WO2015138689A1 - Fused morpholinopyrimidines and methods of use thereof - Google Patents

Fused morpholinopyrimidines and methods of use thereof Download PDF

Info

Publication number
WO2015138689A1
WO2015138689A1 PCT/US2015/020114 US2015020114W WO2015138689A1 WO 2015138689 A1 WO2015138689 A1 WO 2015138689A1 US 2015020114 W US2015020114 W US 2015020114W WO 2015138689 A1 WO2015138689 A1 WO 2015138689A1
Authority
WO
WIPO (PCT)
Prior art keywords
methyl
pyrimido
amine
dihydro
oxazin
Prior art date
Application number
PCT/US2015/020114
Other languages
French (fr)
Inventor
Duane A. Burnett
Matthew Gregory Bursavich
Andrew J. Mcriner
Original Assignee
Forum Pharmaceuticals Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Forum Pharmaceuticals Inc. filed Critical Forum Pharmaceuticals Inc.
Publication of WO2015138689A1 publication Critical patent/WO2015138689A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
    • C07D498/14—Ortho-condensed systems

Definitions

  • This disclosure relates generally to Fused Morpholinopyrimidine compounds. More specifically, the disclosure relates to the use of the Fused Morpholinopyrimidine compounds for the treatment of neurological disease.
  • AD Alzheimer's disease
  • ⁇ -amyloid peptide ⁇
  • APP amyloid precursor protein
  • APP is initially processed by ⁇ -secretase forming a secreted peptide and a membrane bound C99 fragment.
  • the C99 fragment is subsequently processed by the proteolytic activity of ⁇ -secretase. Multiple sites of proteolysis on the C99 fragment lead to the production of a range of smaller peptides ( ⁇ 37-42 amino acids).
  • N-terminal truncations can also be found e.g., ⁇ (4-42).
  • notations ⁇ 40 and ⁇ 42, as used herein, include these N-terminal truncated peptides.
  • the ⁇ peptides Upon secretion, the ⁇ peptides initially form soluble aggregates which ultimately lead to the formation of insoluble deposits and plaques.
  • ⁇ 42 is believed to be the most neurotoxic; the shorter peptides have less propensity to aggregate and form plaques.
  • ⁇ plaques in the brain are also associated with cerebral amyloid angiopathy, hereditary cerebral hemorrhage with amyloidosis, multi infarct dementia, dementia pugilistisca and Down's Syndrome.
  • ⁇ -secretase is an association of four proteins: Aphl, nicastrin, presenilin and Pen- 2 (review De Strooper, Neuron 38:9-12 (2003)). Subjects carrying particular mutations in one of these components, presenilin, show increased ⁇ 42/ ⁇ 40 ratio. These mutations are correlated with early onset familial AD.
  • Inhibition of ⁇ -secretase resulting in the lowering of ⁇ 42 has been investigated by the pharmaceutical community, and numerous inhibitors have been found. See, e.g., Thompson et al. ⁇ Bioorg. Med. Chem. Lett. 2006, 16, 2357-63), Shaw et al. ⁇ Bioorg. Med. Chem. Lett. 2006, 17, 511-16) and Asberom et al. ⁇ Bioorg. Med. Chem. Lett. 2007, 15, 2219-2223).
  • Inhibition of ⁇ -secretase though, is not without side-effects, some of which are due to the ⁇ -secretase complex processing substrates other than C99, e.g., Notch.
  • a more desirable approach is to modulate the proteolytic activity of the ⁇ -secretase complex in a manner that lowers ⁇ 42 in favor of shorter peptides without significantly affecting the activity of ⁇ -secretase on substrates such as Notch.
  • NSAIDs nonsteroidal, anti-inflammatory drugs
  • Flurbiprofen for example Flurbiprofen
  • Other publications that disclose agents said to reduce ⁇ 42 through the modulation of ⁇ -secretase include: WO 2004/074232, WO
  • the invention provides a compound of Formula (I)
  • R is phenyl, C 1 -C 4 alkylene -phenyl, -Ci- C 6 alkyl or 3- to 7-membered monocyclic heterocycle, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH,
  • R is phenyl or C 1 -C 4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH, -CN, -NH 2i -C 1 -C 4 alkyl, halo-substituted C 1 -C 4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci-C 4 alkyl, -N(Ci-C 4
  • R is phenyl or C 1 -C 4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C 1 -C 4 alkyl, -C 1 -C 4 alkoxy, halo-substituted C 1 -C 4 alkyl and halo-substituted C 1 -C 4 alkoxy.
  • Y is piperidinyl or azetidinyl, each of which is
  • Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C 1 -C 4 alkyl, halo-substituted C 1 -C 4 alkyl and -C 1 -C 4 alkoxy.
  • R is phenyl or C 1 -C 4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen or -C 1 -C 4 alkyl; Y is piperidinyl; and Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C 1 -C 4 alkyl, halo-substituted C 1 -C 4 alkyl and -C 1 -C 4 alkoxy.
  • R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C 1 -C 4 alkyl; and Z is pyridinyl which is unsubstituted or substituted with -C 1 -C 4 alkoxy.
  • R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C 1 -C 4 alkyl.
  • R is phenyl which is unsubstituted.
  • Ris phenyl substituted with two or three -F.
  • each -C 1 -C 4 alkyl is methyl and each C 1 -C 4 alkylene is methylene.
  • C 1 -C 4 alkylene is methylene
  • C 1 -C 4 alkylene-phenyl is benzyl.
  • R is phenyl substituted with one methyl.
  • Ris C 1 -C 4 alkylene-phenyl.
  • Ris -Ci-C 6 alkyl In some embodiments, Ris -Ci-C 6 alkyl.
  • Ris 3- to 7-membered monocyclic heterocycle are numbered 3- to 7-membered monocyclic heterocycle.
  • Y is piperidinyl. In some embodiments, Y is azetidinyl.
  • Z is pyridinyl or pyrimidinyl, each of which is
  • Z is pyridinyl substituted with one -CF 3 . In some embodiments, Z is pyridinyl substituted with one methyl. In some embodiments, Z is pyridinyl substituted with one methoxy. In some embodiments, Z is pyrimidinyl which is unsubstituted or substituted with one to three -C 1 -C4 alkyl. In some embodiments, Z is pyrimidinyl substituted with one methyl.
  • the compound of Formula (I) is a compound selected from the list of compounds in Table I.
  • the compound of Formula (I) is selected from the group consisting of: N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-phenyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7-phenyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(o-tolyl)-7,8-dihydro-6H-pyrimido[5,4- b] [ 1 ,4]oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-
  • the compound of Formula (I) is selected from the group consisting of: 7-(3,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(3,5-difluorophenyl)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2- amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxa
  • the invention provides a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H- pyrimido [5 ,4-b]pyrrolo [ 1 ,2-d] [ 1 ,4]oxazin-2-amine; (-)-N-( 1 -(2-methoxypyridin-4- yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8-trimethyl-7,8-dihydro-6H-pyrimido[5,4- b][l,4]oxazin-2-amine; and N
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier or vehicle and an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier or vehicle and an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)- 6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; (-)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2- d][l,4]oxazin-2-amine
  • the invention provides a method for treating a neurodegenerative disease, comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable salt of a compound of Formula (I).
  • the invention provides a method for treating a neurodegenerative disease, comprising administering to a subject in need thereof an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4- yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; (-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4- b]pyrrolo[l,2-d][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8- trimethyl-7,8-dihydro-6H-pyrimid
  • the neurodegenerative disease is panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, or dementia.
  • the invention provides a method for treating Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • the invention provides a method for treating Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)- 6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; (-)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2- d][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8-trimethyl-7,8- dihydro-6H-pyrimido [5
  • the invention provides a method for improving an impaired cognitive function, comprising administering to a subject having impaired cognitive function an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • the invention provides a method for improving an impaired cognitive function, comprising administering to a subject having impaired cognitive function an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2- d][l,4]oxazin-2-amine; (-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro- 6H-pyrimido[5,4-b]pyrrolo[ 1 ,2-d] [ 1 ,4]oxazin-2-amine; N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-7,7,8-trimethyl-7,8-dihydro-6H-
  • the invention provides a method for ameliorating a symptom of Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • the invention provides a method for ameliorating a symptom of Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin- 4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2- amine; (-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H- pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin- 4-yl)-7,7,8-trimethyl-7,8-dihydro-6H
  • Alzheimer's disease is early onset Alzheimer's disease.
  • the subject is a human.
  • the symptom is progressive loss of memory, progressive loss of cognition, progressive loss of reasoning and/or progressive loss of judgment.
  • a compound of Formula (I), Table II, or a pharmaceutically acceptable salt thereof (also referred to herein as a "Fused Morpholinopyrimidine”) is useful for treating, preventing or ameliorating one or more symptoms of a neurodegenerative disease.
  • Exemplary neurodegenerative diseases include, but are not limited to, Alzheimer's disease, early onset Alzheimer's disease, panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, dementia, cognitive impairment, myclonus, seizures, Parkinsonism, extrapyramidal signs (EPS), apraxia, dystonia, dementia with Lewy bodies (DLB), aphasia, visual agnosia, and ataxia.
  • EPS extrapyramidal signs
  • the cognitive function impaired is one or more of attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
  • the subject is 65 years old or older. In other embodiments, the subject is 55 years old or older. In still other embodiments, the subject is 55 years old or younger, or 50 years old or younger.
  • a pharmaceutical composition comprising an effective amount of a Fused Morpholinopyrimidine and a pharmaceutically acceptable carrier or vehicle is useful for treating or preventing a neurodegenerative disease.
  • C 1 -C 4 alkyl refers to a straight chain or branched non-cyclic hydrocarbon having from 1 to 4 carbon atoms, wherein one of the hydrocarbon's hydrogen atoms has been replaced by a single bond.
  • Representative straight chain C 1 -C 4 alkyls include -methyl, -ethyl, -n-propyl and -n-butyl.
  • Representative branched C 1 -C 4 alkyls include
  • C 3 -C 8 monocyclic cycloalkyl refers to a saturated cyclic hydrocarbon having from 3 to 8 carbon atoms.
  • Representative C 3 -C 8 monocyclic cycloalkyls include -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclohexyl, -cycloheptyl and - cyclooctyl.
  • C 1 -C 4 alkylene refers to a straight or branched chain saturated hydrocarbon containing 1-4 carbon atoms, wherein two of the hydrocarbon's hydrogen atoms have been replaced by a single a bond.
  • Representative C 1 -C 4 alkylene groups include, methylene, ethylene, n-propylene, isopropylene, n-butylene and isobutylene.
  • C 1 -C 4 alkoxy refers to a C 1 -C 4 alkyl-O- group wherein the C 1 -C 4 alkyl is as defined above.
  • Examples of C 1 -C 4 alkoxy include, but are not limited to, methoxy, trifluoromethoxy, ethoxy, propoxy or butoxy.
  • halo-substituted C 1 -C 4 alkoxy refers to a C 1 -C 4 alkoxy group, as defined above, wherein one or more of the C 1 -C 4 alkoxy group's hydrogen atoms have been replaced with -F, -CI, -Br or -I.
  • Examples of a halo-substituted C 1 -C 4 alkoxy include, but are not limited to, -0-CH 2 F, -O-CCI 3 , -O-CF 3 , -0-CH 2 Cl, -0-CH 2 CH 2 Br, -O- CH 2 CH 2 I, -0-CF 2 CF 3 , -0-CH 2 CH 2 CH 2 F, -0-CH 2 CH 2 CH 2 Cl, -0-CH 2 CH 2 CH 2 CH 2 Br, -O- CH 2 CH 2 CH 2 CH 2 I, -0-CH 2 CH(Br)CH 3 , -0-CH 2 CH(Cl)CH 2 CH 3 , -0-CH(F)CH 2 CH 3 , - OCH 2 CF 3 and -0-C(CH 3 ) 2 (CH 2 Cl).
  • amino-substituted C 1 -C 4 alkoxy refers to a C 1 -C 4 alkoxy group, as defined above, wherein one or more of the C 1 -C 4 alkoxy group's hydrogen atoms have been replaced with -NH 2 .
  • amino-substituted C 1 -C 4 alkoxy examples include, but are not limited to, -0-CH 2 NH 2 , -0-CH 2 CH 2 NH 2 , -0-CH(NH 2 )CH 3 , -0-CH 2 CH 2 CH 2 NH 2 , -0-CH 2 CH 2 CH 2 CH 2 NH 2 , -0-CH 2 CH(NH 2 )CH 3 , -0-CH(NH 2 )CH 2 CH 3 and -O- C(CH 3 ) 2 (CH 2 NH 2 ).
  • a "nitrogen-containing 4- to 6-membered nonaromatic heterocycle” refers to a monocyclic 4- to 6-membered nonaromatic monocyclic cycloalkyl group in which one of the cycloalkyl group's ring carbon atoms has been replaced with a nitrogen atom and 0-3 of the cycloalkyl group's remaining ring carbon atoms are independently replaced with a N, O or S atom.
  • the nitrogen-containing 4- to 6-membered nonaromatic heterocycles can be attached via a nitrogen or carbon atom.
  • nitrogen-containing 4- to 6- membered nonaromatic heterocycles include, but are not limited to, azetidinyl, piperidinyl, oxazinyl, morpholinyl, imidazolidinyl, pyrazolidinyl and thiomorpholinyl.
  • a "4- to 6-membered nonaromatic heterocycle” refers to a monocyclic 4- to 6- membered nonaromatic monocyclic cycloalkyl group in which 1-3 of the cycloalkyl group's ring carbon atoms are independently replaced with a N, O or S atom.
  • the 4- to 6-membered nonaromatic heterocycles can be attached via a nitrogen or carbon atom.
  • nitrogen-containing 4- to 6-membered nonaromatic heterocycles include, but are not limited to, azetidinyl, piperidinyl, oxazinyl, morpholinyl, imidazolidinyl, pyrazolidinyl, thiomorpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl and dioxanyl.
  • 3- to 7-membered monocyclic heterocycle refers to a monocyclic 3- to 7-membered aromatic or non-aromatic monocyclic cycloalkyl in which 1-4 of the ring carbon atoms have been independently replaced with a N, O or S atom.
  • the 3- to 7- membered monocyclic heterocycles can be attached via a nitrogen or carbon atom.
  • 3- to 7-membered monocyclic heterocycle group include, but are not limited to, nitrogen-containing 3- to 7-membered monocyclic heterocycles discussed above, tetrahydrofuranyl, dihydrofuranyl, pyranyl, dihydropyranyl, tetrahydropyranyl, thiopyranyl, dihydrothiopyranyl, tetrahydrothiopyranyl, dioxanyl, dithianyl, trithianyl, dioxolanyl, furanyl and thiophenyl.
  • the 3- to 7-membered monocyclic heterocycle is a nitrogen-containing 3- to 7-membered monocyclic heterocycle.
  • the 3- to 7-membered monocyclic heterocycle is fully saturated or partially saturated.
  • heterocycle refers to a 5- or 6-membered aromatic monocyclic heterocycle in which from 1 to 4 of the ring carbon atoms have been independently replaced with a nitrogen atom and 0-4 of the cycloalkyl group's remaining ring carbon atoms have been independently replaced with an O or S atom.
  • the nitrogen-containing 5- to 6-membered aromatic monocyclic heterocycle can be attached via a nitrogen or carbon atom.
  • a 5- to 6-membered aromatic monocyclic heterocycles include, but are not limited to, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, pyrimidinyl, pyrazolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrrolyl, thiazolyl, thiadiazolyl, triazinyl, and
  • halogen or halo as used herein, refer to chlorine, bromine, fluorine or iodine.
  • halo-substituted C 1 -C 4 alkyl refers to a C 1 -C 4 alkyl group, as defined above, wherein one or more of the C 1 -C 4 alkyl group's hydrogen atoms have been replaced with -F, -CI, -Br or -I.
  • Examples of a halo-substituted C 1 -C 4 alkyl include, but are not
  • amino-substituted C 1 -C 4 alkyl refers to a C 1 -C 4 alkyl group, as defined above, wherein one or more of the C 1 -C 4 alkyl group's hydrogen atoms have been replaced with -NH 2 .
  • amino-substituted C 1 -C 4 alkyl include, but are not
  • C 1 -C 4 alkylene-phenyl refers to a C 1 -C 4 alkyl group, as defined above, wherein one of the C 1 -C 4 alkyl group's hydrogen atoms has been replaced with phenyl.
  • hydroxy-substituted C 1 -C 4 alkyl refers to a C 1 -C 4 alkyl group, as defined above, wherein one or more of the C 1 -C 4 alkyl group's hydrogen atoms have been replaced with -OH.
  • Representative examples of a hydroxy-substituted C 1 -C 4 alkyl include, but are not limited to, -CH 2 OH, -CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH, - CH 2 CH 2 CH 2 CH 2 OH, - CH 2 CH 2 CH 2 CH 2 OH,
  • any heteroatom with unsatisfied valences is assumed to have hydrogen atoms sufficient to satisfy the valences.
  • pharmaceutically acceptable carrier or vehicle refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or
  • Each carrier or vehicle must be "acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject.
  • materials which can serve as pharmaceutically acceptable carriers or vehicles include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc;
  • excipients such as cocoa butter and suppository waxes
  • oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil
  • glycols such as butylene glycol
  • polyols such as glycerin, sorbitol, mannitol and polyethylene glycol
  • esters such as ethyl oleate and ethyl laurate
  • agar buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen- free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.
  • salts may form salts which are also within the scope of this invention.
  • Reference to a compound described herein is understood to include reference to salts thereof, unless otherwise indicated.
  • a compound described herein contains both a basic moiety, such as, but not limited to, amine, 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.
  • Salts of the compounds described herein may be formed, for example, by reacting a 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.
  • the compounds described herein which contain a basic moiety may form salts with a variety of organic and inorganic acids.
  • Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, for example, trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides, hydroiodides
  • phenylpropionates e.g., 3-phenylpropionates
  • phosphates picrates, pivalates, propionates
  • salicylates succinates
  • sulfates such as those formed with sulfuric acid
  • sulfonates tartrates
  • thiocyanates toluenesulfonates, such as tosylates, undecanoates and the like.
  • the compounds described herein which contain an acidic moiety may form salts with a variety of organic and inorganic bases.
  • 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 benzathines, dicyclohexylamines, hydrabamines (formed with N,N-bis(dehydroabietyl) ethylenediamine), N-methyl-D- glucamines, N-methyl-D-glycamides, t-butyl amines and salts with amino acids, such as arginine, lysine and the like.
  • Basic nitrogen-containing groups may be quaternized with agents, such as lower alkyl halides (e.g., methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.
  • agents such as lower alkyl halides (e.g., methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl and diamyl sulfates),
  • Prodrugs and solvates of the compounds described herein are also contemplated herein.
  • the term "prodrug” as employed herein denotes a compound that, upon
  • Solvates of the compounds described herein include, for example, hydrates.
  • Compounds described herein are, subsequent to their preparation, preferably isolated and purified to obtain a composition containing an amount by weight equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 97%, equal to or greater than 98%, or equal to or greater than 99% of the compounds
  • All stereoisomers of the present compounds are contemplated within the scope of this invention.
  • Individual stereoisomers of the compounds described herein may, for example, be substantially free of other isomers (e.g., as a pure or substantially pure optical isomer having a specified activity), or may be admixed, for example, as racemates or with all other, or other selected,
  • the chiral centers of the compounds described herein may have the S or R configuration as defined by the International Union of Pure and Applied Chemistry (IUPAC) 1974 Recommendations.
  • the racemic forms can be resolved by physical methods, such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives or separation by chiral column chromatography.
  • the individual optical isomers can be obtained from the racemates by any suitable method, including without limitation, conventional methods, such as, for example, salt formation with an optically active acid followed by crystallization.
  • a particular enantiomer of a compound described herein may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers.
  • the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.
  • Isomeric mixtures containing any of a variety of isomer ratios may be utilized in accordance with the present invention. For example, where only two isomers are combined, mixtures containing 50:50, 60:40, 70:30, 80:20, 90: 10, 95:5, 96:4, 97:3, 98:2, 99: 1, or 100:0 isomer ratios are all contemplated by the present invention. Those of ordinary skill in the art will readily appreciate that analogous ratios are contemplated for more complex isomer mixtures.
  • stable refers to compounds which possess stability sufficient to allow manufacture and which maintain the integrity of the compound for a sufficient period of time to be detected and preferably for a sufficient period of time to be useful for the purposes detailed herein.
  • the present invention also includes isotopically labeled compounds, which are identical to the compounds disclosed herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes that can be incorporated into compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine and chlorine, such as 2 H, 3 H, 13 C, U C, 14 C, 15 N, 18 0, 17 0, 31 P, 32 P, 35 S, 18 F and 36 C1, respectively.
  • isotopically labeled compounds can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.
  • substituent When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
  • an effective amount refers to any amount that is necessary or sufficient for achieving or promoting a desired outcome, e.g., for treating, preventing, or ameliorating a symptom of a neurodegenerative disease. In some instances an effective amount is a therapeutically effective amount. A therapeutically effective amount is any amount that is necessary or sufficient for promoting or achieving a desired biological response in a subject. The effective amount for any particular application can vary depending on such factors as the disease or condition being treated, the particular agent being administered, the size of the subject, or the severity of the disease or condition. [0083] As used herein, “treat” or “treating” includes stopping the progression and/or reducing or ameliorating a symptom of a neurodegenerative disease, for example, improving cognitive function.
  • the term "subject" refers to a vertebrate animal.
  • the subject is a mammal.
  • the subject is a human.
  • the subject is a non-human vertebrate animal, including, without limitation, non-human primates, laboratory animals, livestock, domesticated animals and non- domesticated animals.
  • Non-limiting examples of subject include a mammal, e.g., a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, and non-human primate, such as a monkey, chimpanzee, baboon or rhesus.
  • the subject is a human.
  • tautomeric forms for example, as an amide or imino ether.
  • the scope of this disclosure is meant to include all such tautomeric forms.
  • a tetrazole may exist in two tautomeric forms, 1-H tetrazole and a 2-H tetrazole. This is depicted in the figure below. This example is not meant to be limiting in the scope of tautomeric forms.
  • Fused Morpholinopyrimidines i.e., compounds according to Formula (I), the compounds in Table II, and pharmaceutically acceptable salts thereof, as well as methods for preparing the compounds and using the compounds to treat one or more neurodegenerative diseases, e.g., reducing a symptom of Alzheimer's disease (such as improving cognitive function).
  • the compounds of the disclosure are believed to be gamma secretase modulators (GSMs), i.e., compounds that act to shift the relative levels of ⁇ peptides produced by ⁇ -secretase.
  • GSMs gamma secretase modulators
  • the compounds alter the relative levels of ⁇ peptides produced by ⁇ -secretase, for example the level of ⁇ 42 peptide, without significantly changing the total level of ⁇ peptides produced.
  • R, Y and Z are as defined above for the compounds of Formula (I).
  • R is phenyl or C 1 -C 4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH, -CN, -NH 2 , -C 1 -C 4 alkyl, halo-substituted C 1 -C 4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci-C 4 alkyl, -N(Ci-C 4
  • R is phenyl or C 1 -C 4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C 1 -C 4 alkyl, -C 1 -C 4 alkoxy, halo-substituted C 1 -C 4 alkyl and halo-substituted C 1 -C 4 alkoxy.
  • Y is piperidinyl or azetidinyl, each of which is
  • Z is pyridinyl or pyrimidinyl, each of which is
  • R is phenyl or C 1 -C 4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen or -C 1 -C 4 alkyl; Y is piperidinyl; and Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C 1 -C 4 alkyl, halo-substituted C 1 -C 4 alkyl and -C 1 -C 4 alkoxy.
  • R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C 1 -C 4 alkyl; and Z is pyridinyl which is unsubstituted or substituted with -C 1 -C 4 alkoxy.
  • R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C 1 -C 4 alkyl. [0097] In some embodiments, R is phenyl which is unsubstituted. In some embodiments, Ris phenyl substituted with two or three -F.
  • each -C 1 -C 4 alkyl is methyl and each C 1 -C 4 alkylene is methylene.
  • C 1 -C 4 alkylene is methylene
  • C 1 -C 4 alkylene-phenyl is benzyl.
  • R is phenyl substituted with one methyl.
  • Ris C 1 -C 4 alkylene-phenyl.
  • Ris -Ci-C 6 alkyl In some embodiments, Ris -Ci-C 6 alkyl.
  • Ris 3- to 7-membered monocyclic heterocycle [0104] In some embodiments, Ris 3- to 7-membered monocyclic heterocycle.
  • Y is piperidinyl. In some embodiments, Y is azetidinyl.
  • Z is pyridinyl or pyrimidinyl, each of which is
  • Z is pyridinyl substituted with one -CF 3 . In some embodiments, Z is pyridinyl substituted with one methyl. In some embodiments, Z is pyridinyl substituted with one methoxy. In some embodiments, Z is pyrimidinyl which is unsubstituted or substituted with one to three -C 1 -C 4 alkyl. In some embodiments, Z is pyrimidinyl substituted with one methyl.
  • Z is attached to Y at the 4 position of Y. In some embodiments, Z is attached to Y at the 4 position of Z. In some embodiments, Z is attached to Y at the 1 position of Z.
  • halogen is -F. In some embodiments, halogen is -CI.
  • the compound of Formula (I) is a compound selected from the compounds in Table I.
  • a Fused Morpholinopyrimidine is a compound selected from the compounds in Table II.
  • Schemes 1-3 represent general synthetic schemes for manufacturing Fused Morpholinopyrimidines. These schemes are illustrative and are not meant to limit the possible techniques one skilled in the art may use to manufacture compounds disclosed herein. Different methods will be evident to those skilled in the art. Additionally, the various steps in the synthesis may be performed in an alternate sequence or order to give the desired compound(s).
  • a compound of formula 5 is reacted with methyl amine under basic conditions to provide a compound of formula 6.
  • the compound of formula 6 is then reacted with BBr 3 to provide a compound of formula 7.
  • the compound of formula 7 is reacted using, for example, a compound of formula 8, to provide a compound of formula 9.
  • the compound of formula 9 is then cyclized to from a compound of formula 10.
  • the compound of formula 10 is reacted with a compound of formula 4 to provide a Fused Morpholinopyrimidine.
  • the present disclosure provides pharmaceutical compositions for treating, preventing, or ameliorating a symptom of a neurodegenerative disease in a subject having a neurodegenerative disease, wherein the pharmaceutical composition comprises a therapeutically effective amount of a Fused Morpholinopyrimidine, and a pharmaceutically acceptable carrier or vehicle.
  • Fused Morpholinopyrimidines are provided in the form of pharmaceutically acceptable salts. These salts can be prepared in situ during the final isolation and purification of the compounds described herein, or by separately reacting a purified compound described herein in its free base or acid form with a suitable organic or inorganic acid or base, and isolating the salt thus formed.
  • Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, laurylsulphonate, ammonium, amine salts and the like. See, for example, Berge, et al, (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66: 1-19.
  • the pharmaceutically acceptable salts of Fused Morpholinopyrimidines include the conventional nontoxic salts or acid salts of the compounds, e.g., from non-toxic organic or inorganic acids.
  • such conventional nontoxic salts include those derived from inorganic acids, such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids, such as acetic, butionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric,
  • a suitable dose of a Fused Morpholinopyrimidine will be in the range of 0.01 to 100 mg per kilogram body weight of the recipient per day or in the range of 0.2 to 10 mg per kilogram body weight per day.
  • the desired dose can be administered once daily, but may be dosed as two, three, four, five, six or more sub-doses administered at appropriate intervals throughout the day.
  • the concentration of compounds included in compositions used in the methods described herein can range from about 1 nM to about 100 ⁇ . Effective doses are believed to range from about 10 picomole/kg to about 100 micromole/kg.
  • a Fused Morpholinopyrimidine can be administered as the sole active agent, or in combination with other known therapeutics to be beneficial in the treatment of
  • the administering physician can provide a method of treatment that is prophylactic or therapeutic by adjusting the amount and timing of drug administration on the basis of observations of one or more symptoms (e.g., motor or cognitive function as measured by standard clinical scales or assessments) of the disease being treated.
  • symptoms e.g., motor or cognitive function as measured by standard clinical scales or assessments
  • composition has been formulated in an acceptable carrier, it can be placed in an appropriate container and labeled for treatment of an indicated condition.
  • administration of a Fused Morpholinopyrimidine would include, e.g., instructions concerning the amount, frequency, and method of administration.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy.
  • the amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, the particular mode of administration.
  • the amount of active ingredient, which can be combined with a carrier or vehicle material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of 100%, this amount will range from about 1% to about 99% of active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.
  • the compounds and pharmaceutical compositions described herein can be employed in combination therapies, that is, the compounds and pharmaceutical compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures.
  • the particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved.
  • Methods of preparing these formulations or compositions include the step of bringing into association a compound described herein with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound described herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
  • a Fused Morpholinopyrimidine When administered as pharmaceuticals to humans and animals, it can be given per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably, 0.5%> to 90%>) of active ingredient in combination with a pharmaceutically acceptable carrier.
  • compositions described herein can be administered in a variety of dosage forms including, but not limited to, a solid dosage form, a liquid dosage form, an oral dosage form, a parenteral dosage form, an intranasal dosage form, a
  • suppository a lozenge, a troche, a buccal dosage form, a controlled release dosage form, a pulsed release dosage form, an immediate release dosage form, an intravenous solution, a suspension or combinations thereof.
  • compositions described herein suitable for oral administration can be in the form of capsules, cachets, pills, tablets, caplet, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in- water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouthwashes and the like, each containing a predetermined amount of a compound described herein as an active ingredient.
  • the dosage can be an oral dosage form that is a controlled release dosage form.
  • a Fused Morpholinopyrimidine can also be administered as a bolus, electuary or paste.
  • the active ingredient is mixed with one or more pharmaceutically acceptable carriers and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium carbonate, and sodium starch glycolate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol, glycerol monostearate, and poly
  • absorbents such as kaolin and bentonite clay
  • lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof
  • coloring agents such as kaolin and bentonite clay
  • talc calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof
  • coloring agents such as kaolin and bentonite clay.
