubstituted or substituted by 1 to 3 RZ substituents independently selected from the group consisting of fluoro, Ci-3 alkyl, Ci-3 fluoroalkyl or C 1-3 alkoxy;

R³ is selected from the group consisting of:

wherein R⁷ and R¹¹ are independently selected from hydrogen, a halo, -Ci-4 alkyl, cyclopropyl, and -CF?; and R¹⁰, R¹² and R¹³ are independently selected from hydrogen and a halo;

R³ is selected from:

(a) a -C4-7 cycloalkyl, wherein the C4-7 cycloalkyl of R³ is unsubstituted or substituted with a -C1-3 alkyl, a heteroaryl, or fluoro;

(b) a heterocyclyl that is comprised of:

(1) seven carbon atoms and one N atom.

(2) six carbon atoms and (i) one O atom or (ii) one each of O and N, or

(3) five carbon atoms and one each of O and N, and wherein the heterocyclyl of R⁵ is unsubstituted or substituted with a -C1-3 alkyl, a heteroaryl, or fluoro; and

wherein:

Y¹ is C(H). C(F). N or O;

Y² is C(H), C(F), or N;

Y³ is C(H) or N;

R⁴ is selected from hydrogen, a -C4-6 cycloalkyl, a heterocyclyl, and a heteroaryl; wherein the C4-6 cycloalkyl, heterocyclyl and heteroaryl of R⁴ is unsubstituted or substituted with 1 to 3 R4a substituents independently selected from the group consisting of fluoro, C1-3 alkyl, C1-3 fluoroalkyl, C1-3 alkoxy. C1-3 fluoroalkoxy, and C3-6 cycloalkyl, heterocyclyl and heteroaryl;

R⁶ is selected from the group consisting of hydrogen, acetyl, -SO2(CH3), and fluoro; wherein when Y1 is O, then R6 is absent;

R9a and R9b are independently H. fluoro, or C]-3 alkyl; or, alternatively, R9a and R9b. together with the carbon atom to which they are attached, form a C3-6 cycloalkyl; and

R⁸ is hydrogen or fluoro; s is 0 or 1; and z = 1 or 2.

[0010] All structural Formulas, embodiments and classes thereof described herein include the pharmaceutically acceptable salts of the compounds defined therein. Reference to the compounds of Formula (I) herein encompass the compounds of Formulas (II) and (III), and all embodiments and classes thereof. Reference to the compounds of this invention as those of a specific formula or embodiment, e.g. Formula (I), or embodiments thereof, or any other generic structural formula or specific compound described or claimed herein, is intended to encompass the specific compound or compounds falling within the scope of the Formula or embodiment, including salts thereof, particularly pharmaceutically acceptable salts, solvates (including hydrates) of such compounds and solvated salt forms thereof, where such forms are possible, unless specified otherwise.

[0011] The invention also encompasses pharmaceutical compositions comprising an effective amount of a compound of the invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

DETAILED DESCRIPTION

[0012] The present disclosure is directed to compounds of Formula (I), which exhibit activity as agonists of TREM2 receptor. Also provided herein are pharmaceutical preparations comprising any of these compounds and a pharmaceutically acceptable carrier. Further provided herein are methods of treatment and prevention of a neurodegenerative disorder comprising the administration of any of these compounds to a subject. Also provided herein are methods of treatment and prevention of a condition associated with a loss of function of human TREM2 in a subject. COMPOUNDS OF THE DISCLOSURE

[0013] In one embodiment, the compound of Formula (I) has the Formula (II)

wherein:

X³ is CH₂ or O;

R¹ is a -Ci-3 alkyl or -OC1-3 alkyl;

R² is a -C1-3 alkyl; or, alternatively, R¹ and R² are optionally taken together with their intervening atoms to form a C5-8 cycloalkyl or 5- to 8-membered heterocyclyl group, and wherein the C5-8 cycloalkyl or 5- to 8-membered heterocyclyl group is unsubstituted or independently substituted with one or two fluoro or methyl;

R⁷ is a halo; and

R⁴ is selected from hydrogen and a heteroaryl that is comprised of:

(1) three carbon atoms and two N atoms.

(2) four carbon atoms and two N atoms, or

(3) five carbon atoms and one N atom, wherein the heteroaryl of R⁴ is unsubstituted or substituted with methyl or -OCH3.

[0014] In some embodiments, the compounds of Formula (I) and Formula (II) are bicyclic. In a sub-embodiment of these embodiments are compounds of Formula (I) and Formula (II), or a pharmaceutically acceptable salt thereof, wherein R¹ or R² are methyl. In such a sub-embodiment are compounds such as the below exemplary compound

pharmaceutically acceptable salt thereof. [0015] In some embodiments, the compounds of Formula (I) and Formula (II) are tricyclic. In a sub-embodiment of these embodiments are compounds of Formula (I) and Formula (II), or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are taken together to form a C5-6 cycloalkyl or or a 5- to 6-membered heterocyclyl that is unsubstituted or substituted with one or two fluoro. In a sub-embodiment, R¹ and R² are taken together to form a C5-6 cycloalkyl that is unsubstituted. In a sub-embodiment. R¹ and R² are taken together to form a 5- to 6-membered heterocyclyl containing a single O atom. In such sub-embodiments are compounds such as the below exemplary compounds:

pharmaceutically acceptable salt thereof.

[0016] In a sub-embodiment of these tricyclic compound embodiments are compounds of Formula (I) and Formula (II), or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are taken together to form a spiro, caged or bridged C5-6 cycloalkyl. In such sub-embodiments are compounds such as the below exemplary compounds:

pharmaceutically acceptable salt thereof. [0017] In some embodiments are compounds of Formula (I) wherein

In a sub-embodiment of these embodiments, R¹² and R¹¹ are each hydrogen, such that R³ is

[0018] In some embodiments are compounds of Formula (I) and Formula (II), wherein R⁷ is chloro. In some embodiments are compounds of Formula (I) and Formula (II), wherein R¹⁰ is fluoro.

[0019] In some embodiments are compounds of Formula (1), wherein

In a sub-embodiment, Y¹ is O, Y² is N, and Y³ is CH. In this and other sub-embodiments, R⁶ is absent.

[0020] In some embodiments are compounds of Formula (I), wherein R⁵ is a 4- to 8-membered cycloalkyl or heterocyclyl. In some embodiments. R⁵ is a 4- to 8-membered spiro, caged or bridged 4- to 8-membered cycloalkyd or heterocyclyl.

[0021] In some embodiments are compounds of Formulas (I)-(III), wherein R⁴ is heteroary l, such as a 5- to 6-membered heteroaryl. In a sub-embodiment of these embodiments, R⁴ is a pyrazole that is unsubstituted or substituted at the 1 -position with a -Ci-io alkyl.

[0022] In certain embodiments, the compound of Formula (I) has the Formula (III)

wherein:

Xl is N or C(F);

X4 is N or C(H); wherein when Xl is N then X4 is C(H), and when X4 is N then Xl is C(F);

R¹ and R² are independently H or Ci-4 alkyl, or alternatively, R¹ and R², together with the carbon atoms to which they are attached, form a ring CZ; wherein ring CZ is:

(i) non-aromatic, partially unsaturated 5- to 6-membered mono- or a 6- to 8- membered bicyclic cycloalky l; or

(ii) 5- to 6-membered heterocycloalkyl, wherein said 5- to 6-membered heterocycloalkyl is non-aromatic and partially unsaturated and contains one heteroatom selected from the group consisting of N, O and S; wherein ring CZ is unsubstituted or substituted by 1 to 3 RZ substituents independently selected from the group consisting of fluoro, Ci-3 alkyl, Ci-3 fluoroalkyl or C 1-3 alkoxy;

R , R11. R12, and R13 are independently H or halo;

Y² is C(H), C(F), or N;

R⁴ is:

(i) a 5- to 6-membered heteroaryl containing 1 to 2 heteroatoms independently selected from the group consisting of N, O, and S; or

(ii) a 5- to 6-membered saturated heterocyclyl containing 1 O atom; wherein R⁴ is unsubstituted or substituted by 1 to 3 R4a substituents independently- selected from the group consisting of fluoro, C1-3 alkyl, C1-3 fluoroalkyl, C1-3 alkoxy. C1-3 fluoroalkoxy, and C3-6 cycloalkyl;

R9a and R9b are independently H, fluoro, or Ci-3 alkyl; or, alternatively, R9a and 9b. together with the carbon atom to which they are attached, form a C3-6 cycloalkyl; and s is 0 or 1.

[0023] In some embodiments of the compound of Formula (III), Xl is N and X4 is C(H).

[0024] In certain embodiments of the compound of Formula (III),

(i) R¹ and R² are both methyl; or

(ii) R¹ and R² form a 5-membered heterocyclyl containing one O atom.

[0025] In specific embodiments of the compound of Formula (III), s is 0. [0026] In some embodiments of the compound of Formula (III), Y² is C(H). In some other embodiments of the compound of Formula (III), Y² is N.

[0027] In certain embodiments of the compound of Formula (III), R⁴ is unsubstituted or substituted tetrahydrofuryl, pyrazolyl, or pyridinyl.

[0028] In some embodiments of the compound of Formula (III),

[0029] In other certain embodiments of the compound of Formula (III).

wherein t is 0, 1, 2, or 3.

[0030] In other certain embodiments of the compound of Formula (III),

above is C(H). In another specific embodiment, Y² in the moieties above is N.

[0031] In specific embodiments of the compound of Formula (III),

[0032] In one embodiment, in the compound of Formula (I)

X¹ and X² are independently N or CF;

R¹ is a -Ci-4 alkyl, -N(CH?)₂. -(CH₂)z-OH. or -O(CH₂)_Z;

R² is a -Ci-4 alkyl;

R³ is selected from the group consisting of:

wherein R⁷ and R¹¹ are independently selected from hydrogen, a halo, -Ci-4 alkyl, cyclopropyl, and -CF?; and R¹⁰ and R¹² are independently selected from hydrogen and a halo; R⁵ is selected from:

(a) a -C4-7 cycloalkyl,

(b) a heterocyclyl that is comprised of

(1) seven carbon atoms and one N atom,

(2) six carbon atoms and (i) one O atom or (ii) one each of O and N,

(3) five carbon atoms and one each of O and N, or

wherein:

Y¹ is CH, CF, N or O;

Y² is CH or N;

Y³ is CH or N;

R⁴ is selected from hydrogen, a -C4-6 cycloalkyl, a heterocyclyl, and a heteroaryl.

R⁶ is selected from hydrogen, acetyl, -SChlCHs), and fluoro, and

R⁹ is hydrogen or methyl; wherein any cycloalkyl or heterocyclyl moiety of R⁵ is unsubstituted or substituted with a -C1-3 alkyl, a heteroaryl. or fluoro, and wherein any heteroaryl moiety of R⁵ is unsubstituted or substituted with methyl, a heterocyclyl, one or two fluoro; and

R⁸ is hydrogen or fluoro, wherein z = 1 or 2, wherein R¹ and R² are optionally taken together with one or more intervening atoms to form a cycloalkyl or heterocyclic group, and wherein any such cycloalkyl or heterocyclic group is unsubstituted or substituted with one or two fluoro atoms or methyl. [0033] In an embodiment of this invention are compounds of Formula (I) having structural Formula (II), wherein:

X³ is CH2 or O;

R¹ is a -C1-3 alkyl or -OC1-3 alkyl;

R² is a -C1-3 alkyl;

R⁷ is a halo; and

R⁴ is selected from hydrogen and a heteroaryl that is comprised of:

(1) three carbon atoms and two N atoms,

(2) four carbon atoms and tw o N atoms, or (3) five carbon atoms and one N atom, wherein any such heteroaryl is unsubstituted or substituted with methyl or -OCH3, wherein R¹ and R² are optionally taken together with their intervening atoms to form a C5-8 cycloalkyl or C5-8 heterocyclyl group, and wherein any such cycloalkyl or heterocyclyl group is unsubstituted or substituted with one or two fluoro atoms or methyl.

[0034] In some embodiments, the compounds of Formulas (I)-(III) are bicyclic. In some embodiments, the compounds of Formulas (I)-(III) are tricyclic.

[0035] In some embodiments, X¹ and X² are each N.

[0036] In particular embodiments, the compound is selected from the group of Example Nos. 1- 1 - 1-11, 2-1 - 2-17, 3-1 - 3-50, 4-1 - 4-40, 5-1 - 5-34, 6-1, 7-1, 8-1. and 9-1. In specific embodiments, the compound is selected from group of Example Nos. the compound is selected from the group of Example Nos. 1-1 - 1-1 1, 2-1 - 2-17, 4-1 - 4-40, 5-1 - 5-34, 6-1, 7-1, 8-1, and 9-1.

[0037] In some embodiments, provided herein are pharmaceutically acceptable salts of any of the disclosed compounds (e.g, hydrochloride salts). In other embodiments, the compounds provided herein are isolated trifluoroacetate salts. In some embodiments, the phase of any of the disclosed compounds is crystalline. In some embodiments, the phase of any of the disclosed compounds is amorphous.

[0038] In some embodiments, any of the disclosed compounds exhibit high potency in activating the TREM2 receptor, e.g., the human TREM2 receptor. Activation of the TREM2 receptor may be measured using any of several assays known in the art. For example, activation of TREM2 may be measured using a phosphory lation of Spleen Tyrosine Kinase (pSYK) assay. In addition to the pSYK assay, activation of TREM2 may be measured in vivo (e.g., in a subject) using other biomarkers including, but not limited to, interaction of TREM2 with adaptor protein DAP12, pDAP12 elevation, TREM2 clustering, and cytokine elevation, e.g., IP10 or CCL4 cytokine elevation. Several of these biomarkers, such as DAP12, constitute proteins that are activated in the TREM2 pathway downstream of, and following binding of a ligand to, the TREM2 receptor. Potencies of activation may be expressed as IC50 or EC50. Activation of TREM2 may be measured in vitro.

TREATMENT OF NEURODEGENERATIVE DISORDERS

[0039] The disclosed compounds and pharmaceutical compositions are useful for treatment and/or prevention of one or more neurodegenerative disorders. For instance, they may be useful for treatment or prevention of one or more of Alzheimer’s disease, Parkinson’s disease. frontotemporal dementia, demyelination disorder, multiple sclerosis, Huntington’s disease, amyotrophic lateral sclerosis (ALS). tauopathy disease, Nasu-Hakola disease, or adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP). In some embodiments, the disclosed compounds and compositions are useful for treatment of Alzheimer’s Disease. In some embodiments, the disclosed compounds and compositions are useful for treatment of dementia, such as frontotemporal dementia.

[0040] In some aspects, any of the disclosed compounds and compositions may be administered to a subject suffering from Alzheimer’s Disease. In various embodiments, the subject is human. Administration of these compositions may improve cognitive and/or functional benefit by stabilizing amyloid negative status or further modulating multiple biomarkers of Alzheimer’s disease pathology or progression in the subject. In certain embodiments, the disclosed compositions promote the stabilization amyloid negative status in the subject. Modulation of biomarkers of Alzheimer’s disease pathology or progression in the subject may be observed by measuring any of the following biomarkers in a sample of the subject's blood, plasma and/or cerebrospinal fluid: Ab42/40 ratio, pTau and/or total Tau, NfL, GFAP, soluble Trem2 (sTrem2) and YKL-40. In some embodiments, any of the disclosed compounds improve (i.e., reduce) the subject’s Ab42/40 ratio, pTau and/or total Tau, NfL, GFAP, sTrem2 and/or YKL-40 in plasma or cerebrospinal fluid (CSF). In some embodiments, administration of any of the disclosed compounds improve the subject's Ab42/40 ratio. In some embodiments, administration of any of the disclosed compounds reduce the sTREM2 in the subject’s CSF.

[0041] Modulation of the Ab42/40 ratio, pTau and/or total Tau in a subject may be measured using imaging markers, including positron emission tomography (PET) of amyloid or tau protein, respectively. Accordingly, in some embodiments, following administration of any of the disclosed compositions, an amyloid PET and/or tau PET of the subject or subject’s sample is performed.

[0042] Accordingly, further provided herein are methods of treatment and prevention comprising administration of any of the disclosed compounds. Further provided herein are uses of these compounds as medicaments for treatment or prophylaxis of a neurodegenerative disorder, such as Alzheimer’s Disease. In certain embodiments, a compound of Formula (I), or pharmaceutically acceptable salt thereof, is used in the preparation of a medicament for: (a) therapy (e.g., of the human body), (b) medicine, (c) activation of TREM2 receptor, (d) treatment or prevention of Alzheimer’s Disease and/or neurological symptoms thereof, (e) treatment or prevention of dementia, or (I) treatment, prevention of, or delay in the onset or progression of Alzheimer’s Disease, dementia, and/or neurological symptoms thereof. In certain embodiments, a compound of Formula (II), or pharmaceutically acceptable salt thereof, is used in the preparation of a medicament for: (a) therapy (e.g. of the human body), (b) medicine, (c) activation of TREM2 receptor, (d) treatment or prevention of Alzheimer’s Disease and/or neurological symptoms thereof, (e) treatment or prevention of dementia, or (f) treatment, prevention of, or delay in the onset or progression of Alzheimer’s Disease, dementia, and/or neurological symptoms thereof. In certain embodiments, a compound of Formula (III), or pharmaceutically acceptable salt thereof, is used in the preparation of a medicament for: (a) therapy (e.g., of the human body), (b) medicine, (c) activation of TREM2 receptor, (d) treatment or prevention of Alzheimer’s Disease and/or neurological symptoms thereof, (e) treatment or prevention of dementia, or (f) treatment, prevention of, or delay in the onset or progression of Alzheimer’s Disease, dementia, and/or neurological symptoms thereof. In these uses, the compounds of the present invention can optionally be employed in combination with one or more other active agents, such as an amyloid-beta targeting therapy or tau targeting therapy.

[0043] Further provided herein are methods for the treatment or prevention of a condition associated with a loss of function of human TREM2 in a subject (e.g., a human subject). In some embodiments, the condition associated with a loss of function of human TREM2 is dementia or cognitive impairment associated with Alzheimer’s Disease. In some embodiments, this condition is cognitive impairment associated with Parkinson’s Disease. In some embodiments, this condition is cognitive impairment associated with frontotemporal dementia, demyelination disorder, multiple sclerosis, Huntington’s disease, amyotrophic lateral sclerosis (ALS). tauopathy disease, Nasu-Hakola disease, or adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP).

[0044] The disclosure further provides methods of administering any of the disclosed compounds or compositions to a subject. In some embodiments, the disclosed methods may bring about in the subject (i) a reduction in amyloid plaques, (ii) an elevation in disease-associated microglia (DAM) mRNA expression, (iii) an elevation in lipid metabolism, (iv) an elevation in DAM chemotaxis, DAM proliferation, DAM pro-inflammatory cytokine secretion, or DAM phagocytic activity, and/or (v) a reduction in dystrophic neurites. In some embodiments, the disclosed methods bring about a reduction in amyloid plaques. In some embodiments, the presently disclosed methods bring about an elevation in disease-associated microglia (DAM) mRNA expression. In some embodiments, the presently disclosed methods bring about an elevation in lipid metabolism in the subject. In some embodiments, the disclosed methods bring about an elevation in DAM chemotaxis. DAM proliferation, DAM pro-inflammatory cytokine secretion, or DAM phagocytic activity, and/or a reduction in dystrophic neurites. [0045] In some aspects, provided herein are methods for the treatment or prophylaxis of abnormal motor symptoms associated with Parkinson's disease (including but not limited to bradykinesia, rigidity and resting tremor). Another embodiment provides a method for the treatment or prophylaxis of abnormal non-motor symptoms associated with Parkinson’s disease (including but not limited to cognitive dysfunction, autonomic dysfunction, emotional changes and sleep disruption), Lewy body dementia, and L-Dopa induced dyskinesias.

[0046] In some aspects, provided herein are methods for the treatment or prophylaxis of Alzheimer’s disease, mild cognitive impairment, the transition from mild cognitive impairment to Alzheimer’s disease, tauopathy disorders characterized by hyperphosphorylation of tau such as argyrophilic grain disease, Picks disease, corticobasal degeneration, progressive supranuclear palsy, inherited frontotemporal dementia, and Parkinson’s disease linked to chromosome 17. Additional indications include neuroinflammation, including neuroinflammation associated with of microglial inflammatory responses associated with multiple sclerosis, HIV-induced dementia, ALS, ischemic stroke, traumatic brain injury and spinal cord injury.

[0047] In some embodiments, any of the presently described compounds, compositions and methods provide a reduction in the likelihood or severity of symptoms of Alzheimer’s Disease in one or more subjects. In some embodiments, any of these compounds, compositions, and methods may provide a partial or complete reduction/inhibition of one or more symptoms. Any of the disclosed compounds, compositions, and methods may provide a partial or complete activation of the TREM2 receptor. Any of the disclosed compounds, compositions, and methods may provide a partial or complete reversal of a loss of function of human TREM2 in a subject. Any of the disclosed methods may provide an improvement in cognitive function following administration of any of the disclosed compounds.

DEFINITIONS

[0048] Listed below are definitions of various terms used herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0049] The terms used herein have their ordinary meaning and the meaning of such terms is independent at each occurrence thereof. That notwithstanding and except where stated otherwise, the following definitions apply throughout the specification and claims. Chemical names, common names, and chemical structures may be used interchangeably to describe the same structure. These definitions apply regardless of whether a term is used by itself or in combination with other terms, unless otherwise indicated. Hence, the definition of “alkyl’" applies to “alkyd"’ as well as the “alkyl"’ portions of “hydroxyalky l,” “haloalkyl,” “-O-alkyl,” etc.

[0050] As used herein, and throughout this disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0051] A “subject” is a human or non-human mammal. In one embodiment, a subject is a human. In another embodiment, a subject is a primate. In another embodiment, a subject is anon- human primate, e.g.. a monkey. In some embodiments, a subject is a rhesus monkey. In still another embodiment, a subject is a rodent, such as a rat. In some embodiments, the subject is a companion animal. In some embodiments, the subject is a laboratory⁷ animal.

[0052] The term “effective amount” as used herein, refers to an amount of compound and/or an additional therapeutic agent, or a composition thereof that is effective in agonizing the human TREM2 receptor and in producing the desired therapeutic, ameliorative, or preventative effect when administered to a subject suffering from a neurodegenerative disorder. In any of the combination therapies of the present disclosure, an effective amount can refer to each individual agent or to the combination as a whole, wherein the amounts of all agents administered are together effective, but wherein the component agent of the combination may not be present individually in an effective amount.

[0053] The terms “treating” or “treatment” as used herein with respect to a neurodegenerative disorder, includes inhibiting the severity of a neurodegenerative disorder, e.g., arresting or reducing the development of the neurodegenerative disorder or its clinical symptoms; or ameliorating or relieving symptoms of the neurodegenerative disorder, e.g, causing regression of the severity of the neurodegenerative disorder or its clinical symptoms. For example, the disclosed compounds, pharmaceutical compositions, and methods may be useful for arresting or reducing the development of, or relieving symptoms of. Alzheimer’s Disease or neurological conditions associated with Alzheimer’s Disease.

[0054] The terms “preventing,” or “prophylaxis,” as used herein with respect to a neurodegenerative disorder, encompasses impeding the development or progression of clinical symptoms of the disease, disorder, or condition in a mammal that may be exposed to or predisposed to the disease, disorder or condition but does not yet experience or display symptoms of the disease, and the like.

[0055] “Alky l”, as well as other groups having the prefix “alk”, such as alkoxy, and the like, means carbon chains which may be linear or branched, or combinations thereof, containing the indicated number of carbon atoms. For instance, a Ci-6 alkyl means an alkyl group having one (i. e. , methyl) up to 6 carbon atoms (i.e., hexyl). In particular embodiments, linear alkyl groups have 1-6 carbon atoms and branched alkyl groups have 3-7 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl and the like.

[0056] Alkoxy” and “alkyl-O-” are used interchangeably and refer to an alkyd group linked to oxygen. “Haloalkoxy” means an alkoxy that is mono-or multiple-halo-substituted. The bond to the parent group is through the oxygen atom of the group.

[0057] “Cycloalkyl” means a cyclic hydrocarbon radical. Unless otherwise specified, “cycloalkyl” refers to a saturated cycloalkyl. In particular embodiments, the cycloalkyl group has 3-12 carbon atoms, forming 1-3 carbocyclic rings, wherein cyclic systems having 2-3 rings can be fused. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, and the like. In certain embodiments, e.g., the cycloalkyl is fused to a hetereraryl ring, the cycloalkyl may be a non-aromatic, partially unsaturated ring.

[0058] “Fluoroalkyl” includes mono-substituted as well as multiple fluoro-substituted alky 1 groups, up to perfluoro substituted alkyl. For example, fluoromethyl, 1,1 -difluoroethyl, trifluoromethyl or 1,1,1,2,2-pentafluorobutyl are included. The bond to the parent group is through one of the carbon atoms of the alkyl component.

[0059] Bicyclic ring system” refers to two joined rings. “Tricyclic ring system” refers to three joined rings. “Tetracyclic ring system” refers to four joined rings. Heterocyclic and cycloalkyl rings may be fused, i.e., share two adjacent atoms, or “spirocyclic”. i.e., share only a single atom, or “bridged”, i.e., share three or more atoms with two bridgehead atoms being connected by a bridge containing at least one atom. Heteroaryl rings may be fused.

[0060] The term “halo,” as used herein, means -F, -Cl, -Br or -I. A particular class of interest of halo substituents for compounds of Formula (I)-(III), and embodiments thereof, is each of fluoro (— F) and chloro (-C1). The term “haloalkyl” refers to an alkyl group as defined above in which one or more of the hydrogen atoms have been replaced with halo (i.e., -F, -Cl, -Br and/or -I).

[0061] “Fluoroalkyl” includes mono-substituted as well as multiple fluoro-substituted alkyd groups, up to perfluoro substituted alkyl. For example, fluoromethyl, 1,1 -difluoroethyl, trifluoromethyl or 1,1,1,2,2-pentafluorobutyl are included. “Fluoroalkoxy” includes monosubstituted as well as multiple fluoro-substituted “alkoxy” groups as previously defined.

[0062] The term “substituted” means that one or more hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom’s normal valency under the existing circumstances is not exceeded, and that the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. By “stable compound’ or “stable structure” is meant a compound that is sufficiently robust to survive isolation to a useful degree of urity from a reaction mixture, and formulation into an efficacious therapeutic agent. When any substituent or variable (e.g., R¹) occurs more than one time in any constituent or in Formula (I)-(III), its definition on each occurrence is independent of its definition at every other occurrence, unless otherwise indicated. It should also be noted that any carbon as well as heteroatom with unsatisfied valences in the text, schemes, examples and tables herein is assumed to have the sufficient number of hydrogen atom(s) to satisfy the valences.

[0063] When a functional group in a compound is termed "protected", this means that the group is in modified form to preclude undesired side reactions at the protected site when the compound is subjected to a reaction. Suitable protecting groups will be recognized by those with ordinary skill in the art as well as by reference to standard textbooks such as. for example, T. W. Greene et al.. Protective Groups in Organic Synthesis (1991), Wiley, New York.

[0064] When a moiety is noted as being “optionally substituted” in Formulas (I)-(III) or any embodiment thereof, it means that Formulas (I)-(III) or ), or the embodiment thereof. encompasses both compounds that are substituted with the noted substituent (or substituents) on the moiety and compounds that do not contain the noted substituent (or substituents) on the moiety (i.e., wherein the moiety is unsubstituted). As one example, when R1 is a Ci-io alkyl group that can be optionally substituted with halo, then R1 can be Ci-io alkyl or Ci-io haloalkyl. [0065] When any variable (e.g., Rl, R^x, R^Y) occurs more than one time in any constituent or in Formulas (I)-(III), or in any other formula depicting and describing compounds of the present invention, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds. Unless expressly stated to the contrary, substitution by a named substituent is permitted on any atom in a ring (e.g. , cycloalkyl, aryl, or heteroaryl) provided such ring substitution is chemically allowed and results in a stable compound.

[0066] Unless expressly stated to the contrary, all ranges cited herein are inclusive of the recited endpoints and independently combinable. For example, the range of “between about 0.5 and about 95 percent” is inclusive of the endpoints (about) 0.5 percent and (about) 95 percent, and all intermediate values. The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value: they are sufficiently imprecise to include values approximating these ranges and/or values.

[0067] As used herein, "heteroaryl” or “heteroaromatic ring” refers to aromatic monocyclic, bicyclic, tricyclic, or tetracyclic ring structures in which one or more atoms in the ring, the heteroatom(s), is an element other than carbon. Heteroatoms are typically O. S, or N atoms. Examples of heteroaryl groups include pyrazolyl, oxadiazolonyl, pyridinyl, pyrimidinyl, pyrrolyl, pyridazinyl, isoxazolyl, thiazolyl, oxazolyl, indolyl, benzoxazolyl, benzothiazolyl, and imidazolyl.

[0068] As used herein, a “heteroaromatic” ring is a carbon-containing ar l ring may contain 1, 2, 3 or 4 heteroatoms. For instance, a “heteroaromatic” ring may contain one or more nitrogen atoms (e.g., 1 to 3 nitrogen atoms), one or more oxygen atoms, or one or more sulfur atoms. Heteroaromatic rings may be herein expressed using subscripts that denote the total number of atoms making up the ring. In some embodiments, a heteroaromatic ring may have 5-12 ring atoms wherein each atom is selected from carbon, nitrogen, oxygen and sulfur. For instance, a 6- membered heteroaryl substituent may contain 4 carbon atoms and two oxygen atoms.

[0069] Heterocycloalkyl” or “heterocyclic ring” or “heterocycle” or “heterocyclyl” (when the ring is bonded to a parent moiety) means a non-aromatic monocyclic, bicyclic, tricyclic or tetracyclic ring system comprising about 3 to about 17 ring atoms, preferably about 3 to about 10 ring atoms, in which one or more of the atoms in the ring system is an element other than carbon, for example, nitrogen, oxygen, phosphorus or sulfur, alone or in combination. There are no adjacent oxygen and/or sulfur atoms present in the ring system. In some embodiments, heterocycloalkyls contain about 5 to about 6 ring atoms. The prefix aza, oxa, phospha or thia before the heterocyclyl root name means that at least a nitrogen, oxygen, phosphorus or sulfur atom respectively is present as a ring atom. Non-limiting examples of suitable monocyclic heterocyclyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1 ,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, and the like.

“Spiroheterocycloalkyl” refers to a fused ring system in which the rings share only a single atom and at least one of the rings is a heterocycloalkyl.

[0070] In some embodiments, the compounds disclosed herein contain a heteroaryl substituent containing one nitrogen atom. It is also to be understood that any range cited herein includes within its scope all of the sub-ranges within that range. Thus, for example, a “heterocyclic” ring is intended to include as aspects thereof, heterocyclic rings containing 2 to 4 heteroatoms, 3 or 4 heteroatoms, 1 to 3 heteroatoms. 2 or 3 heteroatoms, 1 or 2 heteroatoms, 1 heteroatom. 2 heteroatoms. 3 heteroatoms, or 4 heteroatoms. For instance, the ring may contain one or more nitrogen atoms (e.g, 1 to 3 nitrogen atoms), one or more oxygen atoms, or one or more sulfur atoms. Any of the cycloalkyl, heterocyclyl, aryl and heteroaryl groups described herein may be optionally substituted with one or more groups. As used herein, “optionally substituted with one to five groups” is intended to include as aspects thereof, the cycloalkyl, heterocyclyl. aryl or heteroaryl substituted with 1 to 5 substituents, 2 to 5 substituents, 3 to 5 substituents, 4 to 5 substituents, 5 substituents, 1 to 4 substituents, 2 to 4 substituents, 3 to 4 substituents, 4 substituents, 1 to 3 substituents, 2 to 3 substituents, 3 substituents, 1 to 2 substituents, 2 substituents, and 1 substituent. Likewise, as used herein, ‘'optionally substituted with one to three groups” is intended to include as aspects thereof, the cycloalkyl, heterocyclyl, aryl or heteroaryl substituted with 1 to 3 substituents, 2 to 3 substituents, 3 substituents, 1 to 2 substituents.

[0071] As used herein, the term “composition” is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results from combination of the specified ingredients in the specified amounts.

[0072] The term “salt(s)”, as used herein, denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. In addition, when a compound contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions ("inner salts") may be formed and are included within the term "salt(s)" as used herein. Compounds can be administered in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to a salt which is not biologically or otherwise undesirable (e.g.. is neither toxic nor otherwise deleterious to the recipient thereof).

[0073] The compounds of Formulas (I)-(III), and pharmaceutically acceptable salts thereof, which contain one or more basic groups, i.e., groups which can be protonated, can be used according to the invention in the form of their acid addition salts with inorganic or organic acids as, for example but not limited to, salts with hydrogen chloride, hydrogen fluoride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, benzenesulfonic acid, methanesulfonic acid, /7-toluenesulfonic acid, naphthalenedisulfonic acids, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfaminic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, etc. [0074] In some embodiments, one or more N atoms (e.g., an N atom in a heteroaryl or heterocyclyl ring, or an N atom in an NH2 group) of any of the compounds of Formula (I) is protonated in a salt form. In some embodiments, an N atom that is a ring member of a heteroaryl or heterocyclyl ring of any of the compounds of Formulas (I)-(III) is protonated in a salt form. In some embodiments, this N atom is protonated in a salt form with trifluoroacetate. If the compounds of Formulas I and/or II simultaneously contain acidic and basic groups in the molecule the invention also includes, in addition to the salt forms mentioned, inner salts or betaines (zwitterions). Salts can be obtained from the compounds of Formulas I and/or II by customary methods which are known to the person skilled in the art, for example by combination with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange from other salts. The present disclosure also includes all salts of the compounds of Formulas (I)-(III) which, owing to low physiological compatibility, are not directly suitable for use in pharmaceuticals but which can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.

[0075] The present disclosure encompasses any composition comprised of a compound of Formulas I and/or II or a compound that is a salt thereof, including for example but not limited to, a composition comprised of said compound associated together with one or more additional molecular and/or ionic component(s) which may be referred to as a “co-crystal.” The term “co- crystaf’ as used herein refers to a solid phase (which may or may not be cry stalline) wherein two or more different molecular and/or ionic components (generally in a stoichiometric ratio) are held together by non-ionic interactions including but not limited to hydrogen-bonding, dipole-dipole interactions, dipole-quadrupole interactions or dispersion forces (van der Waals). There is no proton transfer between the dissimilar components and the solid phase is neither a simple salt nor a solvate. A discussion of co-crystals can be found, e.g., in Aitipamula et al., Crystal Growth and Design, 2012, 12 (5), pp. 2147-2152.

[0076] Compounds of the present invention may exist in amorphous form and/or one or more cry stalline forms, and as such all amorphous and cry stalline forms and mixtures thereof of the compounds of Formulas (I)-(III) are intended to be included within the scope of the present invention. In addition, some of the compounds of the instant invention may form solvates with water (i.e., a hydrate) or common organic solvents. Such solvates and hydrates, particularly the pharmaceutically acceptable solvates and hydrates, of the instant compounds are likewise encompassed within the scope of this invention, along with un-solvated and anhy drous forms. Accordingly, the compounds within the generic structural formulas, embodiments and specific compounds described and claimed herein encompass salts, all possible stereoisomers and tautomers, physical forms (e.g., amorphous and crystalline forms), solvate and hydrate forms thereof and any combination of these forms, as well as the salts thereof.

Salts. Solvates and Stereoisomers

[0077] Solvates of the disclosed compounds of Formulas (I)-(III) are contemplated herein. One or more compounds of the invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms. "Solvate" means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves vary ing degrees of ionic and covalent bonding, including hydrogen bonding. In certain situations, the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate" encompasses both solution-phase and isolatable solvates. Non-limiting examples of solvates include ethanolates, methanolates, and the like. A "hydrate" is a solvate wherein the solvent molecule is water.

[0078] One or more compounds of Formulas (I)-(III) may optionally be converted to a solvate. Preparation of solvates is generally known. Thus, for example, M. Caira et al., J. Pharmaceutical Sci., 93(3), 601-611 (2004) describe the preparation of the solvates of the antifungal fluconazole in ethyl acetate as well as from water. Similar preparations of solvates, hemisolvate, hydrates and the like are described by E. C. van Tonder et al., AAPS PharmSciTech.. 5(1). article 12 (2004); and A. L. Bingham et al., Chem. Commun., 603-604 (2001). A typical, non-limiting, process involves dissolving the compound in desired amounts of the desired solvent (organic or water or mixtures thereol) at a higher than room temperature, and cooling the solution at a rate sufficient to form crystals which are then isolated by standard methods. Analytical techniques such as, for example IR spectroscopy, show the presence of the solvent (or water) in the crystals as a solvate (or hydrate).

[0079] The compounds of Formulas (I)-(III) can form salts which are also within the scope of this invention. In some embodiments, the salt is a pharmaceutically acceptable salt. In another embodiment, the salt is other than a pharmaceutically acceptable salt. Salts of the compounds of Formulas (I)-(III) may be formed, for example, by reacting the compound with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.

[0080] Exemplary acid addition salts include acetates, ascorbates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, fumarates, hydrochlorides, hydrobromides, hydroiodides, lactates, maleates, methanesulfonates, naphthalenesulfonates, nitrates, oxalates, phosphates, propionates, salicylates, succinates, sulfates, tartarates, thiocyanates, toluenesulfonates (also known as tosylates) and the like. Additionally, acids which are generally considered suitable for the formation of pharmaceutically useful salts from basic pharmaceutical compounds are discussed, for example, by P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley -VCH; S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food & Drug Administration, Washington, D.C. on their website). These disclosures are incorporated herein by reference thereto.

[0081] Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts with organic bases (for example, organic amines) such as dicyclohexylamine, t-butyl amine, choline, and salts with amino acids such as arginine, lysine and the like. Basic nitrogencontaining groups may be quartemized with agents such as lower alkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfates), long chain halides (e.g., decyl, lauryl, and stearyl chlorides, bromides and iodides), arylalkyl halides (e.g., benzyl and phenethyl bromides), and others.

[0082] All such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.

[0083] This disclosure includes individual diastereomers, particularly epimers, i.e., compounds having the same chemical formula but which differ in the spatial arrangement around a single atom. This disclosure also includes mixtures of diastereomers, particularly mixtures of epimers, in all ratios. This disclosure encompasses compounds of Formulas (I)-(III) having either the R or S stereo-configuration at an asymmetric center and at any additional asymmetric centers that may be present in a compound of Formula (I), as w ell as stereo-isomeric mixtures thereof.

Embodiments of this disclosure also include a mixture of enantiomers enriched with 51% or more of one of the enantiomers, including for example 60% or more, 70% or more, 80% or more, or 90% or more of one enantiomer. A single epimer is preferred. An individual or single enantiomer refers to an enantiomer obtained by chiral synthesis and/or using generally known separation and purification techniques, and which may be 100% of one enantiomer or may contain small amounts (e.g., 10% or less) of the opposite enantiomer. Thus, individual enantiomers are a subject of this disclosure in pure form, both as levorotatory⁷ and as dextrorotatory⁷ antipodes, in the form of racemates and in the form of mixtures of the two enantiomers in all ratios. In the case of a cis/trans isomerism this disclosure includes both the cis form and the trans form as well as mixtures of these forms in all ratios.

[0084] The preparation of individual stereoisomers can be carried out, if desired, by separation of a mixture by customary' methods, for example by chromatography or cry stallization, by the use of stereochemically uniform starting materials for the synthesis or by stereoselective synthesis. Optionally a derivatization can be carried out before a separation of stereoisomers. The separation of a mixture of stereoisomers can be carried out at an intermediate step during the synthesis of a compound of Formulas (I)-(III), or it can be done on a final racemic product. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing a stereogenic center of known configuration. Alternatively, absolute stereochemistry may be determined by Vibrational Circular Dichroism (VCD) spectroscopy analysis. The present disclosure includes all such isomers, as well as salts, solvates (which includes hydrates), and solvated salts of such racemates, enantiomers, diastereomers and tautomers and mixtures thereof. [0085] Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (<?.g., chiral auxiliary such as a chiral alcohol or Mosher’s acid chloride), separating the diastereomers and converting (e.g, hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Sterochemically pure compounds may also be prepared by using chiral starting materials or by employing salt resolution techniques. Also, some of the compounds of Formulas (I)-(III) may be atropisomers (e.g.. substituted biaryls) and are considered as part of this invention. Enantiomers can also be directly separated using chiral chromatographic techniques. [0086] It is also possible that the compounds of Formulas (I)-(III) may exist in different tautomeric forms, and all such forms are embraced within the scope of the invention. For example, each of the keto/enol and imine/enamine tautomeric forms of the disclosed compounds are encompassed w ithin embodiments of the disclosed compounds that depict either form individually. As another example, both the hydroxypyridine and pyridinone forms of oxosubstituted pyridine substituents are encompassed within embodiments of the disclosed compounds that depict either form individually.

[0087] Unless otherwise indicated, all stereoisomers (for example, geometric isomers, optical isomers and the like) of the present compounds (including those of the salts, solvates, hydrates, esters and prodrugs of the compounds as well as the salts, solvates and esters of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this invention. If a compound incorporates a double bond or a fused ring, both the cis- and transforms, as well as mixtures, are embraced within the scope of the invention.

[0088] When a substituent on a chiral carbon atom is depicted without specific stereochemistry (by using a straight-line bond to a chiral center), it is to be understood that both the alpha and beta configurations of said substituent group are to be considered part of the present invention. It is understood that a chiral center in a compound may exist in the S or R absolute configuration, or as a mixture of both. Within a molecule, each bond drawn as a straight line from a chiral center includes both the R and stereoisomers as well as mixtures thereof. An asterisk denotes a stereocenter in a single configuration, either R or S. Absolute stereochemistry of separate stereoisomers in the examples and intermediates may not have been determined unless stated otherwise in an example or explicitly in the nomenclature. Otherwise, for compounds in the Examples that contain a chiral center, isomer mixtures may have been separated, yielding the particular stereoisomer(s) depicted.

[0089] Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present invention can have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms "salt", "solvate" and the like, is intended to apply equally to the salt and solvate of enantiomers, stereoisomers, rotamers, tautomers or racemates of the disclosed compounds.

[0090] In the compounds of Formulas (I)-(III), the atoms may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is meant to include all suitable isotopic variations of the compounds of generic Formulas (I)-(III). For example, different isotopic forms of hydrogen (H) include protium 6¹ H ) and deuterium (²H). Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may provide certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples. Isotopically-enriched compounds of Formulas (I)-(III) can be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the Schemes and Examples herein using appropriate isotopically-enriched reagents and/or intermediates. In one embodiment, a compound of Formulas (I)-(III) has one or more of its hydrogen atoms replaced with deuterium.

[0091] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well-known and commonly employed in the art.

[0092] As used herein, the articles “a’' and “an” refer to one or to more than one (i. e.. to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0093] As used herein, the term “about” when modifying a quantitative term refers to plus or minus 10% of the value it modifies (rounded up to the nearest whole number if the value is not sub-dividable, such as a number of molecules).

[0094] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 100 mg to 1500 mg” is inclusive of the endpoints, 100 mg and 1500 mg, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and/or values.

[0095] As used herein, the term “comprising” may include the embodiments “consisting of’ and “consisting essentially of.” The terms “comprise(s),” “include(s),” “having,” “has,” “may,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named ingredients/steps and permit the presence of other ingredients/steps. However, such description should be construed as also describing compositions or processes as “consisting of’ and “consisting essentially of’ the enumerated components, which allows the presence of only the named components or compounds, along with any acceptable carriers or fluids, and excludes other components or compounds.

TREM2 AGONISM AND NEURODEGENERATIVE CONDITIONS

[0096] Impairment in TREM2 receptor function has been linked to several human diseases. For instance, mutations in both TREM2 and DAP 12 have been linked to the autosomal recessive disorder Nasu-Hakola Disease, which is characterized by bone cysts, muscle wasting and demyelination phenotypes. Guerreiro et al. 2013. Variants in the TREM2 gene have been linked to increased risk for Alzheimer’s disease (AD) and other forms of dementia including frontotemporal dementia. (Jonsson et al. 2013, Guerreiro & Lohmann et al. 2013, and Jay & Miller et al. 2015.) In particular, the R47H variant has been identified in genome-wide studies as being associated with increased risk for late-onset AD with an overall adjusted odds ratio (for populations of all ages) of 2.3, second only to the strong genetic association of ApoE to AD. The R47H mutation resides on the extracellular IgV-set domain of the TREM2 protein and has been shown to impact lipid binding and uptake of apoptotic cells and amyloid-P (Wang et al. 2015; Yeh et al. 2016). suggestive of a loss-of-function linked to disease. Further, postmortem comparison of AD patients’ brains with and without the R47H mutation are supportive of a novel loss-of-microglial barrier function for the carriers of the mutation, with the R47H carrier microglia putatively demonstrating a reduced abi 1 i ty to compact plaques and limit their spread. Yuan et al. 2016. Impairment in microgliosis has been reported in animal models of prion disease, multiple sclerosis, and stroke, suggesting that TREM2 may play an important role in supporting microgliosis in response to pathology' or damage in the CNS. Ulrich and Holtzman 2016.

[0097] In addition, knockdown of TREM2 has been shown to aggravate a-synuclein-induced inflammatory responses in vitro and exacerbate dopaminergic neuron loss in response to adeno- associated viral (AAV) vectors encoding synuclein in vivo (a model of Parkinson's disease), suggesting that impaired microglial TREM2 signaling exacerbates neurodegeneration by modulating microglial activation states. Guo et al. 2019. A variety of animal models also suggest that Toll-Like Receptor (TLR) signaling is important in the pathogenesis of Rheumatoid Arthritis (RA) via persistent expression of pro-inflammatory cytokines by macrophages. Signaling through TREM2/DAP12 inhibits TLR responses by reducing MAPK (Erkl/2) activation, suggesting that TREM2 activation may act as a negative regulator of TLR driven RA pathogenesis. Signaling through DAP10 and DAP12 also elicits recruitment of PI3K and SYK, which drive multiple downstream events resulting in Ca²⁺ mobilization and activation of MAPK- mediated cascades as well as other pathways.

[0098] In view of the data indicating that deficiency in TREM2 activity affects macrophage and microglia function, the TREM2 agonist compounds disclosed herein are of particular use in treating, preventing, and/or reducing the severity of disorders such as those described below, and in neurodegenerative disorders more generally.

[0099] TREM2 consists of a single-pass transmembrane domain, an extracellular stalk region, and extracellular immunoglobulin variable-like (IgV) domain responsible for ligand interaction (Kleinberger et al. Sci TranslMed, 2014). As TREM2 does not possess intracellular signal transduction-mediating domains, biochemical analysis has illustrated that interaction with adaptor proteins DAP 10 and DAP 12 mediate downstream signal transduction following ligand recognition (Peng et al. Sci Signal 2010; Jay et al. Mol Neurodegener , 2017). TREM2/DAP12 complexes in particular act as a signaling unit that can be characterized as pro-activation on microglial phenotypes in addition to peripheral macrophages and osteoclasts (Otero et al. J Immunol, 2012; Kobayashi et al. JNeurosci, 2016; Jaitin et al., Cell, 2019. In the CNS, signaling through TREM2 has been studied in the context of ligands such as phospholipids, cellular debris, apolipoproteins, and myelin (Wang et al. Cell, 2015; Kober and Brett, J Mol Biol, 2017;

Shirotani et al., Sci Rep, 2019). In mice lacking functional TREM2 expression or expressing a mutated form of the receptor, a core observation is blunted microglial responses to insults such as oligodendrocyte demyelination, stroke-induced tissue damage in the brain, and proteotoxic inclusions in vivo (Cantoni et al., Acta Neuropathol, 2015, Wu et al., Mol Brain, 2017).

[0100] In rodent models where TREM2 expression levels are elevated, brain amyloid pathology in the 5XF AD transgenic mice displayed reduced plaque volume and altered morphology (Lee et al. Neuron. 2018). The changes in immunohistological markers relating to brain amyloid pathology were also accompanied by an attenuated presence of dystrophic neurites when TREM2 was overexpressed. Id. As such, pharmacological agonism of TREM2 has gained interest in treating or preventing neurodegenerative disorders and conditions.

[0101] Genetic variation in the TREM2 locus has been associated with late onset Alzheimer's disease (“LOAD”) in human genome-wide association studies, linking a loss-of-receptor function to a gain in disease risk (Jonsson et al. N Engl J Med 2013, Sims et al. Nat Genet 2017). Genetic variation of other genes selectively expressed by microglia in the CNS, for example, CD33, PLCg2 and MS4A4A/6A have reached genome-wide significance for their association with LOAD risk (Hollingworth et al. Nat Genet 2011, Sims et al. Nat Genet 2017. Deming et al. Sci TranslMed 2019). Together, these genetic findings link together in a putative biochemical circuit that highlights the importance of microglial innate immune function in LOAD. Additionally, increase or elevation in the soluble form of TREM2 (“sTREM2”) in the cerebrospinal fluid (CSF) of human subjects is associated with disease progression and emergence of pathological hallmarks of LOAD including phosphorylated Tau (Suarez-Calvet et al. Mol Neurodegener 2019). Furthermore, natural history and human biology studies indicate that baseline sTREM2 levels in the CSF can stratify the rate of temporal lobe volume loss and episodic memory' decline in longitudinally monitored cohorts (Ewers et al. Sci Transl Med 2019). Additional studies have revealed that CSF sTREM2 levels are elevated in the earlier course of AD and attenuated in the dementia stage (Liu et al., 2018), and are elevated following brain amyloidosis, CSF phosphorylation, and total tau protein increases (Halaas et al., 2020).

[0102] Accordingly, provided herein are methods of administering any of the disclosed compounds, or a pharmaceutically acceptable salt thereof, for treatment or prevention of LOAD. Further provided herein are methods for treatment or prevention of a condition associated with a loss of function of human TREM2, human DAP12 or human DAP10. Further provided herein are methods for reducing the amount of sTREM2 in the CSF of a subject.

[0103] In addition to human genetic evidence supporting a role of TREM2 in LOAD, homozygous loss-of-function mutations in TREM2 are causal for an early onset dementia syndrome known as Polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy (“PLOSL’') or Nasu-Hakola disease (“NF ID") (Golde et al. Alzheimers Res Tuer 2013. Dardiotis et al. Neurobiol Aging 2017). This progressive neurodegenerative disease ty pically manifests in the third decade of life and is pathologically characterized by loss of myelin in the brain concomitant with gliosis, unresolved neuroinflammation, and cerebral atrophy . Typical neuropsychiatric presentations are often preceded by osseous abnormalities, such as bone cysts and loss of peripheral bone density (Bianchin et al. Cell Mol Neurobiol 2004; Madry et al. Clin Orthop Relat Res 2007, Bianchin et al. Nat Rev Neural 2010). Given that osteoclasts of the myeloid lineage are also known to express TREM2, the PLOSL-related symptoms of wrist and ankle pain, sw elling, and fractures indicate that TREM2 may act to regulate bone homeostasis through defined signaling pathways that parallel the microglia in the CNS (Paloneva et al. J Exp Med 2003, Otero et al. J Immunol 2012). The link between TREM2 function and PLOSL/NHD and other adult-onset leukoencephalopathies has illustrated the importance of the receptor in sustaining key aspects of myeloid cell function in the human body. [0104] Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP), previously recognized as hereditary diffuse leukoencephalopathy with axonal spheroids (HDLS) or pigmentary orthochromatic leukodystrophy (POLD). is an autosomal-dominant central nervous system disease that manifests in the form of variable behavioral, cognitive and motor function changes in patients suffering from the disease. ALSP is characterized by patchy cerebral white matter abnormalities visible by magnetic resonance imaging. However, the clinical symptoms and MRI changes are not specific to ALSP and are common for other neurological conditions, including Nasu-Hakola disease (NHD) and AD, making diagnosis and treatment of ALSP very difficult. Recent studies have discovered that ALSP is a Mendelian disorder in which patients carry a heterozygous loss of function mutation in the kinase domain of CSF-1R, suggesting a reduced level of signaling on the macrophage colony-stimulating factor (M-CSF) / CSF-1R axis. CSF-1R is a macrophage colony-stimulating factor (M-CSF), which regulates the survival, proliferation, differentiation and function of mononuclear phagocytic cells, including microglia of the central nervous sy stem.

[0105] Accordingly, provided herein are methods of administering any of the disclosed compounds, or a pharmaceutically acceptable salt thereof, for treatment or prevention of PLOSL. Further provided herein are methods of administering any of the disclosed compounds for treatment or prevention of NHD. Further provided herein are methods of administering any of the disclosed compounds for treatment or prevention of adult-onset leukoencephalopathy, with or without axonal spheroids and pigmented glia. Further provided herein are methods of administering any of the disclosed compounds for treatment or prevention of ALSP. [0106] Further provided herein are methods of administering any of the disclosed compounds, or a pharmaceutically acceptable salt thereof, for treatment or prevention of argyrophilic grain disease. Picks disease, corticobasal degeneration, progressive supranuclear palsy, inherited frontotemporal dementia, Parkinson’s disease linked to chromosome 17, HIV-induced dementia or neuroinflammation.

[0107] In addition to the CNS, TREM2 is expressed in myeloid lineage cells of the liver. In some embodiments, these compounds, or a pharmaceutically acceptable salt thereof, may be useful to treat or prevent a liver disease associated with impaired TREM2 function. In some embodiments, these compounds are useful to treat or prevent alcoholic liver disease (ALD) or non-alcoholic steatohepatitis (NASH).

[0108] In various embodiments of any of the disclosed methods, the subject suffers from the disorder or condition being treated. In various embodiments, the subject is human.

[0109] In other embodiments, the subject is a non-human mammal. In some embodiments, the subject is a companion animal. In some embodiments, the subject is a laboratory animal. In some embodiments, the subject is a rodent. In some embodiments, the subject is a non-human primate.

FORMULATIONS

[0110] When administered to a subject, any of the disclosed compounds of Formulas (I)-(III), or a pharmaceutically acceptable salt thereof, may be administered as a component of a composition that comprises a pharmaceutically acceptable carrier. The present disclosure provides pharmaceutical compositions comprising an effective amount of at least one of the disclosed compounds and a pharmaceutically acceptable carrier. In the pharmaceutical compositions and methods of the present invention, the active ingredients will typically be administered in admixture with suitable carrier materials selected with respect to the intended form of administration, i.e., oral tablets, capsules (either solid-filled, semi-solid filled or liquid filled), powders for constitution, oral gels, elixirs, dispersible granules, syrups, suspensions, and the like, and consistent with conventional pharmaceutical practices. For example, adapted for oral administration in the form of tablets or capsules, any of the disclosed compounds may be combined with any pharmaceutically acceptable inert carrier. Solid form preparations include powders, tablets, dispersible granules, capsules, sachets and suppositories. Tablets, powders, sachets and capsules may be suitable for oral administration. Powders and tablets may be comprised of between about 0.5 and about 95 percent of any of the disclosed pharmaceutical compositions. [OHl] Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, com starch, or alginic acid; binding agents, for example starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncoated, or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as gly ceryl monostearate or glyceryl distearate may be employed. They may also be coated to form osmotic therapeutic tablets for control release. Oral tablets may also be formulated for immediate release, such as fast melt tablets or wafers, rapid dissolve tablets or fast dissolve films. [0112] Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive oil.

[0113] Moreover, when desired or needed, suitable binders, glidants, lubricants, disintegrating agents and coloring agents may also be incorporated in the composition, particularly in formulations for oral administration. The compositions may be formulated for extended or controlled release. In other embodiments, the compositions are formulated for immediate or modified release.

[0114] In particular embodiments, the compositions of the present invention may be formulated in extended dosing, or sustained release, forms to provide a rate-controlled release of any one or more of the components or active ingredients to optimize therapeutic effects, i.e.. TREM2 activation. Suitable dosage forms for sustained release include long-acting injectable and implant dosage forms. Other suitable dosage forms for sustained release include layered tablets containing layers of varying disintegration rates or controlled release polymeric matrices impregnated with the active components and shaped in tablet form or capsules containing such impregnated or encapsulated porous polymeric matrices.

[0115] Solid preparations suitable for oral administration (e.g, powders, pills, capsules and tablets) can be prepared according to techniques known in the art and can employ such solid excipients as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like. These preparations may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Liquid preparations suitable for oral administration (e.g., suspensions, syrups, elixirs and the like) can be prepared according to techniques known in the art and can employ any of the usual media such as water, glycols, oils, alcohols and the like.

[0116] In some embodiments, any of the disclosed pharmaceutical compositions comprise pharmaceutically acceptable carriers that are suitable or adapted for administration to the subject by injection. In some embodiments, these carriers are adapted for long-action injection. In some embodiments, these carriers are liquid form preparations that include solutions, suspensions, emulsions, or nano-emulsions for intramuscular or subcutaneous administration. In some embodiments, any of the disclosed pharmaceutical compositions are adapted for long-acting injectable formulations.

[0117] Any of the disclosed compositions may comprise pharmaceutically acceptable carriers that are suitable or adapted for administration parenterally, including subcutaneous, intravenous, intramuscular, intraperitoneal or intrastemal injection, or other infusion techniques (one or more injections or infusions may be administered at each dosing interval as needed to deliver the appropriate amount of active agent), in the form of a unit dosage of a pharmaceutical composition containing an effective amount of the compound and conventional pharmaceutically acceptable carriers, adjuvants and vehicles for the treatment of a subject suffering from a neurodegenerative disorder. The compositions may also be administered parenterally via an implantable drug delivery composition or device adapted to provide an effective amount of the compound over an extended period of time. In some embodiments, the composition is administered parenterally once per month, once per every three months, once per every six months, or once per every twelve months.

[0118] In some embodiments, the disclosed compositions are adapted for intramuscular administration. In some embodiments, the disclosed compositions are adapted for subcutaneous administration. In some embodiments, the disclosed compositions are adapted for intravenous administration. In some embodiments, the disclosed compositions are adapted for intraperitoneal administration. In some embodiments, the disclosed compositions are adapted for intracerebroventricular (ICV), intrathecal, or intracistemal administration. In some embodiments, the disclosed compositions may be adapted for inhalation spray, intranasal, vaginal, rectal, sublingual, buccal or topical routes of administration.

[0119] Parenteral compositions can be prepared according to techniques known in the art. These compositions may employ sterile water as a carrier and optionally other ingredients. A continuous dosing regimen may be used for subjects suffering from a neurodegenerative disease, such as Alzheimer’s Disease. Any of the disclosed pharmaceutical preparations for parenteral inj ection may comprise solutions, suspensions or emulsions that may include water, a suspending agent, a viscosity modifier, a tonicity modifier, and/or a pH modifier.

[0120] Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral or parenteral administration. Such liquid forms include solutions, suspensions, emulsions and nano-emulsions. Parenteral compositions can be prepared according to techniques known in the art and typically employ sterile water as a carrier and optionally other ingredients, such as a solubility aid. Injectable solutions can be prepared according to methods known in the art wherein the carrier comprises a saline solution, a glucose solution or a solution containing a mixture of saline and glucose. Implantable compositions can be prepared according to methods known in the art wherein the carrier comprises the active chemical ingredient with polymers and suitable excipients, or utilizing an implantable device for drug delivery. Further description of methods suitable for use in preparing pharmaceutical compositions for use in the present disclosure and of ingredients suitable for use in said compositions is provided in Remington - The Science and Practice of Pharmacy, 22nd Edition, published by Pharmaceutical Press and Philadelphia College of Pharmacy’ at University of the Sciences. 2012, ISBN 978 0 85711-062-6 and prior editions.

[0121] Formulations of compounds of Formula (I) that result in drug supersaturation and/or rapid dissolution may be utilized to facilitate oral drug absorption. Formulation approaches to cause drug supersaturation and/or rapid dissolution include, but are not limited to, nanoparticulate systems, amorphous systems, solid solutions, solid dispersions, and lipid systems. Such formulation approaches and techniques for preparing them are known in the art. For example, solid dispersions can be prepared using excipients and processes as described in reviews (e.g., A.T.M. Serajuddin, J Pharm Sci, 88: 10. pp. 1058-1066 (1999)). Nanoparticulate systems based on both attrition and direct synthesis have also been described in reviews such as Wu et al. (F. Kesisoglou, S. Panmai, Y. Wu, Advanced Drug Delivery Reviews, 59:7 pp. 631-644 (2007)).

[0122] The compounds of Formula (I) may be administered in a dosage range of, e.g., 1 to 20 mg/kg, or 1 to 10 mg/kg. or about 5 mg/kg of mammal (e.g., human) body weight per day, or at other time intervals as appropriate, in a single dose or in divided doses. The compounds of Formula (I) may be administered in a dosage range of 0.001 to 2000 mg per day in a single dose or in divided doses. Examples of dosage ranges are 0.01 to 1500 mg per day, or 0.1 to 1000 mg per day, administered orally or via other routes of administration in a single dose or in divided doses. [0123] For oral (e.g., tablets or capsules) or other routes of administration, the dosage units may contain 100 mg to 1500 mg of the active ingredient, for example but not limited to 0. 1 mg to about 1500 mg of the active ingredient, for example but not limited to 0.1, 0.25, 0.5, 1, 2, 2.5, 5, 10, 15, 20, 25, 40, 50, 75, 100, 125, 150, 175, 200, 225, 250, 500, 1000, 1250, or 1500 milligrams of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. Furthermore, the compound may be formulated in oral formulations for immediate or modified release such as extended or controlled release. When the compound of Formula (I) is administered as a salt, reference to an amount of the compound in milligrams or grams is based on the free form (i.e., the non-salt form) of the compound.

[0124] Daily administration can be via any suitable route of administration but is preferably via oral administration and can be a single dose or more than one dose at staggered times (divided daily doses) within each 24-hour period. Each dose may be administered using one or multiple dosage units as appropriate. In some embodiments, the disclosed compounds and compositions are administered once daily. In some embodiments, the disclosed compounds and compositions are administered twice daily.

[0125] The specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability⁷ and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, the effect of other drugs the subject is taking, the severity of the particular condition, and the host undergoing therapy. In some cases, depending on the potency of the compound or the individual response, it may be necessary to deviate upwards or downwards from the given dose. The amount and frequency of administration will be regulated according to the judgment of the attending clinician considering such factors.

[0126] The compounds of this invention are also useful in the preparation and execution of screening assays for agonists of TREM2. Furthermore, the compounds of this invention may be useful in establishing or determining the binding site of other TREM2 agonists.

[0127] Additional embodiments of the present disclosure include the follow ing:

(a) a pharmaceutical composition comprising an effective amount of a compound of Formulas (I)-(III) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, and

(b) a pharmaceutical composition which comprises the product prepared by combining (e.g, mixing) an effective amount of a compound of Formulas (I)-(III) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. [0128] Additional embodiments of the present disclosure include each of the pharmaceutical compositions, methods and uses set forth in the preceding paragraphs, wherein the compound of Formula (I) or its salt employed therein in substantially pure. With respect to a pharmaceutical composition comprising a compound of Formulas (I)-(III) or its salt and a pharmaceutically acceptable carrier and optionally one or more excipients, it is understood that the term “substantially pure” is in reference to a compound of Formulas (I)-(III) or its salt per se.

COMBINATION THERAPIES

[0129] In some aspects, the present methods for treating or preventing a neurodegenerative disorder can further comprise the administration of one or more additional therapeutic agents that are not any of the disclosed compounds.

[0130] Examples of additional therapeutic agents that the compounds of this disclosure may also be combined with include, without limitation, treatments for Alzheimer’s disease, Parkinson's disease, rheumatoid arthritis, Nasu-Hakola disease, frontotemporal dementia, multiple sclerosis, demyelination disorder, Huntington’s disease, amyotrophic lateral sclerosis (ALS). tauopathy disease, adult-onset leukoencephalopathy, argyrophilic grain disease, Picks disease, corticobasal degeneration, progressive supranuclear palsy, HIV -induced dementia, or neuroinflammation.

[0131] Accordingly, in one embodiment, the present invention provides methods for treating a neurodegenerative disorder in a subject, the method comprising administering to the subject: (i) at least one compound of Formulas (I)-(III) (which may include two or more different compounds), or a pharmaceutically acceptable salt thereof, and (ii) at least one additional therapeutic agent that is other than any of the disclosed compounds, wherein the amounts administered are together effective to treat or prevent a neurodegenerative disorder.

[0132] In some embodiments, the additional therapeutic agent is a tau targeting therapy. In some embodiments, the additional therapeutic agent is an amyloid-0-targeting therapy. Administration of a tau targeting therapy or an amyloid-P targeting therapy has been shown to improve cognitive function, reverse (partially) the neurodegenerative effects of Alzheimer’s Disease, and/or treat or prevent Alzheimer’s Disease. Accordingly, provided herein are methods of administering any of the disclosed compounds wherein the methods further comprise a step of administering a tau targeting therapy or an amyloid-P targeting therapy to the subject. Such methods may be used to promote the treatment or prevention of a neurodegenerative disorder in a subject. Such methods may be used to promote the treatment or prevention of AD in a subject. [0133] When administering a combination therapy of the invention to a subject, therapeutic agents in the combination, or a pharmaceutical composition or compositions comprising therapeutic agents, may be administered in any order such as, for example, sequentially, concurrently, together, simultaneously and the like. The amounts of the various actives in such combination therapy may be different amounts (different dosage amounts) or same amounts (same dosage amounts). Thus, for non-limiting illustration purposes, a compound and an additional therapeutic agent may be present in fixed amounts (dosage amounts) in a single dosage unit (e.g., a capsule, a tablet and the like). In some embodiments, any of the disclosed compounds or a pharmaceutically acceptable salt thereof, is administered orally, and the additional therapeutic agent is further administered orally. When administered orally, the compound and the other agent(s) may be administered simultaneously (i.e.. in separate compositions one right after the other) or sequentially.

[0134] In some embodiments, a disclosed compound or a pharmaceutically acceptable salt thereof may be administered up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5, hours, 6 hours, 7 hours, 8 hours, 9 hours. 10 hours, 11 hours, 12 hours, 13 hours. 14 hours, 15 hours, 16 hours, 17 hours, or 18 hours before the therapeutic agent. In other embodiments, a disclosed compound or a pharmaceutically acceptable salt thereof may be administered up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5, hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, or 18 hours following the therapeutic agent.

[0135] In one embodiment, at least one compound is administered during a time when the additional therapeutic agent(s) exert their prophylactic or therapeutic effect, or vice versa. [0136] In another embodiment, at least one compound and the additional therapeutic agent(s) are administered in doses commonly employed when such agents are used as monotherapy. In another embodiment, at least one compound and the additional therapeutic agent(s) are administered in doses lower than the doses commonly employed when such agents are used as monotherapy.

[0137] In some embodiments, the additional therapeutic agent(s) is present in a pharmaceutical composition. In one embodiment, this composition is suitable for subcutaneous administration. In another embodiment, this composition is suitable for intramuscular administration. In another embodiment, this composition is suitable for oral administration. In still another embodiment, this composition is suitable for intravenous administration.

[0138] The at least one compound and the additional therapeutic agent(s) can act additively or synergistically. A synergistic combination may allow the use of lower dosages of one or more agents and/or less frequent administration of one or more agents of a combination therapy. A lower dosage or less frequent administration of one or more agents may lower toxicity of therapy without reducing the efficacy of therapy.

[0139] The doses and dosage regimen of the other agents used in the combination therapies of the present invention for the treatment or prevention of a neurodegenerative disorder may be determined by the attending clinician, taking into consideration the approved doses and dosage regimen in the package insert; the age. sex and general health of the subject; and the type and severity of the neurodegenerative disease or neurological symptoms thereof. When administered in combination, the compound and the other agent(s) may be administered simultaneously (i.e., in the same composition or in separate compositions one right after the other) or sequentially. This is particularly useful when the components of the combination are given on different dosing schedules, e.g., one component is administered once daily and another component is administered every six hours, or when the pharmaceutical compositions are different, e.g., one is a tablet and one is a capsule. A kit comprising the separate dosage forms is therefore advantageous.