  • compositions can also comprise buffering agents.
  • Solid compositions of a similar type can also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
  • a tablet can be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets can be prepared using a binder (for example, gelatin or hydroxybutylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent.
  • Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
  • Tablets, and other solid dosage forms of the pharmaceutical compositions described herein can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They can also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxybutylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres.
  • compositions can be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions, which can be dissolved in sterile water, or some other sterile injectable medium immediately before use.
  • These compositions can also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner.
  • embedding compositions which can be used include polymeric substances and waxes.
  • the active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.
  • Capsules for oral use include hard gelatin capsules in which the active ingredient is mixed with a solid diluent, and soft gelatin capsules, wherein the active ingredients is mixed with water or an oil, such as peanut oil, liquid paraffin or olive oil.
  • Dragee cores are provided with suitable coatings.
  • suitable coatings may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
  • Dyestuffs or pigments can be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
  • Liquid dosage forms for oral administration of the compounds described herein include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isobutyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, butylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • cyclodextrins e.g., hydroxybutyl- -cyclo
  • Suspensions in addition to the active compounds, can contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.
  • suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.
  • the oral compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • compositions for oral use can be obtained through combination of a Fused Morpholinopyrimidine with a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable additional compounds, if desired, to obtain tablets or dragee cores.
  • carbohydrate or protein fillers that include, but are not limited to, sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose, such as methyl cellulose, hydroxypropylmethyl-cellulose or sodium carboxymethylcellulose; and gums including arabic and tragacanth; as well as proteins, such as gelatin and collagen.
  • disintegrating or solubilizing agents can be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate.
  • Aqueous suspensions can contain a Fused Morpholinopyrimidine in admixture with excipients suitable for the manufacture of aqueous suspensions.
  • excipients include a suspending agent, such as sodium carboxymethylcellulose, methylcellulose,
  • hydroxypropylmethylcellulose sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents, such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethylene oxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensation product of ethylene oxide with a partial ester derived from fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate).
  • a naturally occurring phosphatide e.g., lecithin
  • the aqueous suspension can also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, aspartame or saccharin.
  • preservatives such as ethyl or n-propyl p-hydroxybenzoate
  • coloring agents such as a coloring agent
  • flavoring agents such as sucrose, aspartame or saccharin
  • sweetening agents such as sucrose, aspartame or saccharin.
  • Formulations can be adjusted for osmolarity.
  • Oil suspensions can be formulated by suspending a Fused Morpholinopyrimidine in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin; or a mixture of these.
  • the oil suspensions can contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol.
  • Sweetening agents can be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose.
  • These formulations can be preserved by the addition of an antioxidant, such as ascorbic acid.
  • an injectable oil vehicle see Minto, J. Pharmacol. Exp. Ther. 281 :93-102, 1997.
  • the pharmaceutical formulations can also be in the form of oil-in- water emulsions.
  • the oily phase can be a vegetable oil or a mineral oil, described above, or a mixture of these.
  • Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum
  • tragacanth naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate, and
  • condensation products of these partial esters with ethylene oxide such as polyoxyethylene sorbitan mono-oleate.
  • the emulsion can also contain sweetening agents and flavoring agents, as in the formulation of syrups and elixirs.
  • Such formulations can also contain a demulcent, a preservative, or a coloring agent.
  • a Fused Morpholinopyrimidine can be administered parenterally, such as intravenous (IV) or intramuscular (IM) administration.
  • IV intravenous
  • IM intramuscular
  • formulations for administration will commonly comprise a solution of a Fused
  • Morpholinopyrimidine dissolved in a pharmaceutically acceptable carrier can be achieved by direct injection of the pharmaceutical composition comprising the Fused Morpholinopyrimidine or by the use of infusion pumps.
  • the pharmaceutical compositions can be formulated in solid form and re-dissolved or suspended immediately prior to use. Lyophilized forms are also included.
  • the injection can be, for example, in the form of a bolus injection or continuous infusion (e.g., using infusion pumps) of pharmaceutical composition.
  • compositions suitable for parenteral administration comprise one or more compounds described herein in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
  • the acceptable vehicles and solvents that can be employed for formulation and/or reconstitution are water (e.g., water for injection) and Ringer's solution, an isotonic sodium chloride.
  • sterile fixed oils can conventionally be employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid can likewise be used in the preparation of injectables.
  • These solutions are sterile and generally free of undesirable matter.
  • These formulations may be sterilized by conventional, well known sterilization techniques such as gamma-radiation or electron beam sterilization.
  • the formulations can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like.
  • concentration of a Fused Morpholinopyrimidine in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight, and the like, in accordance with the particular mode of
  • the formulation can be a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension.
  • This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, such as a solution of 1,3-butanediol.
  • a Fused Morpholinopyrimidine can be administered by introduction into the central nervous system of the subject, e.g., into the cerebrospinal fluid of the subject.
  • the formulations for administration will commonly comprise a solution of the Fused Morpholinopyrimidine dissolved in a pharmaceutically acceptable carrier.
  • the Fused Morpholinopyrimidine is introduced intrathecally, e.g., into a cerebral ventricle, the lumbar region, or the cisterna magna.
  • the pharmaceutical composition comprising a Fused Morpholinopyrimidine is administered into a subject intrathecally.
  • the term "intrathecal administration” is intended to include delivering a pharmaceutical composition comprising a Fused Morpholinopyrimidine directly into the cerebrospinal fluid of a subject, by techniques including lateral cerebroventricular injection through a borehole or cisternal or lumbar puncture or the like (described in Lazorthes et al., Advances in Drug Delivery Systems and Applications in Neurosurgery, 1991, 18:143-192 and Omaya et al., Cancer Drug Delivery, 1984, 1 : 169-179, the contents of which are incorporated herein by reference).
  • lumbar region is intended to include the area between the third and fourth lumbar (lower back) vertebrae.
  • cisterna magna is intended to include the area where the skull ends and the spinal cord begins at the back of the head.
  • cervical ventricle is intended to include the cavities in the brain that are continuous with the central canal of the spinal cord.
  • the pharmaceutical composition is administered by injection into the cisterna magna, or lumbar area of a subject.
  • a Fused Morpholinopyrimidine can be formulated as a depot preparation. Such long acting formulations may be administered by implantation or transcutaneous delivery (e.g., subcutaneously or intramuscularly), intramuscular injection or a transdermal patch.
  • the compounds may be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
  • the absorption of the drug in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.
  • One strategy for depot injections includes the use of polyethylene oxide-polybutylene oxide copolymers, wherein the vehicle is fluid at room temperature and solidifies at body temperature.
  • Injectable depot forms can be made by forming microencapsule matrices of the subject compounds in biodegradable polymers, such as polylactide-polyglycolide.
  • the rate of drug release can be controlled.
  • biodegradable polymers include poly (orthoesters) and poly (anhydrides).
  • Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissue.
  • the compounds are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane,
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.
  • penetrants appropriate to the barrier to be permeated are used in the formulation.
  • penetrants are generally known in the art.
  • Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of the
  • release delivery systems are available and known to those of ordinary skill in the art. They include polymer base systems such as poly(lactide-glycolide), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acid and polyanhydrides.
  • Microcapsules of the foregoing polymers containing drugs are described in, for example, U.S. Pat. No. 5,075,109.
  • Delivery systems also include non-polymer systems that are: lipids including sterols, such as cholesterol, cholesterol esters and fatty acids or neutral fats, such as mono-, di- and tri-glycerides; hydrogel release systems; silastic systems; peptide based systems; wax coatings; compressed tablets using conventional binders and excipients;
  • partially fused implants include, but are not limited to: (a) erosional systems in which an agent described herein is contained in a form within a matrix, such as those described in U.S. Pat. Nos. 4,452,775, 4,675,189, and 5,736,152, and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer, such as described in U.S. Pat. Nos. 3,854,480, 5,133,974 and 5,407,686.
  • pump-based hardware delivery systems can be used, some of which are adapted for implantation.
  • a method for treating a neurodegenerative disease comprising administering to a subject an effective amount a pharmaceutical composition comprising an effective amount of a Fused Morpholinopyrimidine.
  • the method for treating a neurodegenerative disease is a method for reducing or ameliorating a symptom of the neurodegenerative disease.
  • a method for reducing or ameliorating a symptom of a neurological disease comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
  • Ameliorating or reducing the symptoms can be manifested in a variety of ways, for example, by improvement in cognitive function. Such improvement can be assessed relative to the cognitive function of the subject prior to being treated or being administered a Fused Morpholinopyrimidine or a pharmaceutical composition comprising an effective amount of a Fused Morpholinopyrimidine.
  • a method for preventing a neurological disease comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
  • a method for stopping progression of a neurological disease comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
  • Exemplary symptoms of neurological disease that can be reduced or ameliorated by administration of a Fused Morpholinopyrimidine include, but are not limited to, loss of memory, loss of cognition, loss of reasoning and/or loss of judgment. The loss of each of memory, cognition, reasoning and/or judgment can be progressive or sudden. Dementia is an exemplary symptom of neurodegenerative disease. Administration of a Fused Morpholinopyrimidine
  • Morpholinopyrimidine can reduce or improve one or more of these symptoms.
  • Exemplary cognitive functions that can be improved by administration of a Fused Morpholinopyrimidine are attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
  • the neurodegenerative disease is Alzheimer's disease. In some embodiments, the neurodegenerative disease is early onset Alzheimer's disease. In some embodiments, the early onset Alzheimer's disease is autosomal dominant early onset Alzheimer's disease.
  • the subject is 65 years or older. In some embodiments, the subject is 55 years old or younger, or 50 years old or younger. In some embodiments, the subject is older than 55 years and younger than 65 years. In some embodiments, the subject is older than 55 years.
  • the neurodegenerative disease is panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, or dementia.
  • the neurodegenerative disease is cognitive impairment, myclonus, seizures, Parkinsonism, extrapyramidal signs (EPS), apraxia, dystonia, dementia with Lewy bodies (DLB), aphasia, visual agnosia, or ataxia.
  • the subject has impaired cognitive function including one or more of attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
  • the subject has a mutation in at least one gene selected from PSEN1, PSEN2 and APP.
  • the mutation in PSEN1, PSEN2 or APP is a missense mutation.
  • the invention provides a method for treating or
  • a symptom of neurodegenerative disease e.g., Alzheimer's disease
  • the method comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
  • the increased level of ⁇ 42 in cerebrospinal fluid can be detected relative to the level of ⁇ 42 in cerebrospinal fluid of a healthy subject.
  • the invention provides a method for lowering ⁇ 42 concentration in a subject, the method comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
  • the subject has an elevated ⁇ 42 concentration relative to a healthy subject.
  • the invention provides a method for preventing increase of ⁇ 42 concentration in a subject, the method comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
  • kits that can simplify the administration of an Fused
  • the kit can comprise one or more containers filled with one or more of the ingredients of the pharmaceutical compositions described herein.
  • kits comprises a unit dosage form of a Fused Morpholinopyrimidine.
  • the unit dosage form is a container, which can be sterile, containing an effective amount of a Fused Morpholinopyrimidine and a pharmaceutically acceptable carrier or vehicle.
  • the kit can further comprise a label or printed instructions instructing the use of the Fused Morpholinopyrimidine to treat or prevent a neurodegenerative disease.
  • the kit can also further comprise a unit dosage form of another prophylactic or therapeutic agent, for example, a container containing an effective amount of the other prophylactic or therapeutic agent.
  • the kit comprises a container containing an effective amount of a Fused Morpholinopyrimidine and an effective amount of another prophylactic or therapeutic agent.
  • other prophylactic or therapeutic agents include, but are not limited to, those listed above.
  • reaction mixture was heated to 50 °C and stirred for 5 h. After consumption of the starting materials (monitored by TLC), the reaction was filtered through celite and concentrated in vacuo. The crude material was washed with hexane:ether: EtOAc (1 : 1 : 1, 2 x 30 mL) to afford tert-butyl (l-(2-methylpyridin-4-yl) piperidin-4-yl) carbamate (1.4 g, 41%) as a brown solid.
  • the reaction mixture was heated to 120 °C and stirred for 12 h in a sealed tube. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4% MeOH:CH 2 Cl 2 and further purified by preparative HPLC
  • Racemic compound of Example 26 was separated using a Chiralpak IA (250 x 20 mm, 5 ⁇ (15 mg loading; 0.1 % DEA in n-hexane; CH 2 Cl 2 :MeOH (80:20); (75 :25) as the mobile phase) to provide the compound of Example 26A Fraction I (-) and the compound of Example 26B Fraction II (+).
  • Example 26A The compound of Example 26A was produced as described in Example 26.
  • Example 26B The compound of Example 26B was produced as described in Example 26.
  • Example 29 Racemic compound of Example 29 was separated using a Chiralpak IA (250 x 20 mm, 5 ⁇ (50 mg loading; 0.1 % DEA in n-hexane; CH 2 Cl 2 :MeOH (50: 50); (75: 25) as the mobile phase) to provide the compound of Example 29A Fraction I (-) and the compound of Example 29B Fraction II (+).
  • Example 29 A Chiralpak IA (250 x 20 mm, 5 ⁇ (50 mg loading; 0.1 % DEA in n-hexane; CH 2 Cl 2 :MeOH (50: 50); (75: 25) as the mobile phase) to provide the compound of Example 29A Fraction I (-) and the compound of Example 29B Fraction II (+).
  • Example 29 A Chiralpak IA
  • Example 29 A The compound of Example 29 A was produced as described in Example 29.
  • Example 29B The compound of Example 29B was produced as described in Example 29.
  • Racemic compound of Example 31 was separated using a Chiralpak ADH (250 x 20 mm, 5 ⁇ 30 mg loading; (A) 0.1 % DEA in n-hexane (B) IPA (A:B : 70:30) as the mobile phase to provide the compound of Example 31 A Fraction I (-) and the compound of Example 3 IB Fraction II (+).
  • Example 31 A The compound of Example 31 A was produced as described in Example 31.
  • Example 3 IB The compound of Example 3 IB was produced as described in Example 31.
  • Racemic compound of Example 32 was separated using a Chiralpak ADH (250 x 20 mm, 5 ⁇ 70 mg loading; (A) 0.1 % DEA in n-hexane (B) EtOH:MeOH (50:50) (A:B : 70:30) as the mobile phase) to provide the compound of Example 32A Fraction I (+) and the compound of Example 32B Fraction II (-).
  • Example 32A The compound of Example 32A was produced as described in Example 32.
  • Example 32B The compound of Example 32B was produced as described in Example 32.
  • Racemic compound of Example 33 was separated using a Chiralpak IB (250 x 20 mm, 5 ⁇ (20 mg loading; (A) 0.1 % DEA in n-hexane (B) CH 2 Cl 2 :MeOH (80:20) (A:B : 90: 10) as the mobile phase) to provide the compound of Example 33A Fraction I (+) and the compound of Example 33B Fraction II (-).
  • Example 33A Chiralpak IB (250 x 20 mm, 5 ⁇ (20 mg loading; (A) 0.1 % DEA in n-hexane (B) CH 2 Cl 2 :MeOH (80:20) (A:B : 90: 10) as the mobile phase) to provide the compound of Example 33A Fraction I (+) and the compound of Example 33B Fraction II (-).
  • Example 33 A The compound of Example 33 A was produced as described in Example 33.
  • Example 33B The compound of Example 33B was produced as described in Example 33.
  • Racemic compound of Example 34 was separated using a Chiralpak IB (250 x 20 mm, 5 ⁇ (20 mg loading; (A) 0.1 % DEA in n-hexane (B) CH 2 Cl 2 :MeOH (50:50) (A:B :
  • Example 34A The compound of Example 34A was produced as described in Example 34.
  • Example 34B The compound of Example 34B was produced as described in Example 34.
  • reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with CH 2 C1 2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo.
  • Racemic compound of Example 35 was separated using a Chiralpak IB (250 x 20 mm, 5 ⁇ (30 mg loading; (A) 0.1 % DEA in n-hexane (B) CH 2 Cl 2 :MeOH (80:20) (A:B : 90: 10) as the mobile phase) to provide the compound of Example 35A Fraction I (+) and the compound of Example 35B Fraction II (-).
  • Example 35 A The compound of Example 35 A was produced as described in Example 35.
  • Example 35B The compound of Example 35B was produced as described in Example 35.
  • LC-MS 432.5 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 ⁇ ); RT 3.36 min. 5 mM NH 4 OAc: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 ⁇ ); RT 1.62 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
  • Racemic compound of Example 37 was separated using a Chiralpak IA (250 x 20 mm, 5 ⁇ (35 mg loading; 0.1 % DEA in n-hexane: CH 2 Cl 2 :MeOH (50:50); (A:B: 75:25) as the mobile phase) to provide the compound of Example 37 A Fraction I (+) and the compound of Example 37B Fraction II (-).
  • Example 37A The compound of Example 37A was produced as described in Example 37.
  • Example 37B The compound of Example 37B was produced as described in Example 37.
  • Racemic compound of Example 38 was separated using a Chiralpak IB (250 x 20 mm, 5 ⁇ (30 mg loading; (A) 0.1 % DEA in n-hexane (B) CH 2 Cl 2 :MeOH (80:20) (A:B :
  • Example 38A The compound of Example 38A was produced as described in Example 38.
  • Example 38B The compound of Example 38B was produced as described in Example 38.
  • Racemic compound of Example 39 was separated using a Chiralpak IA (250 x 20 mm, 5 ⁇ (50 mg loading; 0.1 % DEA in n-hexane: CH 2 Cl 2 :MeOH (50:50); (A:B : 85: 15) as the mobile phase) to provide the compound of Example 39 A Fraction I (+) and the compound of Example 39B Fraction II (-).
  • Example 39A The compound of Example 39A was produced as described in Example 39.
  • Example 39B The compound of Example 39B was produced as described in Example 39.
  • Racemic compound of Example 40 was separated using a Chiralpak-IA (250 x 20 mm, 5 ⁇ (50 mg loading; 0.1 % DEA in rc-hexane: CH 2 Cl 2 :MeOH (50:50); (A:B : 85: 15) as the mobile phase to provide the compound of Example 40A Fraction I (+) and the compound of Example 40B Fraction II (-).
  • Chiralpak-IA 250 x 20 mm, 5 ⁇ (50 mg loading; 0.1 % DEA in rc-hexane: CH 2 Cl 2 :MeOH (50:50); (A:B : 85: 15) as the mobile phase to provide the compound of Example 40A Fraction I (+) and the compound of Example 40B Fraction II (-).
  • Example 40A The compound of Example 40A was produced as described in Example 40.
  • Example 40B The compound of Example 40B was produced as described in Example 40.
  • Racemic compound of Example 41 was separated using a Chiralpak-IB (250 x 20 mm, 5 ⁇ (20 mg loading; 0.1 % DEA in n-hexane: CH 2 Cl 2 :MeOH (50:50); (A:B : 90: 10) as the mobile phase to provide the compound of Example 41 A Fraction I (+) and the compound of Example 4 IB Fraction II (-).
  • Example 41 A The compound of Example 41 A was produced as described in Example 41.
  • Example 4 IB The compound of Example 4 IB was produced as described in Example 41.
  • Racemic compound of Example 42 was separated using a Chiralpak IB (250 x 20 mm, 5 ⁇ (30 mg loading; 0.1 % DEA in n-hexane: CH 2 Cl 2 :MeOH (50:50); (90: 10) as the mobile phase) to provide the compound of Example 42A Fraction I (+) and the compound of Example 42B Fraction II (-).
  • Example 42A The compound of Example 42A was produced as described in Example 42.
  • Example 42B The compound of Example 42B was produced as described in Example 42.

Abstract

The present disclosure relates to Fused Morpholinopyrimidines, pharmaceutical compositions comprising an effective amount of a Fused Morpholinopyrimidine and methods for using a Fused Morpholinopyrimidine in the treatment of a neurodegenerative disease, comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.

Description

FUSED MORPHOLINOP YRIMIDINE S AND METHODS OF USE THEREOF
1. RELATED APPLICATIONS
[0001] This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application Serial No. 61/952,704, filed on March 13, 2014 and to U.S. Provisional
Application Serial No. 62/054,673, filed on September 24, 2014, the entire disclosures of which are incorporated by reference herein in their entirety.
2. FIELD
[0002] This disclosure relates generally to Fused Morpholinopyrimidine compounds. More specifically, the disclosure relates to the use of the Fused Morpholinopyrimidine compounds for the treatment of neurological disease.
3. BACKGROUND
[0003] Alzheimer's disease (AD) is the most prevalent form of dementia. It is a neurodegenerative disease that is associated (though not exclusively) with aging. The disease is clinically characterized by a progressive loss of memory, cognition, reasoning and judgment that leads to an extreme mental deterioration and ultimately death. The disease is pathologically characterized by the deposition of extracellular plaques and the presence of neurofibrillary tangles. The plaques are considered to play an important role in the pathogenesis of the disease. They mainly consist of fibrillar aggregates of β-amyloid peptide (Αβ), which are products of the amyloid precursor protein (APP). APP is initially processed by β-secretase forming a secreted peptide and a membrane bound C99 fragment. The C99 fragment is subsequently processed by the proteolytic activity of γ -secretase. Multiple sites of proteolysis on the C99 fragment lead to the production of a range of smaller peptides (Αβ 37-42 amino acids). N-terminal truncations can also be found e.g., Αβ (4-42). For convenience, notations Αβ40 and Αβ42, as used herein, include these N-terminal truncated peptides. Upon secretion, the Αβ peptides initially form soluble aggregates which ultimately lead to the formation of insoluble deposits and plaques. Αβ42 is believed to be the most neurotoxic; the shorter peptides have less propensity to aggregate and form plaques. Αβ plaques in the brain are also associated with cerebral amyloid angiopathy, hereditary cerebral hemorrhage with amyloidosis, multi infarct dementia, dementia pugilistisca and Down's Syndrome. [0004] γ-secretase is an association of four proteins: Aphl, nicastrin, presenilin and Pen- 2 (review De Strooper, Neuron 38:9-12 (2003)). Subjects carrying particular mutations in one of these components, presenilin, show increased Αβ42/Αβ40 ratio. These mutations are correlated with early onset familial AD. Inhibition of γ-secretase resulting in the lowering of Αβ42 has been investigated by the pharmaceutical community, and numerous inhibitors have been found. See, e.g., Thompson et al. {Bioorg. Med. Chem. Lett. 2006, 16, 2357-63), Shaw et al. {Bioorg. Med. Chem. Lett. 2006, 17, 511-16) and Asberom et al. {Bioorg. Med. Chem. Lett. 2007, 15, 2219-2223). Inhibition of γ-secretase, though, is not without side-effects, some of which are due to the γ-secretase complex processing substrates other than C99, e.g., Notch. A more desirable approach is to modulate the proteolytic activity of the γ-secretase complex in a manner that lowers Αβ42 in favor of shorter peptides without significantly affecting the activity of γ-secretase on substrates such as Notch.
[0005] Compounds that have shown modulation of γ-secretase include certain nonsteroidal, anti-inflammatory drugs (NSAIDs), for example Flurbiprofen, (Stock et al, Bioorg. Med. Chem. Lett. 2006, 16, 2219-2223). Other publications that disclose agents said to reduce Αβ42 through the modulation of γ-secretase include: WO 2004/074232, WO
2005/054193, Perreto et al, Journal of Medicinal Chemistry 2005, 48, 5705-20, WO
2005/108362, WO 2006/008558, WO 2006/021441, WO 2006/041874, WO 2006/045554, WO 2004/110350, WO 2006/043964, WO 2005/115990, EP 1847524, WO 2007/116228, WO 2007/110667, WO 2007/124394, EP 184752, EP 1849762, WO 2007/125364, WO 2009/086277.
4. SUMMARY
[0006] It is understood that any of the embodiments described below can be combined in any desired way, and that any embodiment or combination of embodiments can be applied to each of the aspects described below, unless the context indicates otherwise.
[0007] In one aspect, the invention provides a compound of Formula (I)
Figure imgf000003_0001
(I) or a pharmaceutically acceptable salt thereof, wherein: R is phenyl, C1-C4 alkylene -phenyl, -Ci- C6 alkyl or 3- to 7-membered monocyclic heterocycle, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH,
-CN, -NH2, -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci-C4 alkyl, -N(Ci-C4 alkyl)(C(0)(Ci-C4 alkyl)), -C(0)NH-Ci-C4 alkyl, -C(0)N(Ci-C4 alkyl)2, hydroxy-substituted C1-C4 alkyl, -S(0)2-Ci-C4 alkyl, -S(0)2-halo- substituted CrC4 alkyl, -S(0)2-NH-Ci-C4 alkyl, -S(0)2-N(d-C4 alkyl)2, -NH-S(0)2-Ci-C4 alkyl, -N(Ci-C4 alkyl)-S(0)2-Ci-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkoxy, 3- to 7- membered monocyclic heterocycle, C3-C8 monocyclic cycloalkyl and -C(0)NH2; Y is 4- to 6- membered nonaromatic heterocycle, which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C1-C4 alkoxy, halo- substituted C1-C4 alkoxy, -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino-substituted C1-C4 alkoxy, (C1-C4 alkyl)2N-Ci-C4 alkoxy, -NH-C1-C4 alkyl, -CN, -OH and -NH2; and Z is nitrogen- containing 5- to 6-membered aromatic monocyclic heterocycle which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -NH2, -OH, -C1-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkyl, halo-substituted C1-C4 alkoxy and 3- to 7-membered monocyclic heterocycle.
[0008] In some embodiments, R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH, -CN, -NH2i -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci-C4 alkyl, -N(Ci-C4
alkyl)(C(0)(Ci-C4 alkyl)), -C(0)NH-Ci-C4 alkyl, -C(0)N(Ci-C4 alkyl)2, hydroxy-substituted C1-C4 alkyl, -S(0)2-Ci-C4 alkyl, -S(0)2-halo-substituted C1-C4 alkyl, -S(0)2-NH-Ci-C4 alkyl, -S(0)2-N(Ci-C4 alkyl)2, -NH-S(0)2-Ci-C4 alkyl, -N(C C4 alkyl)-S(0)2-Ci-C4 alkyl, -C C4 alkoxy, halo-substituted C1-C4 alkoxy, 3- to 7-membered monocyclic heterocycle, C3-C8 monocyclic cycloalkyl and -C(0)NH2.
[0009] In some embodiments, R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C1-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkyl and halo-substituted C1-C4 alkoxy.
[0010] In some embodiments, Y is piperidinyl or azetidinyl, each of which is
unsubstituted. [0011] In some embodiments, Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, halo-substituted C1-C4 alkyl and -C1-C4 alkoxy.
[0012] In some embodiments, R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen or -C1-C4 alkyl; Y is piperidinyl; and Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, halo-substituted C1-C4 alkyl and -C1-C4 alkoxy.
[0013] In some embodiments, R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C1-C4 alkyl; and Z is pyridinyl which is unsubstituted or substituted with -C1-C4 alkoxy.
[0014] In some embodiments, R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C1-C4 alkyl.
[0015] In some embodiments, R is phenyl which is unsubstituted. In some embodiments, Ris phenyl substituted with two or three -F.
[0016] In some embodiments, each -C1-C4 alkyl is methyl and each C1-C4 alkylene is methylene.
[0017] In some embodiments, C1-C4 alkylene is methylene.
[0018] In some embodiments, C1-C4 alkylene-phenyl is benzyl.
[0019] In some embodiments, R is phenyl substituted with one methyl.
[0020] In some embodiments, Ris C1-C4 alkylene-phenyl.
[0021] In some embodiments, Ris -Ci-C6 alkyl.
[0022] In some embodiments, Ris 3- to 7-membered monocyclic heterocycle.
[0023] In some embodiments, Y is piperidinyl. In some embodiments, Y is azetidinyl.
[0024] In some embodiments, Z is pyridinyl or pyrimidinyl, each of which is
unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, -CF3 and -C1-C4 alkoxy.
[0025] In some embodiments, Z is pyridinyl substituted with one -CF3. In some embodiments, Z is pyridinyl substituted with one methyl. In some embodiments, Z is pyridinyl substituted with one methoxy. In some embodiments, Z is pyrimidinyl which is unsubstituted or substituted with one to three -C1-C4 alkyl. In some embodiments, Z is pyrimidinyl substituted with one methyl.
[0026] In one embodiment, the compound of Formula (I) is a compound selected from the list of compounds in Table I.
[0027] In one aspect, the compound of Formula (I) is selected from the group consisting of: N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-phenyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7-phenyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(o-tolyl)-7,8-dihydro-6H-pyrimido[5,4- b] [ 1 ,4]oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(o- tolyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; 8-methyl-N-(l-(6- methylpyrimidin-4-yl)piperidin-4-yl)-7-(3,4,5-trifiuorophenyl)-7,8-dihydro-6H-pyrimido[5,4- b][l,4]oxazin-2-amine; (+)-8-methyl-N-(l-(6-methylpyrimidin-4-yl)piperidin-4-yl)-7-(3,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; 7-(3,5- difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(3,5-difluorophenyl)-N-(l-(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7-(3,4,5-trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (-)-N- (l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8-dihydro- 6H-pyrimido[5,4-b][l,4]oxazin-2-amine; 8-methyl-7-phenyl-N-(l-(2- (trifluoromethyl)pyridin-4-yl)piperidin-4-yl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2- amine; (+)-8-methyl-7-phenyl-N-( 1 -(2-(trifluoromethyl)pyridin-4-yl)piperidin-4-yl)-7,8- dihydro-6H-pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-7-benzyl-8-methyl-N-( 1 -(6- methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2- amine; (+)-7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7-(4- fluorophenyl)-8-methyl-N-(l -(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7- (4-fluorophenyl)-N-( 1 -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7- (4-fluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7-(2, 4-difluorophenyl)-N-(l-(2- methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7-(2, 4- difluorophenyl)-8-methyl-N-( 1 -(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7-(2-(trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2- amine; (+)-N-(l -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2-(trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine ; (+)-7-(2, 4- difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7-benzyl-N-(l-(2-methoxypyridin- 4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine (-)-7- benzyl-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4^ b][l,4]oxazin-2-amine; (+)-7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)- 8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(2,4-dichlorophenyl)- N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4- b][l,4]oxazin-2-amine; (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(2,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(2-chloro-4- fluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-7-(4-fluoro-2-(trifluoromethyl)phenyl)-N-( 1 -(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l^ amine; (+)-7-(5-chloro-2-(trifluoromethyl)phenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4- yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(5-fluoro-2- (trifluoromethyl)phenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dih 6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(2-chloro-4,5-difluorophenyl)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazm^ amine; (+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7- (3,4,5-trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-8-methyl- N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(o-tolyl)-7,8-dihydro-6H- pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(3,5-difluorophenyl)-8-methyl-N-((3R,6S)-6-(2- methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2- amine; (+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7- (2,4,5-trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (-)-8-methyl-N- ((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(2,4,5-trifluorophenyl)-7,8- dihydro-6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine; N-( 1 -(2-methoxypyridin-4-yl)piperidin-4- yl)-8-methyl-7-(trifluoromethyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine and (- )-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7- (trifluoromethyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine.