EXAMPLES

[0140] The following examples are meant to be illustrative and should not be construed as further limiting. The contents of the figures and all references, patents, and published patent applications cited throughout this application are expressly incorporated herein by reference. [0141] The abbreviations used herein have the following tabulated meanings. Abbreviations not tabulated below have their meanings as commonly used unless specifically stated otherwise.

Abbreviations

GENERAL SYNTHETIC SCHEMES

[0142] The following reaction schemes and Examples illustrate methods which may be employed for the synthesis of the compounds of structural Formula (I) described in this invention. These reaction schemes and Examples are provided to illustrate the invention and are not to be construed as limiting the invention in any manner. All substituents are as defined above unless indicated otherwise. Several strategies based upon synthetic transformations known in the literature of organic synthesis may be employed for the preparation of the compounds of structural Formulas (I)-(III).

[0143] The compounds of the present disclosure can be prepared according to the procedures of the following Examples, using appropriate materials. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the invention. The Examples further illustrate details for the preparation of the compounds of the present disclosure. Those skilled in the art will readily understand that known variations of protecting groups, as well as of the conditions and processes of the following preparative procedures, can be used to prepare these compounds. It is also understood that whenever a particular chemical reagent is not commercially available, such a chemical reagent can be readily prepared following one of numerous methods described in the literature. Scheme 1-1

[0144] A general synthetic approach to some of the chemical matter is outlined in Scheme 1-1. In the first step, the ring formation is done through condensation with substituted 3-oxobutanoate under acidic reaction condition. In step two, amine can be installed via a palladium-catalyzed C- N coupling. In step three, the installation of the R³ is done through a Liebeskind-Srogl cross coupling reaction.

Scheme 1-2

[0145] Scheme 1-2 describes a general synthetic method to a chemical matter to install R³ group via Liebeskind Srogl cross coupling first then palladium-catalyzed C-N coupling or Sx Ar reaction.

Scheme 1-3

[0146] In Scheme 1-3, the amine can be installed through SxAr reaction then condensation reaction with substituted 3-oxobutanoate under acidic reaction condition. Lastly the installation of the R³ is done through Liebeskind Srogl cross coupling reaction.

Scheme 1-4

[0147] Scheme 1-4 describes the Liebeskind Srogl cross coupling on the monocyclic core, followed by SxAr with an amine and condensation reaction as the last step. Scheme 1-5

[0148] In Scheme 1-5, the C-C bond transformation can be achieved on either the bicyclic or tricyclic core through C-C bond sp2-sp3 cross coupling chemistry.

Scheme 1-6

[0149] Scheme 1-6 demonstrates an alternative way of C-C bond formation through Suzuki sp2-sp2 coupling, followed by reduction of the alkene double bond to afford the substituted linker compound.

Scheme 1-7

[0150] Scheme 1-7 describes the synthesis of the dihydrofuran tricyclic core through hetero aryl amine condensation with a corresponding keto-ester, followed by ester hydrolysis and cyclization to afford the tricyclic core. Next, sequential C-C coupling reactions are used to install R3 and the R3S/R4S linker substituent.

Scheme 1-8

[0151] An alternative way to synthesize the dihydrofuran linker compound is described in

Scheme 1-8 utilizing a two-steps Suzuki C-C cross coupling and reduction sequence.

SYNTHESIS OF COMMON INTERMEDIATES

Intermediate 1-1. Preparation of ( )-2-(l-methyl-177-pyrazol-4-yl)morpholine from SFC separation

rac. Peak 1

[0152] The (S)-2-(l -methyl- I H-pyrazol-4-yl)morpholine was obtained from commercially available racemic 2-(l -methyl- l/7-pyrazol-4-yl)morpholine and separated as Peak 1 by chiral SFC (tR = 3.5 min), while Peak 2 (tR = 4.5min) was (7?)-2-(l-methyl-lF7-pyrazol-4- yl)morpholine. The SFC separation conditions: Column & Dimensions: IG, 21x250 mm, 5 pm. UV Wavelength: 215 nm; Flow Rate: 80 ml/min; Modifier: 20% MeOH w/ 0.1% NH4OH; Outlet Pressure: 100 bar; Diluent: MeOH.

Intermediate 1-2, Preparation of 8-chloro-2.3-dimethyl-6-(methylthio)-4H-pyrimido[L6- a]pyrimidin-4-one

[0153] To a 40 mL vial containing 6-chloro-2-(methylthio)pyrimidin-4-amine (0.40 g, 2.3 mmol) and ethyl 2-methyl-3-oxobutanoate (1.6 g, 11 mmol) was added PPA (4.6 mL). The resulting mixture was heated in a hot plate at 130 °C overnight (caution: gas releases in first couple of hours). The reaction mixture was cooled to RT, and carefully quenched with 100 mL of sat. NaHCO? aq. (or until pH slightly above 7) and extracted with DCM (50 mL) two times. The combined organic fractions were washed with brine (40 mL) and dried over anhydrous MgSCL. The filtrate was concentrated under reduced pressure and the crude residue was purified by column chromatography on silica gel column, eluting with 10-30% EtOAc in Hexanes to give 8- chloro-2,3-dimethyl-6-(methylthio)-477-pyrimido[l,6-a]pyrimidin-4-one. MS (ESI) m/z: calc’d for C10H10CIN3OS [M+H]⁺: 256.0, found: [M+H]⁺: 256.0

Intermediate 1-3; Preparation of 3-chloro-l-(methylthio)-7.8- dihvdrocyclopentard1pyrimido[1.6-a1pyrimidin-9 -one

[0154] To a 40 mL vial containing 6-chloro-2-(methylthio)pyrimidin-4-amine (0.40 g, 2.3 mmol) and 2-(Methoxycarbonyl)cyclopentan-l-one (1.50 g, 10 mmol) was added PPA (4.6 mL). The resulting mixture was heated in a hot plate at 130 °C for 2 h (caution: gas releases). Upon cooling, the reaction mixture was treated with DCM (10 mL) and carefully neutralized with sat. NaHCCh aq. (120 mL) until the pH was slightly above 7. The organic layer was extracted with DCM (60 mL) two times and the combined organic extracts were then concentrated. The resulting residue was purified by column chromatography on silica gel column, eluting with 0% to 10% EtOAc in DCM to give the desired product, along with the 6-chloro-2- (methylthio)pyrimidin-4-amine starting material. The mixture was treated with 3 mL of EtOAc. The resulting precipitate was collected through filtration to yield 3-chloro-l-(methylthio)-7,8- dihydrocyclopenta[<7|pyrimido[l,6-a]pyrimidin-9(6LZ)-one which was used directly in the next step without further purification. MS (ESI) m/z: calc’d for C11H10CIN3OS [M+H]⁺: 268.0, found: [M+H]⁺: 268.1

Intermediate 1-4; Preparation of 3-chloro-l-(methylthio)-6.7.8.9-tetrahvdro-107f-pyrimido[6.1- bl quinazolin- 10-one

[0155] To a 40 mL vial containing 6-chloro-2-(methylthio)pyrimidin-4-amine (0.44 g, 2.5 mmol) and ethyl 2-oxocyclohexane-l -carboxylate (1.8 g, 11 mmol) was added PPA (5.0 mL). The resulting mixture was heated in a hot plate at 130 °C for 2 h (caution: gas releases). The reaction mixture was neutralized by the addition with sat. Nal ICO? aq. (120 mL, or until the pH was above 7) and extracted with DCM (60 mL) two times. The organic extracts were washed with brine (50 mL), dried over anhydrous MgSO-i. and concentrated. The crude product was treated with Et2O and the precipitate was collected to yield 3-chloro-l-(methylthio)-6,7,8,9- tetrahydro-1077-pyrimido[6,l -A| quinazolin- 10-one, which was used in the next step without further purification. MS (ESI) m/z: calc'd for C12H12CIN3OS [M+H]⁺: 282.0. found: [M+H]L 282.1

Intermediate 1-5: Preparation of (_lS)-6-(2-(l -methyl- 17f-pyrazol-4-yl)morpholino)-2-

[0156] To a 20 mL microwave vial containing 6-chloro-2-(methylthio)pyrimidin-4-amine (400 mg, 2.28 mmol) and (<S)-2-(l -methyl- 17f-pyrazol-4-yl)morpholine (476 mg, 2.85 mmol) in DMF (2.3 mL) was added triethylamine (1.15 g. 11.4 mmol). The reaction mixture was stirred in a microwave reactor at 170 °C for 30 min, then at 160° C for another 10 min. Upon completion, the reaction mixture was diluted w ith water (4 mL) and extracted with EtOAc (3 mL x2). The resulting concentrated residue was purified by column chromatography on silica gel, eluting with 40% to 60% EtOAc / EtOH (3/1) in hexanes to give the desired product (5)-6-(2-(l-methyl-lH- pyrazol-4-yl)morpholino)-2-(methylthio)pyrimidin-4-amine. MS (ESI) m/z: calc’d for CisHisNeOS [M+H]⁺: 307.1, found: [M+H]⁺: 307.1

Intermediate 1-6: Preparation of (5)-2-(4-chloro-2-fluorophenyl)-6-(2-(l -methyl- pyrazol-4-

yl)morpholino)pyrimidin-4-amine

Step 1 : Preparation of 6-chloro-2-(4-chloro-2-fluorophenyl)pyrimidin-4-amine

[0157] In a 20 mL microwave vial containing 6-Chloro-2-(methylsulfanyl)pyrimidin-4-amine (200 mg, 1.14 mmol), (4-chloro-2- fluorophenyl)boronic acid (397 mg, 2.28 mmol) was added Pd2(dba)3 (104.3 mg, 113.9 pmol) and tri(2-furyl)phosphine (52.9 mg, 227.8 pmol). The vial was purged with Ar three times and then THF (9.5 mL) was added. It was then purged with Ar three times again, sealed and heated in a microwave reactor at 100 °C for 14 min. Upon completion, the reaction mixture was diluted with water (10 mL), sat. NaHCCh aq. (5 mL) was added and the aqueous layer extracted with EtOAc (10 mL x 2). The organic extracts were then concentrated. The residue was purified by column chromatography on silica gel, eluting with 30% to 60% EtOAc / EtOH (3/1) in hexanes to yield desired product 6-chloro-2-(4-chloro-2- fluorophenyl)pyrimidin-4-amine, also containing starting material 6-chloro-2- (methylthio)pyrimidin-4-amine. It was used for next step directly without further purification. MS (ESI) m/z: calc’d for CioHeCUFNs [M+H]⁺: 258.0, found: [M+H]⁺: 258.1.

Step 2: Preparation of (S)-2-(4-chloro-2-fluorophenyl)-6-(2-(l -methyl- 17/-pyrazol-4- yl)morpholino)pyrimidin-4-amine

[0158] To a 2 mL microwave vial containing 6-chloro-2-(4-chloro-2-fluorophenyl)pyrimidin-4- amine (50.0 mg, 0.19 mmol) and (S)-2-(l -methyl- lf/-pyrazol-4-yl)morpholine (40.5 mg, 0.24 mmol) in DMF (0.5 mL) was added tri ethylamine (98.0 mg, 0.97 mmol). The reaction mixture was stirred in a microwave reactor at 150 °C for 22 min. Upon completion, the reaction mixture was diluted with water (2 mL) and extracted with EtOAc (3 mL x 2). The residue was purified by column chromatography on silica gel, eluting with 30%-60% EtOAc / EtOH (3/1) in hexanes to give (S)-6-(2-( 1 -methyl- 1 //-py razol-4-y I )morpholino)-2-(methyl thio)py nmidin-4-amine. MS (ESI) m/z: calc’d for CisHisCIFNeO [M+H]⁺: 389.1, found: [M+H]⁺: 389.1.

Intermediate 1-7; Preparation of 6-chloro-8-(methylthio)-1.3-dihvdro-10E7-furo[3.4-

Step 1 : Preparation of methyl 4-((6-chloro-2-(methylthio)pyrimidin-4-yl)amino)-2.5- dihvdrofuran-3-carboxylate

1 2

[0159] To a stirred mixture of 6-chloro-2-(methylthio)pyrimidin-4-amine (5 g, 28.5 mmol) was added methyl 4-oxotetrahydrofuran-3-carboxylate (6.15 g, 42.7 mmol) in AcOH (50 mL), then the mixture was stirred at 100 °C for 6 h. LCMS showed the reaction was completed and the desired target was formed. The mixture was filtered and the solid was concentrated under reduced pressure to give the crude methyl 4-((5-chloro-3-(methylthio)pyrazin-2-yl)amino)-2,5- dihydrofuran-3-carboxylate, which w as used as is without further purification.

MS (ESI) m/z: calc’d for C11H13CIN3O3S [M+H]⁺: 302.1/304, found: [M+H]⁺ 302.0/304.0

'H NMR (400 MHz, DMSO-rfc) 5 ppm 9.83 (s, 1 H), 7.23 (s, 1 H), 5.29 (t, J=3.93 Hz, 2 H), 4.66 (t, J=3.99 Hz, 2 H), 3.73 (s, 3 H), 2.48 (s, 3 H)

Step 2: Preparation of 4-((6-chloro-2-(methylthio)pyrimidin-4-yl)amino)-2,5-dihydrofuran-3- carboxylic acid

[0160] To a stirred mixture of methyl 4-((6-chloro-2-(methylthio)pyrimidin-4-yl)amino)-2,5- dihydrofuran-3-carboxylate (2.7 g, 8.95 mmol) in THF (54.2 ml) was added NaOH (0.477 g, 11.93 mmol) in water (50 mL). The mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction mixture was quenched with water (50 mL) and extracted with CH2CI2 (50 mL*3). The aqueous phase was adjusted to pH 1-3 with 2M HC1 and extracted with DCM (50 mL*3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 4-((6-chloro-2- (methylthio)pyrimidin-4-yl)amino)-2,5-dihydrofuran-3-carboxylic acid, which was used without further purification.

MS (ESI) m/z: calc’d for C10H11CIN3O3S [M+H]⁺: 288.0/290.0, found: [M+H]⁺ 288.0/290.0. Step 3: Preparation of 6-chloro-8-(methylthio)-1.3-dihvdro-10H-furor3.4-d]pyrimido[1.6- al pyrimidin- 10-one

3 4

[0161] To a stirred mixture of 4-((6-chloro-2-(methylthio)pyrimidin-4-yl)amino)-2,5- dihydrofuran-3-carboxylic acid (1.2 g, 4.17 mmol) in MeCN (15 ml) were added POCh (1.749 ml, 18.77 mmol) and TEA (3.49 ml, 25.02 mmol) at 0 °C, and the mixture was stirred at 80 °C for 1 h. Upon completion, the reaction mixture was quenched with water (100 mL) and extracted with CH2CI2 (50 mL*3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was treated with MTBE (20 mL) and stirred for 15 min at 25 °C, filtered and the filtrate was concentrated under reduced pressure to give 6-chloro-8-(methylthio)-L3-dihydro-10L7-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one, which was used without further purification.

MS (ESI) m/z: calc’d for C10H9CIN3O2S [M+H]⁺: 270.0/272.0, found: [M+H]⁺ 270.0/272.0.

Intermediate 1-8: Preparation of 4-(4-bromotetrahvdro-2/7-pyran-2-yl)-l-cvclopropyl-l/7- pyrazole

[0162] Three reactions on the same scale were run in parallel. To a 40 mL vial containing 1- cyclopropylpyrazole-4-carbaldehyde (1.00 g, 7.3 mmol) and but-3-en-l-ol (583 mg, 0.70 mL, 8.08 mmol) was added DCM (12 mL). The reaction was cooled to 0 °C, then followed by the addition of HBr in acetic acid (3.9 mL, 33% wt, 22.0 mmol). The reaction mixture was stirred at room temperature for 0.5 h. The three reaction mixtures were combined and diluted with DCM (20 mL) and carefully quenched with sat. NaHCCh (aq.) until the solution become slightly basic. Then the mixture was extracted with 2x50 mL DCM. The organic extracts were combined and concentrated. The crude residue was purified by column chromatography on silica gel eluting with 10% to 25% to 40% EtOAc in Hexanes to give the desired product 4-(4-bromotetrahydro- 2/7-pyran-2-yl)-l-cyclopropyl-l/7-pyrazole. The product was submitted for SFC purification using the following condition to yield the isomers as Peak 1 at 3.76 min, Peak 2 at 4.36 min and Peak 3 (mixture of two isomers) at 5.03 min. Peak 3 mixture was further separated through SFC. 1st SFC condition: Column & Dimensions: IG, 21x250 mm, 5 pm UV Wavelength: 215 nm Flow Rate: 90 ml/min, Modifier: 20% MeOH w/ 0.1% NH4OH, Outlet Pressure: 100 bar, Sample Amount: 5800 mg, Diluent: 1: 1 MeOH/CHsCN. Diluent Volume: 40 ml, Injection Volume: 0.3 ml; Instrument: Sepiatec 3

[0163] The Peak 3 mixture sample from above was separated via a second SFC run to yield one of the isomers as Peak 3-1 at 2.7 min and Intermediate 1-8 (Peak 3-2) at 3.3 min.

2nd SFC condition: Column & Dimensions: IB-N, 21x250 mm, 5 pm; UV Wavelength: 215 nm; Flow Rate: 70 ml/min; Modifier: 15% MeOH w/ 0. 1% NH4OH; Outlet Pressure: 100 bar; Instrument: Sepiatec 2

Intermediate 1-9; Preparation of 4-iodo-2-(tetrahydrofuran-3-yl)tetrahvdro-277-pyran

Step 1 Step 2

Step 1 : Preparation of 2-(tetrahvdrofuran-3-yl)tetrahvdro-2E7-pyran-4-ol

[0164] To a solution of tetrahydrofuran-3-carbaldehyde (1 g, 9.99 mmol) and but-3-en-l-ol (0.504 g, 6.99 mmol) in DCM (13 mL) at 0 °C was added TFA (5.94 mL, 80 mmol) dropwise. The solution was slowly warmed to 20 °C for 12 h. After LCMS showed the reaction was completed, the reaction mixture was poured into aq. NaHCChto adjust pH~8 and extracted with DCM (30 mLx3). The organic layer was dried over MgSCh. After filtration and concentration, the crude product was purified by flash silica gel chromatography (0-20% EtOAc/Petroleum ether gradient) to give 2-(tetrahydrofuran-3-yl)tetrahydro-2F7-pyran-4-ol.

Step 2: Preparation of 4-iodo-2-(tetrahydrofuran-3-yl )tetrahydro-277-pyran

[0165] To a mixture of 2-(tetrahydrofuran-3-yl)tetrahydro-2//-pyran-4-ol (1 g, 5.81 mmol) and Imidazole (0.514 g, 7.55 mmol). PPhs (1.828 g, 6.97 mmol) in DCM (25 mL) was added h (1.768 g, 6.97 mmol) at 0 °C. The resulting mixture was stirred at 25 °C for 2 h. The solvent was removed under reduced pressure and the residue was dissolved in water (10 mL) and EtOAc (10 mL). The organic layer was separated and the aqueous layer was re-extracted with EtOAc (10 mL*3) and the combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0-10% ethyl acetate/petroleum ether gradient) to give 4-iodo-2- (tetrahy drofuran-3 -y l)tetrahy dro-277-py ran.

MS (ESI) m/z: calc’d for CyHie [M+H]⁺: 283.0, found [M+H]⁺: 283.0.

Intermediate I- 10: Preparation of 8-chloro-6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-4F7- pyrimido[1.6-a1pyrimidin-4-one

[0166] A microwave vial containing 8-chloro-2,3-dimethyl-6-(methylthio)-4E7-pyrimido[l,6- a]pyrimidin-4-one (320 mg, 1.25 mmol), (4-chloro-2-fluorophenyl)boronic acid (273 mg, 1.56 mmol), Tris(dibezylideneacetone)dipalladium (115 mg, 125 pmol) and Phosphine, tri-2-furanyl- (58. 1 mg, 250 pmol) was purged with Ar three times, then THF (6.3 mL) was added. The reaction mixture was purged with Ar three times again. It was sealed and heated in a microwave reactor at 100 °C for 7 min. The reaction mixture was cooled to room temperature, diluted with DCM and filtered through CELITE. The filter cake was rinsed with DCM several times. The filtrate was combined together and diluted with water, followed by sat. NaHCO? aq. solution and extracted with EtOAc. The organic extracts were then concentrated. The residue was purified by column chromatography on silica gel, eluting with 20-60% EtOAc in hexanes to yield 8-chloro- 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-4E7-pyrimido[l,6-a]pyrimidin-4-one as desired product. MS (ESI) m/z: calc’d for C15H11CI2FN3O [M+H]⁺: 338.0, found: [M+H]⁺: 338.0.

[0167] Intermediates shown in Intermediates Example Table Int-1 below, were prepared according to procedures analogous to those outlined above using appropriate starting materials. Table Int-1: Intermediate Examples

Intermediate II-l: Preparation of 2,2-difluoro-6-(2-methylpyridin-4-yl)morpholine

Step 1: Synthesis of (2-methylpyridin-4-yl)methanol

[0168] To a solution of 2-methylisonicotinic acid (40 g, 292 mmol) in THF (400 mL) was added BH3.DMS (55.4 mL, 554 mmol) at 0 °C and the resulting mixture was stirred at 25 °C for 16 h. The mixture was quenched by MeOH (40 mL) slowly and concentrated to give (2- methylpyridin-4-yl)methanol as a desired product.

Step 2: Synthesis of 2-methylisonicotinaldehyde

[0169] To a solution of (2-methylpyridin-4-yl)methanol (40 g, 325 mmol) in DCE (400 mL) was added IBX (182 g, 650 mmol) and the resulting mixture was stirred at 25 °C for 0.5 h. Then the resulting mixture was stirred at 80 °C for 2 h. The mixture was filtered and concentrated to give crude 2-methylisonicotinaldehyde as a desired product.

Step 3: Synthesis of l-(2-methylpyridin-4-yl)-2-nitroethan-l-ol

[0170] To a solution of 2-methylisonicotinaldehyde (40 g, 330 mmol) in nitromethane (81 mL, 1502 mmol) was added TEA (92 mL, 660 mmol) at 0 °C and the resulting mixture was stirred at 25 °C for 1 h. Upon completion, the reaction solution was filtered and concentrated under vacuum to give a crude residue. The residue was purified by flash silica gel chromatography (eluent of 40% ethyl acetate/petroleum ether) to give l-(2-methylpyridin-4-yl)-2-nitroethan-l-ol as a desired product.

MS (ESI) m/z: calc'd for CsHnN2O3⁺[M+H]⁺: 183.1, found [M+H]⁺: 183.1

Step 4: Synthesis of 2-amino-l-(2-methylpyridin-4-yl)ethan-l-

Pd/C, H₂ (30 psi) - ►

THF, 30 °C, 16 h

[0171] A mixture of l-(2-methylpyridin-4-yl)-2-nitroethan-l-ol (31 g, 170 mmol) and Pd-C (9.5 g, 8.93 mmol) in MeOH (150 mL) and AcOH (1 mL) was stirred at 30 °C under Eb 30 psi for 16 h to give black mixture. Upon completion, the mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude 2-amino-l-(2-methylpyridin-4-yl)ethan-l- ol as a desired product, which was not further purified.

MS (ESI) m/z: calc’d for C₈Hi3N₂O⁺[M+H]⁺: 153.1, found [M+H]⁺: 153.2

Step 5: Synthesis of 2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethan-l-ol

[0172] A solution of 2-amino-l-(2-methylpyridin-4-yl)ethan-l-ol (25 g, 164 mmol)), benzaldehyde (52.3 g, 493 mmol), AcOH (1 mL) in MeOH (200 mL) was stirred at 0 °C for 0.5 h, then sodium cyanoborohydride (20.64 g, 329 mmol) was added. The mixture was stirred at 25 °C for 1 h. Upon completion, the reaction mixture was poured into H2O (200 mL), and the mixture was extracted with dichloromethane (3 x 200 mL). The combined organic fractions were washed with brine (150 mL), dried over Na₂SO4, filtered and the solvent was evaporated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 40% ethyl acetate/petroleum ether gradient) to give 2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethan- l-ol as a desired product.

MS (ESI) m/z: calc’d for C₂₂H₂5N₂O⁺[M+H]⁺: 333.2, found [M+H]⁺:333.2

Step 6: Synthesis of 2-(2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethoxy)-2.2-difluoroacetic acid

[0173] To a solution of 2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethan-l-ol (30 g. 90 mmol), TBAI (3.33 g, 9.02 mmol) and sodium 2-chloro-2,2-di fluoroacetate (17.88 g, 1 17 mmol) in THF (300 mL) was added NaH (7.22 g, 180 mmol) at 0 °C, and the resulting mixture was allowed to warm to 65 °C, then stirred at 65 °C for 3 d. Upon completion, the mixture was treated with water (200 mL) and EtOAc (500 mL). The mixture was concentrated to give a crude residue. The residue was purified by flash silica gel chromatography (eluent of 30% ethyl acetate/EtOH gradient) to give 2-(2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethoxy)-2,2-difluoroacetic as a desired product.

MS (ESI) m/z: calc’d for C₂4H₂₅F₂N₂O3⁺[M+H]⁺: 427.2, found [M+H]⁺:427. I

Step 7: Preparation of 2-(2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethoxy)-2.2-difluoroacetic acid

[0174] To a solution of 2-(2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethoxy)-2,2- difluoroacetic acid (34 g, 71.8 mmol) in MeOH (300 mL) was added H2SO4 (11.47 mL, 215 mmol) at 0 °C, and the resulting mixture was stirred at 25 °C for 16 h under N₂. Upon completion, the reaction mixture was quenched with aqueous sodium hydrogen carbonate to adjust the pH= 9 and extracted with EtOAc (200 mL*3). The combined organic phases were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give crude methyl 2-(2-(dibenzylamino)-l-(2-methylpyridin-4-yl)ethoxy)-2,2-difluoroacetate.

MS (ESI) m/z: calc’d for C₂₃H₂7F₂N₂O3⁺[M+H]⁺: 441.2, found [M+H]⁺:441.1 Step 8: Synthesis of 2.2-difluoro-6-(2-methylpyridin-4-yl)morpholin-3-one

[0175] To a solution of methyl 2-(2-(di benzyl amino)- l-(2-methylpy ridin-4-yl)ethoxy)-2, 2- difluoroacetate (20 g, 45.4 mmol) in DCM (350 mL) was added NIS (102 g, 454 mmol) at 25 °C under N2, and the resulting mixture was stirred at 25 °C for 20 h under N2. Upon completion, the reaction mixture was quenched with sat. Na2SOs (1000 mL) and extracted with DCM (500 mL*3). The combined organic phases were washed with brine (200 mL), dried over anhydrous Na2SOr, fdtered and concentrated under reduced pressure to give a crude residue. The residue was purified by flash silica gel chromatography (eluent of 80% ethyl acetate/petroleum ether gradient) to give 4-benzyl-2,2-difluoro-6-(2-methylpyridin-4-yl)morpholin-3-one and 2,2- difluoro-6-(2-methylpyridin-4-yl)morpholin-3-one.

MS (ESI) m/z: calc’d for CIOHIIF₂N₂02⁺[M+H]⁺ 229.1 , found [M+H]⁺ 229.1

Step 9: Synthesis of 2.2-difluoro-6-(2-methylpyridin-4-yl)morpholine

[0176] A mixture of 2,2-difluoro-6-(2-methylpyridin-4-yl)morpholin-3-one (2.5 g, 11.0 mmol) and BH3.DMS (2. 191 mL, 21.91 mmol) in THF (20 mL) was stirred at 0 °C for 16 h. Upon completion, the mixture was quenched by MeOH (5 mL) and then stirred at 50 °C for 1 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a crude residue. The residue was purified by Prep-HPLC (MeCN/water using 0.1% TFA modifier) to give 2,2-difluoro-6-(2-methylpyridin-4-yl)morpholine.

MS (ESI) m/z: calc’d for CioHi3F₂N₂0⁺[M+H]⁺ 215.1, found [M+H]⁺ 215.1

Intermediate II-2; Preparation of 4-(4-iodotetrahydro-2H-pyran-2-yl)-2-methylpyridine

[0177] To a solution of 4-(4-bromotetrahydro-2H-pyran-2-yl)-2-methylpyridine (300 mg, 1.171 mmol) in acetone (5 ml) was added sodium iodide (527 mg, 3.51 mmol). The mixture was stirred at 80 °C for 2h. Upon completion, the reaction mixture was concentrated in vacuo to give the crude residue, which was purified by flash column chromatography on silica gel (ethyl acetate in petroleum ether from 0% to 40%) to afford 4-(4-iodotetrahydro-2H-pyran-2-yl)-2-methylpyridine as a desired product.

MS (ESI) m/z: calc'd for CiiHisINO⁺ [M+H]⁺ 304.0, found [M+H]⁺ 304.1.

Intermediate II-3: Preparation of (cis )-2-methyl-6-(2-metliylpyridin-4-yl (morpholine

Step 1 : Synthesis of 4-( 1 -ethoxy vinyl)-2-methylpyridine

,

[0178] 4-bromo-2-methylpyridine (2.4 kg, 13.95 mol) was stirred into 1,4-dioxane (26.4 L), and TributyK I -ethoxy vinyl)tin (5.78 Kg, 16.00 mol, 1.15 equiv) was added to obtain a mixture degassed with argon, followed by the addition of tetrakis (triphenylphosphine) palladium (800.0 g, 692.28 mmol, 0.05 equiv). The resulting mixture was heated to 110 °C for 6 hours, then filtered and washed with ethyl acetate. The combined filtrate was concentrated under reduced pressure to obtain crude 4-(l -ethoxy vinyl)-2-methylpyri dine. The raw material was used directly for the next step without further purification.

Step 2: Synthesis of l-(2-methylpyridin-4-yl) ethan-l-one

[0179] To a stirred material of crude 4-(l-ethoxyvinyl)-2-methylpyridine (13 kg) at 0 °C was added HC1 (26 L, 2 V, 6M) and the resulting solution was stirred at 25 °C for 2 h. The reaction mixture was quenched with sodium bicarbonate solution and extracted with ethyl acetate. The combined organic layers were washed with water, brine, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure. The crude product was purified by flash silica gel chromatography (eluent of 0-30% Ethyl acetate/Petroleum ether) to give l-(2-methylpyridin- 4-yl) ethan-l-one as a desired product.

Step 3: Synthesis of 2-bromo-l-(2-methylpyridin-4-yl)ethan-l-one

[0180] A mixture of l-(2-methylpyridin-4-yl)ethan-l-one (450 g, 3.33 mol), DIPEA (538 g, 4.16 mol) and TMSOTf (888 g, 3.99 mol) in DCM (4.5 L, 10V) was stirred at 0-5 °C for 0.5 h, followed by the addition of NBS (711 g, 3.99 mol) and stirred at 0-5 °C for an additional 0.5 h. The reaction was quenched with aqueous NaHCCL (8%, 10V, 4.5 L). Separate the organic layer, concentrate under vacuum and use the crude product directly in the next step.

Step 4: Synthesis of 2-(benzyl(2-hvdroxypropyl)amino)-l-(2-methylpyridin-4-yl)ethan-l-one

[0181] A mixture of 2-bromo-l-(2-methylpyridin-4-yl)ethan-l-one hydrobromide (4) (712 g, 3.33 mol) and l-(benzylamino)propan-2-ol (550 g, 3.33 mol), N,N-Diisopropylethylamine (645 g, 4.99 mol) in THF (4500 mL) was stirred at 10 °C for 1 h. Upon completion, the reaction mixture was quenched with water (4500 mL) and extracted with EtOAc (2000 mL*3). The combined organic phases were washed with brine (2000 mL). dried over anhydrous Na2SO4. filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 100% Ethyl acetate/Petroleum ether gradient) to give 2-(benzyl(2- hydroxypropyl)amino)-l-(2-methylpyridin-4-yl)ethan-l-one as a desired product.

Step 5: Synthesis of 4-benzyl-2-methyl-6-(2-methylpyridin-4-yl)-3.4-dihvdro-2H-l,4-oxazine

[0182] TMSOTf (1.25 kg, 5.63 mol) was added to 2-(benzyl (2 -hydroxypropyl) amino)-l-(2- methylpyridin-4-yl) ethan-l-one (5) (560 g, 1.88 mol) and DCE (11.2 L, 20V) at 0 °C. The resulting mixture was heated to 90 °C for 16 h, then quenched with sodium bicarbonate (8%, 5.6 L, 10 V) and extracted with 10% methanol in dichloromethane. The organic layer was washed with brine (10%. 5.6 L, 10V), dried on anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 4-benzyl-2-methyl-6-(2-methylpyridin-4-yl)-3,4-dihydro-2H-l ,4- oxazine, which was used directly for the next step without additional purification.

Step 6: Synthesis of (cis)-2-methyl-6-(2-methylpyridin-4-yl)morpholine

[0183] The solution of 4-benzyl-2-methyl-6-(2-methylpyridin-4-yl)-3,4-dihydro-2H-l,4- oxazine (6) (600 g, 2. 14 mol) in MeOH (6 L) was added Pd(OH)2 (240 g. 40% wt) and ammonium formate (675 g, 10.7 mol). The mixture was stirred at 85 °C under N2 atmosphere for 1 h. Upon completion, the reaction was filtered and the filtrate was concentrated in vacuo, then purified by prep HPLC (MeCN/water using 0.04% NH3H2O + 10 mM NH4HCO3) to give (cis)-2- methyl-6-(2-methylpyridin-4-yl)morpholine as a racemic mixture which was separated through SFC. MS (ESI) m/z: calc'd for CIIHI₇N₂O⁺ [M+H]⁺ =193, found [M+H]⁺ =193.

[0184] After 2 rounds of SFC, 0A and 0B were obtained. 0A was used for compound synthesis.