[0028] In one aspect, the compound of Formula (I) is selected from the group consisting of: 7-(3,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(3,5-difluorophenyl)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2- amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-N-(l-(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4- b] [ 1 ,4]oxazin-2-amine; (+)-7-(4-fluorophenyl)-N-( 1 -(2-methoxypyridin-4-yl) piperidin-4-yl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-7-(2, 4- difluorophenyl)-N-(l -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2- amine; (+)-7-benzyl-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-7-(2-chlorophenyl)-N-( 1 -(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(2,4- dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7-(2,4,5-trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7- (2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(5-fluoro-2-(trifluoromethyl)phenyl)- N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4- b][l,4]oxazin-2-amine; (+)-7-(2-chloro-4,5-difluorophenyl)-N-(l-(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-8- methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(3,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(3,5- difluorophenyl)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)- 7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-8-methyl-N-((3R,6S)-6-(2- methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(2,4,5-trifiuorophenyl)-7,8-dihydro-6H- pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine; and N-( 1 -(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7-(trifluoromethyl)-7,8-dihydro-6H^
[0029] In one aspect, the invention provides a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H- pyrimido [5 ,4-b]pyrrolo [ 1 ,2-d] [ 1 ,4]oxazin-2-amine; (-)-N-( 1 -(2-methoxypyridin-4- yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8-trimethyl-7,8-dihydro-6H-pyrimido[5,4- b][l,4]oxazin-2-amine; and N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8'-methyl-6',8'- dihydrospiro[cyclopentane-l,7'-pyrimido[5,4-b][l,4]oxazin]-2'-amine, or a pharmaceutically acceptable salt thereof.
[0030] In one aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or vehicle and an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
[0031] In one aspect, the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or vehicle and an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)- 6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; (-)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2- d][l,4]oxazin-2-amine
N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8-trimethyl-7,8-dihydro-6H-pyrimido[5,4- b][l,4]oxazin-2-amine; and N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8'-methyl-6',8'- dihydrospiro[cyclopentane-l,7'-pyrimido[5,4-b][l,4]oxazin]-2'-amine, or a pharmaceutically acceptable salt thereof.
[0032] In one aspect, the invention provides a method for treating a neurodegenerative disease, comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable salt of a compound of Formula (I).
[0033] In one aspect, the invention provides a method for treating a neurodegenerative disease, comprising administering to a subject in need thereof an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4- yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; (-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4- b]pyrrolo[l,2-d][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8- trimethyl-7,8-dihydro-6H-pyrimido[5,4-b][l ,4]oxazin-2-amine; and N-(l-(2-methoxypyridin- 4-yl)piperidin-4-yl)-8'-methyl-6\8'-dihydrospiro[cyclopentane-l,7'-pyrimido[5,4- b][l,4]oxazin]-2'-amine, or a pharmaceutically acceptable salt thereof.
[0034] In some embodiments, the neurodegenerative disease is panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, or dementia.
[0035] In one aspect, the invention provides a method for treating Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
[0036] In one aspect, the invention provides a method for treating Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)- 6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; (-)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2- d][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-7,7,8-trimethyl-7,8- dihydro-6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine; and N-( 1 -(2-methoxypyridin-4- yl)piperidin-4-yl)-8'-methyl-6',8'-dihydrospiro[cyclopentane-l,7'-pyrimido[5,4- b][l,4]oxazin]-2'-amine, or a pharmaceutically acceptable salt thereof.
[0037] In one aspect, the invention provides a method for improving an impaired cognitive function, comprising administering to a subject having impaired cognitive function an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
[0038] In one aspect, the invention provides a method for improving an impaired cognitive function, comprising administering to a subject having impaired cognitive function an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2- d][l,4]oxazin-2-amine; (-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro- 6H-pyrimido[5,4-b]pyrrolo[ 1 ,2-d] [ 1 ,4]oxazin-2-amine; N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-7,7,8-trimethyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; and N-(l -(2-methoxypyridin-4-yl)piperidin-4-yl)-8'-methyl-6',8'-dihydrospiro[cyclopentane- 1 ,7'- pyrimido[5,4-b][l,4]oxazin]-2'-amine, or a pharmaceutically acceptable salt thereof.
[0039] In one aspect, the invention provides a method for ameliorating a symptom of Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
[0040] In one aspect, the invention provides a method for ameliorating a symptom of Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound seletected from the group consisting of (+)-N-(l-(2-methoxypyridin- 4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H-pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2- amine; (-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-6a,7,8,9-tetrahydro-6H- pyrimido[5,4-b]pyrrolo[l,2-d][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin- 4-yl)-7,7,8-trimethyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; and N-(l-(2- methoxypyridin-4-yl)piperidin-4-yl)-8'-methyl-6',8'-dihydrospiro[cyclopentane-l,7'- pyrimido[5,4-b][l,4]oxazin]-2'-amine, or a pharmaceutically acceptable salt thereof.
[0041] In some embodiments, Alzheimer's disease is early onset Alzheimer's disease.
[0042] In some embodiments, the subject is a human.
[0043] In some embodiments, the symptom is progressive loss of memory, progressive loss of cognition, progressive loss of reasoning and/or progressive loss of judgment.
[0044] A compound of Formula (I), Table II, or a pharmaceutically acceptable salt thereof (also referred to herein as a "Fused Morpholinopyrimidine") is useful for treating, preventing or ameliorating one or more symptoms of a neurodegenerative disease.
[0045] Exemplary neurodegenerative diseases include, but are not limited to, Alzheimer's disease, early onset Alzheimer's disease, panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, dementia, cognitive impairment, myclonus, seizures, Parkinsonism, extrapyramidal signs (EPS), apraxia, dystonia, dementia with Lewy bodies (DLB), aphasia, visual agnosia, and ataxia.
[0046] In some embodiments, the cognitive function impaired is one or more of attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
[0047] In some embodiments, the subject is 65 years old or older. In other embodiments, the subject is 55 years old or older. In still other embodiments, the subject is 55 years old or younger, or 50 years old or younger.
[0048] A pharmaceutical composition comprising an effective amount of a Fused Morpholinopyrimidine and a pharmaceutically acceptable carrier or vehicle is useful for treating or preventing a neurodegenerative disease.
[0049] The details of the invention are set forth in the accompanying description below.
[0050] All patents and publications cited in this specification are hereby incorporated by reference in their entirety.
5. DETAILED DESCRIPTION
5.1 Definitions and Abbreviations
[0051] The following are definitions of terms used in the present specification. The initial definition provided for a group or term herein applies to that group or term throughout the present specification individually or as part of another group, unless otherwise indicated. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.
[0052] The term "C1-C4 alkyl" as used herein, refers to a straight chain or branched non-cyclic hydrocarbon having from 1 to 4 carbon atoms, wherein one of the hydrocarbon's hydrogen atoms has been replaced by a single bond. Representative straight chain C1-C4 alkyls include -methyl, -ethyl, -n-propyl and -n-butyl. Representative branched C1-C4 alkyls include
-isopropyl, -sec-butyl, -isobutyl and -tert-butyl.
[0053] The term "C3-C8 monocyclic cycloalkyl" as used herein, refers to a saturated cyclic hydrocarbon having from 3 to 8 carbon atoms. Representative C3-C8 monocyclic cycloalkyls include -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclohexyl, -cycloheptyl and - cyclooctyl.
[0054] The term "C1-C4 alkylene" as used herein, refers to a straight or branched chain saturated hydrocarbon containing 1-4 carbon atoms, wherein two of the hydrocarbon's hydrogen atoms have been replaced by a single a bond. Representative C1-C4 alkylene groups include, methylene, ethylene, n-propylene, isopropylene, n-butylene and isobutylene.
[0055] The term "C1-C4 alkoxy" as used herein, refers to a C1-C4 alkyl-O- group wherein the C1-C4 alkyl is as defined above. Examples of C1-C4 alkoxy include, but are not limited to, methoxy, trifluoromethoxy, ethoxy, propoxy or butoxy.
[0056] The term "halo-substituted C1-C4 alkoxy" as used herein, refers to a C1-C4 alkoxy group, as defined above, wherein one or more of the C1-C4 alkoxy group's hydrogen atoms have been replaced with -F, -CI, -Br or -I. Examples of a halo-substituted C1-C4 alkoxy include, but are not limited to, -0-CH2F, -O-CCI3, -O-CF3, -0-CH2Cl, -0-CH2CH2Br, -O- CH2CH2I, -0-CF2CF3, -0-CH2CH2CH2F, -0-CH2CH2CH2Cl, -0-CH2CH2CH2CH2Br, -O- CH2CH2CH2CH2I, -0-CH2CH(Br)CH3, -0-CH2CH(Cl)CH2CH3, -0-CH(F)CH2CH3, - OCH2CF3 and -0-C(CH3)2(CH2Cl).
[0057] The term "amino-substituted C1-C4 alkoxy" as used herein, refers to a C1-C4 alkoxy group, as defined above, wherein one or more of the C1-C4 alkoxy group's hydrogen atoms have been replaced with -NH2. Examples of amino-substituted C1-C4 alkoxy include, but are not limited to, -0-CH2NH2, -0-CH2CH2NH2, -0-CH(NH2)CH3, -0-CH2CH2CH2 NH2, -0-CH2CH2CH2CH2NH2, -0-CH2CH(NH2)CH3, -0-CH(NH2)CH2CH3 and -O- C(CH3)2(CH2NH2).
[0058] A "nitrogen-containing 4- to 6-membered nonaromatic heterocycle" refers to a monocyclic 4- to 6-membered nonaromatic monocyclic cycloalkyl group in which one of the cycloalkyl group's ring carbon atoms has been replaced with a nitrogen atom and 0-3 of the cycloalkyl group's remaining ring carbon atoms are independently replaced with a N, O or S atom. The nitrogen-containing 4- to 6-membered nonaromatic heterocycles can be attached via a nitrogen or carbon atom. Representative examples of nitrogen-containing 4- to 6- membered nonaromatic heterocycles include, but are not limited to, azetidinyl, piperidinyl, oxazinyl, morpholinyl, imidazolidinyl, pyrazolidinyl and thiomorpholinyl.
[0059] A "4- to 6-membered nonaromatic heterocycle" refers to a monocyclic 4- to 6- membered nonaromatic monocyclic cycloalkyl group in which 1-3 of the cycloalkyl group's ring carbon atoms are independently replaced with a N, O or S atom. The 4- to 6-membered nonaromatic heterocycles can be attached via a nitrogen or carbon atom. Representative examples of nitrogen-containing 4- to 6-membered nonaromatic heterocycles include, but are not limited to, azetidinyl, piperidinyl, oxazinyl, morpholinyl, imidazolidinyl, pyrazolidinyl, thiomorpholinyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl and dioxanyl. [0060] The term "3- to 7-membered monocyclic heterocycle"" as used herein, refers to a monocyclic 3- to 7-membered aromatic or non-aromatic monocyclic cycloalkyl in which 1-4 of the ring carbon atoms have been independently replaced with a N, O or S atom. The 3- to 7- membered monocyclic heterocycles can be attached via a nitrogen or carbon atom.
Representative examples of a 3- to 7-membered monocyclic heterocycle group include, but are not limited to, nitrogen-containing 3- to 7-membered monocyclic heterocycles discussed above, tetrahydrofuranyl, dihydrofuranyl, pyranyl, dihydropyranyl, tetrahydropyranyl, thiopyranyl, dihydrothiopyranyl, tetrahydrothiopyranyl, dioxanyl, dithianyl, trithianyl, dioxolanyl, furanyl and thiophenyl. In one embodiment, the 3- to 7-membered monocyclic heterocycle is a nitrogen-containing 3- to 7-membered monocyclic heterocycle. In another embodiment, the 3- to 7-membered monocyclic heterocycle is fully saturated or partially saturated.
[0061] The term "nitrogen-containing 5- to 6-membered aromatic monocyclic
heterocycle" as used herein, refers to a 5- or 6-membered aromatic monocyclic heterocycle in which from 1 to 4 of the ring carbon atoms have been independently replaced with a nitrogen atom and 0-4 of the cycloalkyl group's remaining ring carbon atoms have been independently replaced with an O or S atom. The nitrogen-containing 5- to 6-membered aromatic monocyclic heterocycle can be attached via a nitrogen or carbon atom. Representative examples of a 5- to 6-membered aromatic monocyclic heterocycles include, but are not limited to, imidazolyl, isothiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, pyrimidinyl, pyrazolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrrolyl, thiazolyl, thiadiazolyl, triazinyl, and
triazolyl. Unless otherwise indicated, the nitrogen-containing 5- to 6-membered aromatic monocyclic heterocycle is unsubstituted.
[0062] The terms "halogen" or "halo" as used herein, refer to chlorine, bromine, fluorine or iodine.
[0063] The term "halo-substituted C1-C4 alkyl" as used herein, refers to a C1-C4 alkyl group, as defined above, wherein one or more of the C1-C4 alkyl group's hydrogen atoms have been replaced with -F, -CI, -Br or -I. Examples of a halo-substituted C1-C4 alkyl include, but are not
limited to, -CH2F, -CCI3, -CF3, -CH2C1, -CH2CH2Br, -CH2CH2I, -CF2CF3, -CH2CH2CH2F,
-CH2CH2CH2C1, -CH2CH2CH2CH2Br, -CH2CH2CH2CH2I, -CH2CH(Br)CH3, -CH2CH(C1 )CH2CH3, -CH(F)CH2CH3, -CH2CF3 and -C(CH3)2(CH2C1). [0064] The term "amino-substituted C1-C4 alkyl" as used herein, refers to a C1-C4 alkyl group, as defined above, wherein one or more of the C1-C4 alkyl group's hydrogen atoms have been replaced with -NH2. Examples of amino-substituted C1-C4 alkyl include, but are not
limited to, -CH2NH2, -CH2CH2NH2, -CH(NH2)CH3, -CH2CH2CH2NH2, -CH2CH(NH2)CH3, -CH2CH(NH2)CH2CH3, -CH(NH2)CH2CH3 and -C(CH3)2(CH2NH2).
[0065] The term "C1-C4 alkylene-phenyl" as used herein, refers to a C1-C4 alkyl group, as defined above, wherein one of the C1-C4 alkyl group's hydrogen atoms has been replaced with phenyl.
[0066] The term "hydroxy-substituted C1-C4 alkyl" as used herein, refers to a C1-C4 alkyl group, as defined above, wherein one or more of the C1-C4 alkyl group's hydrogen atoms have been replaced with -OH. Representative examples of a hydroxy-substituted C1-C4 alkyl include, but are not limited to, -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, - CH2CH2CH2CH2OH,
-CH2CH(OH)CH3, -CH2CH(OH)CH2CH3, -CH(OH)CH2CH3 and -C(CH3)2(CH2OH).
[0067] Unless otherwise indicated, any heteroatom with unsatisfied valences is assumed to have hydrogen atoms sufficient to satisfy the valences.
[0068] The phrase "pharmaceutically acceptable carrier or vehicle" as used herein, refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or
transporting the Fused Morpholinopyrimidine from one organ, or portion of the body, to another organ, or portion of the body. Each carrier or vehicle must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. Some examples of materials which can serve as pharmaceutically acceptable carriers or vehicles include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc;
excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as butylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen- free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.
[0069] The compounds described herein may form salts which are also within the scope of this invention. Reference to a compound described herein is understood to include reference to salts thereof, unless otherwise indicated. The term "salt(s)", as employed herein, denotes acidic and/or basic salts formed with inorganic and/or organic acids and bases. In addition, when a compound described herein contains both a basic moiety, such as, but not limited to, amine, 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. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, e.g., in isolation or purification steps which may be employed during preparation. Salts of the compounds described herein may be formed, for example, by reacting a 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.
[0070] The compounds described herein which contain a basic moiety, such as, but not limited to, an amine or a pyridine or imidazole ring, may form salts with a variety of organic and inorganic acids. Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, for example, trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides, hydroiodides, hydroxyethanesulfonates (e.g., 2- hydroxyethanesulfonates), lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonates), nicotinates, nitrates, oxalates, pectinates, persulfates,
phenylpropionates (e.g., 3-phenylpropionates), phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), sulfonates, tartrates, thiocyanates, toluenesulfonates, such as tosylates, undecanoates and the like.
[0071] The compounds described herein which contain an acidic moiety, such as, but not limited to, a carboxylic acid, may form salts with a variety of organic and inorganic bases. 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 benzathines, dicyclohexylamines, hydrabamines (formed with N,N-bis(dehydroabietyl) ethylenediamine), N-methyl-D- glucamines, N-methyl-D-glycamides, t-butyl amines and salts with amino acids, such as arginine, lysine and the like. Basic nitrogen-containing groups may be quaternized with agents, such as lower alkyl halides (e.g., methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others.
[0072] Prodrugs and solvates of the compounds described herein are also contemplated herein. The term "prodrug" as employed herein denotes a compound that, upon
administration to a subject, undergoes chemical conversion by metabolic or chemical processes to yield a compound described herein, or a salt and/or solvate thereof. Solvates of the compounds described herein include, for example, hydrates.
[0073] Compounds described herein are, subsequent to their preparation, preferably isolated and purified to obtain a composition containing an amount by weight equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 97%, equal to or greater than 98%, or equal to or greater than 99% of the compounds
("substantially pure" compounds), which is then used or formulated as described herein. Such "substantially pure" compounds described herein are also contemplated herein as part of the present invention.
[0074] All stereoisomers of the present compounds (for example, those which may exist due to asymmetric carbons on various substituents), including enantiomeric forms and diastereomeric forms, are contemplated within the scope of this invention. Individual stereoisomers of the compounds described herein may, for example, be substantially free of other isomers (e.g., as a pure or substantially pure optical isomer having a specified activity), or may be admixed, for example, as racemates or with all other, or other selected,
stereoisomers. The chiral centers of the compounds described herein may have the S or R configuration as defined by the International Union of Pure and Applied Chemistry (IUPAC) 1974 Recommendations. The racemic forms can be resolved by physical methods, such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives or separation by chiral column chromatography. The individual optical isomers can be obtained from the racemates by any suitable method, including without limitation, conventional methods, such as, for example, salt formation with an optically active acid followed by crystallization. [0075] If, for instance, a particular enantiomer of a compound described herein is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers. Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.
[0076] All configurational isomers of the compounds described herein are contemplated, either in admixture or in pure or substantially pure form. Certain compounds described herein may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, cis (Z) and trans (E) alkene isomers R- and ^-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent, such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
[0077] Isomeric mixtures containing any of a variety of isomer ratios may be utilized in accordance with the present invention. For example, where only two isomers are combined, mixtures containing 50:50, 60:40, 70:30, 80:20, 90: 10, 95:5, 96:4, 97:3, 98:2, 99: 1, or 100:0 isomer ratios are all contemplated by the present invention. Those of ordinary skill in the art will readily appreciate that analogous ratios are contemplated for more complex isomer mixtures.
[0078] Throughout the specifications, groups and substituents thereof may be chosen to provide stable moieties and compounds. Compounds useful in the treatment, for example, are neurodegenerative disorders. The term "stable", as used herein, refers to compounds which possess stability sufficient to allow manufacture and which maintain the integrity of the compound for a sufficient period of time to be detected and preferably for a sufficient period of time to be useful for the purposes detailed herein.
[0079] Definitions of specific functional groups and chemical terms are described in more detail above. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999, the entire contents of which are incorporated herein by reference.
[0080] In some embodiments, the present invention also includes isotopically labeled compounds, which are identical to the compounds disclosed herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine and chlorine, such as 2H, 3H, 13C, UC, 14C, 15N, 180, 170, 31P, 32P, 35S, 18F and 36C1, respectively. Compounds described herein, or an enantiomer, diastereomer, tautomer, or pharmaceutically acceptable salt or solvate thereof, which contain the aforementioned isotopes and/or other isotopes of other atoms, are within the scope of this invention. Certain isotopically labeled compounds described herein, for example those into which radioactive isotopes, such as 3H and 14C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3H, and carbon- 14, i.e., 14C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes, such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements, and hence, may be preferred in some circumstances. Isotopically labeled compounds can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.
[0081] When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
[0082] As used herein, "effective amount" refers to any amount that is necessary or sufficient for achieving or promoting a desired outcome, e.g., for treating, preventing, or ameliorating a symptom of a neurodegenerative disease. In some instances an effective amount is a therapeutically effective amount. A therapeutically effective amount is any amount that is necessary or sufficient for promoting or achieving a desired biological response in a subject. The effective amount for any particular application can vary depending on such factors as the disease or condition being treated, the particular agent being administered, the size of the subject, or the severity of the disease or condition. [0083] As used herein, "treat" or "treating" includes stopping the progression and/or reducing or ameliorating a symptom of a neurodegenerative disease, for example, improving cognitive function.
[0084] As used herein, the term "subject" refers to a vertebrate animal. In one embodiment the subject is a mammal. In one embodiment the subject is a human. In other embodiments the subject is a non-human vertebrate animal, including, without limitation, non-human primates, laboratory animals, livestock, domesticated animals and non- domesticated animals. Non-limiting examples of subject include a mammal, e.g., a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, and non-human primate, such as a monkey, chimpanzee, baboon or rhesus. In one embodiment, the subject is a human.
[0085] Practitioners of the art will recognize that certain chemical groups may exist in multiple tautomeric forms (for example, as an amide or imino ether). The scope of this disclosure is meant to include all such tautomeric forms. For example, a tetrazole may exist in two tautomeric forms, 1-H tetrazole and a 2-H tetrazole. This is depicted in the figure below. This example is not meant to be limiting in the scope of tautomeric forms.
^t-i JN»H
N ' N
H
[0086] Practitioners of the art will recognize that certain electrophilic ketones, may exist in a hydrated form. The scope of this disclosure is to include all such hydrated forms. For example, a trifluoromethyl ketone may exist in a hydrated form via addition of water to the carbonyl group. This is depicted in the figure below. This example is not meant to be limiting in the scope of hydrated forms.
Figure imgf000020_0001
Abbreviations
[0087] Abbreviations used in the following examples and preparations include:
Αβ Amyloid-beta
Ac Acyl (Me-C(O)-)
ACN Acetonitrile
AD Alzheimer's Disease APP Amyloid Precursor Protein
Aq Aqueous
BINAP 2,2'-Bis(diphenylphosphino -l,r-binaphthyl
Bn Benzyl
Boc Tert-butyloxycarbonyi
BSA Bovine Serum Albumin
Bu Butyl
BuLi Butyllithium
c Cyclo
cBu Cylcobutyl
Cone. Concentrated
cPr Cyclopropyl
CSF Cerebrospinal Fluid
dba Dibenzylideneacetone
DBU l,8-Diazabicyclo[5.4.0]undec-7-ene
DCE 1, 2-Dichloroethane
DCM Dichloromethane
DEA Di-ethylamine
DIAD Diisopropyl Azodicarboxylate
DIPEA N, N-Diisopropylethylamine
DMAP 4-Dimethylaminopyridine
DMF D imethylformamide
DMP Dess-Martin Periodinane
DMSO Dimethyl Sulfoxide
DPPA Diphenoxyphosphoryl Azide
dppf 1, 1 '-Bis(diphenylphosphiiio)ferrocene
ELISA Enzyme -Linked Immuno Sorbent Assay
ESI Electrospray Ionization
Et Ethyl
Et3N Triethylamine
Eq. Equivalent
g Grams(s)
HPLC High Pressure Liquid Chromatography h Hour(s)
IPA Isopropyl Alcohol
ipc Isopyropinocampheyl
iPr Isopropyl
i.v or IV. Intravenous
LAH Lithium Aluminum Hydride
LC-MS Liquid Chromatography-Mass Spectrometry
LC/MS Liquid Chromatography Mass Spectrometry
LG Leaving Group
LRMS Low Resolution Mass Spectrometry
m Multiplet
MeOH Methanol
min Minute(s)
mmol Millimoles
μΐ Microliter
ul Microliter
μπι Micrometer
Ms Mesylate
MS Mass Spectrometry
MW Molecular Weight (all values are ±0.05)
n Normal
N Normal
NBS N-Bromosuccinimide
NMP l-Methylpyrrolidin-2-one
NMR Nuclear Magnetic Resonance
NSAIDS Non-Steroidal Anti-Inflammatory Drugs
o/n Overnight
PMB Para-methoxybenzyl
PBS Phosphate Buffered Saline
Py Pyridine
Ra-Ni Raney Nickel
Rf Retention Factor
RT (or rt) Room Temperature (about 20-25°C) or Retention Time RT Retention Time
s Singlet
sat. Saturated
t Triplet
t-Bu Tertiary Butyl
tert Tertiary
Tf Triflate
TFA Trifluoro acetic Acid
TFAA Trifluoroacetic Anhydride
THF Tetrahydrofuran
TLC Thin Layer Chromatography
TMS Trimethylsilyl
TPP Triphenylphosphine
Ts Tosylate
UPLC Ultra Performance Liquid Chromatography
v/v Volume/volume
wt/v Weight/volume
5.2 Fused Morpholinopyrimidines
[0088] Described below are Fused Morpholinopyrimidines, i.e., compounds according to Formula (I), the compounds in Table II, and pharmaceutically acceptable salts thereof, as well as methods for preparing the compounds and using the compounds to treat one or more neurodegenerative diseases, e.g., reducing a symptom of Alzheimer's disease (such as improving cognitive function). The compounds of the disclosure are believed to be gamma secretase modulators (GSMs), i.e., compounds that act to shift the relative levels of Αβ peptides produced by γ-secretase. In some embodiments, the compounds alter the relative levels of Αβ peptides produced by γ-secretase, for example the level of Αβ42 peptide, without significantly changing the total level of Αβ peptides produced.
[0089] In one aspect, described herein are compounds according to Formula (I), below:
Figure imgf000023_0001
(I)
and pharmaceutically acceptable salts thereof,
wherein R, Y and Z are as defined above for the compounds of Formula (I).
[0090] In some embodiments, R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH, -CN, -NH2, -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci-C4 alkyl, -N(Ci-C4
alkyl)(C(0)(Ci-C4 alkyl)), -C(0)NH-C C4 alkyl, -C(0)N(C C4 alkyl)2, hydroxy-substituted C1-C4 alkyl, -S(0)2-Ci-C4 alkyl, -S(0)2-halo-substituted C1-C4 alkyl, -S(0)2-NH-Ci-C4 alkyl, -S(0)2-N(Ci-C4 alkyl)2, -NH-S(0)2-Ci-C4 alkyl, -N(Ci-C4 alkyl)-S(0)2-Ci-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkoxy, 3- to 7-membered monocyclic heterocycle, C3-C8 monocyclic cycloalkyl and -C(0)NH2.
[0091] In some embodiments, R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C1-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkyl and halo-substituted C1-C4 alkoxy.
[0092] In some embodiments, Y is piperidinyl or azetidinyl, each of which is
unsubstituted.
[0093] In some embodiments, Z is pyridinyl or pyrimidinyl, each of which is
unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, halo-substituted C1-C4 alkyl and -C1-C4 alkoxy.
[0094] In some embodiments, R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen or -C1-C4 alkyl; Y is piperidinyl; and Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, halo-substituted C1-C4 alkyl and -C1-C4 alkoxy.
[0095] In some embodiments, R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C1-C4 alkyl; and Z is pyridinyl which is unsubstituted or substituted with -C1-C4 alkoxy.
[0096] In some embodiments, R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C1-C4 alkyl. [0097] In some embodiments, R is phenyl which is unsubstituted. In some embodiments, Ris phenyl substituted with two or three -F.
[0098] In some embodiments, each -C1-C4 alkyl is methyl and each C1-C4 alkylene is methylene.
[0099] In some embodiments, C1-C4 alkylene is methylene.
[0100] In some embodiments, C1-C4 alkylene-phenyl is benzyl.
[0101] In some embodiments, R is phenyl substituted with one methyl.
[0102] In some embodiments, Ris C1-C4 alkylene-phenyl.
[0103] In some embodiments, Ris -Ci-C6 alkyl.
[0104] In some embodiments, Ris 3- to 7-membered monocyclic heterocycle.
[0105] In some embodiments, Y is piperidinyl. In some embodiments, Y is azetidinyl.
[0106] In some embodiments, Z is pyridinyl or pyrimidinyl, each of which is
unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, -CF3 and -C1-C4 alkoxy.
[0107] In some embodiments, Z is pyridinyl substituted with one -CF3. In some embodiments, Z is pyridinyl substituted with one methyl. In some embodiments, Z is pyridinyl substituted with one methoxy. In some embodiments, Z is pyrimidinyl which is unsubstituted or substituted with one to three -C1-C4 alkyl. In some embodiments, Z is pyrimidinyl substituted with one methyl.
[0108] In some embodiments, Z is attached to Y at the 4 position of Y. In some embodiments, Z is attached to Y at the 4 position of Z. In some embodiments, Z is attached to Y at the 1 position of Z.
[0109] In some embodiments, halogen is -F. In some embodiments, halogen is -CI.
[0110] In some embodiments, the compound of Formula (I) is a compound selected from the compounds in Table I.
Table I. Exemplary Fused Morpholinopyrimidines of Formula (I)
Compound
Structure Name
of Example
Figure imgf000026_0001
1 u (racemic)
Figure imgf000027_0001
Figure imgf000028_0001
Figure imgf000029_0001
Figure imgf000030_0001
oxazin-2-amine
Figure imgf000031_0001
Figure imgf000032_0001
Figure imgf000033_0001
oxazn- -amne
Figure imgf000034_0001
Figure imgf000035_0001
b] [ 1 ,4]oxazin-2-amine
Figure imgf000036_0001
F b] [ 1 ,4]oxazin-2-amine
Figure imgf000037_0001
b] [ 1 ,4]oxazin-2-amine
Figure imgf000038_0001
N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7-
76B N il (trifluoromethyl)-7,8-dihydro- 6H-pyrimido[5,4-b] [ 1 ,4]oxazin- 2-amine
H 1
(-)-8-methyl-N-((3R,6S)-6-(2- methylpyridin-4-yl)tetrahydro-
2H-pyran-3-yl)-7-
77A
(trifluoromethyl)-7,8-dihydro- 6H-pyrimido[5,4-b] [ 1 ,4]oxazin-
H 1 2-amine
(+)-8-methyl-N-((3R,6S)-6-(2- methylpyridin-4-yl)tetrahydro-
2H-pyran-3-yl)-7-
77B
(trifluoromethyl)-7,8-dihydro- 6H-pyrimido[5,4-b] [ 1 ,4]oxazin-
H 1 2-amine
[0111] In some embodiments, a Fused Morpholinopyrimidine is a compound selected from the compounds in Table II.
Table II. Exemplary Fused Morpholinopyrimidines
Figure imgf000039_0001
N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-7,7,8-
83 trimethyl-7,8-dihydro-6H- pyrimido[5,4-b] [1 ,4]oxazin-2- amine
H 1
N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-8'-methyl-
84 6',8'-dihydrospiro[cyclopentane- l,7'-pyrimido[5,4- b] [1 ,4]oxazin]-2'-amine
Methods for Making Fused Morpholinopyrimidines
[0112] Methods useful for making the Fused Morpholinopyrimidines are set forth in the Examples below and generalized in Schemes 1-3 for the compounds of Formula (I).
[0113] Schemes 1-3 represent general synthetic schemes for manufacturing Fused Morpholinopyrimidines. These schemes are illustrative and are not meant to limit the possible techniques one skilled in the art may use to manufacture compounds disclosed herein. Different methods will be evident to those skilled in the art. Additionally, the various steps in the synthesis may be performed in an alternate sequence or order to give the desired compound(s).
Various modifications to these methods may be envisioned by those skilled in the art to achieve similar results to that of the inventors provided below. For example, optional protecting groups can be used as described, for example, in Greene et al., Protective Groups in Organic Synthesis (3r ed. 1999).
Scheme 1 heteroc
Figure imgf000040_0001
[0114] As shown in Scheme 1, a compound of formula 1 is coupled to the compound of formula 2 under standard coupling conditions to provide a compound of formula 3. The compound of formula 3 is deprotected to provide a compound of formula 4. Scheme 2
Figure imgf000041_0001
[0115] As shown in Scheme 2, a compound of formula 5 is reacted with methyl amine under basic conditions to provide a compound of formula 6. The compound of formula 6 is then reacted with BBr3 to provide a compound of formula 7. The compound of formula 7 is reacted using, for example, a compound of formula 8, to provide a compound of formula 9. The compound of formula 9 is then cyclized to from a compound of formula 10. The compound of formula 10 is reacted with a compound of formula 4 to provide a Fused Morpholinopyrimidine.
Scheme 3
Figure imgf000042_0001
Figure imgf000042_0002
13 14
Figure imgf000042_0003
15 16
Figure imgf000042_0004
[0116] As shown in Scheme 3, a compound of formula 5 is reacted with BBr3 to provide a compound of formula 11. The compound of formula 11 is then protected to form compound 12. Enantio-pure compound of formula 13 is reduced with LAH to provide a compound of formula 14. The compound of formula 14 is then coupled with compound 12 to form compound 15. The compound of formula 15 is then deprotected under acidic conditions to provide a compound of formula 16. The compound of formula 16 is then cyclized to form a compound of formula 17. The compound of formula 17 is coupled to a compound of formula 4 to provide an enantiomerically-pure Fused Morpholinopyrimidine at the chiral carbon atom (marked with *). 5.4 Pharmaceutical Compositions Comprising a Fused Morpholinopyrimidine
[0117] In another aspect, the present disclosure provides pharmaceutical compositions for treating, preventing, or ameliorating a symptom of a neurodegenerative disease in a subject having a neurodegenerative disease, wherein the pharmaceutical composition comprises a therapeutically effective amount of a Fused Morpholinopyrimidine, and a pharmaceutically acceptable carrier or vehicle.
[0118] As set out above, in some embodiments, Fused Morpholinopyrimidines are provided in the form of pharmaceutically acceptable salts. These salts can be prepared in situ during the final isolation and purification of the compounds described herein, or by separately reacting a purified compound described herein in its free base or acid form with a suitable organic or inorganic acid or base, and isolating the salt thus formed. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, laurylsulphonate, ammonium, amine salts and the like. See, for example, Berge, et al, (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66: 1-19.
[0119] The pharmaceutically acceptable salts of Fused Morpholinopyrimidines include the conventional nontoxic salts or acid salts of the compounds, e.g., from non-toxic organic or inorganic acids. For example, such conventional nontoxic salts include those derived from inorganic acids, such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids, such as acetic, butionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric,
toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic and the like.
[0120] In general, a suitable dose of a Fused Morpholinopyrimidine will be in the range of 0.01 to 100 mg per kilogram body weight of the recipient per day or in the range of 0.2 to 10 mg per kilogram body weight per day. The desired dose can be administered once daily, but may be dosed as two, three, four, five, six or more sub-doses administered at appropriate intervals throughout the day. [0121] The concentration of compounds included in compositions used in the methods described herein can range from about 1 nM to about 100 μΜ. Effective doses are believed to range from about 10 picomole/kg to about 100 micromole/kg.
[0122] A Fused Morpholinopyrimidine can be administered as the sole active agent, or in combination with other known therapeutics to be beneficial in the treatment of
neurodegenerative diseases. In any event, the administering physician can provide a method of treatment that is prophylactic or therapeutic by adjusting the amount and timing of drug administration on the basis of observations of one or more symptoms (e.g., motor or cognitive function as measured by standard clinical scales or assessments) of the disease being treated.
[0123] Details on techniques for formulation and administration are well described in the scientific and patent literature, see, e.g., Remington: The Science and Practice of Pharmacy (2 Volumes), (22nd Edition, 2012), Pharmaceutical Press ("Remington's"). After a
pharmaceutical composition has been formulated in an acceptable carrier, it can be placed in an appropriate container and labeled for treatment of an indicated condition. For
administration of a Fused Morpholinopyrimidine, such labeling would include, e.g., instructions concerning the amount, frequency, and method of administration.
[0124] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, the particular mode of administration. The amount of active ingredient, which can be combined with a carrier or vehicle material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of 100%, this amount will range from about 1% to about 99% of active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.
[0125] The compounds and pharmaceutical compositions described herein can be employed in combination therapies, that is, the compounds and pharmaceutical compositions can be administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. [0126] Methods of preparing these formulations or compositions include the step of bringing into association a compound described herein with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound described herein with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.
[0127] When a Fused Morpholinopyrimidine is administered as pharmaceuticals to humans and animals, it can be given per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (more preferably, 0.5%> to 90%>) of active ingredient in combination with a pharmaceutically acceptable carrier.
[0128] The pharmaceutical compositions described herein can be administered in a variety of dosage forms including, but not limited to, a solid dosage form, a liquid dosage form, an oral dosage form, a parenteral dosage form, an intranasal dosage form, a
suppository, a lozenge, a troche, a buccal dosage form, a controlled release dosage form, a pulsed release dosage form, an immediate release dosage form, an intravenous solution, a suspension or combinations thereof.
Oral Formulations and Administration
[0129] Pharmaceutical formulations described herein suitable for oral administration can be in the form of capsules, cachets, pills, tablets, caplet, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in- water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouthwashes and the like, each containing a predetermined amount of a compound described herein as an active ingredient. The dosage can be an oral dosage form that is a controlled release dosage form. A Fused Morpholinopyrimidine can also be administered as a bolus, electuary or paste.
[0130] In solid dosage forms described herein for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, sodium carbonate, and sodium starch glycolate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol, glycerol monostearate, and polyethylene oxide-polybutylene oxide copolymer;
absorbents, such as kaolin and bentonite clay; lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and coloring agents. In the case of capsules, tablets and pills, the pharmaceutical
compositions can also comprise buffering agents. Solid compositions of a similar type can also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
[0131] A tablet can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using a binder (for example, gelatin or hydroxybutylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets can be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
[0132] Tablets, and other solid dosage forms of the pharmaceutical compositions described herein, such as dragees, capsules, pills and granules, can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They can also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxybutylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They can be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions, which can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions can also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients. [0133] Capsules for oral use include hard gelatin capsules in which the active ingredient is mixed with a solid diluent, and soft gelatin capsules, wherein the active ingredients is mixed with water or an oil, such as peanut oil, liquid paraffin or olive oil.
[0134] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments can be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
[0135] Liquid dosage forms for oral administration of the compounds described herein include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isobutyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, butylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Additionally, cyclodextrins, e.g., hydroxybutyl- -cyclodextrin, may be used to solubilize compounds.
[0136] Suspensions, in addition to the active compounds, can contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, and mixtures thereof.
[0137] Besides inert diluents, the oral compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
[0138] Pharmaceutical preparations for oral use can be obtained through combination of a Fused Morpholinopyrimidine with a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable additional compounds, if desired, to obtain tablets or dragee cores. Suitable solid excipients in addition to those previously mentioned are carbohydrate or protein fillers that include, but are not limited to, sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose, such as methyl cellulose, hydroxypropylmethyl-cellulose or sodium carboxymethylcellulose; and gums including arabic and tragacanth; as well as proteins, such as gelatin and collagen. If desired, disintegrating or solubilizing agents can be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate.
[0139] Pharmaceutical preparations for oral use can be presented as aqueous or liposome formulations. Aqueous suspensions can contain a Fused Morpholinopyrimidine in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include a suspending agent, such as sodium carboxymethylcellulose, methylcellulose,
hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents, such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethylene oxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensation product of ethylene oxide with a partial ester derived from fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous suspension can also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, aspartame or saccharin. Formulations can be adjusted for osmolarity.
[0140] Oil suspensions can be formulated by suspending a Fused Morpholinopyrimidine in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin; or a mixture of these. The oil suspensions can contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents can be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose. These formulations can be preserved by the addition of an antioxidant, such as ascorbic acid. As an example of an injectable oil vehicle, see Minto, J. Pharmacol. Exp. Ther. 281 :93-102, 1997. The pharmaceutical formulations can also be in the form of oil-in- water emulsions. The oily phase can be a vegetable oil or a mineral oil, described above, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum
tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate, and
condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan mono-oleate. The emulsion can also contain sweetening agents and flavoring agents, as in the formulation of syrups and elixirs. Such formulations can also contain a demulcent, a preservative, or a coloring agent.
Parenteral Formulations and Administration
[0141] In another embodiment, a Fused Morpholinopyrimidine can be administered parenterally, such as intravenous (IV) or intramuscular (IM) administration. The
formulations for administration will commonly comprise a solution of a Fused
Morpholinopyrimidine dissolved in a pharmaceutically acceptable carrier. Administration of a Fused Morpholinopyrimidine to any of the above mentioned sites can be achieved by direct injection of the pharmaceutical composition comprising the Fused Morpholinopyrimidine or by the use of infusion pumps. The pharmaceutical compositions can be formulated in solid form and re-dissolved or suspended immediately prior to use. Lyophilized forms are also included. The injection can be, for example, in the form of a bolus injection or continuous infusion (e.g., using infusion pumps) of pharmaceutical composition.
[0142] Pharmaceutical compositions suitable for parenteral administration comprise one or more compounds described herein in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
[0143] Among the acceptable vehicles and solvents that can be employed for formulation and/or reconstitution are water (e.g., water for injection) and Ringer's solution, an isotonic sodium chloride. In addition, sterile fixed oils can conventionally be employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid can likewise be used in the preparation of injectables. These solutions are sterile and generally free of undesirable matter. These formulations may be sterilized by conventional, well known sterilization techniques such as gamma-radiation or electron beam sterilization. The formulations can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, toxicity adjusting agents, e.g., sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of a Fused Morpholinopyrimidine in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight, and the like, in accordance with the particular mode of
administration selected and the subject's needs. For IV administration, the formulation can be a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally-acceptable diluent or solvent, such as a solution of 1,3-butanediol.
[0144] In some embodiments, a Fused Morpholinopyrimidine can be administered by introduction into the central nervous system of the subject, e.g., into the cerebrospinal fluid of the subject. The formulations for administration will commonly comprise a solution of the Fused Morpholinopyrimidine dissolved in a pharmaceutically acceptable carrier. In certain aspects, the Fused Morpholinopyrimidine is introduced intrathecally, e.g., into a cerebral ventricle, the lumbar region, or the cisterna magna.
[0145] In some embodiments, the pharmaceutical composition comprising a Fused Morpholinopyrimidine is administered into a subject intrathecally. As used herein, the term "intrathecal administration" is intended to include delivering a pharmaceutical composition comprising a Fused Morpholinopyrimidine directly into the cerebrospinal fluid of a subject, by techniques including lateral cerebroventricular injection through a borehole or cisternal or lumbar puncture or the like (described in Lazorthes et al., Advances in Drug Delivery Systems and Applications in Neurosurgery, 1991, 18:143-192 and Omaya et al., Cancer Drug Delivery, 1984, 1 : 169-179, the contents of which are incorporated herein by reference). The term "lumbar region" is intended to include the area between the third and fourth lumbar (lower back) vertebrae. The term "cisterna magna" is intended to include the area where the skull ends and the spinal cord begins at the back of the head. The term "cerebral ventricle" is intended to include the cavities in the brain that are continuous with the central canal of the spinal cord. In some embodiments, the pharmaceutical composition is administered by injection into the cisterna magna, or lumbar area of a subject.
Depot Formulations and Administration
[0146] A Fused Morpholinopyrimidine can be formulated as a depot preparation. Such long acting formulations may be administered by implantation or transcutaneous delivery (e.g., subcutaneously or intramuscularly), intramuscular injection or a transdermal patch. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
[0147] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. One strategy for depot injections includes the use of polyethylene oxide-polybutylene oxide copolymers, wherein the vehicle is fluid at room temperature and solidifies at body temperature.
[0148] Injectable depot forms can be made by forming microencapsule matrices of the subject compounds in biodegradable polymers, such as polylactide-polyglycolide.
Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissue.
Intranasal Formulations and Administration
[0149] For administration by inhalation, the compounds are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.
Other Formulations and Modes of Administration [0150] For transmucosal administration (e.g., buccal, rectal, nasal, ocular, etc.), penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
[0151] Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of the
compounds, increasing convenience to the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art. They include polymer base systems such as poly(lactide-glycolide), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acid and polyanhydrides.
Microcapsules of the foregoing polymers containing drugs are described in, for example, U.S. Pat. No. 5,075,109. Delivery systems also include non-polymer systems that are: lipids including sterols, such as cholesterol, cholesterol esters and fatty acids or neutral fats, such as mono-, di- and tri-glycerides; hydrogel release systems; silastic systems; peptide based systems; wax coatings; compressed tablets using conventional binders and excipients;
partially fused implants; and the like. Specific examples include, but are not limited to: (a) erosional systems in which an agent described herein is contained in a form within a matrix, such as those described in U.S. Pat. Nos. 4,452,775, 4,675,189, and 5,736,152, and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer, such as described in U.S. Pat. Nos. 3,854,480, 5,133,974 and 5,407,686. In addition, pump-based hardware delivery systems can be used, some of which are adapted for implantation.
5.5 Treatment, Prevention or Amelioration of Symptoms of a Neurodegenerative
Disease
[0152] In another aspect, a method for treating a neurodegenerative disease is described, comprising administering to a subject an effective amount a pharmaceutical composition comprising an effective amount of a Fused Morpholinopyrimidine.
[0153] In some embodiments, the method for treating a neurodegenerative disease is a method for reducing or ameliorating a symptom of the neurodegenerative disease.
[0154] In some embodiments, a method for reducing or ameliorating a symptom of a neurological disease is described, comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine. Ameliorating or reducing the symptoms can be manifested in a variety of ways, for example, by improvement in cognitive function. Such improvement can be assessed relative to the cognitive function of the subject prior to being treated or being administered a Fused Morpholinopyrimidine or a pharmaceutical composition comprising an effective amount of a Fused Morpholinopyrimidine.
[0155] In some embodiments, a method for preventing a neurological disease is described, comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
[0156] In some embodiments, a method for stopping progression of a neurological disease is described, comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
[0157] Exemplary symptoms of neurological disease that can be reduced or ameliorated by administration of a Fused Morpholinopyrimidine include, but are not limited to, loss of memory, loss of cognition, loss of reasoning and/or loss of judgment. The loss of each of memory, cognition, reasoning and/or judgment can be progressive or sudden. Dementia is an exemplary symptom of neurodegenerative disease. Administration of a Fused
Morpholinopyrimidine can reduce or improve one or more of these symptoms.
[0158] Exemplary cognitive functions that can be improved by administration of a Fused Morpholinopyrimidine are attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
[0159] In one embodiment, the neurodegenerative disease is Alzheimer's disease. In some embodiments, the neurodegenerative disease is early onset Alzheimer's disease. In some embodiments, the early onset Alzheimer's disease is autosomal dominant early onset Alzheimer's disease.
[0160] In some embodiments, the subject is 65 years or older. In some embodiments, the subject is 55 years old or younger, or 50 years old or younger. In some embodiments, the subject is older than 55 years and younger than 65 years. In some embodiments, the subject is older than 55 years.
[0161] In some embodiments, the neurodegenerative disease is panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, or dementia. [0162] In some embodiments, the neurodegenerative disease is cognitive impairment, myclonus, seizures, Parkinsonism, extrapyramidal signs (EPS), apraxia, dystonia, dementia with Lewy bodies (DLB), aphasia, visual agnosia, or ataxia.
[0163] In some embodiments, the subject has impaired cognitive function including one or more of attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
[0164] In some embodiments, the subject has a mutation in at least one gene selected from PSEN1, PSEN2 and APP. In some embodiments, the mutation in PSEN1, PSEN2 or APP is a missense mutation.
[0165] In some embodiments, the invention provides a method for treating or
ameliorating a symptom of neurodegenerative disease (e.g., Alzheimer's disease) in a subject with an increased level of Αβ42 in cerebrospinal fluid, the method comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine. In such subject, the increased level of Αβ42 in cerebrospinal fluid can be detected relative to the level of Αβ42 in cerebrospinal fluid of a healthy subject.
[0166] In some embodiments, the invention provides a method for lowering Αβ42 concentration in a subject, the method comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine. In some embodiments, the subject has an elevated Αβ42 concentration relative to a healthy subject.
[0167] In some embodiments, the invention provides a method for preventing increase of Αβ42 concentration in a subject, the method comprising administering to a subject in need thereof an effective amount of a Fused Morpholinopyrimidine.
5.6 Kits
[0168] Described herein are kits that can simplify the administration of an Fused
Morpholinopyrimidine to a subject. The kit can comprise one or more containers filled with one or more of the ingredients of the pharmaceutical compositions described herein.
Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. [0169] A typical kit comprises a unit dosage form of a Fused Morpholinopyrimidine. In one embodiment, the unit dosage form is a container, which can be sterile, containing an effective amount of a Fused Morpholinopyrimidine and a pharmaceutically acceptable carrier or vehicle. The kit can further comprise a label or printed instructions instructing the use of the Fused Morpholinopyrimidine to treat or prevent a neurodegenerative disease. The kit can also further comprise a unit dosage form of another prophylactic or therapeutic agent, for example, a container containing an effective amount of the other prophylactic or therapeutic agent. In one embodiment, the kit comprises a container containing an effective amount of a Fused Morpholinopyrimidine and an effective amount of another prophylactic or therapeutic agent. Examples of other prophylactic or therapeutic agents include, but are not limited to, those listed above.
[0170] The representative examples which follow are intended to help illustrate the invention, and are not intended to, nor should they be construed to, limit the scope of the invention. Indeed, various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including the examples which follow and the references to the scientific and patent literature cited herein. It should further be appreciated that the contents of those cited references are incorporated herein by reference to help illustrate the state of the art. The following examples contain additional information, exemplification and guidance which can be adapted to the practice of this invention in its various embodiments and equivalents thereof. The examples do not limit the scope of the invention described in the claims.
6. EXAMPLES Example 1
Synthesis of l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000056_0001
Synthesis of tert-Butyl (l-(2-methoxypyridin-4-yl) piperidin-4-yl) carbamate
[0171] To a stirred solution of tert-butyl piperidin-4-ylcarbamate (1.2 g, 6.38 mmol) in toluene (80 mL) was added 4-bromo-2-methoxypyridine (1.3 g, 6.38 mmol), sodium tert- butoxide (1.8 g, 19.14 mmol) and Xantphos (220 mg, 0.06 mmol). The mixture was purged under an argon atmosphere for 20 min, then Pd2(dba)3 (73 mg, 0.12 mmol) was added. The reaction mixture was heated to 90 °C and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (30 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified through silica gel column chromatography using 30% EtOAc:hexanes to afford tert-butyl (l-(2-methoxypyridin-4-yl) piperidin-4-yl) carbamate (1 g, 53%) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.77-7.75 (m, 1H), 6.83-6.82 (m, 1H), 6.52 (d, 1H), 6.08 (s, 1H), 3.80-3.75 (m, 5H), 3.47 (s, 1H), 2.86 (t, 2H), 1.75-1.73 (m, 2H), 1.38-1.30 (m, 11H); LC-MS: 308.3 (M+l); (column; X- Bridge C-18 (50 3.0 mm, 3.5 μ); RT 3.14 min. 0.05% aq TFA: ACN; 0.80 mL/min); TLC: 45% EtOAc:hexanes (Rf. 0.3).
Synthesis of l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride
[0172] To a stirred solution of tert-Butyl (l-(2-methoxypyridin-4-yl) piperidin-4-yl) carbamate (1 g, 3.25 mmol) in EtOAc (15 mL) was added 4M HCI-EtOAc (2 mL) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo. The crude material was washed with ether (2 x 20 mL) to afford l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (950 mg) as an off- white solid which was used for the next step without further purification. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.34 (bs, 2H), 7.90-7.89 (m, 1H), 6.88-6.87 (m, 1H), 6.50 (s, 1H), 4.26 (d, 2H), 4.02 (s, 3H), 3.40-3.39 (m, 1H), 3.24-3.19 (m, 2H), 2.06- 2.04 (m, 2H), 1.60-1.54 (m, 2H); LC-MS: 208.1 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μ); RT 0.52 min. 0.05% aq TFA: ACN; 0.80 mL/min) TLC: 5% MeOH:CH2Cl2 (R/. 0.1).
Example 2
Synthesis of l-(2-chloropyridin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000057_0001
Synthesis of tert-Butyl (4-(2-chloropyridin-4-yl) cyclohexyl) carbamate
[0173] To a stirred solution of 2-chloro-4-fluoropyridine (1.5 g, 11.40 mmol) in NMP (15 mL) was added tert-Butyl piperidin-4-ylcarbamate (2.7 g, 13.68 mmol) and DIPEA (1.9 ml, 14.82 mmol) and purged under an argon atmosphere for 15 min. The reaction mixture was heated 150 °C and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 50% EtOAc:hexanes to afford tert-butyl (4-(2-chloropyridin-4-yl) cyclohexyl) carbamate (2.3 g, 65%) as a brown solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.90-7.89 (m, 1H), 6.83-6.79 (m, 2H), 3.85 (d, 2H), 3.49 (br s, 1H), 2.94 (t, 2H), 1.76-1.73 (m, 2H), 1.37 (s, 9H), 1.33-1.30 (m, 3H); LC-MS: 312 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 2.55 min. 0.05% aq TFA in water: ACN; 0.80 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.2).
Synthesis of l-(2-chloropyridin-4-yl) piperidin-4-amine hydrochloride
[0174] To a stirred solution of tert-Butyl (4-(2-chloropyridin-4-yl) cyclohexyl) carbamate (2.1 g, 3.20 mmol) in EtOAc (25 mL) was added 4M HCl-EtOAc (4 mL) at 0 °C. The reaction was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was neutralized with a saturated NaHCOs solution (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(2-chloropyridin-4-yl) piperidin-4-amine hydrochloride (950 mg, 67%) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.99-7.97 (m, 2H), 7.95-7.94 (m, 1H), 6.88 (s, 1H), 6.86-6.85 (m, 1H), 3.99 (d, 2H), 3.31-3.29 (m, 1H), 2.96 (t, 2H), 1.91 (d, 2H), 1.47-1.45 (m, 2H); LC-MS: 212.2 (M+l); (column; Eclipse XDB C18 (150 4.6 mm, 5 μπι); RT 7.92 min. 0.05% aq TFA: ACN; 1.00 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.1).
Example 3
Synthesis of l-(pyrimidin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000058_0001
Synthesis of tert-butyl (l-(2-chloropyrimidin-4-yl) piperidin-4-yl) carbamate
[0175] To a stirred solution of 2, 4-dichloropyrimidine (2 g, 13.42 mmol) in DMF (20 mL) was added triethyl amine (2.03 g, 20.13 mmol) and tert-butyl piperidin-4-ylcarbamate (2.9 g, 14.76 mmol). The reaction mixture was stirred for 18 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified through silica gel column chromatography using 50% EtOAc:hexanes to afford tert-butyl (l-(2- chloropyrimidin-4-yl) piperidin-4-yl) carbamate (2.2 g, 53%) as an off-white solid. 1H-NMR (CDCls, 400 MHz): δ 8.03-8.02 (m, 1H), 6.40-6.39 (m, 1H), 4.49-4.29 (m, 3H), 3.74 (br s, 1H), 3.09-3.02 (m, 2H), 2.07-2.04 (m, 2H), 1.50 (s, 9H), 1.38-1.25 (m, 2H); LC-MS: 313.2 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 3.21 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.6).
Synthesis of tert-butyl (1 -(pyrimidin-4-yl) piperidin-4-yl) carbamate
[0176] To a stirred solution of tert-butyl (l-(2-chloropyrimidin-4-yl) piperidin-4-yl) carbamate (2 g, 6.39 mmol) in EtOH (300 mL) was added 10% Pd/C (300 mg) and triethylamine (646 mg, 6.39 mmol). The reaction mixture was stirred for 5 h under a ¾ atmosphere (balloon pressure). After consumption of the starting materials (monitored by TLC), the mixture was filtered through celite and concentrated in vacuo. The crude material was purified through silica gel column chromatography using 50% EtOAc:hexanes to afford tert-butyl (l-(pyrimidin-4-yl) piperidin-4-yl) carbamate (1 g, 56%) as an off-white solid. 1H- NMR (DMSO-<¾, 400 MHz): δ 8.45 (s, 1H), 8.14-8.12 (m, 1H), 6.85-6.80 (m, 2H), 4.28-4.25 (m, 2H), 3.54 (br s, 1H), 3.02-2.95 (m, 2H), 1.79-1.76 (m, 2H), 1.35 (s, 9H), 1.32-1.23 (m, 2H); LC-MS: 279 (M+1); (column; X-Bridge C18 (50 3.0 mm, 3.5 μπι); RT 2.83 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.2).
Synthesis of 1 -(pyrimidin-4-yl) piperidin-4-amine hydrochloride
[0177] To a stirred solution of tert-butyl (l-(pyrimidin-4-yl) piperidin-4-yl) carbamate (1 g, 3.59 mmol) in EtOAc (10 mL) was added 4M HCI in EtOAc (2 mL) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was filtered through celite and concentrated in vacuo to afford l-(pyrimidin-4-yl) piperidin-4-amine hydrochloride (700 mg, 94%) as a brown solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 8.8 (s, 1H), 8.35-8.33 (m, 2H), 7.24-7.22 (m, 1H), 4.14- 4.12 (m, 2H), 3.40-3.30 (m, 3H), 2.11-2.06 (m, 2H), 1.62-1.54 (m, 2H); LC-MS: 179.2 (M+1); (column; Deltapak C-4 (150 3.9 mm, 5 μπι); RT 2.02 min. 0.05% aq TFA: ACN; 1.00 mL/min). TLC: 10% MeOH:DCM (Rf. 0.1).
Example 4
Synthesis of l-(6-methylpyrimidin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000059_0001
Synthesis of tert-butyl (l-(6-methylpyrimidin-4-yl) piperidin-4-yl) carbamate [0178] To a stirred solution of 4-chloro-6-methylpyrimidine (1 g, 7.77 mmol) in 1, 4- dioxane (30 mL) was added diisopropylethylamine (1.35 g, 10.50 mmol) and tert-butyl piperidin-4-ylcarbamate (1.7 g, 8.55 mmol). The reaction mixture was heated to 150 °C and stirred for 3 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified through silica gel column chromatography using 5%
MeOH:DCM to afford tert-butyl (l-(6-methylpyrimidin-4-yl) piperidin-4-yl) carbamate (1.75 g, 77%) as an off-white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 8.34 (s, 1H), 6.69 (s, 1H), 4.28-4.24 (m, 2H), 3.54-3.52 (m, 1H), 3.00-2.93 (m, 2H), 2.23 (s, 3H), 1.77-1.75 (m, 2H), 1.38 (s, 9H), 1.30-1.22 (m, 3H); LC-MS: 293.3 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 3.32 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50%
EtOAc:hexanes (Rf. 0.2).