1^st SFC separation method :

Instrument: Prep-SFC-350-2

Column: AD (5*25 cm, 10 pm) Mobile phase: A for CO2 and B for EtOH (0. 1% 7M NEE-MeOH)

Gradient: B%=30% isocratic elution mode

Flow rate: 180 ml/min

Monitor wavelength: 220 nm

Column temperature: 25 °C

Retention time for 0A: 2.89 min and OB: 3.78 min.

2^nd SFC separation method :

Instrument: Prep-SFC-150-1

Column: IK (3*25 cm, 5 pm)

Mobile phase: A for CO2 and B for MeOH (0. 1% 7M NEh-MeOH)

Gradient: B%=20% isocratic elution mode

Flow rate: 100 ml/min

Monitor wavelength: 220 nm

Column temperature: 25 °C

Retention time for 0A: 3.10 min and for 0B 4. 18 min.

Intermediate II-4: Preparation of 4,4,5, 5-tetraethyl-2-(6-(tetrahvdrofuran-3-yl)-3.6-dihvdro-2H- pyran-4-yl)-1.3.2-dioxaborolane

Step 1: Synthesis of 6-(tetrahvdrofuran-3-yl)-3,6-dihvdro-2H-pyran-4-yl trifluoromethanesulfonate

[0185] To a solution of tetrahydrofuran-3-carbaldehyde (1 g, 9.99 mmol) in DCM (20 mL) under N2 in three-necked flask was added but-3-yn-l-ol (1.050 g. 14.98 mmol) and dropwise trifluoromethanesulfonic acid (2.396 mL, 30.0 mmol) at 0 °C. The mixture was stirred at 25 °C for 16 h. Upon completion, the mixture was quenched with sat. aq. NaHCCL (200 mL) and extracted with DCM (100 mL * 3). The combined organic fractions were washed with brine (200 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 0-40% ethyl acetate/petr oleum ether gradient) to give 6-(tetrahydrofuran-3-yl)-3,6-dihydro-2H-pyran-4-yl trifluoromethanesulfonate as a desired product.

MS (ESI) m/z: calc’d for CioHi4F₃0₅S⁺ [M+H]⁺: 303.1, found [M+H]⁺: 303.1.

Step 2: Synthesis of 4,4.5.5-tetraethyl-2-(6-(tetrahydrofuran-3-yl)-3.6-dihydro-2H-pyran-4-yl)-

1.3.2-dioxaborolane

[0186] To a solution of 6-(tetrahydrofuran-3-yl)-3,6-dihydro-2H-pyran-4-yl trifluoromethanesulfonate (1 g, 3.31 mmol) in 1,4-dioxane (8 ml) was added 1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.242 g, 0.331 mmol), 4, 4, 4', 4', 5, 5, 5', 5'- octaethyl-2,2'-bi(l,3,2-dioxaborolane) (1.454 g, 3.97 mmol) and potassium acetate (0.649 g, 6.62 mmol). Then the mixture was purged with N2 three times and stirred for 1 h at 80 °C. Upon completion, the mixture was filtered and the filtrate was concentrated. The crude product was purified by flash silica gel chromatography (eluent of 0-40% EtOAc/Petroleum ether gradient) to afford 4,4,5,5-tetraethyl-2-(6-(tetrahydrofuran-3-yl)-3,6-dihydro-2H-pyran-4-yl)-l,3,2- dioxaborolane as a desired product.

MS (ESI) m/z: calc'd for Ci9H₃₄BO4⁺ [M+H]⁺: 337.1, found [M+H]⁺: 337.1.

Intermediate II-5: Preparation of 6-(2.4-difluorophenyl)-2.3-dimethyl-8-(4.4.5.5-tetramethyl-

1.3.2-dioxaborolan-2-yl)-47/-pyrimido[1.6-a]pyrimidin-4-one

[0187] A 8 mL dram vial was charged with 8-chloro-6-(2,4-difluorophenyl)-2,3-dimethyl-4H- pyrimido[l,6-a]pyrimidin-4-one (100 mg, 311 pmol). potassium acetate (91.5 mg, 933 pmol), 4,4,4',4',5,5,5',5^,-octamethyl-2,2'-bi(l,3,2-dioxaborolane) (1 18 mg, 466 pmol), and PdCh(dppf)-CH2C12 adduct (25.4 mg, 31.1 pmol). The vial was sealed, and the atmosphere was exchanged for inert. Then 1,4-Dioxane (2.8 mL) was added and the solution was heated at 80 °C for 1 hour. Upon completion, the reaction was cooled down and quenched with H2O. The resulting suspension was transferred into a separately’ funnel with EtOAc (30 mL)/ Water (30 mL). The organic layer was separated and subsequently washed with brine (2 x 30 mL). The aqueous phase was backwashed with EtOAc. The combined organic phase was dried over MgSO4 and concentrated. The crude 6-(2,4-difluorophenyl)-2,3-dimethyl-8-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-47/-pyrimido[L6-c?]pyrimidin-4-one was immediately used in the next step without further purification. MS (ESI) m/z: calc’d for C21H23BF2N3O3 [M+H3O- pinacol]⁺: 332, found: [M+H]⁺: 332.

SYNTHESIS OF EXEMPLARY COMPOUNDS

EXAMPLES 1-1 and 1-2

Preparation of 1-1 (S)-6-(4-chloro-2-fluorophenyl)-2.3-dimethyl- 8-(2-(l -methyl- pyrazol-4-

yl)morpholino)-477-pyrimido[1.6-<2lpyrimidin-4-one and 1-2 (R) 6-(4-chloro-2-fluorophenyl)- 2,3-dimethyl- 8-(2-(T-methyl-17/-pyrazol-4-yl)morpholino)-42/-pyrimido|T.6-a1pyrimidin-4-one

Step 1 : Preparation of 2.3-dimethyl-8-(2-(l-methyl-17/-pyrazol-4-yl)morpholino)-6-(methylthio)- 4//-pyrimido|T.6-fl]pyriniidin-4-one

[0188] A 20 mL vial containing 8-chl oro-2, 3-dimethyl-6-(methylthio)-477-pyrimido[ 1,6- a]pyrimidin-4-one (130.0 mg, 0.51 mmol) (intermediate 1-2), 2-(l -methyl- l //-pyrazol-4- yl)morpholine (170.0 mg, 1.02 mmol), and cesium carbonate (331.3 mg, 1.02 mmol) in 1,4- Di oxane (2.5 mL) was purged with argon followed by the addition of RuPhos Pd G4 (64.85 mg. 0.076 mmol). The reaction was then purged with Ar and heated in a hot plate at 100 °C for 1 h. The reaction was cooled to room temperature, water (5 mL) was added, and the mixture was extracted with DCM (5 mL) two times. The combined organic extracts were washed with brine (3 mL) and dried over anhydrous MgSCh. The filtrate was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel column, eluting with 10-70% EtOAc / EtOH (3: 1) in hexanes to give 2,3-dimethyl-8-(2-(l-methyl-17/-pyrazol-4- yl)morpholino)-6-(methylthio)- 47f-pyrimido[1.6-a]pyrimidin-4-one. MS (ESI) m/z: calc’d for C18H22N6O2S [M+H]⁺: 387.1, found: [M+H]⁺: 387.2

Step 2: Preparation of 1-1 (5) 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl- 8-(2-( 1 -methyl- 1//- pyrazol-4-yl)morpholino)-47/-pyrimido[L6-a1pyrimidin-4-one and 1-2 (R) 6-(4-chloro-2- fluorophenyl)-2.3-dimethyl- 8-(2-(l -methyl- l/7-pyrazol-4-yl)morpholino)-47f-pyrimido[ 1.6- alpyrimidin-4-one

[0189] To a 2 mL microwave vial containing CuTc (44.4 mg, 0.23 mmol), (4-chloro-2- fluorophenyl)boronic acid (40.6 mg. 0.23 mmol) and 2,3-dimethyl-8-(2-(l-methyl-U/-pyrazol-4- yl)morpholino)-6-(methylthio)-4//-pyrimido[l,6-a]pyrimidin-4-one (30.0 mg, 0.078 mmol) was added 1,4-Dioxane (0.8 mL). The solution was purged with Ar followed by the addition of Pd(PPhs)4 (17.9 mg, 0.016 mmol). The vial was then purged with Ar again and heated in a microwave reactor at 120 °C for 7 min. The reaction mixture was diluted with water (2 mL) and extracted with DCM (2 mL) two times. The organic extracts were then stirred with 300 mg of Silia MetS ® (Si-Thiol) at room temperature, filtered and concentrated. The residue was diluted with DMSO (3 mL) and was purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl- 8-(2-(l-methyl-l/7-pyrazol-4- yl)morpholino)-477-pyrimido[l,6-«]pyrimidin-4-one as a racemic mixture, which was separated by chiral SFC separation to afford 1-1 (5) 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl- 8-(2-(l- methyl-l/7-pyrazol-4-yl)morpholino)-47f-pyrimido[l,6-a]pyrimidin-4-one (Peak 1; tR = 3.2 min) and 1-2 (R) 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-(2-(l-methyl-177-pyrazol-4- yl)morpholino)-477-pyrimido[l,6-a]pyrimidin-4-one (Peak 2; tR = 5.3 min). MS (ESI) m/z: calc’d for C23H22CIFN6O2 [M+H]⁺: 469.1. found: [M+H]⁺: 469.1

Chiral SFC separation conditions: Column & Dimensions: CCO F4, 21x250 mm, 5 pm. UV Wavelength: 215 nm; Flow Rate: 80 ml/min; Modifier: 50% MeOH w/ 0. 1% NH4OH; Outlet Pressure: 100 bar; Diluent: 1: 1 MeOH/CFLCN.

Example 1-1 Peak 1: 'H NMR (600 MHz, DMSO-O S 7.74 (s, 1H). 7.66 (t. J= 8. 1 Hz. 1H), 7.47 > 7.45 (d, J= 9.7 Hz, 2H), 7.39 (dd, J= 8.3, 1.7 Hz, 1H), 6.57 (s, 1H), 4.53 (s, 1H), 4.34 - 4.18 (m, 2H), 3.99 (d, J= 10.8 Hz, 1H), 3.82 (s, 3H), 3.68 (m, 1H), 3.16 (m, 1H), 3.11 (dd, J = 13.1, 10.6 Hz, 1H), 2.28 (s, 3H). 1.91 (s, 3H).

Example 1-2 Peak 2: 'H NMR (600 MHz, DMSO-tie) 87.7^ 4 (s, 1H), 7.66 (t, J= 8.0 Hz, 1H), 7.46 - 7.45 (m, 2H), 7.39 (dd, J= 8.3, 1.8 Hz, 1H), 6.56 (s, 1H), 4.53 (s, 1H), 4.34 - 4.22 (m, 2H), 3.99 (d, J= 11.1 Hz, 1H), 3.82 (s, 3H), 3.68 (s, 1H), 3.18 - 3.14 (m, 1H), 3.11 (dd, J= 13.1, 10.5 Hz, 1H), 2.28 (s, 3H). 1.91 (s, 3H).

EXAMPLE 1-3 and 1-4

Preparation of 1-3 (R or S')-6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-(3-(2-methylpyrimidin-4- yl)piperidin-l-yl)-47f-pyrimido[1.6-a]pyrimidin-4-one and 1-4 (S or A)-6-(4-chloro-2- fluorophenyl)-2.3-dimethyl-8-(3-(2-methylpyrimidin-4-yl)piperidin-l-yl)-4/f-pyrimido[1.6-

6<|pyrimidin-4-one

Step 1 : Preparation of 8-chloro-6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-4//-pyrimido[L6- g|pyrimidin-4-one

[0190] A 20 mL microwave vial containing 8-chloro-2,3-dimethyl-6-(methylthio)-47/- pyrimido[l,6-a]pyrimidin-4-one (320 mg, 1.25 mmol), (4-chloro-2-fluorophenyl)boronic acid (273 mg. 1.56 mmol), Pd2(dba)3 (115 mg, 125 pmol) and tri(2-furyl)phosphine (58.1 mg, 250 pmol) was purged with Ar \ three times. Then THF (6.3 mL) was added and the reaction was heated in a micro wave reactor at 100 °C for 7 min. Upon completion, the reaction mixture was cooled to RT and diluted with DCM (10 mL), then filtered through Celite. The filter cake was rinsed with DCM three times. The organic extracts were combined and diluted with water. Saturated NaHCCL aq. (3 mL) was added, then extracted with EtOAc (5 mL) three times. The organic extracts were then concentrated. The residue was purified by column chromatography on silica gel, eluting with 20-60% EtOAc in hexanes to yield desired product 8-chloro-6-(4-chloro- 2-fluorophenyl)-2,3-dimethyl-47/-pyrimido[L6-a]pyrimidin-4-one. Step 2: Preparation of 1-3 (R or S')-6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-(3-(2- methylpyrimidin-4-yl)piperidin-l-yl)-477-pyrimido[1.6-a1pyrimidin-4-one and 1-4 (S or R)-6-(4- chloro-2-fluorophenyl)-2.3-dimethyl-8-(3-(2-methylpyrimidin-4-yl)piperidin-l-yl)-47/- pyrimido[1.6-a]pyrimidin-4-one

[0191] To a 20 mL vial were added 8-chloro-6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-477- pyrimido[l,6-a]pyrimidin-4-one (60.0 mg, 177 pmol), RuPhos Pd G4 (22.6 mg, 26.6 pmol) and cesium carbonate (260 mg, 0.8 mmol). The vial was purged with Ar three times then 1,4- dioxane (1.2 mL) was added. After purging with Ar again, the reaction was heated in a hot plate at 100 °C for 3 h. Upon completion, the reaction mixture was cooled to RT, diluted with water and extracted with DCM. The organic extracts were concentrated and the crude residue diluted with DMSO. It was purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield desired product rac-6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-(3-(2-methylpyrimidin- 4-yl)piperidin-l-yl)-477-pyrimido[l,6-a]pyrimidin-4-one. The racemic mixture was further separated by chiral SFC separation to yield 1-3 (R or S)-6-(4-chloro-2-fluorophenyl)-2,3- dimethyl-8-(3-(2-methylpyrimidin-4-yl)piperidin-l-yl)-477-pyrimido[1.6-a]pyrimidin-4-one (Peak 1; tR = 3.9 min) and 1-4 (S or 7?)-6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-(3-(2- methylpyrimidin-4-yl)piperidin-l-yl)-477-pyrimido[l,6-a]pyrimidin-4-one (Peak 2; tR = 6.5 min). MS (ESI) m/z: calc’d for C25H24CIFN6O [M+H]⁺: 479.1, found: [M+H]⁺: 479.1 Example 1-3 Peak 1: 'H NMR (600 MHz, DMSO-t*) <5 8.63 (d, J= 5. 1 Hz, 1H), 7.68 (s, 1H), 7.52 (d, J= 10.0 Hz. 1H), 7.43 (d. J= 8.1 Hz. 1H), 7.31 (s, 1H), 6.60 (s, 1H), 4.38 (s, 2H). 2.96 (s, 1H), 2.59 (s, 3H), 2.31 (s, 3H), 2.06 (d, .7= 10.0 Hz, 1H), 1.91- 1.85 (m, 5H), 1.63 (s, 1H). Example 1-4 Peak 2: ‘H NMR (600 MHz, DMSO-Je) 8.62 (d, J= 5.1 Hz, 1H), 7.64 (s, 1H), 7.45 (d, J= 10.1 Hz, 1H), 7.39 (d, J= 8.2 Hz, 1H), 7.30 (s, 1H), 6.49 (s, 1H). 4.47 - 4.35 (m, 2H), 3.14 (t, J= 12.1 Hz, 1H), 2.91 (s. 1H), 2.59 (s, 3H), 2.27 (s, 3H), 2.03 (d, J= 12.4 Hz, 1H). 1.90 (s, 3H), 1.87-1.80 (m, 13.3 Hz, 2H), 1.61 (s, 1H).

Chiral SFC separation conditions: Column & Dimensions: CCO F4, 21x250 mm, 5 pm UV Wavelength: 215 nm; Flow Rate: 70 rnL/min; Modifier: 30% MeOH w/0. 1%NH4OH; Outlet Pressure: 100 bar; Sample Amount: 9 mg; Diluent: 1 : 1 MeOH/CHsCN

[0192] Examples shown in Example Table 1 below, were prepared according to procedures analogous to those outlined in Examples 1-3 and 1-4 above using appropriate starting materials. Table 2: Examples 1-5 and 1-6

Example 1-7 & 1-8

Preparation of 1-7 (R or S) 8-(2.2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2.4- difluorophenyl)-2.3-dimethyl-4H-pyrimido[1.6-a]pyrimidin-4-one & 1-8 (S or R) 8-(2.2- difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2.4-difluorophenyl)-2.3-dimethyl-4H- pyrimido[1.6-a1pyrimidin-4-one

Step 1 : Synthesis of 8-(2.2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2.4- difluorophenyl)-2.3-dimethyl-4H-pyrimido[ 1.6-a|pyrimidin-4-one

[0193] To a solution of 8-chloro-6-(2,4-difhrorophenyl)-2,3-dimethyl-4H-pyrirnido[l,6- a]pyrimidin-4-one (50 mg. 0.155 mmol), CsF (70.8 mg. 0.466 mmol) and 2,2-difluoro-6-(2- methylpyridin-4-yl)morpholine (49.9 mg, 0.233 mmol) in DMSO (1 mL) was added DIPEA (0.081 mL, 0.466 mmol) in a glove box. The resulting mixture was stirred at 80 °C for 10 min. LCMS showed the starting material was consumed and desired compound was found. The mixture was filtered and the filtrate was purified by Pre-HPLC (MeCN/water using TFA modifier) to give 8-(2,2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2,4-difluorophenyl)- 2,3-dimethyl-4H-pyrimido[l,6-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc’d for C25H₂2F4N₅O2⁺[M+H]⁺ 500.2, found [M+H]⁺ 500.2

Step 2: Synthesis of 1-7 (R or S) 8-(2.2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2.4- difluorophenyl)-2.3-dimethyl-4H-pyrimido[1.6-a]pyrimidin-4-one & 1-8 (S or R) 8-(2.2- difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2.4-difluorophenyl)-2.3-dimethyl-4H- pyrimido) 1 ,6-al py rimidin-4-one

[0194] The 8-(2,2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2,4-difluorophenyl)-2.3- dimethyl-4H-pyrimido[l,6-a]pyrimidin-4-one (15 mg, 0.030 mmol) was resolved by Chiral-SFC DAICEL CHIRALCEL OD (250 mm*30 mm, 10 urn); Condition CCh-EtOH (0.1% NH3H2O); Begin B 45 End B 45 Gradient; Time (min) 10 100% B Hold Time 10; Flow Rate (mL/min) 80; Injections 40) to give (R or S) 8-(2,2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2,4- difluorophenyl)-2,3-dimethyl-4H-pyrimido[l,6-a]pyrimidin-4-one (tr = 2.906 min) as the first eluting peak, and (S' or R) 8-(2,2-difluoro-6-(2-methylpyridin-4-yl)morpholino)-6-(2,4- difluorophenyl)-2,3-dimethyl-4H-pyrimido[l,6-a]pyrimidin-4-one (tr = 3.516 min) as the second eluting peak.

1-7: MS (ESI) m/z: calc’d for C25H₂2F4N₅O2⁺[M+H]⁺ 500.2, found [M+H]⁺ 500.2

'H NMR (CD3OD. 400 MHz) 5 8.49 (d. J=5.25 Hz. 1 H). 7.69 - 7.80 (m. 1 H), 7.44 - 7.52 (m, 1 H), 7.36 - 7.42 (m, 1 H), 7.10 - 7.16 (m, 1 H), 6.96 (s, 1 H), 6.47 (s, 1 H), 5.34 - 5.44 (m, 1 H), 4.48 - 4.60 (m, 2 H), 3.70 - 3.80 (m, 1 H), 3.33 - 3.36 (m, 1 H), 2.59 (s, 3 H), 2.38 (s, 3 H), 2.02 (s, 3 H)

1-8: MS (ESI) m/z: calc’d for C25H22F₄N₅O2⁺[M+H]⁺ 500.2, found [M+H]⁺ 500.2

^XH NMR (CD3OD. 400 MHz) 5 8.49 (d. J=5.13 Hz. 1 H). 7.67 - 7.79 (m. 1 H). 7.47 (s. 1 H). 7.36 - 7.42 (m, 1 H), 7.08 - 7.16 (m, 1 H), 6.95 - 7.04 (m, 1 H), 6.45 - 6.49 (m, 1 H), 5.45 (s, 1 H), 4.51 - 4.60 (m, 2 H), 3.69 - 3.83 (m, 1 H), 3.36 (s, 1 H), 2.58 - 2.60 (m, 3 H), 2.38 (s, 3 H), 2.02 (s, 3 H) [0195] Examples shown below were prepared according to procedures analogous to those outlined for Examples 1-7 and 1-8 using the appropriate starting materials.

Table 3: Examples 1-9 to 1-11

EXAMPLE 2-1 and 2-2

Preparation of 2-1 (5)- 1 -(4-chloro-2-fluorophenyl)-3-(2-( 1 -methyl- 177-pyrazol-4-yl)morphohno)-

7.8-dihvdrocvclopentard1pyrimido[1.6-a1pyrimidin-9(67/)-one and 2-2 (7?)-l-(4-chloro-2- fluorophenyl)-3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-7,8- dihvdrocvclopentard1pyrimido[1.6-a1pyrimidin- one

Step 1 : Preparation of 3-(2-(l-methyl-17/-pyrazol-4-yl)morpholino)-l-(methylthio)-7,8- dihvdrocvclopentak/lpyrirnidol 1 .6-fl|pyrimidin-9(6/7)-one [0196] A 20 mL vial containing 3-chloro-l-(methylthio)-7,8- dihydrocyclopenta[t/|pyrimido[l,6-a]pyrimidin-9(6/7)-one (100 mg, 0.37 mmol) (intermediate I- 3), 2-( 1 -methyl- l/7-pyrazol-4-yl)morpholine (125 mg, 0.75 mmol), and cesium carbonate (243 mg, 0.75 mmol) in 1,4-Dioxane (2.5 mL) was purged with Ar followed by addition of RuPhos Pd G4 (31.8 mg, 0.037 mmol). The reaction mixture was purged with Ar and heated in a hot plate at 100 °C for 1 h. The reaction mixture was cooled to room temperature, diluted with water (5 mL) and extracted with DCM (4 mL) two times. The organic extracts were washed with brine (4 mL), dried over anhydrous MgSCh. and concentrated._The residue was purified by column chromatography on silica gel column, eluting with 30-70% EtOAc / EtOH (3/1) in hexanes to give 3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-l-(methylthio)-7,8- dihydrocyclopenta[<7]pyrimido[l,6-a]pyrimidin-9(6/7)-one. MS (ESI) m/z: calc’d for C19H22N6O2S [M+H]⁺: 399.1, found: [M+H]⁺: 399.2.

Step 2: Preparation of 2-1 (5)-l-(4-chloro-2-fluorophenyl)-3-(2-(l-methyl-177-pyrazol-4- yl)morpholino)-7.8- [1.6-a1pyrimidin-9(677)-one and 2-2 (A)-l-(4-

chloro-2-fluorophenyl)-3-(2-(l-methyl-17f-pyrazol-4-yl)morpholino)-7.8- dihydrocyclopentati71pyrimido[1.6-fi'1pyrimidin-9 -one

[0197] To a microwave vial containing ((thiophene-2-carbonyl)oxy)copper (CuTc) (43. 1 mg, 0.226 mmol), (4-chloro-2- fluorophenyl)boronic acid (39.4 mg, 0.226 mmol) and 3-(2-(l-methyl- l#-pyrazol-4-yl)morpholino)-l-(methylthio)-7,8-dihydrocyclopenta[</]pyrimido[l,6- a|pyrimidin-9(67/)-one (30.0 mg, 0.075 mmol) was added 1,4-Dioxane (0.75 mL). The reaction mixture was purged with Ar followed by addition of Pd(PPhs)4 (17.4 mg, 0.015 mmol). The vial was then purged with Ar again and heated in a micro wave reactor at 120 °C for 7 min. The reaction mixture was diluted with water (2 mL) and extracted with DCM (2.5 mL) two times. To the organic extracts were then added thiol-Si scavenger and stirred at room temperature for 10 min. The mixture was filtered and the filtrate was concentrated. The residue was diluted with DMSO (3 mL) and was purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield l-(4-chloro-2-fluorophenyl)-3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-7,8- dihydrocyclopcnta|t/|pyrimido| l .6-o|pyrirnidin-9(6/7)-one as a racemic mixture which was then separated by chiral SFC separation to give the two S and J? enantiomers. MS (ESI) m/z: calc’d for C24H22CIFN6O2 [M+H]⁺: 481.1. found: [M+H]⁺: 481.1.

SFC purification Conditions: Column & Dimensions: CCO F4, 21x250 mm, 5 pm. UV Wavelength: 215 nm; Flow Rate: 70 ml/min; Modifier: 60% MeOH w/ 0. 1% NH4OH; Outlet Pressure: 100 bar; Diluent: 1: 1 MeOH / CH3CN.

Example 2-1 Peak 1 (tR = 3.8 min): (<S’)-l-(4-chloro-2-fluorophenyl)-3-(2-(l-methyl-177-pyrazol- 4-yl)morpholino)-7,8-dihydrocyclopenta[</]pyrimido[l,6-a]pyrimidin-9(677)-one. 'H NMR (600 MHz, DMSO-c/e) 6 1.14 (s, 1H), 7.66 (t, J= 8.0 Hz, 1H), 7.47 - 7.45 (m, 2H), 7.39 (dd, J= 8.3, 1.9 Hz, 1H), 6.63 (s, 1H), 4.53 (s, 1H), 4.4 - 4.2 (b, 2H), 3.99 (d, J= 10.8 Hz, 1H). 3.82 (s, 3H), 3.68 (d, J= 10.6 Hz, 1H), 3.22 - 3.10 (m, 2H), 2.80 (q, J= 7.9 Hz, 2H), 2.62 (dt. J= 17.5, 7.3 Hz. 2H), 1.99 (p. J = 7.5 Hz. 2H).

Example 2-2 Peak 2 (tR = 6.65 min): (7?)-l-(4-chloro-2-fluorophenyl)-3-(2-(l-methyl-177- pyrazol-4-yl)morpholino)-7,8-dihydrocyclopenta[<7|pyrimido[l,6-a]pyrimidin-9(677)-one. 'H NMR (600 MHz, DMSO-t/e) d 7.74 (s, 1H), 7.66 (t, J= 8.0 Hz, 1H), 7.47 - 7.45 (m, 2H), 7.39 (dd, J= 8.3, 1.9 Hz, 1H), 6.64 (s, 1H). 4.53 (s, 1H). 4.39 - 4. 18 (m. 2H) 3.99 (d, J = 11.1 Hz, 1H), 3.82 (s, 3H), 3.67 (d, ,7= 12.3 Hz, 1H), 3.23 - 3.10 (m, 2H), 2.80 (q, .7= 7.8 Hz, 2H), 2.70 - 2.57 (m, 2H), 1.99 (p, J= 7.5 Hz, 2H).

EXAMPLES 2-3 and 2-4

Preparation of 2-3 (5)- 1 -(4-chloro-2-fluorophenyl)-3-(2-( 1 -methyl- 177-pyrazol-4-yl)morpholino)- 6.7.8.9-tetrahvdro-1077-pyrimidoI6.1-h]quinazolin-10-one and 2-4 (7?)-l-(4-chloro-2- fluorophenyl)-3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-6.7.8,9-tetrahydro-1077- pyrimido[6,l -hl quinazolin- 10-one

Step 1 : Preparation of 3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-l-(methylthio)-6,7.8.9- tetrahy dro-1077-pyrimido[6, 1 -61 quinazolin- 10-one

[0198] To a 20 mL vial containing 3-chloro-l-(methylthio)-6.7.8.9-tetrahy dro-1077- pyrimido[6,l-6]quinazolin-10-one (100 mg, 0.355 mmol) (intermediate 1-4), 2-(l-methyl-177- pyrazol-4-yl)morpholine (119 mg, 0.71 mmol), and cesium carbonate (231 mg, 0.71 mmol) was added 1,4-Dioxane (2.4 mL). The reaction mixture was purged with Ar followed by the addition of RuPhos Pd G4 (30.2 mg, 0.036 mmol). The vial was then purged with Ar again and heated in a hot plate at 100 °C for 2 h. The reaction mixture was cooled to rt, diluted with water (6 mL) and extracted with DCM (4 mL) two times. The organic extracts were washed with brine (4 mL), dried over anhydrous MgSO-i. and concentrated. The residue was purified by column chromatography on silica gel column, eluting with 10-70% EtOAc / EtOH (3: 1) in hexanes to give 3-(2-( 1 -methyl- 177-pyrazol-4-yl )morpholino)- 1 -(methylthio)-6,7,8,9-tetrahy dro- 1077- pyrimido[6,l-6]quinazolin-10-one. MS (ESI) m/z: calc’d for C20H24N6O2S [M+H]⁺: 413.2, found: [M+H]⁺: 413.2

Step 2: Preparation of 2-3 (5)-l-(4-chloro-2-fluorophenyl)-3-(2-(l-methyl-177-pyrazol-4- yl)morpholino)-6.7.8.9-tetrahydro-1077-pyrimido[6.1- 61 quinazolin- 10-one and 2-4 (7?)-l-(4- chloro-2-fluorophenyl)-3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-6.7.8.9-tetrahvdro-1077- pyrimido[6.1 -61 quinazolin- 10-one

[0199] To a microwave vial containing CuTc, (4-chloro-2-fluorophenyl)boronic acid (44.4 mg, 0. 255 mmol) and 3-(2-(l-methyl-17/-pyrazol-4-yl)morpholino)-l-(methylthio)-6,7,8,9- tetrahydro-1077-pyrimido[6, 1-6] quinazolin- 10-one (35.0 mg, 0.085 mmol) was added 1,4- Di oxane (1.4 mL). The reaction mixture was purged with Ar followed by the addition of Pd(PPhs)4 (19.6 mg, 0.017 mmol). The vial was purged with Ar again and heated in a microwave reactor at 120 °C for 12 min. Upon cooling, the reaction mixture was diluted with water (5 mL) and extracted with DCM (4 mL x 2). To the combined organic extracts was added Silia MetS ® (Si-Thiol) (20 mg), filtered and concentrated. The resulting residue was diluted with DMSO (3 mL), filtered and purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to afford l-(4-chloro-2-fluorophenyl)-3-(2-(l-methyl-l/7-pyrazol-4-yl)morpholino)- 6,7,8,9-tetrahydro-1077-pyrimido[6,l-ft]quinazolin-10-one as a racemic mixture. The racemic mixture was purified by chiral SFC to give Peak 1 (tR = 4.5 min) as 2-3 (.V)- l -(4-chloro-2- fluorophenyl)-3-(2-(l-methyl-l/7-pyrazol-4-yl)morpholino)-6,7,8,9-tetrahydro-1077- pyrimido[6,l -ft] quinazolin- 10-one and Peak 2 (tR = 8.0 min) as 2-4 (A)-l-(4-chloro-2- fluorophenyl)-3-(2-(l-methyl-l//-pyrazol-4-yl)morpholino)-6,7,8,9-tetrahydro-1077- pyrimido[6,l -ft] quinazolin- 10-one. MS (ESI) m/z: calc’d for C25H24CIFN6O2 [M+H]⁺: 495.2, found: [M+H]⁺: 495.2

SFC purification Conditions: Column & Dimensions: CCO F4, 21x250 mm, 5 pm. UV Wavelength: 215 nm: Flow Rate: 70 ml/min; Modifier: 50% MeOH w/ 0.1% NH4OH; Outlet Pressure: 100 bar; Diluent: 1: 1 MeOH/CFECN.

Example 2-3 Peak 1: (RT = 4.5 min). ^JH NMR (600 MHz, DMSO-Js) ft 7.73 (s, 1H), 7.65 (s, 1H), 7.46 - 7.45 (m, 2H), 7.39 (d, J= 8.2 Hz, 1H), 6.52 (s, 1H), 4.53 (s, 1H). 4.32 - 4.18 (m, 2H), 3.99 - 3.98 (m, 2H), 3.81 (s, 3H), 3.67 (s, 1H), 3.25 - 3.07 (m, 2H), 2.58 - 2.51 (m, 2H), 2.31 (s, 2H), 1.74 - 1.68 (m, 4H).

Example 2-4 Peak 2: (RT = 8.0 min). ^JH NMR (600 MHz, DMSO-Je) ft 7.73 (s, 1H), 7.65 (t, J= 8.0 Hz, 1H), 7.46 - 7.45 (m, 2H), 7.39 (d, J= 8.2 Hz, 1H), 6.53 (s, 1H), 4.53 (s, 1H). 4.27 (m, 2H), 3.99 - 3.77 (m, 2H), 3.81 (s, 3H), 3.68 (s, 1H). 3.22 - 3.08 (m, 2H). 2.58 (s. 2H), 2.31 - 2.30 (m, 2H), 1 .73 - 1 .68 (m, 4H).