Synthesis of l-(6-methylpyrimidin-4-yl) piperidin-4-amine hydrochloride
[0179] To a stirred solution of tert-butyl (l-(6-methylpyrimidin-4-yl) piperidin-4-yl) carbamate (1.75 g, 5.13 mmol) in EtOAc (5 mL) was added 4M HC1 in EtOAc (4.5 mL) at 0 °C. The reaction mixture was warmed to RT and stirred for 6 h. After consumption of the starting materials (monitored by TLC), the mixture was filtered through celite and concentrated in vacuo to afford l-(6-methylpyrimidin-4-yl) piperidin-4-amine hydrochloride (1.1 g, 84%) as a brown solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 8.76 (s, 1H), 8.42 (bs, 2H), 7.19 (br s, 1H), 4.86-4.36 (m, 2H), 3.4-3.25 (m, 3H), 2.40 (s, 3H), 2.11-2.09 (m, 2H), 1.59- 1.56 (m, 2H); LC-MS: 193.2 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 2.13 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.1).
Example 5
Synthesis of l-(2-methylpyrimidin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000061_0001
DIEA, 1 , 4-dioxane
150 °C, 3 h
Synthesis of tert-butyl (l-(2-methylpyrimidin-4-yl) piperidin-4-yl) carbamate
[0180] To a stirred solution of 4-chloro-2-methylpyrimidine (1 g, 7.77 mmol) in 1, 4- dioxane (30 mL) was added diisopropylethylamine (1.35 g, 10.50 mmol) and tert-butyl piperidin-4-ylcarbamate (1.7 g, 8.55 mmol). The reaction mixture was heated to 150 °C and stirred for 3 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified through silica gel column chromatography using 5%
MeOH:DCM to afford tert-butyl (l-(2-methylpyrimidin-4-yl) piperidin-4-yl) carbamate (1.5 g, 66%) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.04-8.02 (m, 1H), 6.83- 6.82 (m, 1H), 6.61-6.60 (m, 1H), 4.27-4.25 (m, 2H), 3.51 (s, 1H), 2.96-2.91 (m, 2H), 2.33 (s, 3H), 1.77-1.74 (m, 2H), 1.37 (s, 9H), 1.28-1.22 (m, 2H); LC-MS: 293.3 (M+l); (column; X- Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 2.90 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.2).
Synthesis of l-(2-methylpyrimidin-4-yl) piperidin-4-amine hydrochloride
[0181] To a stirred solution of tert-butyl (l-(2-methylpyrimidin-4-yl) piperidin-4-yl) carbamate (2.1 g, 7.18 mmol) in EtOAc (5 mL) was added 4M HCI in EtOAc (5 mL) at 0 °C. The reaction mixture was warmed to RT and stirred for 6 h. After consumption of the starting materials (monitored by TLC), the reaction was filtered through celite and concentrated in vacuo to afford l-(2-methylpyrimidin-4-yl) piperidin-4-amine hydrochloride (1 g, 61%) as a brown solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.37 (bs, 2H), 8.29 -7.96 (d, 1H), 7.13-7.12 (d, 1H), 4.91-4.89 (m, 1H), 4.30-4.28m (m, 1H), 3.41-3.34 (m, 2H), 3.16-3.14 (m, 1H), 2.52 (s, 3H), 2.10-2.07 (m, 2H), 1.58-1.56 (m, 2H); LC-MS: 193.2 (M+l); (column; X-Bridge C- 18 (50 3.0 mm, 3.5 μπι); RT 2.06 min. 5 mM NH4OAC: ACN; 0.80 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.1). Example 6
Synthesis of l-(2-methylpyridin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000062_0001
Pd2(dba)3, Toluene
50 °C, 5 h
Synthesis of tert-butyl (l-(2-methylpyridin-4-yl) piperidin-4-yl) carbamate
[0182] To a stirred solution of 4-bromo-2-methylpyridine (2 g, 11.62 mmol) in toluene (200 mL) was added tert-butyl piperidin-4-ylcarbamate (2.56 g, 12.79 mmol), sodium tertiary butoxide (3.35 mg, 34.86 mmol) and Xantphos (403 mg, 0.69 mmol) at 0 °C. The mixture was stirred while purging under an argon atmosphere for 20 min. To the mixture was added Pd2(dba)3 (212 mg, 0.23 mmol) and the mixture was purged under an argon atmosphere for 20 min. The reaction mixture was heated to 50 °C and stirred for 5 h. After consumption of the starting materials (monitored by TLC), the reaction was filtered through celite and concentrated in vacuo. The crude material was washed with hexane:ether: EtOAc (1 : 1 : 1, 2 x 30 mL) to afford tert-butyl (l-(2-methylpyridin-4-yl) piperidin-4-yl) carbamate (1.4 g, 41%) as a brown solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.01-7.99 (m, 1H), 6.83-6.80 (m, 1H), 6.67 (s, lH),6.64-6.60 (m, 1H), 3.87-3.79 (m, 2H), 3.53-3.48 (m, 1H), 2.88-2.83 (m, 2H), 2.30 (s, 3H), 1.79-1.73 (m, 2H), 1.40 (s, 9H), 1.29-1.26 (m, 2H); LC-MS: 292.1 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 2.49 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 40% EtOAc:hexanes (Rf. 0.1).
Synthesis of l-(2-methylpyridin-4-yl) piperidin-4-amine hydrochloride
[0183] To a stirred solution of tert-butyl (l-(2-methylpyridin-4-yl) piperidin-4-yl) carbamate (800 mg, 2.74 mmol) in EtOAc (8 mL) was added 4M HCI in EtOAc (2 mL) at 0 °C. The reaction mixture was stirred for 1 h, then warmed to RT and continued to stir for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was
concentrated in vacuo. The crude material was washed with ether (2 x 20 mL) to afford l-(2- methylpyridin-4-yl) piperidin-4-amine hydrochloride (600 mg, 96%) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.22-8.16 (m, 3H), 7.10-7.07 (m, 2H), 4.21-4.20 (m, 2H), 3.23-2.19 (m, 3H), 2.43 (s, 3H), 2.07-2.02 (m, 2H), 1.57-1.50 (m, 2H); LC-MS: 192.3 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 0.38 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 10% MeOH:CH2Cl2 (R/. 0.1).
Example 7
Synthesis of l-(2-methoxypyridin-4-yl)azetidin-3-amine hydrochloride
NHBoc HCI
Xantphos, Pd(OAc)2
Figure imgf000063_0001
Cs2C03, 1 , 4 dioxane
90 °C, 12 h
Synthesis of tert-butyl (l-(2-methoxypyridin-4-yl) azetidin-3-yl) carbamate
[0184] To a stirred solution of tert-butyl azetidin-3-ylcarbamate (50 mg, 0.29 mmol) in 1, 4-dioxane (5 mL) was added 4-bromo-2-methoxypyridine (50 mg, 0.29 mmol), Xantphos (11.7 mg, 0.02 mmol), and cesium carbonate (236 mg, 0.72 mmol). The mixture was purged under an argon atmosphere for 15 min. To the mixture was added Pd(OAc)2 (1.95 mg, 0.08 mmol) and the reaction mixture was heated to 90 °C and stirred for 12 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo. The crude material was purified by silica gel column chromatography using 40% EtOAc:hexanes to afford tert-butyl (l-(2-methoxypyridin-4-yl) azetidin-3-yl) carbamate (40 mg, 50%) as an off-white solid. 1H-NMR (CD3OD, 400 MHz): δ 7.69-7.67 (m, 1H), 6.09-6.05 (m, 1H), 5.70 (s, 1H), 4.50 (br s, 1H), 4.21-4.19 (m, 2H), 3.80 (s, 3H), 3.75-3.69 (m, 2H), 1.43 (s, 9H); LC- MS: 280.3 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 3.88 min. 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.4).
Synthesis of l-(2-methoxypyridin-4-yl)azetidin-3-amine hydrochloride [0185] To a stirred solution of tert-butyl (l-(2-methoxypyridin-4-yl) azetidin-3-yl) carbamate 210 mg, 0.75 mmol) in EtOAc (3 mL) was added 4M HC1 in EtOAc (0.2 mL) at 0 °C. The reaction mixture was warmed to RT and stirred for 3 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo to afford l-(2- methoxypyridin-4-yl)azetidin-3-amine hydrochloride (200 mg) as an off-white solid. 1H- NMR (DMSO-< 5, 500 MHz): δ 8.99-8.97 (m, 2H), 7.84 (d, 1H), 6.42-6.40 (d, 1H), 6.18 (s, 1H), 4.46-4.40 (m, 2H), 4.29-4.24 (m, 2H), 4.20 (br s, 1H), 4.03 (s, 3H); LC-MS: 180 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 0.49 min. 0.05% TFA in water: ACN; 0.80 mL/min); TLC: 5% MeOH:CH2Cl2 (R/. 0.1).
Example 8
Synthesis of l-(2-methylpyridin-4-yl) azetidin-3-amine
Figure imgf000064_0001
Synthesis of benzyl (l-(2-methylpyridin-4-yl) azetidin-3-yl) carbamate
[0186] To a solution of 4-bromo-2-methylpyridine (598 mg, 2.90 mmol) in 1, 4-dioxane (15 mL) was added benzyl azetidin-3-ylcarbamate (500 mg, 2.90 mmol), Xantphos (132 mg, 0.22 mmol), Pd(OAc)2 (25.3 mg, 0.11 mmol) and Cs2C03 (2.3 g, 7.26 mmol). The reaction mixture was heated to 100 °C and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the mixture was cooled to RT and concentrated in vacuo. The crude material was diluted with water and extracted with EtOAc (2 x 25 mL). The combined organic extracts were washed successively with water (10 mL), brine (10 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography to afford benzyl (l-(2-methylpyridin-4-yl) azetidin-3-yl) carbamate (0.35 g, 40%) as a white solid. 1H-NMR (DMSO d6, 400 MHz): δ 7.96 (d, 2H), 7.38-7.31 (m, 5H), 6.21 (s, 1H), 6.14 (d, 1H), 5.04 (s, 2H), 4.44-4.41 (m, 1H), 4.14 (t, 2H), 3.64 (m, 2H), 2.26 (s, 3H); Mass (ESI): 298 [M+l]; TLC: 30% MeOH:DCM (Rf. 0.3).
Synthesis of l-(2-methylpyridin-4-yl) azetidin-3 -amine
[0187] To a stirred solution of benzyl (l-(2-methylpyridin-4-yl) azetidin-3 -yl) carbamate (0.35 g, 1.17 mmol) in MeOH (20 mL) was added 10%> Pd-C (75 mg). The suspension was stirred for 3 h under ¾ atmosphere (balloon pressure). After consumption of the starting materials (monitored by TLC), the mixture was filtered through celite and washed with MeOH (10 mL). The filtrate was concentrated in vacuo to afford l-(2-methylpyridin-4-yl) azetidin-3 -amine (200 mg) as a white solid which was used for the next step without further purification. 1H-NMR (DMSO d6, 400 MHz): δ 7.96 (d, 1H), 6.19 (s, 1H), 6.12 (d, 1H), 4.12- 4.04 (m, 2H), 3.84-3.81 (m, 1H), 3.48-3.42 (m, 2H), 2.31 (s, 3H); Mass (ESI): 164 [M+l]; TLC: 30% MeOH/DCM (Rf. 0.2).
Example 9
Synthesis of l-(6-methoxypyrimidin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000065_0001
Synthesis of tert-butyl (l-(6-methoxypyrimidin-4-yl) piperidin-4-yl) carbamate
[0188] To a stirred solution of 4-iodo-6-methoxypyrimidine (500 mg, 2.12 mmol) in
NMP (5 mL) was added diisopropylethylamine (383 mg, 2.96 mmol) and tert-butyl piperidin-
4-ylcarbamate (550 mg, 2.75 mmol) and purged with argon for 15 min. The reaction mixture was heated to 130 °C and stirred for 16 h. After consumption of the starting materials
(monitored by TLC), the mixture was diluted with ice cold water (100 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 10-30% EtOAc:hexanes to afford tert-butyl (l-(6-methoxypyrimidin-
4-yl) piperidin-4-yl) carbamate (400 mg, 61%) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.22 (s, 1H), 6.84-6.82 (m, 1H), 6.06 (s, 1H), 4.24-4.21 (m, 2H), 3.81 (s, 3H), 3.51 (br s, 1H), 2.94 (t, 2H), 1.75-1.73 (m, 2H), 1.38 (s, 9H), 1.30-1.22 (m, 2H); LC-MS: 308.9 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.45 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.67 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 30% EtOAc:hexanes (Rf. 0.2).
Synthesis of l-(6-methoxypyrimidin-4-yl) piperidin-4-amine hydrochloride
[0189] To a stirred solution of tert-butyl (l-(6-methoxypyrimidin-4-yl) piperidin-4-yl) carbamate (300 mg, 0.97 mmol) in CH2C12 (6 mL) was added 4N HCI in 1, 4-dioxane (10 mL) and the reaction mixture was stirred for 6 h. After consumption of the starting materials (monitored by TLC), the mixture was filtered and the solid was washed successively with CH2C12 (50 mL) and ether (50 mL) to afford l-(6-methoxypyrimidin-4-yl) piperidin-4-amine hydrochloride (290 mg, 92%) as a yellow solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 8.35 (s, 1H), 8.30-8.28 (m, 3H), 6.29 (s, 1H), 4.45-4.42 (m, 2H), 3.90 (s, 3H), 3.35-3.31 (m, 1H), 3.04 (t, 2H), 2.01-1.98 (m, 2H), 1.53-1.43 (m, 2H); LC-MS: 209.2 (M+1); (column; Eclipse XDB C-18 (150 x 4.6 mm, 5 μπι); RT 4.03 min 0.05% Aq TFA: ACN; 1.0 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.1).
Example 10
Synthesis of l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride
Figure imgf000066_0001
Synthesis of tert-butyl (l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl) carbamate
[0190] To a stirred solution of 4-iodo-2-(trifluoromethyl) pyridine (2 g, 7.32 mmol) in
NMP (20 mL) was added diisopropylethylamine (1.32 g, 10.23 mmol) and tert-butyl piperidin-4-ylcarbamate (1.90 g, 9.50 mmol) and purged with argon for 15 min. The reaction mixture was heated to 130 °C and stirred for 24 h. After consumption of the starting materials
(monitored by TLC), the mixture was diluted with ice cold water (100 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 25% EtOAc:hexanes to afford tert-butyl (l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl) carbamate (1.7 g, 67%) as an off-white solid. 1H-NMR (DMSO- d6, 400 MHz): δ 8.23 (d, 1H), 7.18 (s, 1H), 7.02-7.00 (m, 1H), 6.85 (d, 1H), 3.97-3.93 (m, 2H), 3.57-3.50 (m, 1H), 3.00 (t, 2H), 1.80-1.77 (m, 2H), 1.34 (s, 9H), 1.31-1.30 (m, 2H); LC- MS: 346.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.25 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.2).
Synthesis of l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride
[0191] To a stirred solution of tert-butyl (l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl) carbamate (1.7 g, 4.92 mmol) in CH2C12 (20 mL) was added 2M HC1 in 1 , 4-dioxane (10 mL) and the reaction mixture was stirred for 16 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo. The crude material was washed successively with n-pentane (2 x 10 mL) and ether (2 x 10 mL) to afford l-(2- (trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (1 g, 90%) as a yellow solid. 1H-NMR (DMSO-de, 500 MHz): δ 8.32-8.30 (m, 3H), 7.35 (s, 1H), 7.16-7.14 (m, 1H), 4.18- 4.15 (m, 2H), 3.36-3.32 (m, 1H), 3.10 (t, 2H), 2.02-2.00 (m, 2H), 1.59-1.51 (m, 2H); LC-MS: 246.1 (M+1); (column; Eclipse XDB C-18 (150 4.6 mm, 5 μπι); RT 5.02, 43.20 min 0.05% Aq TFA: ACN; 1.0 mL/min); TLC: 5% MeOH:CH2Cl2 (Rf. 0.1).
Example 11
Synthesis of 2-chloro-8-methyl-7-phenyl-7,8-dihydro-6H-pyrimido[5,4-b] [l,4]oxazine
Figure imgf000067_0001
Synthesis of 2-chloro-5-methoxy-N-methylpyrimidin-4-amine [0192] To a stirred solution of 2,4-dichloro-5-methoxypyrimidine (5 g, 28.0 mmol) in triethylamine (7.8 mL) under an argon atmosphere was added methyl amine (33% in ethanol, 3.9 mL, 42.1 mmol) in a sealed tube at RT and heated to 110 °C for 6 h. After completion of the reaction (monitored by TLC), the reaction was diluted with water. The resultant solid was filtered, washed with cold water and dried to afford 2-chloro-5-methoxy-N-methylpyrimidin- 4-amine (3.85 g, 79%) as a white crystalline solid. LC-MS: 173.8 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 1.51 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 80% EtOAc:hexanes (Rf. 0.3).
Synthesis of 2-chloro-4-(methylamino) pyrimidin-5-ol
[0193] To a stirred solution of 2-chloro-5-methoxy-N-methylpyrimidin-4-amine (6 g, 34.68 mmol) in CH2CI2 (600 mL) under an argon atmosphere at 0 °C was added 1M BBr3 in CH2CI2 (49.2 mL, 520 mmol). The reaction mixture was warmed to RT and stirred for 48 h. After completion of the reaction (monitored by TLC), the reaction was quenched with methanol (100 mL) and volatile components were concentrated in vacuo. To the resultant residue a saturated sodium bicarbonate solution (100 mL) was added and extracted with EtOAc (3 x 200 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-chloro-4-(methylamino) pyrimidin-5-ol (5 g, 92%) as an off-white solid which was used for the next step without further purification. 1H-NMR (DMSO-< 5, 400 MHz): δ 10.2 (brs, 1H), 7.39 (s, 1H), 7.21 (br s, 1H), 2.80 (s, 3H); LC-MS: 159.8 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 0.93 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 90% EtOAc:hexanes (Rf. 0.3).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l -phenylethan-1 -one
[0194] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (2.0 g, 12.57 mmol) in acetonitrile (35 mL) under an argon atmosphere at 0 °C were added cesium carbonate (8.2 g, 25.14 mmol) and 2-bromo-l -phenylethan-1 -one (2.75 g, 13.8 mmol) and stirred for 2 h. After completion of the reaction (monitored by TLC), the reaction was diluted with water (100 mL) and extracted with EtOAc (2 x 150 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20-30% EtOAc:hexanes to afford 2-((2- chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l -phenylethan-1 -one (2.3 g, 66%) as an off- white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 7.81 (s, 1H), 7.41-7.36 (m, 5H), 7.09 (s, 1H), 4.13 (d, IH), 4.00 (d, IH), 2.78 (s, 3H); LC-MS: 277.9 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπτι); RT 3.25 min. 0.05% TFA in water: ACN; 0.80 ml/min); TLC: 80% EtOAc:hexanes (Rf. 0.5).
Synthesis of 2-chloro-8-methyl- 7 -phenyl- 7, 8-dihydro-6H-pyrimido[ 5, 4-b ] [ 1, 4]oxazine
[0195] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l- phenylethan-l-one (2.0 g, 7.22 mmol) in 1, 2-dichloroethane (40 mL) under an argon atmosphere were added NaBH(OAc)3 (3.06 g, 14.44 mmol) and trifluoroacetic acid (0.55 mL, 7.22 mmol) at RT and stirred for 16 h. After completion of the reaction (monitored by TLC), the reaction was diluted with a IN sodium hydroxide solution (50 mL) and extracted with CH2CI2 (2 x 75 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20-30% EtOAc:hexanes to afford 2-chloro-7-(phenyl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine. 1H-NMR (CDC13, 500 MHz): δ 7.70 (s, IH), 7.40-7.36 (m, 3H), 7.19 (d, 2H), 4.60-4.58 (m, IH), 4.23 (d, IH), 4.19 (d, IH), 3.08 (s, 3H); LC-MS: 262.3 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπ ; RT 4.02 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 60% EtOAc:hexanes (Rf. 0.4).
Example 12
Synthesis of 2-chloro-7-(3, 5-difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4- b] [1, 4] oxazine
Figure imgf000069_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(3, 5-difluorophenyl) ethan-l-one
[0196] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (400 mg, 2.51 mmol) in acetonitrile (8 mL) under an argon atmosphere at 0 °C was added cesium carbonate (1.64 g, 5.02 mmol) and 2-bromo-l-(3, 5-difluorophenyl) ethan-l-one (650 mg, 2.76 mmol) and stirred for 2 h. After completion of the reaction (monitored by TLC), the reaction was diluted with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20-30% EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(3, 5-difluorophenyl) ethan-l- one (420 mg, 54%) as an off-white solid. 1H-NMR (DMSO-<¾, 500 MHz): δ 7.83 (s, IH), 7.30-7.27 (m, 2H), 7.19-7.17 (m, 2H), 4.20 (d, IH), 4.00 (d, IH), 2.78 (s, 3H). LC-MS: 314.3 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 4.17 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 60% EtOAc:hexanes (Rf. 0.5).
Synthesis of 2-chloro-7-(3, 5-difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
[0197] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(3, 5-difluorophenyl) ethan-l-one (500 mg, 1.59 mmol) in 1, 2-dichloroethane (10 mL) under an argon atmosphere was added NaBH(OAc)3 (684 mg, 3.18 mmol) and trif uoroacetic acid (0.122 mL, 1.59 mmol) at RT and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with a IN sodium hydroxide solution (50 mL) and extracted with CH2CI2 (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-chloro-7-(3, 5-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (370 mg, 78%>) as a pale yellow solid. 1H-NMR (CDC13, 400 MHz): δ 7.71 (s, IH), 6.81 (t, IH), 6.70 (d, 2H), 4.55-4.54 (m, IH), 4.25-4.19 (m, 2H), 3.10 (s, 3H); LC-MS: 298 (M+l); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μιη); RT 4.13 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 60%
EtOAc:hexanes (Rf. 0.4).
Example 13
Synthesis of 2-chloro-8-methyl-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5,
4-b] [1, 4] oxazine
Figure imgf000071_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(3, 4, 5-trifluorophenyl) ethan-l-one
[0198] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (630 mg, 3.95 mmol) in acetonitrile (5 mL) under an argon atmosphere at 0 °C were added cesium carbonate (1.4 g, 4.13 mmol) and 2-bromo-l-(3, 4, 5-trifluorophenyl) ethan-l-one (1.1 g, 4.3 mmol) and stirred for 2 h. After completion of the reaction (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed with water (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 15% EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(3, 4, 5-trifluorophenyl) ethan-l-one (430 mg, 66%) as an off-white solid. 1H-NMR (DMSO-d6, 400 MHz): δ 7.82 (s, 1H), 7.42-7.35 (m, 2H), 7.30 (s, 1H), 4.20 (d, 1H), 4.01 (d, 1H), 2.70 (s, 3H); TLC: 30% EtOAc:hexanes (Rf. 0.5).
Synthesis of 2-chloro-8-methyl-7-(3, 4, 5-trifluorophenyl) -7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine
[0199] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(3, 4, 5-trifluorophenyl) ethan-l-one (480 mg, 1.45 mmol) in 1 ,2-dichloroethane (8 mL) under an argon atmosphere was added NaBH(OAc)3 (614 mg, 2.90 mmol) and trifluoroacetic acid (0.11 mL, 1.45 mol) at 0 °C. The reaction mixture was warmed to RT and stirred for 32 h. After completion of the reaction (monitored by TLC), the reaction was quenched with a IN sodium hydroxide solution (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was washed with ether (2 x 10 mL) to afford 2-chloro-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (300 mg, 66%>) as an off- white solid. 1H-NMR (CDCI3, 400 MHz): δ 7.74 (s, 1H), 6.84-6.80 (m, 2H), 4.52-4.51 (m, 1H), 4.25-4.16 (m, 2H), 3.12 (s, 3H); LC-MS: 316.2 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπ ; RT 4.31 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 30% EtOAc:hexanes (Rf. 0.3).
Example 14
Synthesis of 2-chloro-8-methyl-7-(o-tolyl)-7,8-dihydro-6H-pyrimido[5,4-b] [l,4]oxazine
Figure imgf000072_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(o-tolyl) ethan-l-one
[0200] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (100 mg, 0.62 mmol) in CH3CN (2 mL) under an argon atmosphere at 0 °C were added cesium carbonate (409 mg, 1.25 mmol) and 2-bromo-l-(o-tolyl) ethan-l-one (120 mg, 0.69 mmol) and stirred for 2 h. After completion of reaction (monitored by TLC), the reaction was diluted with a saturated sodium bicarbonate solution (20 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20-50%
EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(o-tolyl) ethan-l-one (120 mg, 66%) as an off-white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 7.87 (s, 1H), 7.52 (d, 1H), 7.30-7.19 (m, 3H), 7.08 (s, 1H), 4.25 (d, 1H), 3.95 (d, 1H), 2.79 (s, 3H), 2.30 (s, 3H); TLC: 60% EtOAc:hexanes (Rf. 0.4).
Synthesis of 2-chloro-8-methyl-7-(o-tolyl)-7 , 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine [0201] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(o- tolyl) ethan-l-one (1 g, 3.43 mmol) in 1, 2-dichloroethane (20 mL) under an argon atmosphere was added NaBH(OAc)3 (1.45 g, 6.87 mmol) and trifluoroacetic acid (0.26 mL, 3.43 mmol) at RT and stirred for 16 h. After completion of reaction (monitored by TLC), the mixture was diluted with a IN sodium hydroxide solution (100 mL) and extracted with CH2CI2 (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column
chromatography using 10-20% EtOAc:hexanes to afford 2-chloro-8-methyl-7-(o-tolyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (700 mg, 74%) as a pale yellow solid. 1H-NMR (CDC13, 400 MHz): δ 7.80 (s, 1H), 7.25-7.19 (m, 3H), 6.97 (d, 1H), 4.88-4.86 (m, 1H), 4.27- 4.23 (m, 1H), 4.12-4.09 (m, 1H), 3.03 (s, 3H), 2.40 (s, 3H); LC-MS: 276.3 (M+l); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 4.28 min. 0.05% Aq TFA:ACN; 0.80 ml/min); TLC: 60% EtOAc:hexanes (Rf. 0.5).
Example 15
Synthesis of 2-chloro-7-(4-fluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1,
4] oxazine
Figure imgf000073_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluorophenyl) ethan-l- one
[0202] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (1.5 g, 9.43 mmol) in CH3CN (20 mL) was added cesium carbonate (6.1 g, 18.86 mmol) and stirred for 10 min at 0 °C. 2-Bromo-l-(4-fluorophenyl) ethan-l-one (2.25 g, 10.37 mmol) was added and the reaction mixture was stirred for 3 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic layers were washed with water (30 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was washed with ether: n- hexane (1 : 1, 2 x 5 mL) to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4- fluorophenyl) ethan-l-one (2 g, 71%) as an off-white solid. TLC: 40% EtOAc:hexanes (R/. 0.6).
Synthesis of 2-chloro-7-(4-fluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
[0203] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4- fluorophenyl) ethan-l-one (2 g, 6.75 mmol) in 1, 2-dichloroethane (20 mL) was added trifluoroacetic acid (770 mg, 6.75 mmol) and stirred for 10 min. Sodium
triacetoxyborohydride (2.86 g, 13.51 mmol) was added and the reaction mixture was stirred for 72 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a IN sodium hydroxide solution (30 mL) and extracted with CH2CI2 (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and
concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20% EtOAc:hexanes to afford 2-chloro-7-(4-fluorophenyl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (1.8 g, 96%) as a pale yellow oil. 1H-NMR (CDC13, 400 MHz): δ 7.70 (s, 1H), 7.18-7.02 (m, 4H), 4.53-4.51 (m, 1H), 4.22-4.20 (m, 1H), 4.12-4.08 (m, 1H), 3.08 (s, 3H); LC-MS: 279.8 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπτι); RT 3.53 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 40% EtOAc:hexanes (Rf. 0.6).
Example 16
Synthesis of 7-benzyl-2-chloro-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000074_0001
RT, 20 h
Figure imgf000074_0002
Synthesis of l-bromo-3-phenylpropan-2-one [0204] To a stirred solution of 2-phenylacetic acid (5 g, 36.76 mmol) in CH2CI2 (50 mL) was added oxalyl chloride (4.6 g, 36.76 mmol) and DMF (2 drops) at 0 °C. The reaction mixture was warmed to RT and continued to stir for 2 h. After consumption of acid
(monitored by TLC), the mixture was concentrated in vacuo. The crude material was dissolved in ether and cooled to 0 °C. A solution of CH2N2 in ether was added and the reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo. To a stirred solution of the crude material in THF (20 mL) was added a solution of 48% aq HBr (5 mL) at 0 °C. The reaction mixture was warmed to RT and stirred for 20 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a sodium bicarbonate solution (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were washed successively with a sodium bicarbonate solution (50 mL) and water (50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to afford l-bromo-3-phenylpropan-2-one (6 g, 76%) as a viscous oil. 1H-NMR (CDC13, 400 MHz): δ 7.38-7.30 (m, 3H),. 7.27-7.20 (m, 2H), 3.92 (s, 2H), 3.89 (s, 2H); TLC: 10% EtOAc:hexanes (Rf. 0.6).
Synthesis of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-3-phenylpropan-2-one
[0205] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (3 g, 18.8 mmol) in acetonitrile (30 mL) was added potassium carbonate (12.2 g, 37.60 mmol) followed by 1- bromo-3-phenylpropan-2-one (4.4 g 20.70 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with ice cold water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo . The crude material was triturated with n-hexane : CH2C12 (1 : 1, 2 x 50 mL) to afford 1 - ((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-3-phenylpropan-2-one (3 g, 55%) as a pale brown solid. 1H-NMR (CDC13, 500 MHz): δ 7.70 (s, IH), 7.39-7.30 (m, 3H), 7.29-7.27 (m, 2H), 4.02 (d, IH), 2.62 (d, IH), 3.21 (d, IH), 3.19 (s, 3H), 2.96 (d, IH); LC-MS: 291.9 (M+); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.37 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.8).