EXAMPLE 2-5

Preparation of 2-5 (5)-l-(2.4-difluorophenyl)-3-(2-(l-methyl-U7-pyrazol-4-yl)morpholino)-

6.7.8.9-tetrahydro-10ft7-pyrimidol6.1-ft]quinazolin-10-one

Step 1 : Preparation of (_lS)-3-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-l-(methylthio)-6.7.8.9- tetrahy dro-107f-pyrimido[6, 1 -61 quinazolin- 10-one

[0200] To a 20 mL vial containing 3-chloro-l-(methylthio)-6.7.8.9-tetrahydro-10EZ- pyrimido[6, 1-6] quinazolin- 10-one (100.0 mg. 0.355 mmol) (intermediate 1-4). GS')-2-( l -methyl- l//-pyrazol-4-yl)morpholine (59.3 mg, 0.355 mmol) (intermediate 1-1), and cesium carbonate (231.3 mg, 0.710 mmol) was added 1,4-Di oxane (2.4 mL). The vial was purged with Ar followed by the addition of RuPhos Pd G4 (30.2 mg, 0.035 mmol). The reaction mixture was then purged with Ar and heated in a hot plate at 100 °C for 3 h. The reaction mixture was cooled, diluted with water (6 mL) and extracted with DCM (5 mL) two times. The organic extracts were dried over anhydrous MgSCL then concentrated. The residue w as purified by column chromatography on silica gel column, eluting with 30-70% EtOAc / EtOH in hexanes to provide (<S)-3-(2-(l-methyl- lEZ-pyrazol-4-yl)morpholino)-l-(methylthio)-6,7,8,9-tetrahydro-10EZ-pyrimido[6,l-6]quinazolin- 10-one. MS (ESI) m/z: calc’d for C20H24N6O2S [M+H]⁺: 413.2, found: [M+H]⁺: 413.2

Step 2: Preparation of 2-5 (S)-l-(2.4-difluorophenyl)-3-(2-(l-methyl-17/-pyrazol-4- y l)morpholino)-6,7.8,9-tetrahy dro- 10/7-pyri mi do [6,1-61 quinazolin- 10-one

[0201] To a microwave vial containing (S)-3-(2-(l-methyl-17f-pyrazol-4-yl)morphohno)-l- (methylthio)-6,7,8,9-tetrahydro-1077-pyrimido[6,l-6]quinazolin-10-one (20.0 mg, 0.049 mmol), 2,4-difluorophenylboronicacid (19.1 mg, 0.12 mmol), Pd2(dba)3 and tri(2-furyl)phosphine (7 mg, 0.03 mmol) purged with Ar was added dry THF (1 mL). The vial was purged with Ar again. The reaction mixture was then heated in a microwave reactor at 100 °C for 7 min. The reaction mixture was cooled to room temperature, diluted with water (3 mL) and extracted with DCM (2 mL) two times. To the organic extracts was then added Silia MetS ® (Si-Thiol) (50 mg) and stirred at room temperature for about 10 min. The mixture was filtered, and the filtrate was concentrated. The residue was diluted with DMSO (2.5 mL), filtered to remove solids and purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to afford 2-5 (S)-3-(2-(l-methyl-lT7-pyrazol-4-yl)morpholino)-l-(methylthio)-6,7,8,9-tetrahydro-10T7- pyrimido[6, 1-6] quinazolin- 10-one. MS (ESI) m/z: calc'd for C25H24F2N6O2 [M+H]⁺: 479.2, found: [M+H]⁺: 479.0. 'H NMR (600 MHz, DMSO-d6) 6 7.77 (m, 2H), 7.48 (s, 1H), 7.39 (t, J = 9.9 Hz. 1H), 7.27 (t, J= 8.6 Hz, 1H), 6.58 (s, 1H), 4.58 (s, 3H). 4.06 (d, J= 11.4 Hz. 2H), 3.83 (s, 3H), 3.71 (s, 1H), 3.37 (s, 2H), 2.64 (m, 2H), 2.30 (s, 2H), 1.73 (m, 4H).

EXAMPLES 2-6 and 2-7

Preparation of 2-6 (R or S)-l-(4-chloro-2-fluorophenyl)-3-(2-(2-methylpyridin-4-yl)morpholino)-

6.7.8.9-tetrahvdro-10T7-pyrimido[6.1-6]quinazolin-10-one and 2-7 l-(4-chloro-2-

fluorophenyl)-3-(2-(2-methylpyridin-4-yl)morpholino)-6.7.8.9-tetrahvdro-107/-pyrimido[6.1- b quinazolin- 10-one

Step 1 : Synthesis of 3-chloro-l-(4-chloro-2-fluorophenyl)-6.7.8.9-tetrahvdro-107f-pyrimido[6.1- 61 quinazolin- 10-one

[0202] A 2 mL microwave vial containing 3-chloro-l-(methylthio)-6,7,8,9-tetrahydro-10T7- pyrimido[6, 1-6] quinazolin- 10-one (1-4, 50.0 mg, 0.177 mmol), (4-chloro-2-fluorophenyl)boronic acid (61.9 mg, 0.355 mmol). Pd2(dba)s (16.3 mg, 0.018 mmol) and tri(2-furyl)phosphine (8.24 mg, 0.035 mmol) was purged with Ar three times, then THF (1.5 mL) was added. The vial was purged with Ar again and heated in a micro wave reactor at 100 °C for 7 min. The reaction mixture was diluted with water (3 mL) and sat. NaHCCh aq. (1 mL), and extracted with DCM (3 mL x 2). The organic extracts were concentrated and purified by column chromatography on silica gel, eluting with 60%-100% DCM in hexanes to give 3-chloro-l-(4-chloro-2- fluorophenyl)-6,7,8,9-tetrahydro-107/-pyrimido[6,l-6]quinazolin-10-one as a desired product. MS (ESI) m/z: calc’d for C17H12CI2FN3O [M+H]⁺: 364.0. found: [M+H]⁺: 364.0.

Step 2: Synthesis of 2-6 (R or S)-l-(4-chloro-2-fluorophenyl)-3-(2-(2-methylpyridin-4- yl)morpholino)-6.7.8.9-tetrahvdro-10/Z-pyrimido[6.1-6]quinazolin-10-one and 2-7 (S or R)-l-(4- chloro-2-fluorophenyl)-3-(2-(2-methylpyridin-4-yl)morpholino)-6.7.8.9-tetrahvdro-107/- pyrimido[6,l -6] quinazolin- 10-one

[0203] To a 20 mL vial were added 3-chloro-l-(4-chloro-2-fluorophenyl)-6,7,8,9-tetrahydro- 107f-pyrimido [6,1 -6] quinazolin- 10-one (60.0 mg, 0.165 mmol), 2-(2-methylpyridin-4- yl)morpholine (49.9 mg, 0.28 mmol), RuPhos Pd G4 (21.0 mg, 0.025 mmol) and cesium carbonate (161 mg, 0.49 mmol). The vial was purged with Ar three times then 1,4-Dioxane (1.1 mL) was added. The vial was purged with Ar again and heated in a hot plate at 100 °C for 2.5 h. Upon completion, the reaction mixture was cooled to rt, diluted with water (3 mL) and extracted with DCM (3 mL x 2). The organic extracts were treated with Silia MetS ® (Si-Thiol), filtered and concentrated. The residue was diluted with DMSO, filtered and purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield l-(4-chloro-2-fluorophenyl)-3- (2-(2-methylpyridin-4-yl)morpholino)-6,7,8,9-tetrahydro- 107/-pyrimido[6.1 -6J quinazolin- 10-one as a desired product, which was subjected to chiral SFC separation to yield Peak 1 product 2-6 (R or ,S')- 1 -(4-chloro-2-fluorophenyl)-3-(2-(2-methylpyridin-4-yl)morpholino)-6.7.8.9-tetrahydro- lOTApyrimido [6, 1-6] quinazolin- 10-one (tR = 2.6 min) and Peak 2 product as 2-7 (S or R)-l-(4- chloro-2-fluorophenyl)-3-(2-(2-methylpyridin-4-yl)morpholino)-6,7,8,9-tetrahydro-1077- pyrimido[6,l-6]quinazolin-l 0-one (tR = 4.1 min). MS (ESI) m/z: calc’d for C27H25CIFN5O2 [M+H]⁺: 506.2, found: [M+H]⁺: 506.1.

Example 2-6 Peak 1 ’H NMR: *H NMR (600 MHz, DMSO-d6) 6 8.45 (d, J= 5.1 Hz, 1H), 7.68 (m. 1H), 7.47 (d. J= 10.2 Hz, 1H), 7.41 (m. 1H), 7.36 (s, 1H), 7.27 (m. 1H), 6.63 (s, 1H), 4.62 (m, 1H), 4.41 (m, 2H), 4.10 (m, 1H), 3.73 (s, 1H), 3.17 (m, 1H), 3.00 - 2.93 (m, 1H), 2.59 (m, 2H), 2.32 (m, 2H), 1.74 - 1.67 (m, 4H).

Example 2-7 Peak 2 'H NMR: 'H NMR (600 MHz, DMSO-d6) 6 8.44 (d, J= 5. 1 Hz, 1H), 7.67 (m. 1H), 7.47 (dd, J= 10.1, 1.8 Hz, 1H), 7.41 (m, 1H), 7.36 (s, 1H), 7.28 (m, 1H), 6.63 (s, 1H), 4.62 (m, 1H), 4.41 (m. 2H), 4.10 (m, 1H). 3.74 (s. 1H), 3.17 (m, 1H). 2.97 (dd. J= 13.1. 10.7 Hz, 1H), 2.59 (m, 2H), 2.31 (t, J= 5.9 Hz, 2H), 1.74-1.67 (m, 4H).

SFC condition: Column & Dimensions: IH, 21x250 mm, 5 pm UV Wavelength: 215 nm Flow Rate: 70 mL/min Modifier: 30% MeOH w/0.1%NH4OH Outlet Pressure: 100 bar Sample Amount: 15 mg Diluent: 1: 1 MeOH/CHsCN EXAMPLE 2-8

Preparation of 2-8 (5')-9-(4-chloro-2-nuorophenyl)-7-(2-( 1 -methyl- l//-pyrazol-4-yl )morpholino)-

3.4-dihydropyranol3.2-a^flpyrimido[1.6-a1pyrimidin-ll(277)-one

Step 1: Preparation of (_<S)-7-(2-(l-methyl-l/f-pyrazol-4-yl)morpholino)-9-(methylthio)-3.4- dihvdropyrano[3,2-6<npyrimido[L6-a|pyrimidin-l l(27/)-one

[0204] To a 8 mL of vial containing (<S)-6-(2-(l -methyl- l/f-pyrazol-4-y l)morpholino)-2- (methylthio)pyrimidin-4-amine (100 mg, 0.326 mmol) (intermediate 1-5) and ethyl 3- oxotetrahydro-277-pyran-2-carboxylate (169 mg, 0.98 mmol) was added PPA (0.33 mL). The resulting mixture w as heated in a hot plate at 120 °C for 0.5 h. Upon cooling, the reaction mixture was treated with DCM (5 mL) and carefully neutralized with sat. NaHCCL aq. (6 mL) until the pH was slightly above 7. The organic layer was extracted with DCM (5 mL) two times and the combined organic extracts were concentrated. The resulting residue was purified by column chromatography on silica gel column, eluting with 0% to 10% EtOAc in DCM to give desired product (5)-7-(2-(l -methyl- 17f-pyrazol-4-yl)morpholino)-9-(methylthio)-3,4- dihydropyrano[3,2-J]pyrimido[l,6-«]pyrimidin-ll(2/0-one. MS (ESI) m/z: calc'd for C19H22N6O3S [M+H]⁺: 415.1, found: [M+H]⁺: 415.1

Step 2: Preparation of 2-8 (5)-9-(4-chloro-2-fluorophenyl)-7-(2-(l-methyl-l/7-pyrazol-4- yl)morpholino)-3.4-dihydropyrano[3.2-tZ]pyrimido[1.6-a1pyrimidin-l l(2/Z)-one

[0205] In a 2 mL microwave vial containing (<.S)-7-(2-(l-methyl-l//-pyrazol-4-yl)morpholino)- 9-(methylthio)-3.4-dihydropyrano|3.2-c/|pyrimido| 1.6-o|pyrimidin- 1 l(2/Z)-one (25.0 mg, 0.06 mmol), (4-chloro-2-fluorophenyl)boronic acid (21.0 mg, 0.121 mmol), Tris(dibezylideneacetone)dipalladium (5.5 mg, 0.006 mmol) and phosphine, tri-2-furanyl- (2.80 mg, 0.012 mmol) was added THF (0.6 mL). The reaction was then heated in a microwave reactor at 100 °C for 7 min. The reaction mixture was diluted with DCM and treated with 100 mg of Silia MetS® (Si-Thiol). The mixture was stirred at rt for 5 min, then filtered. The filtrate w as concentrated, diluted with water (2 mL) and sat. NaHCOs aq. (1 mL) and extracted with DCM (2 mL x 2). The concentrated residue was diluted with DMSO, filtered and purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield 2-8 (₁S)-9-(4-chloro-2- fluorophenyl)-7-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-3,4-dihydropyrano[3,2- <^pyrimido[l,6-a]pyrimidin-l l(2LZ)-one as a desired product. MS (ESI) m/z: calc’d for C24H22CIFN6O3 [M+H]⁺: 497.1, found: [M+H]⁺: 497.1

‘H NMR (600 MHz, DMSO-d6) d 7.74 (s, 1H), 7.71 (t, J= 8.1 Hz, 1H), 7.52 (d, J= 10.2 Hz, 1H), 7.47 (s, 1H), 7.43 (dd, J= 8.3, 1.8 Hz, 1H), 6.50 (s, 1H), 4.55 (s, 1H), 4.32-4.01 (b, 5H), 3.82 (s, 3H), 3.68 (s, 1H), 3.28 - 3.14 (m, 2H), 2.72 (s, 2H), 2.00 (m, 2H).

EXAMPLE 2-9

Preparation of 2-9 (₁S^T)-l-(4-chloro-2-fluorophenyl)-8.8-difluoro-3-(2-(l-methyl-177-pyrazol-4- yl)morpholino)-6,7.8,9-tetrahy dro- 10/Z-pyrimido [6.1 -Z>] quinazolin- 10-one

[0206] To a 8 mL vial containing (6’)-2-(4-chloro-2-fluorophenyl)-6-(2-(l-methyl-177-pyrazol- 4-yl)morpholino)pyrimidin-4-amine (30.0 mg, 0.077 mmol) (1-6) and ethyl 5,5-difluoro-2- oxocyclohexane-l-carboxylate (159 mg, 0.77 mmol) was added PPA (0.1 mL). The resulting mixture was heated in a hot plate at 100 °C for 45 mins. Upon completion, the reaction was diluted with DCM (5 mL) and carefully quenched with sat. NaHCCh aq. until the solution become slightly basic (pH>7). The organic extract was concentrated, diluted with DMSO (2 mL), filtered and purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to afford 2-9 (5)-l-(4-chloro-2-fluorophenyl)-8,8-difluoro-3-(2-(l-methyl-177-pyrazol-4- yl)morpholino)-6,7,8,9-tetrahydro-1077-pyrimido[6.1-6]quinazolin-10-one as a desired product. MS (ESI) m/z: calc’d for C25H22CIF3N6O2 [M+H]⁺: 531.1, found: [M+H]⁺: 531.2. 'H NMR (600 MHz, DMSO-d6) O' 7.74 (s, 1H), 7.69 (t, J = 7.9 Hz, 1H), 7.51 (d, J= 10.1 Hz, 1H), 7.47 (s, 1H), 7.44 > 7.40 (m, 1H), 6.66 (s, 1H), 4.55 (m, 1H), 4.4 - 4.2 (m, 2H), 4.01 (m, 1H), 3.82 (s, 3H), 3.31 - 3.18 (m. 2H), 3.75 - 3.70 (m, 1H) 2.88 - 2.82 (m. 4H), 2.30 (m, 2H).

[0207] Examples 2-10 through 2-12 below were prepared according to procedures analogous to those outlined in Example 2-9 above using the appropriate starting materials.

Table 4: EXAMPLES 2-10 to 2-12

EXAMPLE 2-13 and 2-14

Preparation of 2-13 8-(2.4-difluorophenyl)-6-((2S. 6R or 2R.6S)-2-methyl-6-(2-methylpyridin-4- yl)morpholino)-1.3-dihydro-10H-furo[3.4-dlpyrimido[1.6-alpyrimidin-10-one & 2-14 8-12,4- difluorophenyl)-6-((2R.6S or 2S.6R) -2-methyl-6-12-methylpyridin-4-yl)morpholino)-1.3- dihydro-10H-furo[3.4-d1pyrimido[1.6-a1pyrimidin-l 0-one

Step 1: Synthesis of 8-(2.4-difluorophenyl)-6-llCIS)-2-methyl-6-(2-methylpyridin-4- yl)morpholino)-1.3-dihydro-10H-furo[3.4-d]pyrimido[1.6-a1pyrimidin-10-one

[0208] To a solution of 6-chloro-8-(2,4-difluorophenyl)-l,3-dihydro-10H-furo[3.4- d]pyrimido[l,6-a]pyrimidin-10-one (50 mg, 0.149 mmol), CsF (67.9 mg, 0.447 mmol) and 2- methyl-6-(2-methylpyridin-4-yl)morpholine (34.4 mg, 0.179 mmol) in DMSO (1 ml) was added DIPEA (0.078 ml, 0.447 mmol) in a glove box. The resulting mixture was stirred at 80 °C for 10 min. Upon completion, the mixture was filtered and the filtrate was purified by Pre-HPLC (MeCN/ water with 0.1% TFA) to give 8-(2,4-difluorophenyl)-6-(2-methyl-6-(2-methylpyridin- 4-yl)morpholino)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one as a desired product.

MS (ESI) m/z: calc'd for C26H24F₂N₅O3⁺ [M+H]⁺ 492.2, found [M+H]⁺ 492.0.

Step 2: Preparation of 2-13 8-(2.4-difluorophenyl)-6-( 6R or 2R,6S)-2-methyi-6-(2-

methylpyridin-4-yl)morpholino)-1.3-dihvdro-10H-furo[3.4-d]pyrimido[1.6-a1pyrimi din- 10-one

& 2-14 8-(2.4-difluorophenyl)-6-( r 2 ,dA)-2-methyl-6-(2-methylpyridin-4-

yl)morpholino)-1.3-dihvdro-10H-furor3.4-d1pyrimidoll.6-a1pyrimidin-l 0-one

[0209] The cis-8-(2,4-difluorophenyl)-6-(2-methyl-6-(2-methylpyridin-4-yl)morpholino)-l,3- dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (50 mg, 0.102 mmol) was resolved by Chiral-SFC (Column DAICEL CHIRALCEL OD (250 mm*30 mm, 10 um); Condition: CO2 iPrOH (0.1% NH3H2O); Begin B 45 End B 45 Gradient; Time (min) 10; 100% B Hold Time 10; Flow Rate (ml/min): 80) to give 2-13 8-(2,4-difluorophenyl)-6-((2S. 6R or 2/ . CS')-2-methyl-6-(2- methylpyridin-4-yl)morpholino)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-l 0-one (ti = 1.370 min) as the first eluting peak, and 2-14 8-(2.4-difluorophenyl)-6-((2/ 6S or 2S,6R)-2- methyl-6-(2-methylpyridin-4-yl)morpholino)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6- a]pyrimidin-10-one (ti = 1.862 mm) as the second eluting peak.

SFC Peak 1: MS (ESI) m'z: calc'd for C26H24F2N5O3’ [M+H] ⁺ 492.0, found [M+H] ⁺ 492.0 'H NMR (400 MHz, CDCh) 5 ppm 8.53 (d, 2=5.01 Hz, 1 H), 7.51 - 7.61 (m, 1 H), 7.23 (s, 1 H), 7.13 - 7.17 (m, 1 H), 6.98 - 7.06 (m, 1 H), 6.82 - 6.90 (m, 1 H), 6.37 (s, 1 H), 4.89 - 5.13 (m, 4 H). 4.56 - 4.64 (m. 1 H). 4.08 - 4.54 (m. 2 H). 3.82 - 3.94 (m. 1 H), 2.84 - 3.00 (m, 2 H), 2.61 (s. 3 H), 1.39 (d, 2=6.08 Hz, 3 H)

SFC Peak 2: MS (ESI) m,z: calc'd for C26H24F2N5O3 [M+H] ⁺ 492.0, found [M+H] ⁺ 492.0 ’H NMR (400 MHz, CDCh) 5 ppm 8.53 (d, 2=5.13 Hz, 1 H), 7.50 - 7.62 (m, 1 H), 7.23 (s, 1 H), 7.13 - 7.18 (m, 1 H), 6.99 - 7.06 (m, 1 H), 6.80 - 6.90 (m, 1 H), 6.37 (s. 1 H). 4.90 - 5.15 (m. 4 H), 4.56 - 4.63 (m, 1 H), 4.03 - 4.54 (m, 2 H), 3.83 - 3.93 (m, 1 H), 2.84 - 2.99 (m, 2 H), 2.61 (s, 3 H), 1.39 (d, .7=6,20 Hz, 3 H)

[0210] Examples 2-15 through 2-17 below were prepared according to procedures analogous to those outlined in Example 2-13 to 2-14 above using the appropriate starting materials.

Table 5: EXAMPLES 2-15 to 2-17

Additional Exemplary Compounds

[0211] Additional exemplary compounds 3-1 through 3-50 are listed in Table 6 below.

[0212] The synthesis of diastereomers of Example 3-29 are described below in Examples 4-1 and 4-2.

EXAMPLES 4-1 and 4-2

Preparation of 4-1 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-((2R,4S or 2S,4R)-2-(2- melhylpyridin-4-yl )tetrahvdro-2//-pyrari-4-yl )-477-pyrirnido| I ,6-a1pyrimidin-4-one and 4-2 6-(4- chloro-2-fluorophenyl)-2,3-dimethyl-8-((2S.4R or 2R.4S)-2-(2-methylpyridin-4-yl)tetrahvdro-

21/-pyran-4-yl)-477-pyrimido[ 1.6-a1pyrimidin-4-one

[0213] To a 20 mL vial containing 4-(4-b romotetrahy dro-2 /-pyran-2-y 1 )-2-methy 1 pyridine (94.7 mg, 369.6 pmol) were added 8-chloro-6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-4/7- pyrimido[l,6-a]pyrimidin-4-one 1-10 (100.0 mg, 295.7 pmol), zinc (58.00 mg, 887.2 pmol), pyridine-2,6-bis(carboximidamide) hydrochloride (8.9 mg, 44.36 pmol), sodium iodide (17.7 mg, 118.3 pmol) and nickel chloride, dimethoxy ethane adduct (9.7 mg, 44.4 pmol). The vial was purged with Ar three times, followed by the addition of DMA (1.5 mL). It was purged with Ar three times again and stirred at rt for a few minutes, then heated in a hot plate at 60 °C for 1 h 15 min. The mixture was cooled to room temperature, then water was added, and the mixture was extracted with DCM. The combined organic extracts were concentrated. The crude residue was purified by reverse phase HPLC (0-100% acetonitrile in water, with NELOH modifier) to yield desired product as mixture of isomers, which was then submitted for SFC chiral separation to yield two major products 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-((2R,4S or 2S.4R)-2-(2- methylpyridin-4-yl)tetrahydro-2/7-pyran-4-yl)-42/-pyrimido[l,6-a]pyrimidin-4-one as Peak 1 4-1 (at 3.4 min SFC) and 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-((2S,4R or 2R,4S)-2-(2- methylpyridin-4-yl)tetrahydro-21/-pyran-4-yl)-477-pyrimido[l ,6-a]pyrimidin-4-one Peak 2 4-2 (at 4.8 mm SFC). MS (ESI) m/z: calc’d for C26H25CIFN4O2 [M+H]⁺: 479.2, found: [M+H]⁺: 479.1.

SFC Column & Dimensions: IH, 21x250 mm, 5 pm; UV Wavelength: 215 nm; Flow Rate: 70 ml/min; Modifier: 20% MeOH w/ 0.1% NH4OH; Outlet pressure: 100 bar.

SFC Peak 1 (4-1): ’H NMR (600 MHz, DMSO-O 5 8.38 (d, J = 5.1 Hz, IH), 7.66 (s, IH), 7.48 (dd, J= 10.2, 1.7 Hz, IH). 7.41 (dd, J= 8.3. 1.6 Hz. IH), 7.26-7.25 (m. 2H), 7.17 (d. J= 5.0 Hz. IH), 4.53 (d, J = 9.7 Hz, IH), 4.20 (dd, J= 11.3, 3.7 Hz, IH), 3.74 - 3.69 (m, IH), 3.19 (ddd, J = 11.9, 8.4, 3.4 Hz, IH), 2.45 (s, 3H), 2.38 (s, 3H), 2.21 (b, IH), 1.99 (s, 3H), 1.94 (b, IH), 1.82 (b, IH), 1.58 (b, IH).

SFC Peak 2 (4-2): 'H NMR (600 MHz, DMSO-Je) 5 8.38 (d, J= 5.1 Hz, IH), 7.66 (s, IH), 7.48 (dd, J= 10.1, 1.8 Hz, IH), 7.41 (dd, J= 8.3, 1.8 Hz, IH), 7.26-7.25 (m, 2H), 7. 17 (d, J= 4.6 Hz, IH), 4.53 (d, J= 9.5 Hz, IH), 4.20 (dd, J= 11.4, 3.6 Hz, IH), 3.71 (td, J= 11.9, 2.1 Hz, IH), 3.23 - 3.16 (m, IH), 2.45 (s, 3H), 2.38 (s, 3H), 2.21 (b, IH), 1.99 (s, 3H), 1.94 (b, IH), 1.82 (b, IH), 1.58 (b, IH).

EXAMPLE 4-3

Preparation of 4-3 6-(4-chloro-2-fluorophenyl)-8-((2R,4S or 2S.4R)-2-(l-cyclopropyl-17f- pyrazol-4-yl)tetrahydro-27/-pyran-4-yl)-2.3-dimethyl-4/7-pyrimido[1.6-a1pyrimidin-4-one

[0214] To a vial containing 8-chloro-6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-47/- pyrimido[l,6-a]pyrimidin-4-one (50.0 mg, 148 pmol) and 4-((2R,4S or 2S,4R)-4- bromotetrahydro-2H-pyran-2-yl)-l-cyclopropyl-lH-pyrazole (Intermediate 1-8) (50.1 mg, 185 pmol) were added sodium iodide (5.5 mg. 37.0 pmol), zinc (19.3 mg. 296 pmol). pyridine-2,6- bis(carboximidamide) hydrochloride (3.0 mg, 14.8 pmol) and nickel chloride, dimethoxyethane adduct (3.3 mg, 14.8 pmol). The mixture was purged with Ar several times, followed by addition of DMA (1 mL). The resulting solution was purged with Ar three times and stirred at rt for a few mins before the reaction mixture was heated at 60 °C for 1 h. Upon completion, the reaction was diluted with DMSO and purified by reverse phase HPLC (0-100% acetonitrile in water, with NH4OH modifier) to yield desired product 4-3 6-(4-chloro-2-fluorophenyl)-8-((2R,4S or 2S,4R)- 2-( I -cyclopropyl- 1 H-pyrazol-4-yl)tetrahydro-2//-pyran-4-yl)-2.3-dimethyl-4H-pyrimido| 1.6- a]pyrimidin-4-one. MS (ESI) m/z: calc’d for C26H26CIFN5O2 [M+H]⁺: 494.0, found: [M+H]⁺: 494.2

^XH NMR (600 MHz, DMSO-O 5 7.74 (s, 1H), 7.69 (t, J= 8. 1 Hz, 1H), 7.49 (d, J= 10.2 Hz, 1H), 7.42 (d, J= 8.2 Hz, 1H), 7.39 (s, 1H), 7.27 (s, 1H), 4.44 (d, J= 11.0 Hz, 1H), 4.06 (d, J = 7.5 Hz, 1H), 3.68 - 3.62 (m, 2H), 3.09 (t, J= 12.0 Hz, 1H), 2.39 (s, 3H), 2.15 (b, 1H), 2.00 (s, 3H), 1.90 (b, 2H), 1.75 (b, 2H), 1.0 - 0.99 (m, 2H), 0.92 (d, J= 5.9 Hz, 2H).

EXAMPLES 4-4, 4-5, 4-6 and 4-7

Preparation of 4-4, 4-5, 4-6 and 4-7 6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-((2S.4R)-2- ((R)-tetrahvdrofuran-3-yl)tetrahvdro-277-pyran-4-yl)-4/f-pyrimido[1.6-a1pyrimidin-4-one. 6-(4- chloro-2-fluorophenyl)-2.3-dimethyl-8-((2R.4S)-2-((S)-tetrahvdrofuran-3-yl)tetrahydro-277- pyran-4-yl )-4H-pyrimido| 1.6-a1pyrimidin-4-one. 6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8- ((2R.4S)-2-((R)-tetrahvdrofuran-3-yl)tetrahvdro-27/-pyran-4-yl)-477-pyrimido[1.6-a1pyrimi din-4- one and 6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-((2S.4R)-2-((S)-tetrahvdrofuran-3- v] )tetrahvdro-2/7-pyran-4-yl)-4H-pyrimidol 1.6-al pyrimidin-4-one.

Step 1 : Preparation of 6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-(2-(tetrahvdrofuran-3- yl )tetrahvdro-2/7-pyran-4-yl )-4H-pyrimido| 1.6-a1pyrimidin-4-one

[0215] A mixture of NiCb DME (16.9 mg, 0.077 mmol) and picolinimidamide hydrochloride (12.0 mg, 0.077 mmol) in DMA (15 mL) in a glove box was stirred at 25 °C for 1 h. Then 8- chloro-6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-4/7-pyrimido[l,6-a]pyrimidin-4-one (130 mg, 0.384 mmol), 4-iodo-2-(tetrahydrofuran-3-yl)tetrahydro-277-pyran (130 mg, 0.461 mmol), TBAI (142 mg, 0.384 mmol) and zinc (101 mg, 1.538 mmol) were added, and the reaction mixture stirred at 40 °C for 1 h. The solvent was removed under reduced pressure and the residue was dissolved in water (20 mL) and EtOAc (20 mL). The organic layer was separated and the aqueous was re-extracted with EtOAc (20 mL*3) and the combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase HPLC (50-100% water and acetonitrile gradient, TFA modifier) to give 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-(2-(tetrahydrofuran- 3-yl)tetrahydro-277-pyran-4-yl)-477-pyrimido[l,6-a]pyrimidin-4-one.

MS (ESI) m/z: calc’d for C24H26CIFN3O3 [M+H]⁺: 459.2/561.2, found [M+H]⁺: 459.1/561.1

Step 2: Preparation 6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-((2S.4R)-2-((R)-tetrahvdrofuran- 3-yl )tetrahvdro-2//-pyran-4-yl )-4//-pyrimido| 1 .6-a1pyrimidin-4-one, 6-(4-chloro-2- nuorophenyl)-2.3-diniethyl-8-((2R.4S)-2-((S)-tetrahvdrofuran-3-yl)tetrahvdro-2//-pyran-4-yl )- 4/7-pyrimido[L6-a1pyrimidin-4-one. 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-((2R.4S)-2- ((R)-tetrahydrofuran-3-yl)tetrahydro-2/7-pyran-4-yl)-47f-pyrimido[ L6-a]pyrimidin-4-one and 6- (4-chloro-2-fluorophenyl)-2.3-dimethyl-8-((2S.4R)-2-((S)-tetrahvdrofuran-3-yl)tetrahvdro-27/- pyran-4-yl )-4//-pyrimido| l .6-a|pyrimidin-4-one

[0216] 6-(4-chloro-2-fluorophenyl)-2,3-dimethyl-8-(2-(tetrahydrofuran-3-yl)tetrahydro-2/7- pyran-4-yl)-477-pyrimido[l,6-a]pyrimidin-4-one (75 mg, 0.164 mmol) was resolved by Chiral- SFC [Column DAICEL CHIRALPAK IG (250 mm x 30 mm, 10 pm); Condition CCh-EtOH (0.1% NH3H2O); Begin B 60% End B 60%, Gradient Time (min): 10; 100% B Hold Time 1, Flow Rate (mL/min): 80) to give 4-4 (SFC Peak 1; Rt = 0.884 min), 4-5 (SFC Peak 2; Rt = 1.121 mm), 4-6 (SFC Peak 3; Rt = 1.535 mm) and 4-7 (SFC Peak 4; Rt = 2. 104 mm).

4-4 (SFC Peak 1): MS (ESI) m/z: calc’d for C24H26CIFN3O3 [M+H]⁺: 459.2/461.2, found [M+H]⁺: 459.1/461.1

'H NMR (400 MHz, CDCh) 5 = 7.58 (t. J= 8.0 Hz. 1H), 7.29 (br dd. J= 1.6, 8.3 Hz. 1H), 7.11 (dd, J= 1.8,10.0 Hz. 1H), 7.02 (s, 1H), 4.17 (br dd, J = 3.9.11.4 Hz, 1H). 3.92 - 3.70 (m. 3H),

3.66 - 3.49 (m, 2H), 3.42 - 3.21 (m, 1H), 3.02 - 2.85 (m, 1H), 2.45 (s, 3H), 2.38 - 2.29 (m, 1H), 2.12 (s, 3H), 2.09 - 2.00 (m, 1H), 1.98 - 1.85 (m, 3H), 1.80 (br d, J= 10.6 Hz, 1H), 1.50 (q, J = 12.1 Hz, 1H)

4-5 (SFC Peak 2): MS (ESI) m/z: calc’d for C24H26CIFN3O3 [M+H]⁺: 459.2/461.2, found [M+H]⁺: 459.1/461.1

'H NMR (400 MHz, CDCh) 5 = 7.58 (t, J= 8.0 Hz, 1H), 7.29 (dd, J= 1.6, 8.4 Hz, 1H), 7.11 (dd, J= 1.8,10.1 Hz, 1H), 7.03 (s, 1H), 4.14 (br dd, J= 3.8,11.1 Hz, 1H), 4.00 - 3.84 (m, 2H), 3.81 -

3.67 (m, 2H), 3.57 (dt, J = 2.0.11.9 Hz, 1H), 3.30 (br t, J= 9.7 Hz, 1H), 2.98 - 2.86 (m, 1H), 2.45 (s, 3H). 2.37 - 2.28 (m, 1H). 2.13 (s. 3H), 2.07 (br d, J = 11.1 Hz. 1H), 2.03 - 1.90 (m, 2H), 1.87 - 1.71 (m, 1H), 1.68 - 1.61 (m, 1H), 1.52 - 1.41 (m, 1H)

4-6 (SFC Peak 3): MS (ESI) m/z: calc’d for C24H26CIFN3O3 [M+H]⁺: 459.2/461.2, found [M+H]⁺: 459.1/461.1

'H NMR (400 MHz, CDCh) 5 = 7.58 (t. J= 8.0 Hz. 1H), 7.32 - 7.28 (m, 1H), 7.11 (dd, J = 1.5,10.0 Hz, 1H), 7.03 (s, 1H), 4.14 (dd, J= 3.9,1 1.5 Hz, 1H), 3.98 - 3.84 (m, 2H), 3.80 - 3.67 (m, 2H), 3.57 (dt, J= 1.8,12.0 Hz, 1H), 3.30 (br t, J= 9.5 Hz, 1H), 3.01 - 2.84 (m, 1H), 2.45 (s, 3H), 2.37 - 2.27 (m, 1H), 2.13 (s, 3H). 2.09 (br s, 1H), 2.03 - 1.91 (m, 2H), 1.85 - 1.72 (m, 1H), 1.68 - 1.61 (m, 1H), 1.46 (q, J= 12.0 Hz, 1H)

4-7 (SFC Peak 4): MS (ESI) m/z: calc’d for C24H26CIFN3O3 [M+H]⁺: 459.2/461.2, found [M+H]⁺: 459.1/461.1

¹H NMR (400 MHz, CDC13) 5 = 7.60 (t. J= 7.9 Hz. 1H), 7.31 (dd, J= 1.7, 8.2 Hz, 1H), 7.13 (dd, J= 1.7.10.1 Hz, 1H), 7.04 (s, 1H). 4.19 (dd. J= 4. 1.11.3 Hz, 1H), 3.93 - 3.82 (m, 2H), 3.81 - 3.74 (m, 1H), 3.66 - 3.52 (m, 2H), 3.43 - 3.28 (m, 1H), 2.98 - 2.85 (m, 1H), 2.46 (s, 3H), 2.41 - 2.31 (m, 1H), 2.14 (s, 3H), 2.10 - 2.02 (m, 1H), 1.97 (br d, J= 12.4 Hz, 2H), 1.94 - 1.86 (m, 1H), 1.84 - 1.72 (m, 1H), 1.55 - 1.46 (m, 1H)

[0217] Examples shown in Table 7 below were prepared according to procedures analogous to those outlined for Examples 4-1 to 4-7 above, using the appropriate starting materials and purification methods. Examples 4-18 and 4-19 were made using Intermediate 1-8.