Synthesis of 7-benzyl-2-chloro-8-methyl-7 , 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine [0206] To a stirred solution of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-3- phenylpropan-2-one (3 g, 10.3 mmol) in 1, 2-dichloroethane (30 mL) under argon was added trifluoroacetic acid (1.1 g, 10.31 mmol) at 0 °C and stirred for 10 min. Sodium
triacetoxyborohydride (4.3 g, 20.62 mmol) was added and the reaction mixture was stirred for 48 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with 1 N sodium hydroxide (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20%
EtOAc:hexanes to afford 7-benzyl-2-chloro-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (1.6 g, 57%) as a white solid. 1H-NMR (CDC13, 500 MHz): δ 7.70 (s, 1H), 7.38- 7.34 (m, 2H), 7.32-7.28 (m, 1H), 7.28 (d, 2H), 4.10 (d, 1H), 3.81 (d, 1H), 3.60-3.58 (m, 1H), 3.14 (s, 3H), 3.10-3.07 (m, 1H), 2.81-2.78 (m, 1H); LC-MS: 275.8 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.61 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.5).
Example 17
Synthesis of 2-chloro-8-methyl-7-(2-(trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido
[5, 4-b] [1, 4] oxazine
Figure imgf000076_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethyl) phenyl) ethan-l-one [0207] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (100 mg, 0.62 mmol) in CH3CN (2 mL) was added cesium carbonate (409 mg, 1.25 mmol) and 2-bromo-l- (2-(trifluoromethyl) phenyl) ethan-l-one (184 mg, 0.69 mmol) and the reaction mixture was stirred for 3 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 10% EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethyl) phenyl) ethan-l-one (100 mg, 46%) as an off-white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 8.03 (d, 1H), 7.90 (d, 1H), 7.89-7.79 (m, 2H), 7.69 (s, 1H), 7.48-7.40 (m, 1H), 5.55 (s, 2H), 2.83 (d, 3H); LC-MS: 345.8 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.41 min. 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 50% EtOAc: hexanes (Rf. 0.7).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethyl) phenyl) ethan-l-ol
[0208] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- (trifluoromethyl) phenyl) ethan-l-one (400 mg, 1.15 mmol) in MeOH (4 mL) was added sodium borohydride (64 mg, 1.73 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 10%
EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- (trifluoromethyl) phenyl) ethan-l-ol (250 mg, 62%) as an off-white solid. LC-MS: 347.8 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.30 min. 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.6).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethyl) phenyl) ethyl methanesulfonate
[0209] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- (trifluoromethyl) phenyl) ethan-l-ol (550 mg, 1.58 mmol) in CH2CI2 (6 mL) was added successively triethyl amine (0.26 mL, 1.90 mmol) and methane sulfonyl chloride (180 mg, 1.58 mmol) at 0 °C. The reaction mixture was warmed to room temperate and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was filtered. The filtrate was diluted with water (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was recrystallized from diisopropylether (2 x 5 mL) to afford 2-((2- chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethyl) phenyl) ethyl methanesulfonate (500 mg, 74%) as an off-white solid. LC-MS: 425.7 (M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.63 min. 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 40% EtOAc:hexanes (Rf. 0.6).
Synthesis of 2-chloro-8-methyl-7 -(2-(trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazine
[0210] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- (trifluoromethyl) phenyl) ethyl methanesulfonate (650 mg, 1.52 mmol) in DMF (6 mL) was added potassium carbonate (316 mg, 2.29 mmol). The reaction mixture was heated to 80 °C and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo. The crude material was dissolved in water (50 mL) and extracted with ether (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 10% EtOAc:hexanes to afford 2-chloro-8-methyl-7-(2- (trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (100 mg, 19%>) as an off-white solid. 1H-NMR (CDC13, 400 MHz): δ 7.77-7.71 (m, 2H), 7.59-7.51 (m, 1H), 7.49-7.45 (m, 1H), 7.19 (d, 1H), 5.02-5.00 (m, 1H), 4.30-4.25 (m, 1H), 4.18-4.12 (m, 1H), 3.02 (s, 3H); LC-MS: 330.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.55 min. 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 40% EtOAc:hexanes (Rf. 0.7).
Example 18
Synthesis of 2-chloro-7, 8-dimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000079_0001
Synthesis of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) propan-2-one
[0211] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (200 mg, 1.25 mmol) in CH3CN (4 mL) was added successively l-bromopropan-2-one (243 mg, 1.38 mmol) and cesium carbonate (815 mg, 2.50 mmol) at 0 °C and stirred for 4 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with a saturated sodium carbonate solution (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 15-20%
EtOAc:hexanes to afford l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) propan-2-one (220 mg, 82%) as an off-white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 7.70 (s, 1H), 6.41 (s, 1H), 4.01 (d, 1H), 3.89 (d, 1H), 3.00 (s, 3H), 1.38 (s, 3H); LC-MS: 216(M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.18 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 60% EtOAc:hexanes (Rf. 0.2).
Synthesis of 2-chloro-7, 8-dimethyl-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine
[0212] To a stirred solution of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) propan-2-one (200 mg, 0.93 mmol) in 1, 2-dichloroethane (4 mL) was added sodium triacetoxyborohydride (395 mg, 1.86 mmol) followed by trifluoroacetic acid (0.07 mL, 0.93 mmol) and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with 1 N sodium hydroxide (20 mL) and extracted with CH2CI2 (2 x
20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 10-15%) EtOAc:hexanes to afford 2-chloro-7, 8-dimethyl-7, 8-dihydro-6H-pyrimido [5,
4-b] [1, 4] oxazine (110 mg, 59%) as an off-white solid. 1H-NMR (CDC13, 400 MHz): δ 7.63
(s, 1H), 4.01 (s, 2H), 3.61-3.58 (m, 1H), 3.14 (s, 3H), 1.31 (d, 3H); LC-MS: 200 (M+l);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.49 min. 0.05% Aq TFA: ACN;
0.8 mL/min); TLC: 60% EtOAc:hexanes (Rf. 0.4). Example 19
Synthesis of 2-chloro-7-(4-fluoro-2-methylphenyl)-8-methyl-7, 8-dihydro-6H-pyrimido
[5, 4-b] [1, 4] oxazine
Figure imgf000080_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2-methylphenyl) ethan-l-one
[0213] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (100 mg, 0.62 mmol) in CH3CN (2 mL) was added cesium carbonate (409 mg, 1.25 mmol) followed by 2- bromo-l-(4-fluoro-2-methylphenyl) ethan-l-one (159 mg, 0.69 mmol) and stirred for 3 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2-methylphenyl) ethan-l-one (120 mg, 62%) as an off-white solid. LC-MS: 309.8 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπτι); RT 3.35 min. 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.7).
Synthesis of 2-chloro-7-(4-fluoro-2-methylphenyl)-8-methyl-7 , 8-dihydro-6H-pyrimido [5, 4- b] [I, 4] oxazine
[0214] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4- fluoro-2-methylphenyl) ethan-l-one (150 mg, 0.48 mmol) in 1, 2-dichloroethane (2 mL) was added sodium triacetoxyborohydride (205 mg, 0.97 mmol) followed by trifluoroacetic acid
(0.1 mL, 0.48 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 16 h.
After consumption of the starting materials (monitored by TLC), the mixture was diluted with a IN sodium hydroxide (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford
2-chloro-7-(4-fluoro-2-methylphenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine and used without further purification. LC-MS: 294 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.31 min. 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.3).
Example 20
Synthesis of 7-chloro-l-methyl-2-(2, 2, 2-trifluoroethyl)-2, 3-dihydro-lH-pyrido [3, 4-b]
[1, 4] oxazine
Figure imgf000081_0001
Synthesis of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-4, 4, 4-trifluorobutan-2-ol
[0215] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (200 mg, 1.25 mmol) in DMF (5 mL) was added cesium carbonate (810 mg, 0.50 mmol) followed by 2-(2, 2, 2-trifluoroethyl) oxirane (160 mg, 1.25 mmol). The reaction mixture was heated to 50 °C and stirred for 6 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20-30%
EtOAc:hexanes to afford l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-4, 4, 4- trifluorobutan-2-ol (220 mg, 62%) as an off-white solid. 1H-NMR (CDC13, 500 MHz): δ 7.53 (s, 1H), 5.68 (br s, 1H), 4.43-4.39 (m, 1H), 4.02-3.99 (m, 2H), 3.04 (s, 3H), 2.90 (br s, 1H), 2.54-2.38 (m, 2H); LC-MS: 285.8 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.63 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 20% EtOAc:hexanes (Rf. 0.6).
Synthesis of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-4, 4, 4-trifluorobutan-2-one
[0216] To a stirred solution of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-4, 4, 4-trifluorobutan-2-ol (400 mg, 1.40 mmol) in CH2CI2 (5 mL) was added Dess-Martin periodinone (893 mg, 2.10 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 6 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with a saturated sodium bicarbonate solution (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were washed with a sodium thoisulfate solution (20 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 30% EtOAc:hexanes to afford l-((2- chloro-4-(methylamino) pyrimidin-5-yl) oxy)-4, 4, 4-trifluorobutan-2-one (220 mg, 55%) as a pale yellow solid. 1H-NMR (CDC13, 400 MHz): δ 7.70 (s, 1H), 4.31 (d, 1H), 4.00 (d, 1H), 3.18 (s, 3H), 2.78-2.63 (m, 2H); LC-MS: 284.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.60 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 40%
EtOAc:hexanes (Rf. 0.4).
Synthesis of 7 -chloro-l-methyl-2-(2 , 2, 2-trifluoroethyl)-2 , 3-dihydro- lH-pyrido [3, 4-b] [I, 4] oxazine
[0217] To a stirred solution of l-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-4, 4, 4-trifluorobutan-2-one (200 mg, 0.70 mmol) in 1, 2-dichloroethane (4 mL) was added sodium triacetoxyborohydride (300 mg, 1.41 mmol) followed by trifluoroacetic acid (80 mg, 0.70 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with a IN sodium hydroxide solution (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 15-20%
EtOAc:hexanes to afford 7-chloro-l-methyl-2-(2, 2, 2-trifluoroethyl)-2, 3-dihydro-lH-pyrido [3, 4-b] [1, 4] oxazine (90 mg, 48%) as an off-white solid. 1H-NMR (CDC13 400 MHz): δ 7.72 (s, 1H), 4.32 (d, 1H), 4.02 (d, 1H), 3.83-3.79 (m, 1H), 3.21 (s, 3H), 2.60-2.40 (m, 2H); LC-MS: 268 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.77 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 50% EtOAc:hexanes (Rf. 0.4).
Example 21
Synthesis of 2-chloro-8-methyl-7-(2-(trifluoromethoxy) phenyl)-7, 8-dihydi
pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000083_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethoxy) phenyl) ethan-l-one
[0218] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (300 mg, 1.88 mmol) in CH3CN (10 mL) was added cesium carbonate (1.22 g, 0.37 mmol) followed by 2- bromo-l-(2-(trifluoromethoxy) phenyl) ethan-l-one (580 mg, 0.20 mmol) at 0 °C and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 10% EtOAc :hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-(trifluoromethoxy) phenyl) ethan-l-one (350 mg, 51%) as an off white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.88 (s, 1H), 7.77 (d, 1H), 7.58 (d, 1H), 7.46 (t, 1H), 7.41 (d, 1H), 7.50 (s, 1H), 4.21 (d, 1H), 4.11 (d, 1H), 2.79 (s, 3H); LC-MS: 362.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπτι); RT 4.25 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 40% EtOAc:hexanes (Rf. 0.7).
Synthesis of 2-chloro-8-methyl-7 -(2-(trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine
[0219] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- (trifluoromethoxy) phenyl) ethan-l-one (350 mg, 0.96 mmol) in 1 ,2-dichloroethane (10 mL) under an argon atmosphere was added sodium triacetoxyborohydride (400 mg, 1.93 mmol) followed by trifluoroacetic acid (0.1 mL, 0.96 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 24 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with IN sodium hydroxide (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was triturated with EtOAc (2 x 10 mL) to afford 2-chloro-8-methyl-7-(2-(trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (220 mg, 66%) as an off white solid. 1H-NMR (CDC13, 400 MHz): δ 7.71 (s, 1H), 7.45-7.30 (m, 3H), 7.08-7.03 (m, 1H), 5.00 (s, 1H), 4.24 (s, 2H), 3.11 (s, 3H); LC-MS: 346.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπ ; RT 4.51 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 30% EtOAc:hexanes (Rf. 0.7).
Example 22
Synthesis of 2-chloro-8-methyl-7-(tetrahydro-2H-pyran-4-yl)-7, 8-dihydro-6H-pyrimido
[5, 4-b] [1, 4] oxazine
Figure imgf000084_0001
Synthesis of2-bromo-l-(tetrahydro-2H-pyran-4-yl) ethan-l-one
[0220] To a stirred solution of tetrahydro-2H-pyran-4-carboxylic acid (1.5 g, 0.01 mmol) in CH2C12 (10 mL) was added oxalyl chloride (1.6 g, 0.01 mmol) and DMF (1 drop) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of acid (monitored by TLC), the mixture was concentrated in vacuo. The crude mateiral was dissolved in ether and cooled to -10 °C. A solution of CH2N2 in ether (20 mL) was added and the reaction mixture was warmed to RT and stirred for 19 h. After consumption of the starting materials (monitored by TLC), the mixture was concentrated in vacuo. To a stirred solution of the crude material in CH2C12 (20 mL) was added a solution of 48% aq HBr (5 mL) at -10 °C. The reaction mixture was warmed to RT and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a sodium bicarbonate solution (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were washed successively with a sodium bicarbonate solution (20 mL) and water (20 mL). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-bromo-l-(tetrahydro-2H-pyran-4-yl) ethan-l-one (2 g, crude) as a yellow solid. 1H- NMR (DMSO- 500 MHz): δ 4.49 (s, 2H), 3.83 (d, 2H), 3.33 (t, 2H), 2.90-2.80 (m, 1H), 1.80-1.70 (m, 2H), 1.57-1.44 (m, 2H); TLC: 30% EtOAc:hexanes (Rf. 0.7).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl)oxy)-l-(tetrahydro-2H-pyran-4-yl) ethan-l-one
[0221] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (3 g, 18.8 mmol) in CH3CN (60 mL) was added cesium carbonate (12.2 g, 37.70 mmol) followed by 2-bromo-
1- (tetrahydro-2H-pyran-4-yl) ethan-l-one (3.9 g 18.8 mmol) at 0 °C and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a sodium carbonate solution (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford
2- ((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(tetrahydro-2H-pyran-4-yl) ethan-l- one (2.5 g) as a yellow solid and used without further purification. LC-MS: 286.1 (M+); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.86 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 30% EtOAc:hexanes (Rf. 0.5).
Synthesis of2-chloro-8-methyl-7-(tetrahydro-2H-pyran-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine
[0222] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l- (tetrahydro-2H-pyran-4-yl) ethan-l-one (2.5 g, 8.70 mmol) in 1 , 2-dichloroethane (50 mL) was added trifluoroacetic acid (1 mL) followed by sodium triacetoxyborohydride (3.7 g, 17.50 mmol) and stirred for 48 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with IN sodium hydroxide (50 mL) (pH>9) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 20-30% EtOAc:hexanes to afford 2-chloro-8-methyl-7-(tetrahydro- 2H-pyran-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1 , 4] oxazine (2 g, 86%>) as a pale yellow solid. LC-MS: 270.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.10 min. 0.05% Aq TFA: ACN; 0.8 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.63 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; TLC: 50%
EtOAc:hexanes (Rf. 0.3).
Example 23 Synthesis of 2-chloro-7-(2-chlorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1,
4] oxazine
Figure imgf000086_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-chlorophenyl) ethan-1- one
[0223] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (4 g, 25.15 mmol) in CH3CN (100 mL) was added cesium carbonate (16.25 g, 50.31 mmol) followed by 2-bromo-l-(2-chlorophenyl) ethan-l-one (12 g, 37.7 mmol) at 0 °C and stirred for 30 min. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 6% EtOAc:hexanes to afford 2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy)-l-(2-chlorophenyl) ethan-l-one (4.3 g, 55%) as a white solid. 1H-NMR (DMSO-<¾, 500 MHz): δ 7.95-7.88 (m, 2H), 7.59-7.41 (m, 3H), 7.38 (s, 1H), 4.37 (d, 1H), 4.05 (d, 1H), 2.78 (s, 3H); TLC: 20% EtOAc:hexanes (Rf. 0.7).
Synthesis of 2-chloro-7-(2-chlorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
[0224] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- chlorophenyl) ethan-l-one (4.3 g, 13.78 mmol) in 1, 2-dichloroethane (100 mL) was added sodium triacetoxyborohydride (6.1 g, 28.94 mmol) followed by trifluoroacetic acid (1.1 mL, 13.78 mmol) and stirred for 36 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a 5% sodium bicarbonate solution (100 mL) and extracted with CH2CI2 (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 12% EtOAc:hexanes to afford 2-chloro-7-(2-chlorophenyl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (1.8 g, 45%) as an off white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.77 (s, 1H), 7.55 (d, 1H), 7.40 (t, 1H), 7.36 (t, 1H), 6.98 (d, 1H), 5.23 (s, 1H), 4.34 (s, 2H), 3.01 (s, 3H); LC-MS: 296.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.30 min. 0.05% Aq TFA: ACN; 0.80 ml/min); TLC: 20% EtOAc:hexanes (Rf. 0.7).
Example 24
Synthesis of (S)-2-chloro-8-methyl-7-phenyl-7,8-dihydro-6H-pyrimido [5,4- b] [l,4]oxazine
Figure imgf000087_0001
Synthesis of 2,4-Dichloropyrimidin-5-ol
[0225] To a solution of 2,4-dichloro-5-methoxypyrimidine (5.30 g, 29.6 mmol) in 1,2- dichloroethane (75 mL) at 0°C was slowly added BBr3 (14 mL, 148 mmol). The reaction mixture was heated to 80 °C for 16 h and monitored by LC/MS until completion of the reaction. The reaction mixture was cooled to 0°C and basified with a IN NaOH solution. The mixture was stirred for 1 h at RT and then acidified with NH4C1. The pH was adjusted between pH 6-7 using AcOH. The layers were separated and the aqueous layer was extracted 3 times with ethyl acetate. The combined organic extracts were dried over Na2S04, filtered and concentrated in vacuo. The crude material was triturated with DCM:hexanes and filtered to afford pure 2,4-dichloropyrimidin-5-ol (2.03 g, 73 %) as a solid. 1H NMR (CDC13, 400 MHz): δ 8.35 (s, 1H), 5.73 (s, 1H); LRMS (ESpos) calcd for C4H2C12N20 [M+H]+: 164.95. found: 165.13.
Figure imgf000088_0001
Acetone, RT
Synthesis of 2,4-dichloro-5-(4-methoxybenzyloxy)pyrimidine
[0226] To a solution of 2,4-dichloropyrimidin-5-ol (3.58 g, 21.7 mmol), K2CO3 (7.5 g, 54.3 mmol) and Nal (325 mg, 2.2 mmol) in acetone (45 mL) at RT was added PMBCI (4.41 mL, 32.5 mmol). The reaction was stirred for 16 h. When the reaction was complete (by LC/MS), the mixture was partitioned between ethyl acetate and water. The layers were separated and the aqueous was extracted 3 times with ethyl acetate. The combined organic extracts were dried over Na2S04, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography (gradient 0-30% EtOAc:hexanes) to afford pure 2,4-dichloro-5-(4-methoxybenzyloxy)pyrimidine (4.48 g, 72 %) as a white solid. 1H NMR (CDC13, 400 MHz): ^ 8.18 (s, 1H), 7.34 (d, J = 8.8 Hz, 2H), 6.93 (d, J = 8.7 Hz, 2H), 5.17 (s, 2H), 3.82 (s, 3H).
Figure imgf000088_0002
Synthesis of (S)-2-(methylamino)-2-phenylethanol
[0227] To a dry flask under N2 atmosphere was added LAH (956 mg, 25.2 mmol), followed by THF (42 mL) and (S)-tert-butyl 2-hydroxy-l-phenylethylcarbamate (3.0 g, 12.6 mmol). The reaction mixture was heated to reflux for 16 h. When the reaction was complete (by LC/MS), a 10 % NaOH aqueous solution was added slowly at 0 °C until no gas evolution was observed. The reaction mixture was stirred at RT for 15 minutes and Na2S04 was added. The reaction mixture was stirred vigorously for 15 min, filtered and washed with THF. The filtrate was concentrated in vacuo to afford (S)-2-(methylamino)-2-phenylethanol (1.91 g) as a light yellow gum which solidified upon standing. 1H NMR (CDC13, 400 MHz): δ 7.38 - 7.33 (m, 2H), 7.31 - 7.26 (m, 3H), 3.72 (dd, J = 10.4, 4.4 Hz, 1H), 3.65 (dd, J = 8.3, 4.4 Hz, 1H), 3.55 (dd, J = 10.4, 8.3 Hz, 1H), 2.35 (s, 3H).
Figure imgf000089_0001
Synthesis of (S)-2-((2-chloro-5-(4-methoxybenzyloxy)pyrimidin-4-yl)(methyl)amino)-2- phenylethanol
[0228] To a mixture of 2,4-dichloro-5-(4-methoxybenzyloxy)pyrimidine (2.04 g, 7.15 mmol) and (S)-2-(methylamino)-2-phenylethanol (1.91 g, 7.49 mmol) in 2-methyl tetrahydrofuran (18 mL) was added triethylamine (2.5 mL, 17.9 mmol). The reaction mixture was stirred at reflux for 16 h. When the reaction was complete (by LC/MS), the mixture was diluted with ethyl acetate. The solution was washed twice with water, brine, dried over Na2S04, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography (gradient 10-80% EtOAc:hexanes) to afford of (S)-2-((2-chloro-5- (4-methoxybenzyloxy)pyrimidin-4-yl)(methyl)amino)-2-phenylethanol (2.14 g, 75 %) as a yellow gummy solid. 1H NMR (CDC13, 400 MHz): δ 7.80 (s, 1H), 7.35 - 7.27 (m, 3H), 7.22 (dd, J = 14.3, 7.9 Hz, 4H), 6.88 - 6.84 (m, 2H), 6.09 (dd, J = 8.9, 5.6 Hz, 1H), 4.96 (s, J = 11.1 Hz, 2H), 4.22 - 4.15 (m, 2H), 3.80 (s, 3H), 2.93 (s, 3H), 2.15 (s, 1H); LRMS (ESpos) calcd for C2iH22ClN303 [M+H]+: 400.13. found: 400.53.
Figure imgf000090_0001
Synthesis of (S)-2-chloro-4-((2-hydroxy-l-phenylethyl)(methyl)amino
[0229] To a mixture of (S)-2-((2-chloro-5-(4-methoxybenzyloxy)pyrimidin-4- yl)(methyl)amino)-2-phenylethanol (2.14 g, 5.35 mmol) at 0 °C in DCM (35 mL) was added trifluoro acetic acid (15mL). When the reaction was complete (by LC/MS), the reaction was concentrated to dryness and co-evaporated several times with toluene. The crude material was partitioned between ethyl acetate and a sodium bicarbonate solution. The layers were separated and the aqueous layer was extracted with ethyl acetate twice. The combined organic layers were dried over Na2S04, filtered and concentrated in vacuo. The residue was purified by normal phase chromatography on silica gel (gradient 0-10% MeOH:DCM) to afford (S)-2- chloro-4-((2-hydroxy-l-phenylethyl)(methyl)amino)pyrimidin-5-ol as a white foam (1.19 g, 80 %). 1H NMR (CDCls, 400 MHz): δ 7.83 (s, 1H), 7.33 (dq, J = 14.2, 7.0 Hz, 4H), 7.1 1 (d, J = 7.1 Hz, 2H), 5.71 (d, J = 10.9 Hz, 1H), 4.36 (t, J = 10.7 Hz, 1H), 4.27 (dd, J = 10.7, 3.8 Hz, 1H), 3.42 (s, 1H), 2.72 (s, 3H); LRMS (ESpos) calcd for Ci3Hi4ClN302 [M+H]+: 280.08. found: 280.46.
Figure imgf000090_0002
Synthesis of (S)-2-chloro-8-methyl- 7 -phenyl- 7, 8-dihydro-6H-pyrimido[ 5, 4-bJ [ 1, 4]oxazine [0230] To a mixture of (S)-2-chloro-4-((2 -hydroxy- 1- phenylethyl)(methyl)amino)pyrimidin-5-ol (600 mg, 2.14 mmol) and triphenylphosphine (616 mg, 2.35 mmol) at 0 °C in DCM (11 mL) was added DIAD (0.46 mL, 2.35 mmol) drop- wise. The reaction was shown to be complete after 30 min (by LC/MS). The mixture was concentrated to dryness and purified by silica gel column chromatography (gradient 0-70% methyl tert-butylether (MTBE):hexanes) to afford (S)-2-chloro-8-methyl-7-phenyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazine (0.2 g, 36 %) as a colourless oil. 1H NMR (CDC13, 400 MHz): δ 7.71 (s, 1H), 7.41 - 7.34 (m, 3H), 7.17 (dd, J = 7.7, 1.6 Hz, 2H), 4.57 (t, J = 3.6 Hz, 1H), 4.27 (dd, J = 11.2, 3.3 Hz, 1H), 4.18 (dd, J = 11.2, 4.0 Hz, 1H), 3.08 (s, 3H); LRMS (ESpos) calcd for Ci3Hi2ClN30 [M+H]+: 262.07. found: 262.42.
Example 25
Synthesis of (R)-2-chlor o-8-methyl-7-phenyl-7,8-dihydro-6H-pyrimido [5,4- b] [l,4]oxazine
Figure imgf000091_0001
Synthesis of (R)-2-(methylamino)-2-phenylethanol
[0231] To a dry flask under nitrogen was charged with LAH (943 mg, 24.9 mmol) then tetrahydrofuran (THF) (40 mL) and (R)-tert-butyl 2-hydroxy-l-phenylethylcarbamate (2.95 g, 12.4 mmol). The reaction mixture was heated to reflux for 16 h. When the reaction was shown to be complete by LC/MS, a 10 %> NaOH aqueous solution was added slowly at 0 °C until no more gas evolution was observed. The reaction mixture was stirred at RT for 15 minutes and Na2S04 was added. The reaction mixture was vigorously stirred for 15 min, filtered and washed with THF. The volatile components were evaporated to afford (R)-2- (methylamino)-2-phenylethanol as a light yellow gum which solidifies upon standing (1.85 g,
98 %). 1H NMR (CDC13, 400 MHz): δ 7.38 - 7.33 (m, 2H), 7.31 - 7.26 (m, 3H), 3.72 (dd, J = 10.4, 4.4 Hz, 1H), 3.65 (dd, J = 8.3, 4.4 Hz, 1H), 3.55 (dd, J = 10.4, 8.3 Hz, 1H), 2.35 (s, 3H).
Figure imgf000092_0001
Synthesis of (R)-2-((2-chloro-5-(4-methoxybenzyloxy)pyrimidin-4-yl)(methyl)amino phenylethanol
[0232] To a mixture of 2,4-dichloro-5-(4-methoxybenzyloxy)pyrimidine (1.2 g, 4.21 mmol) and (R)-2-(methylamino)-2 -phenylethanol (0.67 g, 4.42 mmol) in 2-methyl tetrahydrofuran (10 mL) was added triethylamine (1.5 mL, 10.5 mmol). The reaction mixture was stirred at reflux for 16 h. The reaction was diluted with ethyl acetate, washed twice with water, brine, dried over Na2S04, filtered and concentrated. The crude material was purified by silica gel column chromatography (gradient 10-80% EtOAc:hexanes) to afford (R)-2-((2- chloro-5-(4-methoxybenzyloxy)pyrimidin-4-yl)(methyl)amino)-2-phenylethanol as a yellow gummy solid (852 mg, 51 %). 1H NMR (CDC13, 400 MHz): δ 7.80 (s, 1H), 7.35 - 7.27 (m, 3H), 7.22 (dd, J = 14.3, 7.9 Hz, 4H), 6.88 - 6.84 (m, 2H), 6.09 (dd, J = 8.9, 5.6 Hz, 1H), 4.96 (s, J = 11.1 Hz, 2H), 4.22 - 4.15 (m, 2H), 3.80 (s, 3H), 2.93 (s, 3H), 2.15 (s, 1H); LRMS (ESpos) calcd for C2iH22ClN303 [M+H]+: 400.13. found: 400.53.
Figure imgf000092_0002
Synthesis of (R)-2-chloro-4-((2-hydroxy-l-phenylethyl) (methyl) amino)pyrimidin- 5 -ol [0233] To a mixture of (R)-2-((2-chloro-5-(4-methoxybenzyloxy)pyrimidin-4- yl)(methyl)amino)-2-phenylethanol (850 mg, 2.13 mmol) at 0 °C in dichloromethane (13.5 mL) was added trifluoroacetic acid (6.5 mL). When the reaction was shown to be complete by LC/MS, the mixture was concentrated to dryness and co-evaporated several times with toluene. The crude material was partitioned between ethyl acetate and a NaHC03 solution. The layers were separated and the aqueous layer was extracted with ethyl acetate twice. The combined organic layers were dried over Na2S04, filtered and concentrated in vacuo. The residue was triturated in DCM:hexanes mixture, filtered and dried in vacuo to afford (R)-2- chloro-4-((2-hydroxy-l-phenylethyl)(methyl)amino)pyrimidin-5-ol as a white solid (327 mg, 55 %). 1H NMR (CDC13, 400 MHz): δ 7.83 (s, 1H), 7.33 (dq, J = 14.2, 7.0 Hz, 4H), 7.11 (d, J = 7.1 Hz, 2H), 5.71 (d, J = 10.9 Hz, 1H), 4.36 (t, J = 10.7 Hz, 1H), 4.27 (dd, J = 10.7, 3.8 Hz, 1H), 3.42 (s, 1H), 2.72 (s, 3H); LRMS (ESpos) calcd for d3Hi4ClN302 [M+H]+: 280.08 found: 280.46.