Table 7

Example 4-37, 4-38, 4-39, 4-40:

Preparation of 4-37 6-(2Adifluorophenyl)-2.3-dirnethyl-8-((27?,45) or (2S,4R)-2-(((R or S)- tetrahvdrofuran-3-yl)methyl)tetrahydro-2H-pyran-4-yl)-4H-pyrimido[1.6-a1pyrimidin-4-one & 4- 38, 6-(2.4-difluorophenyl)-2.3-dimethyl-8-((2S,47?) or (2R4S)-2-(((R or 53-tetrahydrofuran-3- yl)methyl)tetrahydro-2H-pyran-4-yl)-4H-pyrimido[1.6-a1pyrimidin-4-one & 4-39, 6-(2,4- difluorophenyl)-2.3-dimethyl-8-(27?.4_<S7 or (2S,4R)-2-((( S or 7?)-tetrahydrofuran-3- yl)methyl)tetrahvdro-2H-pyran-4-yl)-4H-pyrimido[ 1.6-a1pyrimidin-4-one & 4-40, 6-(2,4- difluorophenyl)-2,3-dimethyl-8-((2S, 47?) or (2R,4S) 2-((( S’ or /?)-tetrahvdrofuran-3- yl)methyl)tetrahvdro-2H-Dyran-4-yl)-4H-pyrirnidol 1.6-al pyrimidin-4-one

Step 1: Synthesis of (E)-3-(2-methoxyvinyl)tetrahydrofuran

[0218] To a mixture of (methoxymethyl)triphenylphosphonium bromide (27. 1 g, 69.9 mmol) in THF (330 mL) was added sodium amide (2.73 g, 69.9 mmol), at 25 °C . The mixture was stirred at same temperature for 0.5 h, followed by the addition of tetrahydrofuran-3-carbaldehyde (7 g, 69.9 mmol). The resulting mixture was stirred at the same temperature for 2 h. Upon completion, the solvent was removed under reduced pressure and the residue was dissolved in water (300 mL) and EtOAc (300 mL). The organic layer was separated and the aqueous layer was reextracted with EtOAc (300 mL*3), then the combined organic layers were washed with brine (200 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the (E)-3-(2-methoxyvinyl)tetrahydrofuran as desired product. MS (ESI) m/z: calc'd for C₇Hi3O2⁺ [M+H]+: 129, found: 129.

Step 2: Synthesis of 2-(tetrahydrofuran-3-yl)acetaldehyde

[0219] A mixture of (E)-3-(2-methoxyvinyl)tetrahydrofuran (10 g, 78 mmol) and hydrogen chloride (15 mL, 30.0 mmol) in MeCN (12 mL) was stirred at 25 °C for 16 h. The solvent was removed under reduced pressure and the residue was dissolved in water (200 mL) and EtOAc (200 mL). The organic layer was separated and the aqueous layer was re-extracted with EtOAc (200 mL*3). The combined organic layers were washed with brine (200 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 30% ethyl acetate/petroleum ether gradient) to give 2-(tetrahydrofuran-3-yl)acetaldehyde as a desired product.

Step 3: Synthesis of 6-((tetrahvdrofuran-3-yl)methyl)-3.6-dihvdro-2H-pyran-4-yl trifluoromethanesulfonate

[0220] A mixture of 2-(tetrahydrofuran-3-yl)acetaldehyde (100 mg, 0.876 mmol) and but-3-yn- l-ol (92 mg, 1.314 mmol) and Tf-OH (0.233 mL, 2.63 mmol) in DCM (2 mL) was stirred at 25 °C for 16 h. The solvent was removed under reduced pressure and the residue was dissolved in water (5 mL) and EtOAc (5 mL). The organic layer was separated and the aqueous layer was reextracted with EtOAc (5 mL*3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 25% ethyl acetate/petroleum ether gradient) to give 6-((tetrahydrofuran-3-yl)methyl)-3,6-dihydro-2H-pyran-4-yl trifluoromethanesulfonate as a desired product.

Step 4: Synthesis of 6-(2.4-difluorophenyl)-2.3-dimethyl-8-(6-((tetrahvdrofuran-3-yl)methyl)- 3.6-dihvdro-2H-pyran-4-yl)-4H-pyrimido[1.6-a]pyrimidin-4-one

[0221] A mixture of 6-(2,4-difluorophenyl)-2,3-dimethyl-8-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-4H-pyrimido[l,6-a] pyrimidin-4-one (400 mg, 0.774 mmol), DTBPF Pd G3 (64.9 mg, 0.077 mmol) . K3PO4 (329 mg, 1.549 mmol) and 6-((tetrahydrofuran-3-yl)methyl)-3,6- dihydro-2H-pyran-4-yl trifluoromethanesulfonate (245 mg, 0.774 mmol) in 1,4-dioxane (5 mL) and water (0.500 mL) was degassed and backfilled with N2 (three times). The mixture was heated to 80 °C for 0.5 h. After cooling to 25 °C, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL*3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC (MeCN/water using 0. 1%TFA modifier) to give 6-(2,4- difluorophenyl)-2,3-dimethyl-8-(6-((tetrahydrofuran-3-yl)methyl)-3,6-dihydro-2H-pyran-4-yl)- 4H-pyrimido[l,6-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc’d for C25H26F₂N3O₃ ⁺ [M+H]⁺: 454.1, found [M+H]⁺: 454.1.

Step 5: Synthesis of 6-(2.4-difluorophenyl)-2,3-dimethyl-8-(2-((tetrahvdrofuran-3- yl)methyl)tetrahvdro-2H-pyran-4-yl)-4H-pyrimidol 1.6-a|pyrimidin-4-one

[0222] To a solution of 6-(2,4-difluorophenyl)-2,3-dimethyl-8-(6-((tetrahydrofuran-3- yl)methyl)-3,6-dihydro-2H-pyran-4-yl)-4H-pyrimido[l,6-a]pyrimidin-4-one (180 mg, 0.397 mmol) in THF (5 mL) was added TEA (0.166 mL, 1.191 mmol), Palladium(II) chloride (70.4 mg, 0.397 mmol) under N2. Then tri ethylsilane (0.190 mL, 1.191 mmol) was added via syringe and the resulting mixture was stirred at 25 °C for 24 h. Upon completion, the solvent was removed under reduced pressure and the residue was dissolved in water (10 mL) and EtOAc (10 mL). The organic layer was separated, the aqueous layer was re-extracted with EtOAc (10 mL*3) and the combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC (MeCN/water using 10 mM NH4HCO3) to give 6-(2,4-difluorophenyl)-2,3-dimethyl-8-(2- ((tetrahydrofuran-3-yl)methyl)tetrahydro-2H-pyran-4-yl)-4H-pyrimido[l,6-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc’d for C₂5H₂8F₂N3O3⁺ [M+H]⁺: 456.1, found [M+H]⁺: 456.1.

Step 6: Preparation of 4-37 6-(2.4-difluorophenyl)-2.3-dimethyl-8-((2A,45) or (2S,4R)-2-(((R or

S)-tetrahydrofuran-3-yl)methyl)tetrahydro-2H-pyran-4-yl)-4H-pyrimido[1.6-a1pyrimidin-4-one

& 4-38, 6-(2.4-difluorophenyl)-2.3-dimethyl-8-((2_<S’, 47?) or (2R4S)-2-(((R or M-tetrahvdrofuran- 3-yl)methyl)tetrahvdro-2H-pyran-4-yl)-4H-pyrimido[1.6-a1pyrimidin-4-one & 4-39, 6-(2,4- difluorophenyl)-2.3-dimethyl-8-(27?.45) or (2S,4R)-2-((( S o/' /?)-tetrahvdrofuran-3- yl)methyl)tetrahvdro-2H-pyran-4-yl)-4H-pyrimido[ 1.6-a1pyrimidin-4-one & 4-40, 6-(2.4- difluorophenyl)-2.3-dimethyl-8-((2S, 47?) or (2R,4S) 2-((( S or 7?)-tetrahydrofuran-3- yl)methyl)tetrahvdro-2H-pyran-4-yl)-4H-pyrimido[1.6-a1pyrimidin-4-one

[0223] The 6-(2,4-difluorophenyl)-2,3-dimethyl-8-(2-((tetrahydrofuran-3-yl)methyl)tetrahydro- 2H-pyran-4-yl)-4H-pyrimido[l,6-a]pyrimidin-4-one (40 mg, 0.088 mmol) was resolved by Chiral-SFC (Column DAICEL CHIRALPAK IC (250 mm*30 mm. 5 um); Condition: CO₂-i- PrOH (0. 1%NH3H₂Q); Begin B 45 End B 45; Gradient Time (mm): 5; 100%B Hold Time: 1; Flow Rate (rnL/min): 150) to give 4-37 6-(2,4-difluorophenyl)-2,3-dimethyl-8-((27?,45>) or (2S, 4R)-2-(((R or 5')-tetrahydrofuran-3-y 1 )methyl )tetrahy dro-2H-py ran-4-yl )-4H-pynmido| 1 ,6- a]pyrimidin-4-one_(Rt = 1.807 min) as the first eluting peak, and 4-38 6-(2,4-difluorophenyl)-2,3- dimethyl-8-((2S, 4R) or (2R.4S)-2-(((R or <S’)-tetrahydrofuran-3-yl)methyl)tetrahydro-2H-pyran-4- yl)-4H-pyrimido[l,6-a]pyrimidin-4-one (Rt = 2.042 min) as the second eluting peak, 4-39 6-(2,4- difluorophenyl)-2,3-dimethyl-8-(27?, 4S) or (2S,4R)-2-((( S or //(-telrahy drofuran-3- yl)methyl)tetrahydro-2H-pyran-4-yl)-4H-pyrimido[l,6-a]pyrimidin-4-one (Rt = 2.304 min) as the third eluting peak, and 4-40 6-(2,4-difluorophenyl)-2,3-dimethyl-8-((2<S’, 4R) or (2R,4S) 2-((( S or /?)-tetrahydrofuran-3-yl (methyl )tetrahydro-2H-pyran-4-yl)-4H-pyrimido| 1.6-a|pynmidin-4-one (Rt = 3.307 min) as the fourth eluting peak.

SFC Peak 1; 4-37: MS (ESI) m/z: calc’d for C₂₅H₂₈F₂N₃O₃ ⁺ [M+H]⁺: 456.1. found [M+H]⁺:

456.1.

'H NMR (400 MHz, CDCh) 5 ppm 7.53 - 7.61 (m, 1 H), 6.90 - 7.01 (m, 2 H), 6.72 - 6.79 (m, 1 H), 4.04 - 4.12 (m, 1 H), 3.83 - 3.91 (m, 1 H), 3.76 - 3.82 (m, 1 H), 3.64 - 3.72 (m, 1 H), 3.49 (td, .7=11.95, 2.09 Hz, 1 H), 3.22 - 3.39 (m, 2 H), 2.79 - 2.93 (m, 1 H), 2.37 (s, 3 H), 2.31 - 2.36 (m, 1 H). 2.05 (s, 3 H). 1.84 - 2.01 (m. 3 H). 1.62 - 1.78 (m. 2 H). 1.39 - 1.53 (m. 3 H)

SFC Peak 2; 4-38: MS (ESI) m/z: calc’d for C₂₅H₂₈F₂N₃O₃ ⁺ [M+H]⁺: 456.1, found [M+H]⁺:

456.1.

'H NMR (400 MHz, CDCh) 5 ppm 7.53 - 7.61 (m, 1 H), 6.92 - 6.98 (m, 2 H), 6.72 - 6.79 (m, 1 H), 4.05 - 4.12 (m, 1 H), 3.88 (t, 7=7.87 Hz, 1 H), 3.75 - 3.82 (m, 1 H), 3.65 - 3.72 (m, 1 H), 3.49 (td, 7=11.95, 2.09 Hz, 1 H), 3.32 - 3.43 (m, 1 H), 3.27 (t, 7=8.11 Hz, 1 H), 2.82 - 2.92 (m, 1 H),

2.37 (s, 3 H), 2.29 - 2.36 (m, 1 H), 2.05 (s, 3 H), 1.86 - 2.03 (m, 3 H), 1.61 - 1.80 (m. 2 H), 1.41 - 1.52 (m, 3 H)

SFC Peak 3; 4-39: MS (ESI) m/z: calc’d for C₂₅H₂₈F₂N₃O₃ ⁺ [M+H]⁺: 456.1, found [M+H]⁺:

456.1.

'H NMR (400 MHz, CDCh) 5 ppm 7.53 - 7.61 (m, 1 H), 6.92 - 6.99 (m, 2 H), 6.72 - 6.80 (m, 1 H), 4.05 - 4.11 (m, 1 H), 3.85 - 3.91 (m, 1 H), 3.76 - 3.83 (m, 1 H), 3.65 - 3.73 (m, 1 H), 3.49 (td, 7=11.86, 1.91 Hz, 1 H), 3.32 - 3.42 (m, 1 H), 3.27 (t, 7=8.05 Hz, 1 H), 2.82 - 2.91 (m, 1 H), 2.37 (s, 3 H), 2.27 - 2.35 (m, 1 H), 2.05 (s, 3 H), 1.83 - 2.02 (m, 3 H), 1.59 - 1.65 (m, 2 H), 1.38 - 1.51 (m, 3 H)

SFC Peak 4; 4-40: MS (ESI) m/z: calc’d for C₂₅H₂₈F₂N₃O₃ ⁺ [M+H]⁺: 456.1, found [M+H]⁺:

456.1.

'H NMR (400 MHz, CDCh) 5 ppm 7.60 - 7.69 (m, 1 H), 7.00 - 7.08 (m, 2 H), 6.80 - 6.87 (m, 1 H). 4.12 - 4.19 (m. 1 H). 3.94 (t. 7=7.69 Hz. 1 H). 3.87 (td, 7=8.23, 4.65 Hz, 1 H), 3.72 - 3.80 (m, 1 H), 3.57 (td, 7=11.92, 2.03 Hz, 1 H), 3.33 - 3.48 (m, 2 H), 2.89 - 2.98 (m, 1 H), 2.45 (s, 3 H),

2.38 - 2.44 (m, 1 H), 2.12 (s, 3 H), 1.92 - 2.10 (m, 3 H), 1.67 - 1.75 (m, 2 H), 1.48 - 1.60 (m, 3 H) EXAMPLES 5-1 AND 5-2

Preparation of 5-1 8-(2,4-difluorophenyl)-6-((2S.4R or 2R.4S)-2-(2-methylpyridin-4- yl')tetrahydro-2//-pyran-4-yl)-1.3-dihydro-lQ7f-furor3.4-d1pyrimido[1.6-a1pyrimidin-lQ-one and 5-2 8-(2.4-difluorophenyl)-6-((2R.4S or 2S.4R)-2-(2-methylpyridin-4-yl)tetrahvdro-277-pyran-4- y 1)- 1.3-dihvdro- 1 OH- furo [3,4-dl pyrimidol 1.6-al pyrimidin- 10-one

Step 1 : Preparation of 6-(2-(2-methylpyridin-4-yl)tetrahydro-27/-pyran-4-yl)-8-(methylthio)-l,3- dihydro-10H-furo[3.4-d1pyrimido[1.6-a1pyrimi din-10-one

[0224] A mixture of pyridine-2,6-bis(carboximidamide) hydrochloride (33.3 mg, 0.167 mmol) and nickel(II) chloride ethylene glycol dimethyl ether complex (36.7 mg. 0. 167 mmol) in DMA (1 ml) in glove box was stirred at 25 °C for 0.5 h. Then zinc (291 mg, 4.45 mmol), sodium iodide (66.7 mg, 0.445 mmol), 6-chloro-8-(methylthio)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6- a]pyrimidin-10-one (300 mg, 1.112 mmol) and 4-(4-bromotetrahydro-2H-pyran-2-yl)-2- methylpyridine (342 mg. 1.335 mmol) were added, and the reaction mixture was stirred at 80 °C for 2 h. Upon completion, the reaction mixture was quenched with aqueous ammonium chloride (100 mL) and extracted with EtOAc (50 mL*3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure. The residue was purified by Prep-TLC (silica gel, ethyl acetate/petroleum ether = 1/0, v/v) to give 6-(2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-8-(methylthio)-1.3-dihydro-10H- furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one. MS (ESI) m/z: calc’d for C21H23N4O3S [M+H]⁺: 411.0, found [M+H]⁺: 411.0.

Step 2: Preparation of 8-(2.4-difluorophenyl)-6-(2-(2-methylpyridin-4-yl)tetrahydro-27/-pyran-4- yl)- 1 ,3-dihvdro- 1 OEf-furo [3,4-dl pyrimidof 1.6-a] pyrimidin- 10-one

[0225] A solution of Pd2(dba)3 (4.46 mg, 4.87 pmol), 6-2-(2-methylpyridin-4-yl)tetrahydro-22T- pyran-4-yl)-8-(methylthio)-l,3-dihydro-1077-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (40 mg, 0.097 mmol), copper(I) thiophene-2-carboxylate (55.7 mg, 0.292 mmol), tri(furan-2- yl)phosphane (2.3 mg, 9.74 pmol) and (2,4-difluorophenyl)boronic acid (23.1 mg, 0.146 mmol) in THF (2 mL) was prepared in a glove box. The reaction was heated to 100 °C and stirred for 15 min by microwave. The reaction mixture was quenched with aqueous ammonium chloride (5 mL) and extracted with EtOAc (5 mL*3). The combined organic phases were washed with brine (10 mL), dried over anhydrous NazSO-i. filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC (0-100% water and MeCN; NH4OH modifier) to give 8-(2,4- difluorophenyl)-6-(2-(2-methylpyridin-4-yl)tetrahydro-27/-pyran-4-yl)-L3-dihydro-10H- furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one.

MS (ESI) m/z: calc’d for C26H23F₂N4O3[M+H]⁺: 477.2, found [M+H]⁺ 477.2.

Step 3: Preparation of 5-1 8-(2,4-difluorophenyl)-6-((2S,4R or 2R,4S)-2-(2-methylpyridin-4- yl)tetrahvdro-27/-pyran-4-yl)-l,3-dihydro-1077-furo[3.4-d1pyrimido[L6-a1pyrimi din- 10-one and 5-2 8-(2.4-difluorophenyl)-6-((2R,4S or 2S.4R)-2-(2-methylpyridin-4-yl)tetrahydro-277-pyran-4- yl)- 1.3-dihvdro- 1 O/7-furol 3.4-d I pyrimidol 1.6-al pyrimidin- 10-one

[0226] 8-(2,4-difluorophenyl)-6-(2-(2-methylpyridin-4-yl)tetrahydro-277-pyran-4-yl)- 1 ,3- dihydro-1077-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (15 mg, 0.031 mmol) was resolved by Chiral-SFC [Column: DAICEL CHIRALPAK AD (250 mm x 30 mm, 10 pm); Condition: CO2- iPrOH (0.1% NH3H2O); Begin B 60% End B 60%; Gradient Time (min) 60; 100% B Hold Time: 1; Flow Rate (ml/min): 80] to give 5-1 8-(2,4-difluorophenyl)-6-(2S,4R or 2R,4S)-2-(2- methylpyridin-4-yl)tetrahydro-277-pyran-4-yl)-l,3-dihydro-1077-furo[3,4-d]pyrimido[l,6- a] pyrimidin- 10-one (SFC Peak 1: tr = 1.015 min), and 5-2 8-(2,4-difluorophenyl)-6-(2R,4S or 2S,4R)-2-(2-methylpyridin-4-yl)tetrahydro-277-pyran-4-yl)-l,3-dihydro-10/7-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one (SFC Peak 2; tr = 1.548 min).

5-1 (SFC Peak 1): MS (ESI) m/z: calc’d for C26H23F2N4O3 [M+H]⁺: 477.2/479.2, found [M+H]⁺:477.2/479.2

'H NMR (400 MHz, CHLOROFORM- ) 5 ppm 8.47 (d, 7=5.13 Hz, 1 H), 7.62 - 7.70 (m, 1 H),

7.17 - 7.22 (m.2 H). 7.10 (br d, 7=4.89 Hz, 1 H), 7.01 - 7.08 (m, 1 H), 6.81 - 6.89 (m, 1 H), 5.00 - 5.20 (m, 4 H), 4.50 (d, 7=10.37 Hz, 1 H), 4.33 - 4.41 (m, 1 H), 3.75 - 3.83 (m, 1 H), 3.14 - 3.23 (m, 1 H), 2.57 (s, 3 H), 2.26 - 2.33 (m, 1 H), 1.97 - 2.08 (m, 2 H), 1.72 - 1.78 (m, 1 H)

5-2 (SFC Peak 2): MS (ESI) m/z: calc’d for C26H23F2N4O3 [M+H]⁺: 477.2/479.2, found [M+H]⁺: 477.2/479.2.

'H NMR (400 MHz, CHLOROFORM-7) 5 ppm 8.47 (d, J=5.13 Hz, 1 H), 7.61 - 7.71 (m, 1 H),

7.17 - 7.21 (m, 2 H), 7.10 (d, 7=5.13 Hz, 1 H), 7.02 - 7.08 (m, 1 H), 6.81 - 6.88 (m, 1 H), 5.00 - 5.21 (m, 4 H), 4.50 (d, 7=10.25 Hz, 1 H), 4.33 - 4.40 (m, 1 H), 3.75 - 3.84 (m, 1 H), 3.14 - 3.24 (m, 1 H), 2.57 (s. 3 H). 2.26 - 2.33 (m. 1 H). 1.98 - 2.07 (m. 2 H). 1.71 - 1.78 (m. 1 H)

EXAMPLES 5-3 and 5-4

Preparation of 5-3 6-((2S,4R or 2R.4S)-2-(l-cyclopropyl-17f-pyrazol-4-yl)tetrahvdro-277-pyran-

4-yl)-8-(2.4-difluorophenyl)-1.3-dihydro-1077-furo[3.4-d1pyrimido[1.6-a]pyrimi din- 10-one and

5-46-((2R.4S or 2S.4R)-2-(l-cvclopropyl-177-pyrazol-4-yl)tetrahvdro-27f-pyran-4-yl)-8-(2.4-

Step 1 : Preparation of 6-chloro-8-(2.4-difluorophenyl)-1.3-dihydro-1077-furo[3.4- d1pyrimido[L6-a1pyrimidin-10-one

[0227] To a solution Pd2(dba)3 (50.9 mg, 0.056 mmol), 6-chloro-8-(methylthio)-l,3-dihydro- 1077-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (300 mg, 1.112 mmol), copper(I) thiophene-2- carboxylate (636 mg, 3.34 mmol). tri(furan-2-yl)phosphane (25.8 mg, 0. 111 mmol) and (2,4- difluorophenyl)boronic acid (263 mg, 1.669 mmol) was added THF (10 mL) in a glove box. The reaction was heated to 100 °C for 15 min in a microwave. The reaction mixture was quenched with aqueous ammonium chloride (15 mL) and extracted with EtOAc (15 mL*3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0-30% ethyl acetate/petroleum ether) to give 6-chloro-8-(2,4-difluorophenyl)- 1 ,3-dihy dro- 1 ()//-furo| 3.4-d |pyrimido| 1 ,6-a| pyrimidin- 10-one.

MS (ESI) m/z: calc’d for C15H9CIF2N3O2 [M+H]⁺: 334.0/336.0 found [M+H] ⁺: 334.1/336.1.

Step 2: Preparation of 6-(6-(l-cyclopropyl-177-pyrazol-4-yl)-3.6-dihydro-2//-pyran-4-yl)-8-(2.4- difluorophenyl)-L3-dihvdro-10/7-furo[3.4-d1pyrimido[L6-a1pyrimidin-10-one

[0228] A mixture of Pd (dtbpf)C12 (23.30 mg, 0.036 mmol), Na2CCh (76 mg, 0.715 mmol), 1- cyclopropyl-4-(4-(4,4,5,5-tetraethyl-l,3,2-dioxaborolan-2-yl)-5,6-dihydro-277-pyran-2-yl)-l£7- pyrazole (173 mg, 0.465 mmol) and 6-chloro-8-(2,4-difluorophenyl)-L3-dihydro-107 -furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one (120 mg, 0.357 mmol) in toluene (4 mL) and water (0.400 mL) was degassed and backfilled with N2 (three times). The mixture was heated to 60 °C for 16 h. After cooling to room temperature, the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0-100% ethyl acetate/petroleum ether) to give 6-(6-( I -cyclopropyl- 17/-pyrazol-4-yl)-3.6-dihydro-2/7- pyran-4-yl)-8-(2.4-difluorophenyl)- 1 ,3-dihydro- 107f-furo[3,4-d]pyrimido[ 1 ,6-a]py rimidin- 1 Ci- one.

MS (ESI) m/z: calc'd for C26H22F2N5O3 [M+H]⁺ 490.0, found [M+H]⁺490.1.

Step 3: Preparation of 6-(2-(l-cyclopropyl-177-pyrazol-4-yl)tetrahydro-27/-pyran-4-yl)-8-(2.4- difluorophenyl)-1.3-dihvdro-1077-furo[3.4-d1pyrimido[L6-a1pyrimidin-10-one

[0229] To a solution of 6-(6-(l-cyclopropyl-lF7-pyrazol-4-yl)-3,6-dihydro-2/7-pyran-4-yl)-8- (2, 4-difluorophenyl)-l,3-dihydro-1077-furo[3,4-d]pyrimido[l,6-a]pyrimidin- 10-one (37 mg, 0.076 mmol) and palladium (II) chloride (1.3 mg, 7.56 pmol) in THF (1 mL) was added TEA (0.032 mL. 0.227 mmol) and triethylsilane (0.036 mL, 0.227 mmol) under N2. The resulting mixture was stirred at 0-25 °C for 2 h. The reaction mixture was quenched with water (1 mL) and extracted with EtOAc (1 mL*3). The combined organic phases were washed with brine (1 mL), dried over anhydrous Na2SO4, fdtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give 6-(2-(l-cyclopropyl-l//-pyrazol-4-yl)tetrahydro-277-pyran-4-yl)- 8-(2,4-difluorophenyl)-l ,3-dihydro-l 07/-furo[3,4-d]pyrimido[l ,6-a]pyrimidin- 10-one.

MS (ESI) m/z: calc'd for C26H24F2N5O3 [M+H]⁺ 492.1, found [M+H]⁺ 492.1

Step 4: Preparation of 5-3 6-((2S.4R or 2R.4S)-2-(l-cvclopropyl-l/f-pyrazol-4-yl)tetrahydro-27/- pyran-4-yl)-8-(2.4-difluorophenyl)-1.3-dihydro-107/-furo[3.4-d1pyrimido[1.6-a1pyrimidin-10- one and 5-4 6-((2R,4S or 2S.4R)-2-(l-cvclopropyl-17f-pyrazol-4-yl)tetrahvdro-27if-pyran-4-yl)-8-

(2.4-difluorophenyl)-1.3-dihydro-10//-furo[3.4-d1pyrimidori.6-a1pyrimi din- 10-one

[0230] 6-(2-(l-cyclopropyl-17f-pyrazol-4-yl)tetrahydro-277-pyran-4-yl)-8-(2,4-difluorophenyl)- l,3-dihydro-10/7-furo[3.4-d]pyrimido[l,6-a]pyrimidin-10-one (17 mg, 0.035 mmol) was resolved by Chiral-SFC (Column DAICEL CHIRALPAK IH (250 mm x 30 mm, 10 pm);

Condition: CO2 -iPrOH (0.1% NH3H2O); Begin B 30 End B 30; Gradient Time (min) 5; 100% B Hold Time: 1; Flow Rate (ml/min): 150) to give 5-3 6-((2S,4R or 2R,4S)-2-(l-cyclopropyl-177- pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-8-(2,4-di fluorophenyl)-!, 3-dihydro-10 -furo[3, 4- d]pyrimido[l,6-a]pyrimidin-10-one (SFC Peak 1; tr = 2.616 min), and 5-4 6-((2R,4S or 2S,4R)-2- (l-cyclopropyl-lH-pyrazol-4-yl)tetrahydro-27f-pyran-4-yl)-8-(2,4-difluorophenyl)-l,3-dihydro- 107f-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (SFC Peak 2; ti = 2.818 min).

5-3 (SFC Peak 1): MS (ESI) mz calc'd for C26H24F2N5O3 [M+H] ⁺ 492.2, found [M+H] ⁺ 492.2 'H NMR (400 MHz, CDCh-J) 5 ppm 7.68 (td, 7=8.31, 6.38 Hz, 1 H), 7.47 (s, 2 H), 7.20 (s, 1 H), 7.05 (td, 7=8.17, 2.15 Hz, 1 H), 6.85 (ddd, 7=10.61, 8.58, 2.38 Hz, 1 H), 5.02 - 5.21 (m, 4 H), 4.52 (dd, 7=11.27, 1.73 Hz, 1 H), 4.22 - 4.30 (m, 1 H), 3.76 (td, 7=11.71, 2.68 Hz, 1 H), 3.56 (tt, 7=7.29. 3.74 Hz, 1 H), 3.07 - 3.18 (m, 1 H), 2.29 (br d, 7=13.23 Hz, 1 H), 1.94 - 2.07 (m. 2 H).

1.82 - 1.92 (m, 1 H), 1.07 - 1.13 (m, 2 H), 0.97 - 1.03 (m, 2 H).

5-4 (SFC Peak 2): MS (ESI) m/z: calc'd for C26H24F2N5O3 [M+H]+ 492.2, found [M+H]⁺ 492.2 ^JH NMR (400 MHz, CDCh-7) 5 ppm 7.68 (td, 7=8.29, 6.32 Hz, 1 H), 7.47 (s, 2 H), 7.20 (s, 1 H), 7.05 (td, 7=8.26, 1.97 Hz, 1 H), 6.85 (ddd. 7=10.58, 8.55, 2.32 Hz, 1 H), 5.02 - 5.23 (m, 4 H), 4.52 (dd, 7=1 1.21, 1.67 Hz, 1 H), 4.21 - 4.30 (m, 1 H), 3.76 (td, 7=11.68, 2.74 Hz, 1 H), 3.56 (tt, J=121, 3.76 Hz, 1 H), 3.08 - 3.19 (m, 1 H), 2.29 (br d, 7=12.87 Hz, 1 H), 1.93 - 2.07 (m, 2 H),

1.83 - 1.90 (m, I H), 1.06 - 1.13 (m, 2 H), 0.97 - 1.04 (m, 2 H).

[0231] Examples shown in Table 5 below were prepared according to procedures analogous to those outlined in examples 5-1 to 5-4 above using the appropriate starting materials.

Table 8

Example 5-18 & 5-19 & 5-20 & 5-21

Preparation of 5-18 (R or S)-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2R.4S or 2S.4R)-2-(2- methylpyridin-4-yl)tetrahvdro-2H-pyran-4-yl)-1.3-dihvdro-10H-furor3.4-d1pyrimido[1.6- a] pyrimidin- 10-one & 5-19 (S or R)-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2R.4S or 2S.4R)- )-2-(2-methylpyridin-4-yl)tetrahvdro-2H-pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-d1pyrimidori.6- al pyrimidin- 10-one & 5-20 (S or R)-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2S.4R or 2R.4S)- 2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihydro-10H-furo|3.4-d|pyrimido| 1.6- a]pyrimidin-10-one & 5-21 (R or S)-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2S.4R or 2R.4S)- 2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-d1pyrimido[1.6-

- I l l - al pyrimidin- 10-one

Step 1 : Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-l-methyl-1.3-dihydro-10H-furo[3.4- dlpyrimidol L6-a1pyrimidin-10-one

1 2

[0232] To a solution of Pd2(dba)s (0. 161 g, 0. 176 mmol), (4-chloro-2-fluorophenyl)boronic acid (0.922 g, 5.29 mmol), copper(l) thiophene-2-carboxylate (2.016 g. 10.57 mmol), tri(furan-2- yl)phosphane (0.082 g, 0.352 mmol) in THF (15 mL) was added 6-chloro-l-methyl-8- (methylthio)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (1 g, 3.52 mmol) in glove box. The reaction was heated to 100 °C and stirred for 15 min by micro wave reactor. Upon completion, the mixture was quenched with sat. NaHCCh.aq. (10 mL) and extracted with EtOAc (20 mL * 3). The combined organic fractions were washed with brine (20 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 0-30% EtOAc : EtOH=3 : 1 /Hexane gradient) to give 6-chloro- 8-(4-chloro-2 -fluorophenyl)- l-methyl-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimi din- 10- one as a desired product.

MS (ESI) m/z: calc'd for CieHi 1CI2FN3O2’ [M+H] ⁺ 366, found [M+H] ⁺ 366. Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-l-methyl-6-(2-(2-methylpyridin-4- yl)tetrahvdro-2H-pyran-4-yl)-1.3-dihydro-10H-furo[3.4-d1pyrimido[1.6-a1pyri midin- 10-one

[0233] A mixture of picolinimidamide hydrochloride (0.086 g, 0.546 mmol) and Nickel(II) chloride ethylene glycol dimethyl ether complex (0.120 g, 0.546 mmol) in DMA (2 mL) was stirred at 25 °C for 30 min under N2 atmosphere. Then zinc (0.536 g, 8.19 mmol), tetrabutylammonium iodide (1.513 g, 4.10 mmol), 4-(4-iodotetrahydro-2H-pyran-2-yl)-2- methylpyridine (0.993 g. 3.28 mmol), 6-chloro-8-(4-chloro-2-fluorophenyl)-l-methyl-l,3- dihydro-10H-furo[3.4-d]pyrimido[l,6-a]pyrimidin-10-one (1 g. 2.73 mmol) in DMA (8 mL) were added. The mixture was stirred at 40 °C for 2 h under N2 atmosphere. Upon completion, the reaction mixture was quenched with aqueous ammonium chloride (50 mL) and extracted with EtOAc (20 mL*3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC (water (MeCN/water with 10 mM-NLLHCCh) to give 8-(4-chloro-2-fluorophenyl)- l-methyl-6-(2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one.

MS (ESI) m/z: calc'd for C27H25CIFN4CU [M+H]⁺ 507, found [M+H]⁺ 507

Step 3: Preparation of 5-18 (R or M-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((27?, AS* or 2S.4RY 2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-L3-dihvdro-10H-furo[3.4-d1pyrimido[L6- a] pyrimidin- 10-one & 5-19 (S or 7?)-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2J?,-/S or 2S,4R)- )-2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-L3-dihvdro-10H-furo[3.4-d1pyrimido[L6- al pyrimidin- 10-one & 5-20 (S or 7U-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2S, AR or 2]

2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-L3-dihydro-10H-furo[3.4-d1pyrimido[L6- a] pyrimidin- 10-one & 5-21 (R or M-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2S,47? or 2R.4S}-

2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-L3-dihydro-10H-furo[3.4-d1pyrimido[L6- al pyrimidin- 10-one

[0234] The 8-(4-chloro-2-fluorophenyl)- 1 -methyl-6-(2-(2-methylpyridin-4-yl)tetrahy dro-2H- pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (230 mg, 0.454 mmol) was resolved by Chiral-SFC (Colum DAICEL CHIRALPAK AD: (250 mm * 50 mm, 10 um);

Condition: CCh-i-PrOH; Begin B 60 End B 60; Gradient Time (min): 130; 100%B Hold Time: 1; Flow Rate (ml/min): 70) to give mixture 1 as the first eluting peak, 5-19 [(S' or R)-8-(4-chloro-2- fl uoro phony I)- 1 -methyl -6-((2/<’.4S or 2S. •7R)-)-2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4- yl)-1.3-dihydro-10H-furo[3,4-d]pyrimido[1.6-a]pyrimidin-10-one It. = 1.706 min)] as the second eluting peak, and mixture 2.

5-19: MS (ESI) m/z: calc’d for C27H25C1FN4O2/ [M+H]⁺ 507, found [M+H]⁺ 507.

'H NMR (400 MHz, CDCI3) 5 8.49 (d, 7=5.40 Hz, 1 H), 7.57 - 7.67 (m, 1 H), 7.30 - 7.34 (m, 1 H), 7.27 - 7.29 (m, 1 H), 7.16 - 7.22 (m, 2 H), 7.13 (dd, 7=10.19, 1.85 Hz, 1 H), 5.32 - 5.52 (m, 1 H), 4.93 - 5.09 (m, 2 H), 4.53 (d, 7=10.00 Hz, 1 H), 4.34 - 4.41 (m, 1 H), 3.69 - 3.85 (m, 1 H), 3.13 - 3.27 (m, 1 H), 2.63 (s, 3 H), 2.29 (d, 7=1.00 Hz, 1 H), 2.00 - 2.10 (m, 2 H), 1.72 - 1.74 (m, 1 H), 1.48 - 1.57 (m, 3 H)

[0235] Mixture 1 was resolved by Chiral-SFC (Column DAICEL CHIRALCEL OX (250 mm * 30 mm, 10 um); Condition CO2-EtOH; Begin B 50 End B 50; Gradient Time (min): 50; 100%B Hold Time: 1; Flow Rate (ml/min): 150) to give 5-18: (R or S)-8-(4-chloro-2-fluorophenyl)-l- methyl-6-((2R,4S or 2S,4R)-2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro- 10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (ti = 0.708 min) as the first eluting peak.

5-18: MS (ESI) m/z: calc’d for C₂7H₂5C1FN4O3⁺ [M+H]⁺ 507. found [M+H]⁺ 507.

'H NMR (400 MHz, CDCh): 5 8.49 (d, 7=5.40 Hz, 1 H), 7.58 - 7.66 (m, 1 H), 7.29 - 7.34 (m, 2 H), 7.23 (d, 7=4.60 Hz, 1 H), 7.18 (s, 1 H), 7.13 (d, 7=10.30 Hz, 1 H), 5.34 - 5.52 (m, 1 H), 4.91 - 5.09 (m, 2 H), 4.54 (d, 7=10.80 Hz, 1 H), 4.31 - 4.44 (m, 1 H), 3.76 - 3.83 (m, 1 H), 3.14 - 3.25 (m. 1 H), 2.66 (s. 3 H). 2.31 (d, 7=13.20 Hz. 1 H). 2.02 - 2.10 (m. 2 H). 1.68 - 1.77 (m. 1 H). 1.46 - 1.58 (m, 3 H)

[0236] Mixture 2 was resolved by Chiral-SFC (Column DAICEL CHIRALPAK IC (250 mm * 50 mm, 10 um); Condition EtOH-ACN(Neu); Begin B 55 End B 55; Gradient Time (min): 5; 100%B Hold Time: 1; Flow Rate (rnL/min): 140) to give 5-20: (S’ or ?)-8-(4-chloro-2- fluorophenyl)-l-methyl-6-((2S,4R or 2R,4S)-2-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)- l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-l 0-one (ti = 1 .835 min) as the first eluting peak, and 5-21: (R or S)-8-(4-chloro-2-fluorophenyl)-l-methyl-6-((2S,4A or 2R,4S)-2-(2- methylpyridin-4-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6- a] pyrimidin- 10-one (tr = 3.346 min) as the second eluting peak.

5-20: MS (ESI) m/z: calc'd for C₂7H₂₅F₂N₄O3⁺ [M+H]⁺ 491.1/493. 1 , found [M+H]⁺ 491.1/493.1. 'H NMR (400 MHz, CDCh): 5 8.47 (d, J=5.13 Hz, 1 H), 7.58 - 7.68 (m, 1 H), 7.31 (d, 7=8.30 Hz, 1 H), 7.22 (s, 1 H), 7.17 (s, 1 H), 7.10 - 7.15 (m, 2 H), 5.31 - 5.51 (m, 1 H), 4.91 - 5.11 (m, 2 H), 4.51 (d, 7=9.90 Hz, 1 H), 4.32 - 4.40 (m, 1 H), 3.75 - 3.83 (m, 1 H), 3.13 - 3.24 (m, 1 H), 2.59 (s. 3 H). 2.29 (d, 7=13.00 Hz. 1 H). 1.94 - 2.09 (m. 2 H). 1.67 - 1.78 (m. 1 H). 1.45 - 1.59 (m, 3 H)

5-21: MS (ESI) m/z: calc'd for C₂7H₂₅F₂N4O3⁺ [M+H]⁺ 491.1/493.1, found [M+H]⁺ 491.1/493.1. 'H NMR (400 MHz, CDCh): 5 8.47 (d, 7=4.80 Hz, 1 H), 7.62 (d, 7=1.00 Hz, 1 H), 7.31 (d, 7=7.90 Hz. 1 H). 7.22 (s. 1 H). 7.18 (br s, 1 H), 7.09 - 7.15 (m, 2 H), 5.33 - 5.53 (m, 1 H), 4.92 - 5.10 (m, 2 H), 4.50 (d, 7=10.01 Hz, 1 H), 4.33 - 4.41 (m, 1 H), 3.74 - 3.84 (m, 1 H), 3.13 - 3.24 (m, 1 H), 2.58 (s, 3 H), 2.29 (d, 7=13.50 Hz, 1 H), 1.98 - 2.09 (m, 2 H), 1.69 - 1.78 (m, 1 H), 1.47 - 1.59 (m, 3 H)

Example 5-22, 5-23, 5-24, 5-25

Synthesis of 5-22 8-(2.4-difluorophenyl)-6-((27?,-7S)-2-< or R) tetrahvdrofuran-3-yl)tetrahvdro-

2H-pyran-4-yl)-1.3-dihydro-10H-furor3.4-d]pyrimido[1.6-a1pyrimidin-10-one . 5-23 8-(2,4- difluorophenyl)-6-((27?,4M-2-((7? or S) tetrahvdrofuran-3-yl)tetrahvdro-2H-pyran-4-yl)-1.3- dihydro-10H-furo[3.4-d1pyrimido[ 1.6-alpyrimidin-10-one, 5-24 8-(2.4-difluorophenyl)-6-

((2S, 4R)-2-((S or R) tetrahvdrofuran-3-yl)tetrahvdro-2H-pyran-4-yl)- 1 .3-dihydro-l 0H-furo[3,4- d]pyrimido[1.6-a1PYrimidin-10-one and 5-25 8-(2.4-difluorophenyl)-6-((2S,77?)-2-f(7? or S) tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihydro-10H-furo[3.4-d1pyrimidori.6-

al pyrimidin- 10-one

Step 1 : Synthesis of 8-(2.4-difluorophenyl)-6-(4.4.5.5-tetramethyl-1.3.2-dioxaborolan-2-yl)-l,3- dihvdro-10H-furo[3.4-d1pyri mi do[L6-a1pyrimi din- 10-one

[0237] A mixture of 6-chloro-8-(2,4-difluorophenyl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one (400 mg, 1.192 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'- bi(l,3,2-dioxaborolane) (454 mg, 1.787 mmol), potassium acetate (351 mg, 3.57 mmol) and [l,r-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (87 mg, 0.1 19 mmol) in 1,4- dioxane (5 mL) was degassed and backfilled with N2 (three times). The mixture was heated to 80 °C for 2 h. Upon completion, the reaction mixture was quenched with water (5 mL) and extracted with EtOAc (5 mL*3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure to give 8-(2,4- difluorophenyl)-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one which was not further purified.

MS (ESI) m/z: calc’d for C2iH₂iBF₂N3O4⁺[M+H]⁺: 428.0 found 346.0 [M+H]⁺. Step 2: Synthesis of 8-(2.4-difluorophenyl)-6-(6-(tetrahydrofuran-3-yl)-3.6-dihydro-2H-pyran-4- yD- 1.3-dihvdro- 10H-furo[3.4-d1pyrimido[ 1.6-al pyrimidin- 10-one

[0238] A mixture of 8-(2,4-difluorophenyl)-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-

1.3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (500 mg, 1.170 mmol), dtbpf Pd G3 (98 mg. 0. 117 mmol), K3PO4 (497 mg. 2.341 mmol) and 6-(tetrahydrofuran-3-yl)-3,6- dihydro-2H-pyran-4-yl trifluoromethanesulfonate (425 mg, 1 .404 mmol) in 1 ,4-dioxane (6 mL) and water (0.600 mL) was degassed and backfdled with N2 (three times). The mixture was heated to 80 °C for 0.5 h. Upon completion, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL*3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 0-30% ethyl acetate/petroleum ether) to give 8-(2,4-difluorophenyl)-6-(6-(tetrahydrofuran-3-yl)-3,6-dihydro-2H-pyran-4-yl)-

1.3-dihy dro- 10H-furo[3,4-d]pyrimido[l,6-a]pyrimi din- 10-one.

MS (ESI) m/z: calc’d for C₂4H₂2F₂N3O4⁺[M+H]⁺: 454.0 found 454.0 [M+H]⁺.

Step 3: Synthesis of 8-(2.4-difluorophenyl)-6-(2-(tetrahvdrofuran-3-yl)tetrahvdro-2H-pyran-4- vP- 1.3-dihy dro- 1 OH-furo [ 3 ,4-d] pyrimido [ 1.6-al pyrimidin- 10-one

[0239] To a solution of 8-(2.4-difluorophenyP-6-(6-(tetrahydrofuran-3-yl)-3.6-dihydro-2H- pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (357 mg, 0.512 mmol) and Palladium(II) chloride (9.07 mg, 0.051 mmol) in THF (3 mL) was added TEA (0.214 mL, 1.535 mmol) and triethylsilane (0.245 mL, 1.535 mmol) under N2. The resulting mixture was stirred at 0-25 °C for 2 h. Upon completion, the reaction mixture was quenched with water (3 mL) and extracted with EtOAc (3 mL*3). The combined organic phases were washed with brine (3 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC (water/MeCN with 10 mM NH4HCO3) to give 8-(2,4- difluorophenyl)-6-(2-(tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one.

MS (ESI) m/z: calc'd for C24H₂4F₂N3O4⁺ [M+H]⁺ 456.0. found [M+H]⁺ 456.1

Step 4: Preparation of 5-22 8-(2.4-difluorophenyl)-6-( or 2S,4R)-2-('(S or R)

tetrahydrofuran-3-yl)tetrahvdro-2H-pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-d1pyrimidori.6- al pyrimidin- 10-one . 5-23 8-(2,4-difluorophenyl)-6-((27?,45 or 2S.4R )-2-((R or S) tetrahvdrofuran-3-yl)tetrahvdro-2H-pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-d1pyrimido|T.6- al pyrimidin- 10-one, 5-24 8-(2.4-difluorophenyl)-6-((27?, 45 or 2S,4R }-2-((S or R) tetrahvdrofuran-3-yl)tetrahvdro-2H-pyran-4-yl)-L3-dihydro-10H-furo[3.4-d1pyrimido|T.6- al pyrimidin- 10-one and 5-25 8-(2.4-difluorophenyl)-6-((27?,45 or 2S,4R )-2-((R or S) tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihydro-10H-furo[3.4-d]pyrimido[1.6- a] pyrimidin- 10-one

[0240] The 8-(2,4-difluorophenyl)-6-(2-(tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3- dihydro-10H-furo[3.4-d]pyrimido[l,6-a]pyrimidin-10-one (170 mg, 0.373 mmol) was resolved by Chiral-SFC (Column DAICEL CHIRALPAK AD (250mm*30mm,10um); Condition CO2- EtOH; Begin B 50 End B 50; Gradient Time (min): 50; 100%B Hold Time: 1; Flow Rate (ml/min): 150) to give mixture 1 of first and second peaks, 5-24 [8-(2,4-difluorophenyl)-6- {(2R,4S or 2S,4R )-2-((5 or R) tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihydro-10H- furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (tr = 0.953 min)] as the third eluting peak, and 5-25 [8-(2,4-difluorophenyl)-6-((27?.45 or 2S,4R_)-2-((S or R) tetrahydrofuran-3-yl)tetrahydro-2H- pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (tr = 1.453 min)] as the third eluting peak.

5-24: MS (ESI) m/z: calc'd for C24H24F2N3OC [M+H]⁺ 456.0, found [M+H]⁺ 456.1. 'H NMR (400 MHz, CDCI3) 5 ppm 7.64 - 7.72 (m, 1 H), 7.17 (s, 1 H), 7.03 - 7.10 (m, 1 H), 6.82 - 6.89 (m, 1 H), 5.02 - 5.22 (m, 4 H), 4.13 - 4.19 (m, 1 H), 3.86 - 3.97 (m, 2 H), 3.70 - 3.79 (m, 2 H), 3.54 - 3.63 (m, 1 H), 3.28 - 3.36 (m, 1 H), 2.95 - 3.04 (m, 1 H), 2.29 - 2.39 (m, 1 H), 2.08 - 2.15 (m, 1 H). 1.91 - 2.03 (m, 2 H). 1.78 - 1.89 (m, 1 H), 1.62 - 1.68 (m, 1 H), 1.46 - 1.57 (m, 1 H) 5-25: MS (ESI) m/z: calc'd for C₂4H₂4F₂N3O4⁺ [M+H]⁺ 456.0, found [M+H]⁺ 456.1. 'H NMR (400 MHz, CDCh) 5 ppm 7.64 - 7.72 (m, 1 H), 7.16 (s, 1 H), 7.02 - 7.09 (m, 1 H), 6.82 - 6.89 (m, 1 H), 5.00 - 5.21 (m, 4 H), 4.16 - 4.23 (m, 1 H), 3.81 - 3.91 (m, 2 H), 3.73 - 3.80 (m, 1 H), 3.55 -

3.64 (m, 2 H), 3.31 - 3.37 (m, 1 H), 2.94 - 3.04 (m, 1 H), 2.30 - 2.41 (m, 1 H), 2.01 - 2.12 (m, 1 H). 1.93 - 2.01 (m. 2 H). 1.62 - 1.92 (m. 2 H). 1.50 - 1.59 (m. 1 H)

[0241] Mixture 1 was further resolved by Chiral-SFC (Column Phenomenex-Cellulose-2 (250 mm*50 mm, 10 um); Condition: CO₂- MeOH (0.1%NH3H₂O); Begin B 60 End B 60; Gradient Time (min): 120; 100%B Hold Time: 1; Flow Rate (ml/min): 80) to give 5-23: 8-(2,4- difluorophenyl)-6-((2/?.TS' or 2S.4R_)-2-RR or S) tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4- yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (tr = 0.731 min) as the second eluting peak, and mixture 2.

5-23: MS (ESI) m/z: calc'd for CMHMFZNSOT [M+H]⁺ 456.0, found [M+H]⁺ 456.1

'H NMR (400 MHz, CDCh) 5 ppm 7.64 - 7.71 (m, 1 H), 7.16 (s, 1 H), 7.02 - 7.09 (m, 1 H), 6.81 - 6.89 (m, 1 H), 5.01 - 5.24 (m, 4 H), 4.16 - 4.23 (m, 1 H), 3.81 - 3.91 (m, 2 H), 3.73 - 3.80 (m, 1 H), 3.55 - 3.64 (m, 2 H), 3.31 - 3.38 (m, 1 H), 2.94 - 3.04 (m, 1 H), 2.32 - 2.41 (m, 1 H), 2.01 - 2.12 (m, 1 H), 1.93 - 2.00 (m, 2 H), 1.79 - 1.93 (m, 2 H), 1.50 - 1.57 (m, 1 H).

[0242] Mixture 2 was further resolved by Chiral-SFC (Column DAICEL CHIRALPAK AD (250 mm*30 mm, 10 um); Condition: CO₂-iPrOH: Begin B 40 End B 40; Gradient Time (min): 50; 100%B Hold Time: 1 ; Flow Rate (ml/min): 150) to give 5-22: 8-(2,4-difluorophenyl)-6- (\2R.4S or 2S,4R )-2-((S or R) tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H- furo[3,4-d]pyrimido[l,6-a]pyrimi din-10-one (tr = 0.662 min) as the first eluting peak.

5-22: MS (ESI) m/z: calc'd for CMHMFZNSOC [M+H]⁺ 456.0, found [M+H]⁺ 456.1. 'H NMR (400 MHz, CDCh) 5 ppm 7.64 - 7.73 (m, 1 H), 7.17 (s, 1 H), 7.02 - 7.09 (m, 1 H), 6.82 - 6.88 (m, 1 H), 5.00 - 5.22 (m, 4 H), 4.13 - 4.20 (m, 1 H), 3.86 - 3.97 (m, 2 H), 3.70 - 3.80 (m, 2 H), 3.53 - 3.63 (m, 1 H), 3.28 - 3.36 (m, 1 H), 2.94 - 3.04 (m, 1 H), 2.28 - 2.40 (m, 1 H), 2.07 - 2.15 (m, 1 H), 1.91 - 2.04 (m, 2 H), 1.78 - 1.89 (m, 1 H), 1.64 - 1.69 (m, 1 H), 1.47 - 1.56 (m, 1 H)

Example 5-26, 5-27, 5-28, 5-29

Preparation of 5-26 8-(4-chloro-2-fluorophenyl)-6-( or 2S,4R)-2-((S or 7?)-tetrahydrofuran-

3-yl)tetrahydro-2H-pyran-4-yl)- 1 ,3-dihydro-l 0H-furo[3.4-d1pyrimido[ 1 ,6-alpyrimidin- 10-one, 5- 27 8-(4-chloro-2-fluorophenyl)-6-((2A,45 or 2S,4R)-2-((R or S)-tetrahydrofuran-3-yl)tetrahydro- 2H-pyran-4-yl)-1.3-dihvdro-10H-furor3.4-d1pyrimido[1.6-a1pyrimidin-10-one. 5-28 8-(4-chloro- 2-fluorophenyl)-6-( or 2S,4R)-2-((S or 7?)-tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-

1.3-dihvdro-10H-furo[3,4-d1pyrimido[1.6-a1pyrimidin-10-one and 5-29 8-(4-chloro-2- fluorophenyl)-6-((2R.45 or 2S.4R)-2-((R or 5)-tetrah\ drofuran-3-\'l)tetrah\ dro-2H-p\ ran-4-\'l )-

1.3-dihvdro-10H-furo[3.4-d1pyrimidori.6-a1pyrimidin-10-one

Step 1 : Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-1.3-dihydro-10H-furol3.4- d1pyrimido[l,6-a1pyrimidin-10-one

1 2

[0243] A solution of Pd2(dba)? (119 mg, 0.130 mmol), 6-chloro-8-(methylthio)-l,3-dihydro- 10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (700 mg, 2.60 mmol), copper(I)thiophene-2- carboxylate (1485 mg, 7.79 mmol), tri(furan-2-yl)phosphine (60.3 mg, 0.260 mmol) and (4- chloro-2-fluorophenyl)boronic acid (679 mg, 3.89 mmol) in THF (10 mL) was prepared in glove box. The reaction was heated to 100 °C and stirred for 15 min in a micro wave reactor. The reaction mixture was quenched with aqueous ammonium chloride (15 mL) and extracted with CH2CI2 (15 mL*3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SC>4, fdtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 0-30% ethyl acetate/petroleum ether gradient) to give 6-chloro-8-(4-chloro-2-fluorophenyl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6- a] pyrimidin- 10-one. MS (ESI) m/z: calc’d for Ci5H9ClF2NsO22⁺ [M+H]⁺: 352, found 352 [M+H]⁺ Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(6-(tetrahvdrofuran-3-yl)-3.6-dihydro-2H- pyran-4-yl)

[0244] A mixture of PdC12(dtbpf) (76 mg, 0.116 mmol), Na2COs (247 mg, 2.329 mmol), 4,4,5,5-tetraethyl-2-(6-(tetrahydrofuran-3-yl)-3,6-dihydro-2H-pyran-4-yl)-l,3,2-dioxaborolane (509 mg. 1.514 mmol) and 6-chloro-8-(4-chloro-2-fluorophenyl)-1.3-dihydro-10H-furo[3.4- d]pyrimido[l,6-a]pyrimidin-l 0-one (410 mg, 1. 164 mmol) in toluene (5 mL) and water (0.500 mL) was degassed and backfilled with N2 (three times). The mixture was heated to 60 °C for 1 h. Upon completion, the reaction mixture was quenched with water (5 mL) and extracted with EtOAc (5 mL*3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 0-100% ethyl acetate/petroleum ether gradient) to give 8-(4-chloro-2-fluorophenyl)-6-(6-(tetrahydrofuran-3-yl)-3,6-dihydro-2H-pyran-4-yl)-l,3- dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one. MS (ESI) wz: calc'd for C24H22CIFN3OL [M+HJ⁺ 470.0, found [M+H]⁺ 470.

Step 3: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(tetrahvdrofuran-3-yl)tetrahvdro-2H- pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-dlpyrimido[1.6-a1pyrimidin-l 0-one

[0245] To a solution of 8-(4-chloro-2-fluorophenyl)-6-(6-(tetrahydrofuran-3-yl)-3,6-dihydro- 2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (170 mg, 0.275 mmol) and Palladium(II) chloride (4.88 mg. 0.027 mmol) in THF (2 mL) was added TEA (0. 115 mL, 0.825 mmol) and triethylsilane (0.132 mL, 0.825 mmol) under N2. The resulting mixture was stirred at 0-25 °C for 2 h. Upon completion, the reaction mixture was quenched with water (2 mL) and extracted with EtOAc (2 mL*3). The combined organic phases were washed with brine (2 mL), dried over anhydrous Na₂SO4. filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC (MeCN/water with 0.1%TFA) to give 8-(4-chloro-2- fluorophenyl)-6-(2-(tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one. MS (ESI) m/z: calc'd forC24H24ClFN3O4⁺ [M+H]⁺ 472, found [M+H]⁺ 472.

Step 4: Preparation of 5-26 8-(4-chloro-2-fluorophenyl)-6-((27?, 45 or 2S,4RY2-((S

tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-d1pyrimido[1.6- al pyrimidin- 10-one, 5-27 8-(4-chloro-2-fluorophenyl)-6-((2A, 45 or 2S,4RY2-((R or 5)- tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihvdro-10H-furo[3.4-d1pyrimidorL6- al pyrimidin- 10-one, 5-28 8-(4-chloro-2-fluorophenyl)-6-((2A.45 or 2S.4R )-2-((5 or RY tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihydro-10H-furo[3.4-d1pyrimido|T.6- a] pyrimidin- 10-one and 5-29 8-(4-chloro-2-fluorophenyl)-6-((27?, 45 or 2S,4R Y2-((R or 5)- tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-1.3-dihydro-10H-furo[3.4-d1pyrimidoll.6- al pyrimidin- 10-one

[0246] 8-(4-chloro-2-fluorophenyl)-6-(2-(tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3- dihydro-10H-furo[3.4-d]pyrimido[1.6-a]pyrimidin-10-one (50 mg, 0.106 mmol) was resolved by Chiral-SFC (Column DAICEL CHIRALPAK AD: (250 mm*30 mm, 10 um): Condition: CO2- iPrOH (0.1%NH₃H₂O); Begin B 50 End B 50; Gradient Time (mm): 50; 100%B Hold Time: 1; Flow Rate (ml/min): 150) to give mixture of 8-(4-chloro-2-fluorophenyl)-6-(2-(tetrahydrofuran- 3-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one as the first peak and second eluting peak, 5-28 [8-(4-chloro-2-fluorophenyl)-6-((2R, 4S or 2S,4R )-2- ((5 or 7?)-tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one (tr=1.005min)] as the third eluting peak, and 5-29 [8-(4- chloro-2-fluorophenyl)-6-((25,45 or 2S,4R)-2-((R or 5)-tetrahydrofuran-3-yl)tetrahydro-2H- pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimidin-10-one (tr=1.429 min)J as the fourth eluting peak.

5-28: MS (ESI) m/z: calc’d for C24H24CIFN3O4 [M+H]⁺ 472, found [M+H]⁺472. 'H NMR (400 MHz, CDCh) 6 ppm 7.62 (t, J=8.05 Hz, 1 H), 7.32 (dd, J=8.29, 1.61 Hz, 1 H), 7.19 (s, 1 H), 7.13 (dd, 4=10.07, 1.73 Hz. 1 H). 5.02 - 5.21 (m, 4 H), 4.12 - 4.19 (m, 1 H), 3.85 - 3.97 (m, 2 H), 3.70 - 3.79 (m, 2 H), 3.53 - 3.63 (m, 1 H), 3.27 - 3.35 (m, 1 H), 2.94 - 3.05 (m, 1 H), 2.30 - 2.38 (m, 1 H). 2.06 - 2.13 (m, 1 H), 1.93 - 2.04 (m, 2 H), 1.76 - 1.92 (m, 2 H), 1.46 - 1.53 (m, 1 H)

5-29: MS (ESI) m/z: calc'd for C₂₄H₂₄C1FN3O4⁺ [M+H]⁺ 472, found [M+H]⁺472. 'H NMR (400 MHz, CDCh) 5 ppm 7.61 (t, 7=7.99 Hz, 1 H), 7.32 (dd, 7=8.34, 1.55 Hz, 1 H), 7.18 (s, 1 H), 7.13 (dd, 7=10.01, 1.79 Hz, 1 H), 5.01 - 5.23 (m, 4 H), 4.15 - 4.23 (m, 1 H), 3.81 - 3.91 (m, 2 H), 3.73

- 3.80 (m, 1 H), 3.55 - 3.64 (m, 2 H), 3.30 - 3.38 (m, 1 H), 2.94 - 3.04 (m, 1 H), 2.32 - 2.41 (m, 1 H). 2.02 - 2.11 (m. 1 H). 1.93 - 2.00 (m. 2 H). 1.84 - 1.92 (m. 2 H). 1.50 - 1.57 (m. 1 H) [0247] The mixture from above (22 mg) was further resolved by Chiral-SFC (Column DAICEL CHIRALCEL IG: (250 mm*30 mm, 10 urn); Condition: CCh-iPrOH (0.1%NH3H₂O); Begin B 55 End B 55; Gradient Time (min): 20; 100%B Hold Time: 20; Flow Rate (ml/min): 80) to give 5-26 8-(4-chloro-2-fluorophenyl)-6-((2R,4S or 2S.4R)-2-((S or /<)-tetrahydrofuran-3- yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4-d]pyrimido[l,6-a]pyrimi din-10-one (tr = 0.724 min) as the first eluting peak, and 5-27 8-(4-chloro-2-fluorophenyl)-6-((27?.4S' or 2S,4R)-2- ((R or S)-tetrahydrofuran-3-yl)tetrahydro-2H-pyran-4-yl)-l,3-dihydro-10H-furo[3,4- d]pyrimido[l,6-a]pyrimidin-10-one (tr =0.797 min) as the second eluting peak.

5-26: MS (ESI) m/z: calc'd for C₂₄H₂₄C1FN?O₄ ⁺ [M+H]⁺ 472, found [M+H]⁺472. 'H NMR (400 MHz, CDCh) 5 ppm 7.61 (t, 7=7.93 Hz, 1 H), 7.30 - 7.35 (m, 1 H), 7.18 (s, 1 H), 7.13 (dd, 7=10.07, 1.85 Hz, 1 H), 4.99 - 5.22 (m, 4 H), 4.16 - 4.22 (m, 1 H), 3.81 - 3.91 (m, 2 H), 3.72 - 3.80 (m, 1 H), 3.55 - 3.64 (m, 2 H), 3.30 - 3.38 (m, 1 H), 2.93 - 3.04 (m, 1 H), 2.32 - 2.41 (m, 1 H). 1.98 - 2.11 (m. 2 H). 1.93 - 1.96 (m. 1 H). 1.80 - 1.92 (m. 2 H), 1.52 - 1.59 (m. 1 H) 5-27: MS (ESI) m/z: calc'd for C₂4H₂₄C1FN3O4⁺ [M+H]⁺ 472 found [M+H]⁺472. ‘H NMR (400 MHz, CDCh) 5 ppm 7.59 - 7.64 (m, 1 H), 7.32 (dd, 7=8.23, 1.67 Hz, 1 H), 7.18 (s, 1 H), 7.13 (dd, 7=10.07, 1.85 Hz, 1 H), 5.03 - 5.21 (m, 4 H), 4.12 - 4.19 (m, 1 H), 3.85 - 3.96 (m, 2 H), 3.70

- 3.79 (m, 2 H), 3.53 - 3.63 (m, 1 H), 3.28 - 3.35 (m, 1 H), 2.94 - 3.04 (m, 1 H), 2.29 - 2.38 (m, 1 H), 2.07 - 2.14 (m, 1 H), 1.93 - 2.03 (m, 2 H), 1.77 - 1.92 (m, 2 H), 1.46 - 1.53 (m, 1 H) [0248] Examples shown below were prepared according to procedures analogous to those outlined in examples 5-18 to 5-29 using the appropriate starting materials.