Figure imgf000093_0001
Synthesis of (R)-2-chloro-8-methyl- 7 -phenyl- 7, 8-dihydro-6H-pyrimido[ 5, 4-bJ [ 1, 4]oxazine
[0234] To a mixture of (R)-2-chloro-4-((2-hydroxy-l- phenylethyl)(methyl)amino)pyrimidin-5-ol (325 mg, 1.16 mmol) and triphenylphosphine (335 mg, 1.28 mmol) at 0 °C in dichloromethane (11 mL) was added DIAD (0.25 mL, 1.28 mmol) drop-wise. When the reaction was shown to be complete by LC/MS, the mixture was concentrated to dryness. The crude material was purified by silica gel column
chromatography (gradient 0-70% methyl tert-butylether (MTBE):hexanes) to afford (R)-2- chloro-8-methyl-7-phenyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazine as a colourless oil (0.13 g, 43 %). 1H NMR (CDC13, 400 MHz): δ 7.71 (s, 1H), 7.41 - 7.34 (m, 3H), 7.17 (dd, J = 7.7, 1.6 Hz, 2H), 4.57 (t, J = 3.6 Hz, 1H), 4.27 (dd, J = 11.2, 3.3 Hz, 1H), 4.18 (dd, J = 11.2, 4.0 Hz, 1H), 3.08 (s, 3H); LRMS (ESpos) calcd for Ci3Hi2ClN30 [M+H]+: 262.07. found: 262.42.
Example 26
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-phi
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000094_0001
Synthesis ofN-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-phenyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0235] To a stirred solution of 2-chloro-8-methyl-7-phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (80 mg, 0.30 mmol) in 1, 4-dioxane (2 mL) was added l-(2- methoxypyridin-4-yl) piperidin-4-amine (82 mg, 0.33 mmol), Pd2(dba) (14 mg, 0.01 mmol), (±) BINAP (28 mg, 0.04 mmol), and sodium tertiary butoxide (88 mg, 0.91 mmol). The mixture was degassed under an argon atmosphere for 10 min. The reaction mixture was heated to 120 °C and stirred for 12 h in a sealed tube. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4% MeOH:CH2Cl2 and further purified by preparative HPLC
[Chiralpak IA (250 x 20 mm, 5 μπι) (15 mg loading; 0.1 % DEA in n-hexane; CH2Cl2:MeOH (80: 20); (75: 25 ) as the mobile phase with a Flow rate=15 mL/min] to afford N-(l-(2- methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (50 mg, 38%) as a pale yellow solid. 1H-NMR (DMSO-<¾, 500 MHz): δ 7.78 (d, 1H), 7.44 (s, 1H), 7.40-7.37 (m, 2H), 7.33-7.30 (m, 1H), 7.20 (d, 2H), 6.56-6.55 (m, 1H), 6.26 (br s, 1H), 6.11 (s, 1H), 4.71 (s, 1H), 4.15-4.09 (m, 2H), 3.86-3.83 (m, 3H), 3.76 (s, 3H), 2.95-2.90 (m, 5H), 1.92-1.90 (m, 2H), 1.48-1.42 (m, 2H); Mass (ESI): 433.4 [M+1]; LC-MS: 433.1 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 2.48 min. 0.05% TFA in water: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.56 min. ACN: 0.025% Aq TFA; 0.50 mL/min. TLC: 10%
MeOH:CH2Cl2 (R . 0.5).
[0236] Racemic compound of Example 26 was separated using a Chiralpak IA (250 x 20 mm, 5μιη (15 mg loading; 0.1 % DEA in n-hexane; CH2Cl2:MeOH (80:20); (75 :25) as the mobile phase) to provide the compound of Example 26A Fraction I (-) and the compound of Example 26B Fraction II (+).
Example 26A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-ph
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000095_0001
[0237] The compound of Example 26A was produced as described in Example 26.
Analytical data for product Fraction I (-): 1H NMR (DMSO-d6, 400 MHz): δ 7.68 (d, 1H), 7.44 (s, 1H), 7.38-7.35 (m, 2H), 7.32 (d, 1H), 7.20 (d, 2H), 6.57-6.56 (m, 1H), 6.30-6.28 (m, 1H), 6.10 (s, 1H), 4.74 (s, 1H), 4.23-4.20 (m, 2H), 3.86-3.82 (m, 3H), 2.98-2.95 (m, 5H), 2.78 (s, 3H), 1.93-1.90 (m, 2H), 1.42-1.39 (m, 2H); Mass (ESI): 433.5 [M+l]; LC-MS: 433.6 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.70 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.54 min. ACN: 0.025% TFA (Aq); 0.50 ml/min. Chiral HPLC:RT = 22.06 min (Chiralpak IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane; CH2C12: MeOH (80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 24": -93.00° (c = 0. 25%, DCM); TLC: 10% MeOH:CH2Cl2 (Rf. 0.5).
Example 26B Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-ph
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000096_0001
[0238] The compound of Example 26B was produced as described in Example 26.
Analytical data for product Fraction II (+): 1H NMR (DMSO-d6, 400 MHz): δ 7.78 (d, 1H), 7.42 (s, 1H), 7.36-7.34 (m, 2H), 7.30-7.29 (m, 1H), 7.20 (d, 2H), 6.56-6.54 (m, 1H), 6.30- 6.29 (m, 1H), 6.10 (s, 1H), 4.72-4.71 (m, 1H), 4.12-4.09 (m, 2H), 3.91-3.86 (m, 3H), 3.74 (s, 3H), 2.96 (s, 3H), 2.91-2.89 (m, 2H), 1.94-1.90 (m, 2H), 1.48-1.44 (m, 2H); Mass (ESI): 433.6 [M+1]; LC-MS: 433.7 (M+1); (column; X-select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.69 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 x 2.1mm, 1.7 μπι); RT 1.54 min. ACN: 0.025% TFA (Aq); 0.50 ml/min. Chiral HPLC: RT = 24.93 min (Chiralpak IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane CH2C12: MeOH (80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 25 °°: +81.31° (c = 0. 25%, DCM); TLC: 10% MeOH:CH2Cl2 (R . 0.5).
Example 27
Synthesis of 8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7-phenyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000096_0002
Synthesis of 8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7 -phenyl-'/ ', 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0239] To a dry vial was added a suspension of Pd2(dba)3 (8.7 mg, 0.01 mmol) and (±) BINAP (18 mg, 0.03 mmol) in 1, 4-dioxane (0.25 mL) at RT. The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture 2-chloro-8-methyl-7-phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (50 mg, 0.19 mmol), l-(6-methylpyrimidin-4- yl) piperidin-4-amine hydrochloride (48 mg, 0.21 mmol) and sodium tertiary butoxide (55 mg, 0.57 mmol) in 1, 4-dioxane (0.25 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 24 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (20 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4%
MeOH:CH2Cl2 and further purified by preparative HPLC [Kromasil C18 (250 x 21.2mm 10 μιη) (25 mg loading; n-hexane; CH2Cl2:MeOH (50:50); (85: 10) as the mobile phase with a Flow rate=15 mL/min] to afford 8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7- phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (15 mg, 19%) as a white solid. 1H-NMR (CO3OO-d4, 400 MHz): δ 8.33 (s, 1H), 7.39-7.32 (m, 4H), 7.22 (d, 2H), 6.68 (s, 1H), 4.64-4.62 (m, 1H), 4.42-4.39 (m, 2H), 4.20-4.10 (m, 2H), 4.02-3.97 (m, 1H), 3.18- 3.11 (m, 2H), 3.03 (s, 3H), 2.32 (s, 3H), 2.11-2.08 (m, 2H), 1.51-1.47 (m, 2H); LC-MS: 418.7 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.68 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 2.1 X 50 mm, 1.7 μιη); RT 1.46 min. ACN: 0.025% TFA (Aq); 0.50 mL/min. TLC: 5% MeOH:CH2Cl2 (Rf. 0.2).
Example 28
Synthesis of N-(l-(2-methoxypyridin-4-yl) azetidin-3-yl)-8-methyl-7-phj
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000098_0001
Synthesis ofN-(l-(2-methoxypyridin-4-yl) azetidin-3-yl)-8-methyl-7-phenyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0240] To a dry vial was added a suspension of Pd2(dba)3 (14 mg, 0.01 mmol) and (±) BINAP (28 mg, 0.04 mmol) in 1, 4-dioxane (1 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-phenyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (80 mg, 0.03 mmol), l-(2-methoxypyridin-4-yl) azetidin-3 -amine (60 mg, 0.33 mmol) and sodium tertiary butoxide (88 mg, 0.91 mmol) in 1, 4-dioxane (1 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After
consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by silica gel column chromatography using 2% MeOH:CH2Cl2 to afford N-(l- (2-methoxypyridin-4-yl) azetidin-3-yl)-8-methyl-7-phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (10 mg, 8%) as an off-white solid. 1H-NMR (DMSO-d6, 400 MHz): δ
7.75 (d, 1H), 7.45 (s, 1H), 7.39-7.31 (m, 3H), 7.19 (d, 2H), 7.00 (d, , 1H), 6.08-6.07 (m, 1H), 5.65 (s, 1H), 4.71-4.62 (m, 2H), 4.18-4.12 (m, 4H), 3.75-3.73(m, 2H), 3.72 (s, 3H), 2.94 (s, 3H); Mass (ESI): 405.4 [M+1]; LC-MS: 405.5 (M+1); (column; X-Bridge C-18 (50 3.0 mm, 3.5 μπι); RT 3.01 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC(column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.51 min. ACN: 0.025% TFA (Aq); 0.50 mL/min. TLC: 5% MeOH:CH2Cl2 (Rf. 0.4).
Example 29
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(o-tolyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000099_0001
Synthesis ofN-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(o-tolyl)-7, 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0241] To a dry vial was added a suspension of Pd2(dba)3 (6 mg, 0.007 mmol) and (±) BINAP (13 mg, 0.02 mmol) in 1, 4-dioxane (0.2 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (40 mg, 0.14 mmol), l-(2-methoxypyridin-4- yl)piperidin-4-amine (28 mg, 0.16 mmol) and sodium tertiary butoxide (41 mg, 0.43 mmol) in 1, 4-dioxane (0.2 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After
consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was diluted with water (10 mL) and extracted with 5% MeOH:CH2Cl2 (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 5% MeOH:CH2Cl2 to afford N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (10 mg, 16%) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.80 (d, 1H), 7.41 (s, 1H), 7.21-7.16 (m, 3H), 6.84 (d, 1H), 6.59-6.57 (m, 1H), 6.23 (br s, 1H), 6.12 (s, 1H), 4.93- 4.90 (m, 1H), 4.17-4.03 (m, 2H), 3.86-3.71 (m, 3H), 3.78 (s, 3H), 2.94-2.90 (m, 2H), 2.89 (s, 3H), 2.28 (s, 3H), 1.95-1.90 (m, 2H), 1.50-1.42 (m, 2H); Mass (ESI): 447.5 [M+1]; LC-MS: 447.7 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.90 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.63 min. ACN: 0.025% Aq TFA; 0.5 mL/min. TLC: 10% MeOH:CH2Cl2 (Rf. 0.3).
[0242] Racemic compound of Example 29 was separated using a Chiralpak IA (250 x 20 mm, 5μιη (50 mg loading; 0.1 % DEA in n-hexane; CH2Cl2:MeOH (50: 50); (75: 25) as the mobile phase) to provide the compound of Example 29A Fraction I (-) and the compound of Example 29B Fraction II (+). Example 29 A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(o-tolyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000100_0001
[0243] The compound of Example 29 A was produced as described in Example 29.
Analytical data for product Fraction I (-): 1H-NMR (DMSO-d6, 500 MHz): δ 7.78 (d, 1H), 7.42 (s, 1H), 7.24-7.14 (m, 3H), 6.84 (d, 1H), 6.56-6.52 (m, 1H), 6.22 (d, 1H), 6.11-6.09 (m, 1H), 5.90-5.88 (m, 1H), 4.11-4.02 (m, 2H), 3.91-3.88 (m, 3H), 3.77 (s, 3H), 2.93 (s, 5H), 2.38 (s, 3H), 1.94-1.90 (m, 2H), 1.45-1.38 (m, 2H); Mass (ESI): 447.6 [M+l]; LC-MS: 447.6 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.89 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.64 min. ACN: 0.025% TFA (Aq); 0.50 ml/min. Chiral HPLC: RT = 7.54 min (Chiralpak IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) DCM:MeOH (50:50); flow Rate: 1.0 mL/min); Optical rotation [a]D 19 98: -94.91° (c = 0. 25%, DCM).
Example 29B
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(o-tolyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000101_0001
[0244] The compound of Example 29B was produced as described in Example 29.
Analytical data for product Fraction II (+): 1H-NMR (DMSO-d6, 500 MHz): δ 7.76 (d, 1H), 7.41 (s, 1H), 7.25-7.14 (m, 3H), 6.85 (d, 1H), 6.56-6.52 (m, 1H), 6.23 (d, 1H), 6.11-6.09 (m, 1H), 5.90-5.87 (m, 1H), 4.11-4.03 (m, 2H), 3.90-3.88 (m, 3H), 3.78 (s, 3H), 2.91 (s, 5H), 2.37 (s, 3H), 1.92-1.87 (m, 2H), 1.47-1.40 (m, 2H); Mass (ESI): 447.6 [M+1]; LC-MS: 447.6 (M+1); (column; X-select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.89 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.64 min. ACN: 0.025% TFA (Aq); 0.50 ml/min. Chiral HPLC: RT = 13.04 min (Chiralpak IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) DCM:MeOH (50:50); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +110.25° (c = 0. 25%, DCM).
Example 30
Synthesis of N-(l-(2-methoxypyridin-4-yl) azetidin-3-yl)-8-methyl-7-(o-tolyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000101_0002
Synthesis of N-(l-(2-methoxypyridin-4-yl) azetidin-3-yl)-8-methyl- 7-(o-tolyl)- 7, 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine [0245] To a dry vial was added a suspension of Pd2(dba)3 (6 mg, 0.007 mmol) and (±) BINAP (13 mg, 0.02 mmol) in 1, 4-dioxane (0.2 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (40 mg, 0.14 mmol), l-(2-methoxypyridin-4- yl) azetidin-3 -amine (28 mg, 0.16 mmol) and sodium tertiary butoxide (41 mg, 0.43 mmol) in 1, 4-dioxane (0.2 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After
consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude mixture was diluted with water (10 mL) and extracted with 5% MeOH:CH2Cl2 (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 5% MeOH:CH2Cl2 to afford N-(l-(2-methoxypyridin-4-yl) azetidin-3-yl)-8-methyl-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (7 mg, 12%) as an off-white solid.1H-NMR (DMSO-<¾, 400 MHz): δ 7.73 (d, 1H), 7.41 (s, 1H), 7.22-7.10 (m, 3H), 6.99 (d, 1H), 6.81 (d, 1H), 6.08 (d, 1H), 5.61 (s, 1H), 4.91-4.89 (m, 1H), 4.64-4.60 (m, 1H), 4.19-4.01 (m, 4H), 3.73-3.71 (m, 2H), 3.69 (s, 3H), 2.90 (s, 3H), 2.32 (s, 3H); Mass (ESI): 419.5 [M+1]; LC-MS: 419.6 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.78 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column;
Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.52 min. ACN: 0.025% Aq TFA; 0.5 mL/min. TLC: 10% MeOH:CH2Cl2 (Rf. 0.3).
Example 31
Synthesis of 8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000102_0001
Synthesis of 8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine [0246] To a dry vial was added a suspension of Pd2(dba)3 (7 mg, 0.008 mmol) and (±) BINAP (15 mg, 0.02 mmol) in 1, 4-dioxane (0.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (50 mg, 0.15 mmol), l-(6- methylpyrimidin-4-yl) piperidin-4-amine hydrochloride (40 mg, 0.17 mmol) and sodium tertiary butoxide (46 mg, 0.47 mmol) in 1, 4-dioxane (0.25 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (10 mL) and extracted with CH2C12 (2 x 10 mL). The combined organic extracts were washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by preparative HPLC [Kromasil C 18, 250 x 21.2 mm, 5 μιη) (45 mg loading; CH3CN; 5 mM Aq NH4OAc
(0.01/75, 15/40, 25/10, 35/10) as the mobile phase with a Flow rate=15 mL/min] to afford 8- methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7-(3, 4, 5-trifluorophenyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (8 mg, 10%) as an off- white solid.1H- NMR (CD3OD, 400 MHz): δ 8.30 (s, 1H), 7.41 (s, 1H), 7.01-6.95 (m, 2H), 6.69 (s, 1H), 4.69-4.65 (m, 1H), 4.41-4.39 (m, 2H), 4.12 (s, 2H), 4.02-3.98 (m, 1H), 3.16-3.10 (m, 2H), 3.02 (s, 3H), 2.31 (s, 3H), 2.11-2.08 (m, 2H), 1.53-1.47 (m, 2H); LC-MS: 472.5 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.44 min 5 mM NH4OAc: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB C-18 150 X 4.6 mm, 5 μπι); RT 10.58 min. ACN: 5 mM NH4OAc; 0.5 mL/min. TLC: 5% MeOH:CH2Cl2 (Rf. 0.4).
[0247] Racemic compound of Example 31 was separated using a Chiralpak ADH (250 x 20 mm, 5μ 30 mg loading; (A) 0.1 % DEA in n-hexane (B) IPA (A:B : 70:30) as the mobile phase to provide the compound of Example 31 A Fraction I (-) and the compound of Example 3 IB Fraction II (+).
Example 31 A
Synthesis of (-)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000104_0001
[0248] The compound of Example 31 A was produced as described in Example 31.
Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 8.31 (s, 1H), 7.41 (s, 1H), 7.02-6.99 (m, 2H), 6.69 (s, 1H), 4.69-4.66 (m, 1H), 4.43-4.38 (m, 2H), 4.15 (d, 2H), 4.02-3.98 (m, 1H), 3.19-3.10 (m, 2H), 3.08 (s, 3H), 2.31 (s, 3H), 2.11-2.05 (m, 2H), 1.51- 1.47 (m, 2H); Mass (ESI): 472.4 [M+1]; LC-MS: 472.6 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.55 min. 5 mM Aq NH4OAc: ACN; 0.80 ml/min); UPLC (column; Eclipse-XDB-C-18, 150 4.6 mm, 5 μπι); RT 10.52 min. ACN: 5 mM Aq
NH4OAc; 1.0 ml/min; Chiral HPLC: RT = 10.27 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) IPA (A:B : 70:30; flow Rate: 1.0 mL/min); Optical rotation [<x]D 19-98: -59.92 (c 0.25, DCM); TLC: 5% MeOH:CH2Cl2 (R . 0.4).
Example 3 IB
Synthesis of (+)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000104_0002
[0249] The compound of Example 3 IB was produced as described in Example 31.
Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 8.31 (s, 1H), 7.41 (s, 1H), 7.02-6.99 (m, 2H), 6.69 (s, 1H), 4.69-4.66 (m, 1H), 4.43-4.38 (m, 2H), 4.15 (d, 2H), 4.02-3.98 (m, 1H), 3.19-3.10 (m, 2H), 3.08 (s, 3H), 2.31 (s, 3H), 2.11-2.05 (m, 2H), 1.51-1.47 (m, 2H); Mass (ESI): 472.5 [M+1]; LC-MS: 472.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.56 min. 5 mM Aq NH4OAc: ACN; 0.80 ml/min); UPLC (column; Eclipse-XDB-C-18, 150 4.6 mm, 5 μπι); RT 10.53 min. ACN: 5 mM Aq
NH4OAc; 1.0 ml/min; Chiral HPLC: RT = 12.05 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) IPA (A:B : 70:30; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +64.94 (c 0.25, DCM); TLC: 5% MeOH:CH2Cl2 (R . 0.4).
Example 32
Synthesis of 7-(3, 5-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin- 4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000105_0001
Synthesis of 7-(3, 5-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0250] To a dry vial was added a suspension of Pd2(dba)3 (6 mg, 0.006 mmol) and (±) BINAP (12 mg, 0.02 mmol) in 1, 4-dioxane (0.4 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(3, 5-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (40 mg, 0.13 mmol), l-(6- methylpyrimidin-4-yl) piperidin-4-amine hydrochloride (33 mg, 0.14 mmol) and sodium tertiary butoxide (38 mg, 0.40 mmol) in 1, 4-dioxane (0.4 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was diluted with water (5 mL) and extracted with 5% MeOH:CH2Cl2 (2 x 5 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by preparative HPLC [Kromasil C 18, 250 x 21.2 mm, 5 μπι) (50 mg loading; CH3CN; 0.05% TFA (0.01/90, 15/70, 25/10, 35/10) as the mobile phase with a Flow rate=15 mL/min] to afford 7-(3, 5-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1 , 4] oxazin-2-amine (10 mg, 16%>) as an off-white solid. 1H- NMR (OMSO-d6, 400 MHz): δ 8.32 (s, 1H), 7.46 (s, 1H), 7.16 (t, 1H), 6.88 (d, 2H), 6.70 (s, 1H), 6.28 (d, 1H), 4.78 (s, 1H), 4.30 (d, 2H), 4.18 (d, 1H), 4.10 (d, 1H), 3.92-3.88 (m, 1H), 3.06-2.96 (m, 5H), 2.20 (s, 3H), 1.88 (d, 2H), 1.38 (d, 2H); Mass (ESI): 454.5 [M+1]; LC- MS: 454.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.80 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.60 min. ACN: 0.025% Aq TFA; 0.5 mL/min. TLC: 5% MeOH:CH2Cl2 (R/. 0.2).
[0251] Racemic compound of Example 32 was separated using a Chiralpak ADH (250 x 20 mm, 5μ 70 mg loading; (A) 0.1 % DEA in n-hexane (B) EtOH:MeOH (50:50) (A:B : 70:30) as the mobile phase) to provide the compound of Example 32A Fraction I (+) and the compound of Example 32B Fraction II (-).
Example 32A
Synthesis of (+)-7-(3, 5-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000106_0001
[0252] The compound of Example 32A was produced as described in Example 32.
Analytical data for product Fraction I (+): 1H-NMR (DMSO-< 5, 400 MHz): δ 8.32 (s, 1H), 7.45 (s, 1H), 7.22 (t, 1H), 6.88 (d, 2H), 6.70 (s, 1H), 6.26 (d, 1H), 4.79 (s, 1H), 4.36-4.29 (m, 2H), 4.18 (d, 1H), 4.10 (d, 1H), 3.94-3.86 (m, 1H), 3.07-3.00 (m, 2H), 2.99 (s, 3H), 2.22 (s, 3H), 1.92-1.85 (m, 2H), 1.40-1.32 (m, 2H); Mass (ESI): 454.4 [M+1]; LC-MS: 454.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.77 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.51 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 15.91 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) EtOH:MeOH (50:50) (A:B : 70:30; flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +63.32 (c 0.25, DCM).
Example 32B
Synthesis of (-)-7-(3, 5-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000107_0001
[0253] The compound of Example 32B was produced as described in Example 32.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.32 (s, 1H), 7.45 (s, 1H), 7.22 (t, 1H), 6.88 (d, 2H), 6.70 (s, 1H), 6.26 (d, 1H), 4.79 (s, 1H), 4.37-4.29 (m, 2H), 4.18 (d, 1H), 4.10 (d, 1H), 3.94-3.86 (m, 1H), 3.07-3.00 (m, 2H), 2.99 (s, 3H), 2.22 (s, 3H), 1.94-1.88 (m, 2H), 1.42-1.32 (m, 2H); Mass (ESI): 454.6 [M+1]; LC-MS: 454.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.77 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 x 2.1 mm, 1.7 μπι); RT 1.50 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 24.37 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH:MeOH (50:50) (A:B : 70:30; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 00: -69.96 (c 0.25, DCM).
Example 33
Synthesis of 7-(3, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000108_0001
Synthesis of 7-(3, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7 , 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0254] To a dry vial was added a suspension of Pd2(dba)3 (8 mg, 0.009 mmol) and (±) BINAP (17 mg, 0.02 mmol) in 1, 4-dioxane (0.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(3, 5-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (50 mg, 0.18 mmol), l-(2- methoxypyridin-4-yl) piperidin-4-amine hydrochloride (48 mg, 0.19 mmol) and sodium tertiary butoxide (52 mg, 0.54 mmol) in 1, 4-dioxane (0.25 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (5 mL) and extracted with CH2C12 (2 x 5 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 40% EtOAc:hexanes to afford 7-(3, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4- yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (35 mg, 41%) as an off-white solid.1H-NMR (DMSO-<¾, 400 MHz): δ 7.80 (d, 1H), 7.48 (s, 1H), 7.20 (t, 1H), 6.90 (d, 2H), 6.55 (d, 1H), 6.32 (d, 1H), 6.10 (s, 1H), 4.80 (s, 1H), 4.18-4.12 (m, 2H), 3.90- 3.80 (m, 3H), 3.74 (s, 3H), 2.97-2.90 (m, 5H), 1.92-1.89 (m, 2H), 1.46-1.43 (m, 2H); Mass (ESI): 469.6 [M+1]; LC-MS: 469.6 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.86 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.64 min. ACN: 0.025% Aq TFA; 0.5 mL/min. TLC: 50% EtOAc:hexanes (Rf. 0.4).
[0255] Racemic compound of Example 33 was separated using a Chiralpak IB (250 x 20 mm, 5μ (20 mg loading; (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 90: 10) as the mobile phase) to provide the compound of Example 33A Fraction I (+) and the compound of Example 33B Fraction II (-). Example 33A
Synthesis of (+)-7-(3, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000109_0001
[0256] The compound of Example 33 A was produced as described in Example 33. Analytical data for product Fraction I (+): 1H-NMR (OMSO-d6, 400 MHz): δ 7.77 (d, 1H), 7.45 (s, 1H), 7.21 (t, 1H), 6.90 (d, 2H), 6.53 (d, 1H), 6.28 (d, 1H), 6.10 (s, 1H), 4.79 (s, 1H), 4.20-4.10 (m, 2H), 3.88-3.80 (m, 3H), 3.75 (s, 3H), 2.98 (s, 3H), 2.97-2.90 (m, 2H), 1.90- 1.85 (m, 2H), 1.48-1.40 (m, 2H); Mass (ESI): 454.6 [M+1]; LC-MS: 469.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.82 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.57 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC RT = 24.35 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 90: 10; flow Rate: 1.0 mL/min); Optical rotation [a]D 19 %: +75.02 (c 0.25, DCM); TLC: 50% EtOAc:hexanes (Rf. 0.4).
Example 33B
Synthesis of (-)-7-(3, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000110_0001
[0257] The compound of Example 33B was produced as described in Example 33.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 7.77 (d, 1H), 7.45 (s, 1H), 7.21 (t, 1H), 6.90 (d, 2H), 6.53 (d, 1H), 6.28 (d, 1H), 6.10 (s, 1H), 4.79 (s, 1H), 4.20-4.10 (m, 2H), 3.88-3.80 (m, 3H), 3.75 (s, 3H), 3.98 (s, 3H), 3.97-3.90 (m, 2H), 1.90- 1.85 (m, 2H), 1.48-1.40 (m, 2H); Mass (ESI): 469.4 [M+1]; LC-MS: 469.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.81 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.57 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 29.92 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 90: 10; flow Rate: 1.0 mL/min); Optical rotation [a]D 19": -67.56 (c 0.25, DCM); TLC: 50% EtOAc:hexanes (Rf. 0.4).
Example 34
Synthesis of V-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000110_0002
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine [0258] To a dry vial was added a suspension of Pd2(dba)3 (22 mg, 0.02 mmol) and (±) BINAP (20 mg, 0.03 mmol) in 1, 4-dioxane (0.5 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (50 mg, 0.15 mmol), l-(2- methoxypyridin-4-yl) piperidin-4-amine hydrochloride (42 mg, 0.17 mmol) and sodium tertiary butoxide (61 mg, 0.60 mmol) in 1, 4-dioxane (0.5 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (5 mL) and extracted with EtOAc (2 x 5 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 60% EtOAc:hexanes to afford 50 mgs further purified by preparative HPLC [Kromasil CI 8 (250 x 21.2mm 10 μιη) (40mg loading; n-hexane; CH2Cl2:MeOH (50:50); (85: 15) as the mobile phase with a Flow rate=15 mL/min] to afford N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)- 8-methyl-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (12 mg, 15%) as an off-white solid. 1H-NMR (CD3OD+DMSO-<¾, 400 MHz): δ 7.76 (d, 1H), 7.42 (s, 1H), 7.02 (t, 2H), 6.56 (d, 1H), 6.16 (s, 1H), 4.68 (s, 1H), 4.16 (s, 2H), 3.98-3.92 (m, 3H), 3.84 (s, 3H), 3.10-3.02 (m, 5H), 2.12-2.09 (m, 2H), 1.57-1.54 (m, 2H); Mass (ESI): 487.5 [M+l]; LC-MS: 487.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.88 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μιη); RT 1.64 min. ACN: 0.025% Aq TFA; 0.5 mL/min. TLC: 70%
EtOAc:hexanes (Rf. 0.2).
[0259] Racemic compound of Example 34 was separated using a Chiralpak IB (250 x 20 mm, 5μ (20 mg loading; (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B :
90: 10) as the mobile phase) to provide the compound of Example 34A Fraction I (+) and the compound of Example 34B Fraction II (-).
Example 34A
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000112_0001
[0260] The compound of Example 34A was produced as described in Example 34.