Table 9

EXAMPLE 6-1

Preparation of 6-1 (S)-6-(4-chloro-2-fluorophenyl)-7-fluoro-2.3-dimethyl-8-(2-(l-methyl-177- pyrazol-4-yl)morpholino)-4/7-pyrimido[l,2-a1pyrimidin-4-one

Step 1 : Preparation of (S)-4-chloro-5-fluoro-6-(2-(l-methyl-17/-pyrazol-4- yl)morpholino)pyrimidin-2-amine

[0249] To a microwave vial containing 4,6-dichloro-5-fluoropyrimidin-2-amine (257 mg, 1.41 mmol) and (S)-2-(l-methyl-17/-pyrazol-4-yl)morpholine (236 mg, 1.41 mmol) in DMF (2.8 mL) was added triethylamine (143 mg, 197 pL, 1.41 mmol). The reaction mixture was stirred in a microwave reactor at 120 °C for 7 min. The reaction was cooled to room temperature and used as is in the next step. MS (ESI) m/z: calc’d for C12H15CIFN6O [M+H]⁺: 313.1, found: [M+H]⁺: 313.1

Step 2: Preparation of (S)-5-fluoro-4-(2-(l-methyl-177-pyrazol-4-yl)morpholino)-6- (methylthio)pyrimidin-2-amine

[0250] Sodium methanethiolate (98.9 mg, 1.41 mmol) was directly added to the above reaction mixture, and the reaction heated in a microwave reactor at 140 °C for 13 min. Another 50 mg of NaSMe was added, and again heated in microwave at 140 °C for 5 min. Third batch of 50 mg of NaSMe was added and heated in microwave at 140 °C for another 5 min. Upon completion, the reaction mixture was cooled to rt and 15 mL of water was added. A white precipitate was formed and collected through filtration. The filter cake was dried under high vacuum overnight. It was used in the next step directly. MS (ESI) m/z: calc’d for CisHisFNeOS [M+H]⁺: 325.1, found: [M+H]⁺: 325.1

Step 3: Preparation of (S)-7-fluoro-2.3-dimethyl-8-(2-(l-methyl-17/-pyrazol-4-yl)morpholino)-6- (methylthio)-4//-pyrimido[ 1 .2-a]pyrimidin-4-one [0251] To a vial containing (S)-5-fluoro-4-(2-(l -methyl- l//-pyrazol-4-y l)morpholino)-6- (methylthio)pyrimidin-2-amine (160 mg, 493 pmol) and ethyl 2-methylacetoacetate (356 mg, 2.47 mmol) was added PPA (990 pL). The resulting mixture was heated on a hot plate at 110 °C for 10 min. The reaction was quenched with 20 mL of sat. NaHCCh aq. solution and extracted with DCM. The organic extracts were concentrated and washed with Et20 to yield (S)-7-fluoro- 2,3-dimethyl-8-(2-(l-methyl-lF7-pyrazol-4-yl)morpholino)6-(methylthio)-477-pyrimido[l,2- a]pyrimidin-4-one which is used in the next step directly. MS (ESI) m/z: calc’d for C18H22FN6O2S [M+H]⁺: 405.1, found: [M+H]⁺: 405.2

Step 4: Preparation of 6-1 (S)-6-(4-chloro-2-fluorophenyl)-7-fluoro-2.3-dimethyl-8-(2-(l-methyl- l//-pyrazol-4-yl)morpholino)-477-pyrimido[ 1.2-a1pyrimidin-4-one

[0252] To a microwave vials containing ((thiophene-2-carbonyl)oxy)copper (35.4 mg, 185 pmol), (4-chloro-2-fluorophenyl)boronic acid (25.9 mg, 148 pmol) and (S)-7-fluoro-2,3- dimethyl-8-(2-(l -methyl- I H-py razol-4-y I )morpholino)-6-(methy I thio)-4f/-pyrimido| I ,2- a]pyrimidin-4-one. were added phosphine, tri -2 -furanyl- (5.2 mg. 22.3 pmol) and tris(dibezyhdeneacetone)dipalladium (13.6 mg. 14.8 pmol). The vial was purged with Ar then THF (740 pL) was added. The resulting mixture was purged with Ar again and heated in a microwave reactor at 100 °C for 7 min. The reaction was diluted with DMSO and was purified by reverse phase HPLC (acetonitrile in water, with TFA modifier) to yield (S)-6-(4-chloro-2- fluorophenyl)-7-fluoro-2,3-dimethyl-8-(2-(l-methyl-l/7-pyrazol-4-yl)morpholino)-4F7- pyrimido[l,2-a]pyrimidin-4-one. MS (ESI) m/z: calc’d for C23H22CIF2N6O2 [M+H]⁺: 487.1, found: [M+H]⁺: 487.1

1H NMR (600 MHz, DMSO-d6) 5 7.80 (s, 1H), 7.67 (d, J = 8.5 Hz, 1H), 7.57 - 7.44 (m. 3H), 4.86 (b, 1H), 4.66 (s. 1H), 4.38 (b. 1H), 4.08 (b. 2H), 3.87 - 3.75 (m, 5H), 3.52 (b, 1H), 2.36 (s, 3H), 1.88 (s, 3H).

Example 7-1

Preparation of 7-1 6-(4-chloro-2-fluorophenyl)-2.3-dimethyl-8-((27?, 6S)-2-methyl-6-(2- methylpyridin-4-yl)morpholino)-4H-pyrimido[1.2-a]pyrimidin-4-one

Step 1 : Synthesis of 4-chloro-6-(methylthio)pyrimidin-2-amine

1 2

[0253] To a solution of 4,6-dichloropyrimidin-2-amine (10 g, 61.0 mmol) in DMF (100 mL) was added NaSMe (4.70 g, 67. 1 mmol). The reaction was stirred at 80 °C for 3 h. Upon completion, the mixture was poured into ice water (400 mL) and extracted with EtOAc (300 rnL * 3). The combined organic fractions were washed with brine (300 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 0 - 25% Ethyl acetate/Petroleum ether gradient) to give 4-chloro-6- (methylthio)pyrimidin-2-amine as a desired product.

MS (ESI) m/z: calc’d for C5H7CIN3S+ [M+H]⁺: 176, found [M+H]+: 176.

Step 2: Synthesis of 8-hvdroxy-2.3-dimethyl-6-(methylthio)-4H-pyrimido[L2-a1pyrimidin-4-one

2 3

[0254] A solution of 4-chloro-6-(methylthio)pyrimidin-2-amine (5 g, 28.5 mmol) and ethyl 2- methyl-3-oxobutanoate (8.21 g, 56.9 mmol) in Eaton's reagent (6.78 g, 28.5 mmol) was stirred at 100 °C for 1 h. Upon completion, the reaction mixture was poured into H2O (5 mL), quenched by aq NaHCO? (8 mL), and extracted with EtOAc (10 mL * 3). The combined organic fractions were washed with brine (15 mL), dried over Na2SC>4, filtered and concentrated under reduced pressure. The residue was purified by flash C18 chromatography (eluent of 80% water/MeOH gradient) to give a crude mixture which was triturated with EtOAc (2 * 30 mL). The mixture was filtered and the filtrate cake was dried to give 8-hydroxy-2,3-dimethyl-6-(methylthio)-4H- pyrimido[l,2-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc’d for CioHi2N₃0₂S⁺ [M+H]+: 238.1, found [M+H]+: 237.9.

Step 3: Synthesis of 8-chloro-2,3-dimethyl-6-(methylthio)-4H-pyrimido[L2-a1pyrimidin-4-one

POCI3, DIEA, reflux

[0255] To a solution of 8-hydroxy-2,3-dimethyl-6-(methylthio)-4H-pyrimido[l,2-a]pyrimidin- 4-one (1 g, 4.21 mmol) and DIEA (1.47 mL, 8.43 mmol) in toluene (0.5 mL) was slowly added POCh (0.786 mL, 8.43 mmol) at 0 °C. The reaction mixture was stirred at 100 °C for 2 h. Upon completion, the reaction mixture was poured into H2O (30 mL) and extracted with EtOAc (35 mL * 3). The combined organic fractions were washed with brine (50 mL). dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (eluent of 40% ethyl acetate/petroleum ether gradient) to give 8-chloro-2,3- dimethyl-6-(methylthio)-4H-pyrimido[l,2-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc’d for CioHnCIN30S+[M+H]⁺:256, found [M+H]+ 256.

Step 4: Synthesis of 2.3-dimethyl-8-((2R,6S)-2-methyl-6-(2-methylpyridin-4-yl)morpholino)-6-

(methylthio)-4H-pyrimido[1.2-a]pyrimidin-4-one

4 5

[0256] To a solution of 8-chloro-2,3-dimethyl-6-(methylthio)-4H-pyrimido[l,2-a]pyrimidin-4- one (50 mg, 0.196 mmol) in DMSO (1 mL) was added DIEA (0.068 mL, 0.391 mmol) and

(2R. 6S)-2-methyl-6-(2-methylpyridin-4-yl)morphohne (45.1 mg, 0.235 mmol). The reaction was stirred at 80 °C for 2 h. Upon completion, the reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (10 mL * 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Pre-TLC (DCM/MeOH = 100/10) to give 2.3-dimethyl-8-((2/?.6,S')-2- methyl-6-(2-methylpyridin-4-yl)morpholino)-6-(methylthio)-4H-pyrimido[l,2-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc’d C21H26N5O2S [M+H]⁺: 412. found [M+H]⁺: 412.

Step 5: Synthesis of 7-1 6-(4-chloro-2-riuoroDhenyl )-2.3-dimethyl-8-( -2-methyl-6-(2-

methylpyridin-4-yl)morpholino)-4H-pyrimido[1.2-a1pyrimidin-4-one

[0257] To a mixture of 2,3-dimethyl-8-((27?, AS)-2-methyl-6-(2-methylpyridin-4- yl)morpholino)-6-(methylthio)-4H-pyrimido[l,2-a]pyrimidin-4-one (50 mg, 0.121 mmol) and (4- chloro-2-fluorophenyl)boronic acid (25.4 mg, 0.146 mmol) in THF (1 mL) was added Pd2(dba)3 (11.13 mg, 0.012 mmol), tri(2-furyl)phosphine (2.82 mg, 0.012 mmol) and copper(I) thiophene- 2-carboxylate (30. 1 mg, 0. 158 mmol) at 25 °C, then the mixture was stirred at 100 °C for 0.5 h under N2 atmosphere in the microwave. The reaction mixture was next poured into H2O (5 mL) and extracted with EtOAc (5 mL * 3). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by Pre-HPLC (MeCN/water with 0.1%TFA) to give 7-1 6-(4-chloro-2- fluorophenyl)-2,3-dimethyl-8-((2R,6S)-2-methyl-6-(2-methylpyridin-4-yl)morpholino)-4H- pyrimido[l,2-a]pyrimidin-4-one as a desired product.

MS (ESI) m/z: calc'd for C26H26CIFN5O2 [M+H]⁺: 494, found [M+H]⁺: 494. 'H NMR (400 MHz, CDsOD-d): 5 1.36 - 1.49 (m. 3 H) 1.99 (s. 3 H) 2.43 (s, 3 H) 2.73 - 2.82 (m, 3 H) 2.97 - 3.14 (m.

1 H) 3.17 - 3.28 (m, 1 H) 3.90 - 4.09 (m, 1 H) 4.42 - 4.70 (m, 1 H) 4.97 - 5.03 (m, 1 H) 5.09 - 5.29 (m, 1 H) 7.20 - 7.46 (m, 3 H) 7.51 - 7.79 (m, 1 H) 7.82 - 8.02 (m, 2 H) 8.58 - 8.79 (m, 1 H). Example 8-1

Preparation of 8-1 GS)-l-(4-chloro-2-fluorophenyl)-4-fluoro-3-(2-(l-methyl-lH-pyrazol-4- yl)morpho no)-6.7.8.9-tetrahydro-10H-pyrimido[6.1-b1quinazolin- 10-one

Step 1 : Synthesis of 6-chloro-5-fluoro-2-(methylthio)pyrimidin-4-amine

[0258] A microwave vial containing 4,6-Dichloro-5-fluoro-2-(methylthio)pyrimidine (180.0 mg, 0.845 mmol) and ammonia (2M) in isopropanol (4.2 mL, 2.00 molar, 8.449 mmol) was heated in a microwave reactor at 100 °C for 17 min. Upon completion, the mixture was cooled to rt and 30 mL of water was added. The white precipitate formed which was filtered and dried under high vacuum to yield 6-chloro-5-fluoro-2-(methylthio)pyrimidin-4-amine as desired product. MS (ESI) m/z: calc’d for CsHeCIFNsS [M+H]⁺: 194, found: [M+H]⁺:194.

Step 2: Synthesis of (S)-5-fluoro-6-(2-(l -methyl- lH-pyrazol-4-yl)morpholino)-2- (methylthio)pyrimidin-4-amine

[0259] To a microwave vial containing 6-chloro-5-fluoro-2-(methylthio)pyrimidin-4-amine (105.0 mg. 0.54 mmol) and (S)-2-(l-methyl-lH-pyrazol-4-yl)morpholine (113.3 mg. 0.68 mmol) in DMF (1.1 mL) was added triethylamine (0.38 mL, 5 eq, 2.7 mmol). The reaction mixture was stirred in a microwave reactor at 145 °C for 22 min. Upon completion, the reaction was diluted with water and extracted with EtOAc. The organic phase was concentrated and the crude product treated with 1 mL of DCM. The precipitate formed was collected through filtration and washed with Et20, then dried on high vacuum to yield (S)-5-fluoro-6-(2-(l-methyl-lH-pyrazol-4- yl)morpholino)-2-(methylthio)pyrimidin-4-amine. MS (ESI) m/z: calc’d for CisHisFNeOS [M+H]⁺: 325, found: [M+H]⁺: 325.

Step 3: Synthesis of ( )-4-fluoro-3-(2-(l -methyl- lH-pyrazol-4-yl)morpholino)-l-(methylthio)-

6.7.8,9-tetrahy dro- 1 OH-py rimido [6, 1 -bl quinazolin- 10-one

[0260] To a vial containing (5)-5-fluoro-6-(2-(l -methyl- lH-pyrazol-4-yl)morpholino)-2- (methylthio)pyrimidin-4-amine (30.0 mg, 92.5 pmol) and ethyl 2-oxocyclohexane-l -carboxylate (63.0 mg, 370 pmol) was added PPA (92.5 pL). The resulting mixture was heated on hot plate at 130 °C for 15 min. Upon completion, the reaction mixture was cooled to rt and carefully quenched with 10 mL of sat. NaHCCh aq., then extracted with DCM. The organic fractions were collected and washed with brine, dried (MgSCL), filtered and the solvent was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel, eluting with 20% to 40% to 60% EtOAc / EtOH (3: 1) in hexanes to give (S)-4-fluoro-3-(2-(l-methyl-lH- pyrazol-4-yl)morpholino)- 1 -(methy lthio)-6, 7, 8,9-tetrahy dro-10H-pyrimido[6, 1 -b] quinazolin- 10- one. MS (ESI) m/z: ca ’d for C20H24FN6O2S [M+H]⁺: 431. found: [M+H]⁺: 431.

Step 4: Synthesis of 8-1 -l-(4-chloro-2-fluorophenyl)-4-fluoro-3-(2-(l-methyl-lH-pyrazol-4-

yl)morpholino)-6.7.8.9-tetrahydro-10H-pyrimido[6.1-b1quinazolin-l 0-one

[0261] In a microwave vial containing (S^-4-fluoro-3-(2-(l-methy l-lH-pyrazol-4- y l)morpholino)- 1 -(methy lthio)-6, 7.8,9-tetrahy dro- 1 OH-py rimido [6.1 -b] quinazolin- 10-one (17.0 mg, 0.04 mmol), (4-chloro-2-fluorophenyl)boronic acid (10.3 mg, 0.06 mmol) and Tris(dibezylideneacetone)dipalladium (3.62 mg, 0.004 mmol) was added phosphine, tri-2- furanyl- (1.83 mg, 0.008 mmol). The vial was purged with Ar for three times, then THF (0.4 mL) was added. The reaction mixture was purged with Ar for three times again, then heated in a microwave reactor at 100 °C for 7 min. Upon completion, the reaction mixture was diluted with water and extracted with 2 mL of EtOAc. The organic extracts were concentrated and the crude product was purified by reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield desired product (S)-l-(4-chloro-2-fluorophenyl)-4-fluoro-3-(2-(l-methyl-lH-pyrazol-4- yl)morpholino)-6,7,8,9-tetrahydro-10H-pyrimido[6.1-b]quinazolin-10-one. MS (ESI) m/z: calc’d for C25H24CIF2N6O2 [M+H]⁺: 513. found: [M+H]⁺: 513. 'H NMR (600 MHz. DMSO-d6) 5 7.73 (s, 1H), 7.63 (t, J = 8.1 Hz, 1H), 7.49 (d, J = 10.1 Hz, 1H), 7.44 (s, 1H), 7.41 (d, J = 8.2 Hz, 1H), 4.60 (s, 1H), 4.30 - 4.20 (m, 2H), 3.98 (d, J = 11.3 Hz, 1H), 3.82 (s, 3H), 3.70 - 3.50 (m, 3H), 2.66 (m, 2H), 2.34 (m, 2H), 1.75 - 1.69 (m, 4H).

Example 9-1

Preparation of 9-1 6-(2.4-difluorophenyl)-8-( -4-fluoro-2-((S and

R)tetrahydrofuran-3-yl)tetrahydro-21/-pyran-4-yl)-2.3-dimethyl-47F-pyrimido[ 1 .6-g|p\ rimidin-4- one

Step 1: Synthesis of -2-((R and S)tetrahydrofuran-3-yl)-2.3-dihydro-47f-pyran-4-one

[0262] A 30-mL oven-dried dram vial equipped with a magnetic stir bar was charged with molecular sieves 4A (15 g) and (7?,7?)-Jacobsen catalyst, Tetrafluoroborate (1.046 g. 1.530 mmol). The vial was sealed and purged with N2 for 5 min. The catalyst was dissolved in EtOAc (10.20 mL). This mixture was stirred at rt for 1 h after which tetrahydrofuran-3-carbaldehyde (6.127 g, 61.20 mmol) was added to the vial. The mixture was cooled to 0 °C and stirred an additional 10 min. (E)-((4-methoxybuta-l,3-dien-2-yl)oxy)trimethylsilane (8.787 g, 9.93 mL, 51.00 mmol) was added and the reaction was allowed to warm to 4 °C and allowed to stir at this temperature for 22.5 h. The reaction was then cooled to 0 °C, and CH2CI2 (2 mL) was added, followed by TFA (1.163 g, 785.8 pL, 10.20 mmol). The reaction was allowed to warm to rt and stirred for 10 min. The reaction was then filtered through a plug of silica gel on CELITE and the filter pad was washed with Et20. The filtrate was concentrated to yield crude (27?)-2-((R and S)tetrahydrofuran-3-yl)-2.3-dihydro-477-pyran-4-one which was used without further purification.

Step 2: Synthesis of (2/?)-2-((R and S)tetrahydrofuran-3-yl)tetrahydro-47f-pyran-4-one

[0263] A 500-mL flask equipped with a magnetic stir bar was charged with (2/?)-2- (tetrahydrofuran-3-yl)-2.3-dihydro-477-pyran-4-one (8.6 g, 51 mmol) in EtOAc (0.10 L) followed by palladium on carbon (0.54 g, 5. 1 mmol). The flask was purged with H2 for 10 min and left overnight at 1 atm. Upon completion, the solution was filtered through a plug of CELITE and concentrated. The crude material was purified by distillation under high vacuum affording (27?)- 2-((R and S)tetrahydrofuran-3-yl)tetrahydro-47/-pyran-4-one.

Step 3: Synthesis of (AS)-6-((R and S)tetrahvdrofuran-3-yl)-3.6-dihvdro-277-pyran-4-yl trifluoromethanesulfonate and (27?)-2-((R and S)tetrahydrofuran-3-yl)-3.6-dihvdro-277-pyran-4-yl trifluoromethanesulfonate

[0264] To a stirred solution of diisopropylamine (214 mg, 298 pL, 2. 12 mmol) in THF at 0 °C under an atmosphere of nitrogen was added slowly BuLi (130 mg, 1.27 mL, 1.6 molar, 2.03 mmol). After 10 min, the solution was cooled to -78 °C, and (27?)-2-((R and S)tetrahydrofuran-3- yl)tetrahydro-4//-pyran-4-one (300 mg, 1.76 mmol) in THF was added dropwise to this solution. After 30 min, phenyl triflimide (787 mg, 2.20 mmol) in THF was added dropwise. The final concentration reached 0.2M. The solution was warmed slowly to room temperature overnight. Upon completion, the reaction mixture was quenched with aq. sat. NH4CI. The resulting suspension was transferred into a separatory funnel with EtOAc (50 mL). The organic layer was collected and subsequently washed with brine (2 x 50 mL). The aqueous phase was backwashed with EtOAc. The combined organic phase was dried over MgSO4 and concentrated. The crude material was purified by silica gel chromatography using isocratic eluent 7.5%— 15% of EtOAc:EtOH = 3:1/Hexanes to afford (6S)-6-((R and S)tetrahydrofuran-3-yl)-3,6-dihydro-277- pyran-4-yl trifluoromethanesulfonate and (2 )-2-((R and S)tetrahydrofuran-3-yl)-3,6-dihydro- 27/-pyran-4-yl trifluoromethanesulfonate in 1: 1 ratio. MS (ESI) m/z: calc'd for C10H14F3O6S [M+H]⁺: 303, found: [M+H]⁺: 303.

Step 4: Synthesis of 6-(2.4-di fluoro phen yl )-2,3-dimethyl-8-((27?)-2-((R and S)tetrahydrofuran-3- yl)-3.6-dihydro-277-pyran-4-yl)-47/-pyrimido[1.6-a1pyrimidin-4-one and 6-(2,4-difluorophenyl)-

2.3-dimethyl-8-( -6-((R and S)tetrahydrofuran-3-yl)-3.6-dihydro-27f-pyran-4-yl)-4/7-

pyrimido[L6-a]pyrimidin-4-one

[0265] A vial was charged with crude 6-(2,4-difluorophenyl)-2,3-dimethyl-8-(4,4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)-47/-pyrimido[l,6-a]pyrimidin-4-one (128 mg, 311 pM), DTBPF-Pd-G3 (26.2 mg, 31.1 pmol), K3PO4 (132 mg, 622 pmol), (6S)-6-((R and S)tetrahydrofuran-3-yl)-3,6-dihydro-2/7-pyran-4-yl trifluoromethanesulfonate (112 mg, 373 pmol). The vial was sealed, and the atmosphere was exchanged to inert. Then 1,4-dioxane (2.83 mL) and H2O (283 pL) were added, and the solution was heated at 80 °C for 10 min. Upon completion, the reaction was cooled down and quenched with H2O. The resulting suspension was transferred into a separatory funnel with EtOAc (10 mL)/ Water (10 mL). The organic layer was collected and subsequently washed with brine (2 x 10 mL). The aqueous phase was backwashed with EtOAc. The combined organic phase was dried over MgSO4 and concentrated. The crude material was purified by silica gel flash chromatography using isocratic eluent 25%-30% of EtOAc:EtOH = 3: 1/Hexanes to afford 6-(2,4-difluorophenyl)-2,3-dimethyl-8-((2A)-2-((S and R)tetrahydrofiiran-3-yl)-3.6-dihydro-2//-pyran-4-yl)-4//-pyrimido| l .6-fl|pyrimidin-4-one and 6- (2,4-difluorophenyl)-2,3-dimethyl-8-((6A)-6-((S and R)tetrahydrofuran-3-yl)-3,6-dihydro-27T- pyran-4-yl)-42/-pyrimido[l,6-a]pyrimidin-4-one as a mixture in 1: 1 ratio. MS (ESI) m/z: calc’d for C24H23F2N3O3 [M+H]⁺: 440, found: [M+H]⁺: 440.

Step 5: Synthesis of 9-1 6-(2.4-difluorophenyl)-8-((2A!.4R or 2R,4A)-4-fluoro-2-((S and

R)tetrahvdrofuran-3-yl)tetrahvdro-2//-pyran-4-yl)-2.3-dimethyl-4/7-pyrimido[L6-a1pyrimidin-4- one

[0266] Iron (III) oxalate hydrate (296 mg, 751 pmol) was stirred in water (9. 10 mL) until completely dissolved. The solution was cooled to 0 °C and degassed for 10 min. Selectfluor (645 mg, 1.82 mmol) and acetonitrile (4.55 mL) were added to the reaction mixture. A solution of 6- (2,4-difluorophenyl)-2,3-dimethyl-8-((6A)-6-((R and S)tetrahydrofuran-3-yl)-3,6-dihydro-2/f- pyran-4-yl)-4/7-pyrimido[l,6-«]pyrimidin-4-one (100 mg, 228 pmol) in acetonitrile (4.55 mL) was transferred by pipet to the reaction mixture and half of the NaBLL (94.7 mg, 2.50 mmol) was added to the mixture at 0 °C. After 2 min, the reaction mixture was treated with the second half of NaBEh (94.7 mg, 2.50 mmol). The resulting mixture was stirred for 30 min. Upon completion, the crude material was filtered and purified with reverse phase HPLC (eluting acetonitrile in water, with TFA modifier) to yield 6-(2.4-difluorophenyl)-8-((2/?.4/? or 2R,4S)-4- fluoro-2-((S and R)tetrahydrofuran-3-yl)tetrahydro-277-pyran-4-yl)-2,3-dimethyl-4F7- pyrimido[l,6-a]pyrimidin-4-one. MS (ESI) m/z: calc'd for C24H25F3N3O3 [M+H]⁺: 460, found: [M+H]⁺: 460. 'H NMR (500 MHz, DMSO) 5 7.76 (q, J= 7.9 Hz. 1H), 7.35 - 7.28 (m, 2H), 7.23 (t, J = 8.4 Hz, 1H). 3.96 (m, 1H), 3.82 - 3.64 (m, 3H), 3.64 - 3.46 (m, 3H). 2.39 (s, 3H). 2.26 (s. 3H), 2.01 (s, 3H), 1.98 - 1.70 (m, 4H).

ASSAY DATA

[0267] The compounds of the disclosure, surprisingly and advantageously, exhibit good potency as agonists of TREM2. The potency of the compounds were measured as follows. In vitro measurement of TREM2 activity' using cell-based pSyk (phosphorylation of Spleen Tyrosine Kinase) assay

[0268] Measurement of TREM2 agonist potency was done using a HEK cell line expressing human TREM2 and DAP12 (HEK293T-hTREM2 cells). Binding of small molecules to, and activation of, TREM2 increases the phosphorylation of Syk. The resultant levels of Syk phosphorylation are measured using a commercial AlphaLisa reagent kit. To perform the assay, HEK-hTREM2 cells were plated at 14,000 cells per well in a 384 well plate, in 25 pL of complete grow th media and incubated at 37 °C, 5% CO2 for 20-24 hours.

[0269] Prior to the assay, test compounds were diluted in the 384 well plates in serum-free media and allowed to equilibrate for 30 minutes. Growth media w as removed from cell plates byinversion and flicking, followed by tapping out excess media on blotting paper, and 25 pL of test articles in serum-free media was added to cells. Cells were incubated for 45 minutes at room temperature. After 45 minutes, the serum-free media was removed by the same flicking and tapping procedure above, and 10 pL of lysis buffer were added. Plates were shaken for 20 minutes at 350 RPM at room temperature. After complete lysis. AlphaLISA reagents were added to the lysate using the manufacturer's recommended bead ratios and incubation times, and fluorescence intensity was measured using a BMG LabTech PHERAstar FSX plate reader. Intensities were used to generate Max and Min controls, and % activation w as calculated. Curve fitting was performed using IDBS ABASE software with XE runner, which generates a four parameter fit with variable slope using the log(agonist) vs response, and EC50s were calculated from the curve fit.

[0270] This assay may be used to test any of the compounds described herein to assess and characterize a compound’s ability to act as an agonist of TREM2. Potency (ECso) results from the pSYK assays of the disclosed compounds are provided in Table 6, below: Table 6

[0271] The disclosed subject matter is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0272] All references (e.g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety' for all purposes. Other embodiments are within the follow ing claims.

Claims

CLAIMS What Is Claimed Is:

1. A compound of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

X¹ and X² are independently N, C(F) or C(H);

X4 is N or C(R⁸); wherein when Xl is N then X4 is C(R8), and when X4 is N then Xl is C(F) or C(H);

R¹ is H, -Ci-4 alkyl, -N(CH₃)₂. -(CH₂)_Z-OH, or -O(CH₂)_ZCH₃;

R² is H, -Ci-4 alkyl; or, alternatively, R¹ and R² are optionally taken together with one or more intervening atoms to form a ring C’Z; wherein ring CZ is:

R³ is selected from the group consisting of:

wherein R⁷ and R¹¹ are independently selected from hydrogen, a halo, -Ci-4 alkyl, cyclopropyl, and -CF₃; and R¹⁰, R¹² and R¹³ are independently selected from hydrogen and a halo; R⁵ is selected from:

(a) a -C4-7 cycloalkyl, wherein the C4-7 cycloalkyl of R⁵ is unsubstituted or substituted with a -C1-3 alkyl, a heteroaryl, or fluoro;

(b) a heterocyclyl that is comprised of:

(1) seven carbon atoms and one N atom,

(2) six carbon atoms and (i) one O atom or (ii) one each of O and N, or

wherein:

Y¹ is C(H), C(F), N or O;

Y² is C(H), C(F), or N;

Y³ is C(H) or N;

R⁴ is selected from hydrogen, a -C4-6 cycloalkyl, a heterocyclyl. and a heteroaryl; wherein the C4-6 cycloalkyl, heterocyclyl and heteroaryl of R⁴ is unsubstituted or substituted with 1 to 3 R4a substituents independently selected from the group consisting of fluoro, C1-3 alkyl. C1-3 fluoroalkyl, C1-3 alkoxy. C1-3 fluoroalkoxy, and C3-6 cycloalkyl, heterocyclyl and heteroaryl;

R⁶ is selected from the group consisting of hydrogen, acetyl, -SChCCHs), and fluoro; wherein when Y1 is O. then R6 is absent;

R9a and R9b are independently H, fluoro, or Ci-3 alkyl; or, alternatively, R9a and R9b. together with the carbon atom to which they are attached. form a C3-6 cycloalkyl; and

R⁸ is hydrogen or fluoro; s is 0 or 1; and z = 1 or 2.

2. A compound of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein:

X³ is CH₂ or O;

R¹ is a -Ci-3 alkyl or -OC1-3 alkyl;

R⁷ is a halo; and

R⁴ is selected from hydrogen and a heteroaryl that is comprised of:

(1) three carbon atoms and two N atoms.

(2) four carbon atoms and two N atoms, or

3. The compound of claim 1 or 2. or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are taken together to form a Cs-6 cycloalkyl or a 5- to 6-membered heterocyclyl that is unsubstituted or substituted with one or two fluoro.

4. The compound of any one of claims 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R¹ or R² are methyl.

5. The compound of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, wherein R⁷ is chloro.

6. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) has Formula (III)

wherein:

Xl is N, C(F), or C(H);

X4 is N C(F), or C(H); wherein when Xl is N then X4 is C(H), and when X4 is N then Xl is C(F) or C(H);

R¹ and R² are independently H or Ci-4 alkyl, or alternatively, R¹ and R², together with the carbon atoms to which they are attached, form a ring C'Z; wherein ring CZ is:

(ii) 5- to 6-membered heterocycloalkyl, wherein said 5- to 6-membered heterocycloalkyl is non-aromatic and partially unsaturated and contains one heteroatom selected from the group consisting of N, O and S; wherein ring CZ is unsubstituted or substituted by 1 to 3 RZ substituents independently selected from the group consisting of fluoro, Cl -3 alkyd, Cl -3 fluoroalkyl or Cl -3 alkoxy;

R7, R1 1, R12, and R13 are independently H or halo;

Y² is C(H), C(F), or N;

R⁴ IS:

(ii) a 5- to 6-membered saturated heterocyclyl containing 1 O atom; wherein R⁴ is unsubstituted or substituted by 1 to 3 R4a substituents independently selected from the group consisting of fluoro, C1-3 alkyl, C1-3 fluoroalkyl, C1-3 alkoxy, C1-3 fluoroalkoxy, and C3-6 cycloalkyl;

R9a and R9b are independently H. fluoro, or Cj-3 alkyl; or, alternatively, R9a and R b together with the carbon atom to which they are attached, form a C3-6 cycloalkyl; and s is 0 or 1.

7. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein Xl is N and X4 is C(H).

8. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein

(i) R¹ and R² are both methyl; or

(ii) R¹ and R² form a 5-membered heterocyclyl containing one O atom.

9. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein s is 0.

10. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein Y² is C(H).

11. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein R⁴ is unsubstituted or substituted tetrahydrofuryl, pyrazolyl, or pyridinyl.

12. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein

13. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein

wherein t is 0, 1, 2. or 3.

14. The compound of claim 13 or a pharmaceutically acceptable salt thereof, wherein

15. The compound of claim 13 or 14 or a pharmaceutically acceptable salt thereof, wherein Y2 is C(H).

16. The compound of claim 13 or 14 or a pharmaceutically acceptable salt thereof, wherein Y2 is N.

17. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein

18. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group of Example Nos. 1-1 - 1-11, 2-1 - 2-17, 4-1 - 4-40, 5-1 - 5-34, 6-1, 7-1, 8-1, and 9-1.

19. A pharmaceutical composition comprising an effective amount of the compound of any one of claims 1-18, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

20. The pharmaceutical composition of claim 19, wherein the composition is adapted for oral, injectable, intramuscular, subcutaneous, intravenous, or intraperitoneal administration.

21. The pharmaceutical composition of claim 20, wherein the composition is adapted for oral administration.

22. A method for the activation of TREM2 receptor in a subject which comprises administering to the subject an effective amount of the compound or pharmaceutically acceptable salt thereof of any one of claims 1-18, or pharmaceutical composition of any one of claims 19-21.

23. A method for the treatment or prevention of a condition associated with a loss of function of human TREM2 in a subject, comprising administering to the subject an effective amount of the compound or pharmaceutically acceptable salt thereof of any one of claims 1-18, or pharmaceutical composition of any one of claims 19-21.

24. A method for the treatment or prevention of a neurodegenerative disorder in a subject, comprising administering to the subject an effective amount of the compound or pharmaceutically acceptable salt of any one of claims 1-18, or pharmaceutical composition of any one of claims 19-21.

25. The method of claim 24, wherein the neurodegenerative disorder is Alzheimer’s disease, Parkinson’s disease, frontotemporal dementia, demyelination disorder, multiple sclerosis, Huntington’s disease, amyotrophic lateral sclerosis (ALS), tauopathy disease, Nasu- Hakola disease, or adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP).

26. The method of claim 25, wherein the neurodegenerative disorder is Alzheimer’s Disease.

27. The method of any one of claims 22-26, wherein the subject is human.

28. The method of any one of claims 22-25, further comprising the step of administering a tau targeting therapy or an amyloid-P targeting therapy.

29. The compound of any one of claims 1-18, or a pharmaceutically acceptable salt thereof, for use in the preparation of a medicament for the treatment or prevention of a neurodegenerative disease in a subject.

30. The compound of any one of claims 1-18, or a pharmaceutically acceptable salt thereof, for use in therapy.