Analytical data for product Fraction I (+): 1H-NMR (CD3OD + DMSO (2 drops), 400 MHz): δ 7.75 (d, 1H), 7.41 (s, 1H), 7.02 (t, 2H), 6.54 (d, 1H), 6.17 (s, 1H), 4.69-4.67 (m, 1H), 4.15 (d, 2H), 3.96-3.89 (m, 3H), 3.82 (s, 3H), 3.25 (s, 3H), 3.03-3.00 (m, 2H), 2.10-2.08 (m, 2H), 1.60-1.51 (m, 2H); Mass (ESI): 487.5 [M+1]; LC-MS: 487.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.82 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.65 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 29.09 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 90: 10; flow Rate: 1.0 mL/min); Optical rotation [a]D 19 98: +71.24 (c 0.25, DCM); TLC: 70%
EtOAc:hexanes (Rf. 0.2).
Example 34B
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000112_0002
[0261] The compound of Example 34B was produced as described in Example 34.
Analytical data for product Fraction II (-): 1H-NMR (CD3OD + DMSO (2 drops), 400 MHz):
- I l l - δ 7.75 (d, 1H), 7.41 (s, 1H), 7.03-6.99 (m, 2H), 6.54 (d, 1H), 6.17 (s, 1H), 4.69-4.67 (m, 1H), 4.26 (d, 2H), 3.96-3.89 (m, 3H), 3.82 (s, 3H), 3.25 (s, 3H), 3.03-3.00 (m, 2H), 2.10-2.08 (m, 2H), 1.60-1.51 (m, 2H); Mass (ESI): 487.5 [M+1]; LC-MS: 487.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.82 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.65 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 34.06 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 90: 10; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 00: -54.96 (c 0.25, DCM); TLC: 70%
EtOAc:hexanes (Rf. 0.2).
Example 35
Synthesis of 8-methyl-7-phenyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7,
8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000113_0001
Synthesis of8-methyl-7-phenyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7 , 8- dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0262] To a dry vial was added a suspension of Pd2(dba)3 (17 mg, 0.01 mmol) and (±)
BINAP (36 mg, 0.05 mmol) in 1, 4-dioxane (0.5 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-phenyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (100 mg, 0.38 mmol), l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (118 mg, 0.42 mmol) and sodium tertiary butoxide (110 mg, 1.14 mmol) in 1, 4-dioxane (0.5 mL) was degassed with argon and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4% MeOH:CH2Cl2 to afford 120 mgs further purified by preparative HPLC [Kromasil C18 (250 x 21.2mm 10 μιη) (50 mg loading; CH3CN;0.05% TFA Aq; (0.1/90, 15/70, 25/10, 35/10) as the mobile phase with a Flow rate=15 mL/min] to afford 8-methyl-7-phenyl-N-(l-(2- (trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (52 mg, 29%) as an off-white solid. 1H-NMR (DMSO-<¾, 400 MHz): δ 8.24 (d, 1H), 7.43 (s, 1H), 7.39-7.29 (m, 3H), 7.20-7.18 (m, 3H), 7.04-7.02 (m, 1H), 6.24 (d, 1H), 4.70-4.69 (m, 1H), 4.15-4.10 (m, 2H), 4.08-4.07 (m, 2H), 4.03-4.02 (m, 1H), 3.06 (t, 2H), 2.94 (s, 3H), 1.56-1.93 (m, 2H), 1.53-1.51 (m, 2H);LC-MS: 471.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.07 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μιη); RT 1.83 min. ACN: 0.025% Aq TFA; 0.5 mL/min. TLC: 5% MeOH:DCM (Rf. 0.2).
[0263] Racemic compound of Example 35 was separated using a Chiralpak IB (250 x 20 mm, 5μ (30 mg loading; (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 90: 10) as the mobile phase) to provide the compound of Example 35A Fraction I (+) and the compound of Example 35B Fraction II (-).
Example 35A
Synthesis of (+)-8-methyl-7-phenyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000114_0001
[0264] The compound of Example 35 A was produced as described in Example 35.
Analytical data for Fraction I (+): 1H-NMR (DMSO-d6, 400 MHz): δ 8.25-8.20 (m, 1H), 7.41 (s, 1H), 7.40-7.33 (m, 3H), 7.20-7.18 (m, 3H), 7.04-7.02 (m, 1H), 6.22 (d, 1H), 4.70-4.68 (m, 1H), 4.12-4.09 (m, 2H), 4.01-3.98 (m, 2H), 3.91-3.86 (m, 1H), 3.06 (t, 2H), 2.92 (s, 3H), 1.97-1.89 (m, 2H), 1.51-1.40 (m, 2H); LC-MS: 471.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.10 min. 0.05% TFA in water: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.80 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 20.84 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 90: 10; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +89.72 (c 0.25, DCM).
Example 35B
Synthesis of (-)-8-methyl-7-phenyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000115_0001
[0265] The compound of Example 35B was produced as described in Example 35.
Analytical data for Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.25-8.20 (m, 1H), 7.41 (s, 1H), 7.40-7.33 (m, 3H), 7.20-7.18 (m, 3H), 7.04-7.02 (m, 1H), 6.22 (d, 1H), 4.70-4.68 (m, 1H), 4.12-4.09 (m, 2H), 4.01-3.98 (m, 2H), 3.91-3.86 (m, 1H), 3.06 (t, 2H), 2.92 (s, 3H), 1.97-1.89 (m, 2H), 1.51-1.40 (m, 2H; LC-MS: 471.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.28 min. 0.05% TFA in water: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.80 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 23.42 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 90: 10; flow Rate: 1.0 mL/min); Optical rotation [a]D 19 98: -85.02 (c 0.25, DCM).
Example 36
Synthesis of N-(l-(6-methoxypyrimidin-4-yl) piperidin-4-yl)-8-methyl-7-phi
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000116_0001
Synthesis of N-(l-( 6-methoxypyrimidin-4-yl) piperidin-4-yl)-8-methyl- 7 -phenyl- 7, 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0266] To a dry vial was added a suspension of Pd2(dba)3 (17 mg, 0.01 mmol) and (±) BINAP (36 mg, 0.05 mmol) in 1, 4-dioxane (0.5 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (100 mg, 0.38 mmol), l-(6-methoxypyrimidin- 4-yl) piperidin-4-amine hydrochloride (103 mg, 0.42 mmol) and sodium tertiary butoxide (110 mg, 1.14 mmol) in 1, 4-dioxane (0.5 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4% MeOH:CH2Cl2 and further purified by preparative HPLC [Kromasil CI 8 (250 x 21.2mm 10 μιη) (50mg loading; CH3CN: 0.05% TFA 0.01/90, 15/70, 25/10, 35/10 as mobile phase with a Flow rate =15 mL/min] to afford N- ( 1 -(6-methoxypyrimidin-4-yl) piperidin-4-yl)-8-methyl-7-phenyl-7, 8-dihydro-6H-pyrimido
[5, 4-b] [1, 4] oxazin-2-amine (40 mg, 24%) as a white solid. 1H-NMR (DMSO-<¾, 400 MHz): δ 8.22 (s, 1H), 7.43 (s, 1H), 7.39-7.36 (m, 2H), 7.31 (d, 1H), 7.19 (d, 2H), 6.20 (d, 1H), 6.07 (s, 1H), 4.70 (d, 1H), 4.29-4.26 (m, 2H), 4.14-4.10 (m, 2H), 3.91-3.89 (m, 1H), 3.81 (s, 3H), 3.03-2.76 (m, 2H), 2.94 (s, 3H), 1.92-1.89 (m, 2H), 1.42-1.33 (m, 2H); LC-MS: 434.3 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.89 min. 0.05% Aq TFA:ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μιη); RT 1.66 min. ACN: 0.025% TFA (Aq); 0.50 mL/min; TLC: 5% MeOH:CH2Cl2 (Rf. 0.1).
Example 37 Synthesis of 7-benzyl-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000117_0001
Synthesis of 7-benzyl-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7 , 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0267] To a dry vial was added a suspension of Pd2(dba)3 (34 mg, 0.01 mmol) and (±) BINAP (68 mg, 0.54 mmol) in 1, 4-dioxane (0.5 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 7-benzyl-2-chloro-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.72 mmol), l-(6-methylpyrimidin-4- yl) piperidin-4-amine hydrochloride (167 mg, 0.87 mmol) and sodium tertiary butoxide (279 mg, 2.90 mmol) in 1, 4-dioxane (0.5 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4% MeOH:CH2Cl2 and further purified by preparative HPLC to afford 7-benzyl-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (42 mg, 27%) as a pale yellow solid. LC-MS: 432.5 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.36 min. 5 mM NH4OAc: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.62 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0268] Racemic compound of Example 37 was separated using a Chiralpak IA (250 x 20 mm, 5μιη (35 mg loading; 0.1 % DEA in n-hexane: CH2Cl2:MeOH (50:50); (A:B: 75:25) as the mobile phase) to provide the compound of Example 37 A Fraction I (+) and the compound of Example 37B Fraction II (-).
Example 37A Synthesis of (+)-7-benzyl-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
[0269] The compound of Example 37A was produced as described in Example 37.
Analytical data for product Fraction I (+): 1H-NMR (OMSO-d6, 400 MHz): δ 8.33 (s, 1H), 7.41 (s, 1H), 7.31-7.21 (m, 5H), 6.70 (s, 1H), 6.09 (d, 1H), 4.33-4.28 (m, 2H), 3.89 (d, 2H), 3.73-3.67 (m, 2H), 3.04-3.00 (m, 3H), 2.93 (s, 3H), 2.71-2.67 (m, 1H), 2.21 (s, 3H), 1.91- 1.89 (m, 2H), 1.40-1.30 (m, 2H); Mass (ESI): 432.5 [M+1]; LC-MS: 432.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.64 min. 5 mM NH4OAc: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C- 18, 50 2.1mm, 1.7 μπι); RT 1.63 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 1 1.58 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 75 :25); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +33.13 (c 0.25, DCM).
Example 37B
Synthesis of (-)-7-benzyl-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000118_0002
[0270] The compound of Example 37B was produced as described in Example 37.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.33 (s, 1H), 7.41 (s, 1H), 7.31-7.21 (m, 5H), 6.70 (s, 1H), 6.09 (d, 1H), 4.33-4.28 (m, 2H), 3.89 (d, 2H), 3.73-3.67 (m, 2H), 3.04-3.00 (m, 3H), 2.93 (s, 3H), 2.71-2.67 (m, 1H), 2.21 (s, 3H), 1.91- 1.89 (m, 2H), 1.40-1.30 (m, 2H); Mass (ESI): 432.5 [M+1]; LC-MS: 432.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.61 min. 5 mM NH4OAc: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.63 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 22.98 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 75:25); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: -50.03 (c 0.25, DCM).
Example 38
Synthesis of 7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000119_0001
Synthesis of 7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0271] To a dry vial was added a suspension of Pd2(dba)3 (31 mg, 0.03 mmol) and (±) BINAP (63 mg, 0.10 mmol) in 1, 4-dioxane (1 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(3, 5-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.67 mmol), l-(2- (trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (208 mg, 0.74 mmol) and sodium tertiary butoxide (194 mg, 2.01 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4%
MeOH:CH2Cl2 to afford 7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin- 4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (200 mg, 59%) as a pale yellow solid. LC-MS: 507.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.14 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 x 2.1 mm, 1.7 μπι); RT 1.89 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0272] Racemic compound of Example 38 was separated using a Chiralpak IB (250 x 20 mm, 5μ (30 mg loading; (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B :
90: 10) as the mobile phase) to provide the compound of Example 38A Fraction I (+) and the compound of Example 38B Fraction II (-).
Example 38A
Synthesis of (+)-7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000120_0001
[0273] The compound of Example 38A was produced as described in Example 38.
Analytical data for product Fraction I (+): 1H-NMR (OMSO-d6, 400 MHz): δ 8.21 (d, 1H), 7.44 (s, 1H), 7.20-7.14 (m, 2H), 7.00-6.98 (m, 1H), 6.90-6.88 (m, 2H), 6.28 (d, 1H), 4.78- 4.75 (m, 1H), 4.19-4.08 (m, 4H), 4.00-3.88 (m, 1H), 3.03 (t, 2H), 2.93 (s, 3H), 1.94-1.89 (m, 2H), 1.50-1.40 (m, 2H); LC-MS: 507.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.18 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 x 2.1mm, 1.7 μπι); RT 1.88 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 15.39 min (CHIRALPAK-IB (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +62.08 (c 0.25, DCM).
Example 38B
Synthesis of (-)-7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000121_0001
[0274] The compound of Example 38B was produced as described in Example 38.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.21 (d, 1H), 7.44 (s, 1H), 7.20-7.14 (m, 2H), 7.00-6.98 (m, 1H), 6.90-6.88 (m, 2H), 6.28 (d, 1H), 4.78- 4.75 (m, 1H), 4.19-4.08 (m, 4H), 4.00-3.88 (m, 1H), 3.03 (t, 2H), 2.93 (s„ 3H), 1.94-1.89 (m, 2H), 1.50-1.40 (m, 2H); LC-MS: 507.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.18 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.89 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 17.65 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (80:20) (A:B : 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": -71.68 (c 0.25, DCM).
Example 39
Synthesis of 7-(4-fluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000122_0001
Synthesis of 7-(4-fluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7 , 8- dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazin-2-amine
[0275] To a dry vial was added a suspension of Pd2(dba)3 (33 mg, 0.03 mmol) and (±) BINAP (67 mg, 0.10 mmol) in 1, 4-dioxane (2 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(4-fluorophenyl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.71 mmol), l-(6- methylpyrimidin-4-yl) piperidin-4-amine hydrochloride (150 mg, 0.78 mmol) and sodium tertiary butoxide (205 mg, 2.14 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 130 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4%
MeOH:CH2Cl2 to afford 7-(4-fluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (200 mg, 59%) as an off-white solid. UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.56 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0276] Racemic compound of Example 39 was separated using a Chiralpak IA (250 x 20 mm, 5μιη (50 mg loading; 0.1 % DEA in n-hexane: CH2Cl2:MeOH (50:50); (A:B : 85: 15) as the mobile phase) to provide the compound of Example 39 A Fraction I (+) and the compound of Example 39B Fraction II (-).
Example 39A
Synthesis of (+)-7-(4-fluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000123_0001
[0277] The compound of Example 39A was produced as described in Example 39.
Analytical data for product Fraction I (+): 1H-NMR (DMSO-< 5, 400 MHz): δ 8.35 (s, 1H), 7.42 (s, 1H), 7.26-7.17 (m, 4H), 6.69 (s, 1H), 6.21 (d, 1H), 4.71-4.69 (m, 1H), 4.34-4.27 (m, 2H), 4.11-4.08 (m, 2H), 3.92-3.88 (m, 1H), 3.00 (t, 2H), 2.90 (s, 3H), 2.22 (s, 3H), 1.93-1.88 (m, 2H), 1.41-1.38 (m, 2H); Mass (ESI): 436.4 [M+1]; LC-MS: 436.5 (M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.52 min. 5 mM NH4OAc:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.53 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 23.11 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D 20"01: +76.00 (c 0.25, DCM).
Example 39B
Synthesis of (-)-7-(4-fluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000123_0002
[0278] The compound of Example 39B was produced as described in Example 39.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.35 (s, 1H),
7.42 (s, 1H), 7.26-7.17 (m, 4H), 6.69 (s, 1H), 6.21 (d, 1H), 4.71-4.69 (m, 1H), 4.34-4.27 (m,
2H), 4.11-4.08 (m, 2H), 3.92-3.88 (m, 1H), 3.00 (t, 2H), 2.90 (s, 3H), 2.22 (s, 3H), 1.93-1.88
(m, 2H), 1.41-1.38 (m, 2H); Mass (ESI): 436.4 [M+1]; LC-MS: 436.5 (M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.34 min. 5 mM NH4OAc:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.52 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 32.68 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 85: 15); flow Rate: 1.0 mL/min); Optical rotation [<x]D 19"95: -72.44 (c 0. 25, DCM).
Example 40
Synthesis of 7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7,
8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000124_0001
Synthesis of 7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7 , 8- dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0279] To a dry vial was added a suspension of Pd2(dba)3 (42 mg, 0.04 mmol) and (±) BINAP (85 mg, 0.13 mmol) in 1, 4-dioxane (1.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 7-benzyl-2-chloro-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.90 mmol), l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (281 mg, 1.00 mmol) and sodium tertiary butoxide (262 mg, 2.72 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 2-4%
MeOH:CH2Cl2 to afford 7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin- 4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (200 mg, 45%) as an off-white solid. LC-MS: 485.4 (M+l); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 3.20 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.88 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0280] Racemic compound of Example 40 was separated using a Chiralpak-IA (250 x 20 mm, 5μιη (50 mg loading; 0.1 % DEA in rc-hexane: CH2Cl2:MeOH (50:50); (A:B : 85: 15) as the mobile phase to provide the compound of Example 40A Fraction I (+) and the compound of Example 40B Fraction II (-).
Example 40A
Synthesis of (+)-7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000125_0001
[0281] The compound of Example 40A was produced as described in Example 40.
Analytical data for product Fraction I (+): 1H-NMR (DMSO-< 5, 400 MHz): δ 8.21 (d, 1H), 7.41 (s, 1H), 7.35-7.27 (m, 2H), 7.25-7.20 (m, 3H), 7.19 (s, 1H), 7.02-7.00 (m, 1H), 6.11 (d, 1H), 4.00-3.96 (m, 2H), 3.91-3.82 (m, 2H), 3.71-3.65 (m, 2H), 3.09-3.00 (m, 3H), 2.91 (s, 3H), 2.70-2.67 (m, 1H), 1.93-1.89 (m, 2H), 1.47-1.39 (m, 2H); LC-MS: 485.4 (M+l);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.15 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 x 2.1 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 11.77 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 80:20); flow Rate: 1.0 mL/min); Optical rotation [<x]D 2L41: +44.78 (c 0. 25, DCM).
Example 40B
Synthesis of (-)-7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000126_0001
[0282] The compound of Example 40B was produced as described in Example 40.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.21 (d, IH), 7.40 (s, IH), 7.30-7.27 (m, 2H), 7.25-7.20 (m, 3H), 7.19 (s, IH), 7.02-7.00 (m, IH), 6.11 (d, IH), 4.00-3.96 (m, 2H), 3.80-3.82 (m, 2H), 3.70-3.62 (m, 2H), 3.09-3.00 (m, 3H), 2.91 (s, 3H), 2.70-2.67 (m, IH), 1.93-1.89 (m, 2H), 1.47-1.39 (m, 2H); LC-MS: 485.3 (M+l);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.15 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 15.06 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 80:20); flow Rate: 1.0 mL/min); Optical rotation [<x]D 2L22: -41.69 (c 0. 25, DCM).
Example 41
Synthesis of 7-(4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000126_0002
Synthesis of 7-(4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7 , 8- dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine [0283] To a dry vial was added a suspension of Pd2(dba)3 (66.8 mg, 0.03 mmol) and (±) BINAP (32 mg, 0.10 mmol) in 1, 4-dioxane (1 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(4-fluorophenyl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.71 mmol), l-(2- methoxypyridin-4-yl) piperidin-4-amine (191 mg, 0.78 mmol) and sodium tertiary butoxide (232 mg, 2.14 mmol) in 1, 4-dioxane (1.5 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was purified by silica gel column chromatography using 2% MeOH:CH2Cl2 to afford 7-(4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (180 mg, 37%) as an off-white solid. LC-MS: 451.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.66 min. 0.05% Aq TFA:ACN; 0.8 mL/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.60 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0284] Racemic compound of Example 41 was separated using a Chiralpak-IB (250 x 20 mm, 5μιη (20 mg loading; 0.1 % DEA in n-hexane: CH2Cl2:MeOH (50:50); (A:B : 90: 10) as the mobile phase to provide the compound of Example 41 A Fraction I (+) and the compound of Example 4 IB Fraction II (-).
Example 41 A
Synthesis of (+)-7-(4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000127_0001
[0285] The compound of Example 41 A was produced as described in Example 41. Analytical data for Fraction I (+): 1H-NMR (CD3OD, 400 MHz): δ 7.75 (d, 1H), 7.37 (s, 7.27-7.19 (m, 2H), 7.10 (t, 2H), 6.52-6.50 (m, 1H), 6.12 (s, 1H), 4.63-4.61 (m, 1H), 4.20-4.08 (m, 2H), 3.95-3.87 (m, 3H), 3.80 (s, 3H), 3.08-3.00 (m, 5H), 2.10-2.01 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 451.4 [M+1]; LC-MS: 451.4 (M+1); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 2.78 min. 0.05% Aq TFA:ACN; 0.8 mL/min); UPLC (column;
Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.56 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 18.19 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) CH2Cl2:MeOH (50:50) (90: 10); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: +75.47 (c 0.25, DCM).
Example 41B
Synthesis of (-)-7-(4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000128_0001
[0286] The compound of Example 4 IB was produced as described in Example 41.
Analytical data for Fraction II (-): 1H-NMR (CD3OD, 400 MHz): δ 7.75 (d, 1H), 7.37 (s, 1H), 7.27-7.19 (m, 2H), 7.10 (t, 2H), 6.52-6.50 (m, 1H), 6.12 (s, 1H), 4.63-4.61 (m, 1H), 4.20-4.08 (m, 2H), 3.95-3.87 (m, 3H), 3.80 (s, 3H), 3.08-3.00 (m, 5H), 2.10-2.01 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 451.4 [M+1]; LC-MS: 451.4 (M+1); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 2.78 min. 0.05% Aq TFA: ACN; 0.8 mL/min); UPLC (column;
Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.57 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 20.73 min (CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 90: 10); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": -60.96 (c 0. 25, DCM).
Example 42 Synthesis of 8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000129_0001
Synthesis of8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7 , 8- dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0287] To a dry vial was added a suspension of Pd2(dba)3 (33 mg, 0.03 mmol) and (±) BINAP (68 mg, 0.10 mmol) in 1, 4-dioxane (2 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.72 mmol), l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (220 mg, 0.79 mmol) and sodium tertiary butoxide (210 mg, 2.18 mmol) in 1, 4-dioxane (2 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (20 mL) and extracted with CH2C12 (2 x 25 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 2-5%
MeOH:CH2Cl2 to afford 8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (100 mg, 28%) as an off-white solid. LC-MS: 485.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.16 min. 0.05% Aq TFA:ACN; 0.80 ml/min).
[0288] Racemic compound of Example 42 was separated using a Chiralpak IB (250 x 20 mm, 5μιη (30 mg loading; 0.1 % DEA in n-hexane: CH2Cl2:MeOH (50:50); (90: 10) as the mobile phase) to provide the compound of Example 42A Fraction I (+) and the compound of Example 42B Fraction II (-).
Example 42A Synthesis of (+)-8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000130_0001
[0289] The compound of Example 42A was produced as described in Example 42.
Analytical data for product Fraction I (+): 1H-NMR (CD3OD, 400 MHz): δ 8.20 (d, IH), 7.40 (s, IH), 7.23-7.12 (m, 4H), 7.00-6.92 (m, 2H), 4.94-4.92 (m, IH), 4.20-4.17 (m, IH), 4.09- 3.99 (m, 4H), 3.18 (t, 2H), 2.99 (s, 3H), 2.40 (s, 3H), 2.15-2.10 (m, 2H), 1.61-1.51 (m, 2H); Mass (ESI): 485.5 [M+l]; LC-MS: 485.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.21 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.86 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 12.71 min (CHIRALPAK-IB (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 90:10); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +115.21 (c 0.25, DCM).
Example 42B
Synthesis of (-)-8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000130_0002
[0290] The compound of Example 42B was produced as described in Example 42.
Analytical data for product Fraction II (-): 1H-NMR (CD3OD, 400 MHz): δ 8.21 (d, IH), 7.40 (s, IH), 7.23-7.13 (m, 4H), 7.00-6.92 (m, 2H), 4.94-4.92 (m, IH), 4.20-4.17 (m, IH), 4.09- 3.99 (m, 4H), 3.17 (t, 2H), 2.99 (s, 3H), 2.40 (s, 3H), 2.15-2.10 (m, 2H), 1.61-1.51 (m, 2H); Mass (ESI): 485.5 [M+l]; LC-MS: 485.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.20 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 14.76 min (CHIRALPAK-IB (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 90:10); flow Rate: 1.0 mL/min); Optical rotation [<x]D 19-98: -101.68 (c 0.25, DCM).
Example 43
Synthesis of 7-(4-fluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000131_0001
sealed tube
Synthesis of 7-(4-fluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4- yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0291] To a dry vial was added a suspension of Pd2(dba)3 (41 mg, 0.04 mmol) and (±) BINAP (83 mg, 0.13 mmol) in 1, 4-dioxane (1.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(4-fluorophenyl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.89 mmol), l-(2- (trifluoromethyl)pyridin-4-yl) piperidin-4-amine hydrochloride (276 mg, 0.98 mmol) and sodium tertiary butoxide (257 mg, 2.68 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was diluted with water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 3% MeOH:CH2Cl2 to afford 7-(4-fluorophenyl)-8- methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (120 mg, 27%) as an off-white solid. LC-MS: 489.5 (M+1);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.12 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.72 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0292] Racemic compound of Example 43 was separated using a Chiralpak IA (250 x 20 mm, 5μιη (30 mg loading; 0.1 % DEA in rc-hexane: CH2Cl2:MeOH (50:50); (85: 15) as the mobile phase) to provide the compound of Example 43 A Fraction I (+) and the compound of Example 43B Fraction II (-).
Example 43A
Synthesis of (+)-7-(4-fluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000132_0001
[0293] The compound of Example 43 A was produced as described in Example 43.
Analytical data for product Fraction I (+): 1H-NMR (CD3OD, 400 MHz): δ 8.19 (d, 1H), 7.40 (s, 1H), 7.27-7.21 (m, 2H), 7.20-7.18 (m, 1H), 7.10 (t, 2H), 7.00-6.98 (m, 1H), 4.65-4.63 (m, 1H), 4.18-3.98 (m, 5H), 3.20-3.12 (m, 2H), 3.00 (s, 3H), 2.13-2.10 (m, 2H), 1.61-1.51 (m, 2H); Mass (ESI): 489.5 [M+1]; LC-MS: 489.4 (M+1); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 3.07 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.67 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 18.09 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 85:15); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +76.44 (c 0.25, DCM).
Example 43B Synthesis of (-)-7-(4-fluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000133_0001
[0294] The compound of Example 43B was produced as described in Example 43.
Analytical data for product Fraction II (-): 1H-NMR (CD3OD, 400 MHz): δ 8.19 (d, 1H), 7.40 (s, 1H), 7.26-7.21 (m, 2H), 7.21-7.18 (m, 1H), 7.10 (t, 2H), 7.00-6.98 (m, 1H), 4.65-4.63 (m, 1H), 4.18-3.98 (m, 5H), 3.20-3.12 (m, 2H), 3.00 (s, 3H), 2.13-2.10 (m, 2H), 1.61-1.51 (m, 2H); Mass (ESI): 489.5 [M+1]; LC-MS: 489.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.07 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.68 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 21.11 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 85:15); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: -69.04 (c 0.25, DCM).
Example 44
Synthesis of 7-(2, 4-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000133_0002
Synthesis of 7-(2, 4-dtfluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7 , 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazin-2 -amine
[0295] To a dry vial was added a suspension of Pd2(dba)3 (41 mg, 0.04 mmol) and (±) BINAP (85 mg, 0.13 mmol) in 1, 4-dioxane (1.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(2, 4-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.90 mmol), l-(2- methoxypyridin-4-yl) piperidin-4-amine hydrochloride (242 mg, 0.99 mmol) and sodium tertiary butoxide (262 mg, 2.72 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 3% MeOH:CH2Cl2 to afford 7-(2, 4-difluorophenyl)-N-(l- (2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (140 mg, 33%) as an off-white solid. LC-MS: 469.4 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.80 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.58 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0296] Racemic compound of Example 44 was separated using a Chiralpak-IA (250 x 20 mm, 5μιη (50 mg loading; 0.1 % DEA in n-hexane: CH2Cl2:MeOH (50:50); (A:B : 70:30) as the mobile phase) to provide the compound of Example 44A Fraction I (-) and the compound of Example 44B Fraction II (+).
Example 44A
Synthesis of (-)-7-(2, 4-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000135_0001
[0297] The compound of Example 44 A was produced as described in Example 44.
Analytical data for Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, 1H), 7.40 (s, 1H), 7.10-7.00 (m, 2H), 6.98-6.92 (m, 1H), 6.53 (d, 1H), 6.16 (s, 1H), 4.98-4.97 (m, 1H), 4.29- 4.27 (m, 2H), 3.98-3.90 (m, 3H), 3.82 (s, 3H), 3.07 (s, 3H), 3.06-3.01 (m, 2H), 2.10-2.03 (m, 2H), 1.60-1.51 (m, 2H); Mass (ESI): 469.5 [M+1]; LC-MS: 469.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.89 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.60 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 9.41 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 70:30); flow Rate: 1.0 mL/min); Optical rotation [a]D 19 98: -90.38 (c 0.25, DCM).
Example 44B
Synthesis of (+)-7-(2, 4-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000135_0002
[0298] The compound of Example 44B was produced as described in Example 44.
Analytical data for Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, 1H), 7.40 (s,
1H), 7.10-7.00 (m, 2H), 6.98-6.92 (m, 1H), 6.53 (d, 1H), 6.16 (s, 1H), 4.98-4.97 (m, 1H), 4.29-4.27 (m, 2H), 3.98-3.90 (m, 3H), 3.82 (s, 3H), 3.07 (s, 3H), 3.06-3.01 (m, 2H), 2.10- 2.03 (m, 2H), 1.60-1.51 (m, 2H); Mass (ESI): 469.5 [M+1]; LC-MS: 469.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.90 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.60 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 13.33 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 70:30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: +93.02 (c 0.25, DCM).
Example 45
Synthesis of 8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000136_0001
Synthesis of8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0299] To a dry vial was added a suspension of Pd2(dba)3 (36 mg, 0.04 mmol) and (±)
BINAP (74 mg, 0.11 mmol) in 1, 4-dioxane (1.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.79 mmol), l-(2-
(trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (245 mg, 0.87 mmol) and sodium tertiary butoxide (229 mg, 2.38 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for
16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and
LCMS), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL).
The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 2%
MeOH:CH2Cl2 to afford 8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7- (3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1 , 4] oxazin-2-amine (150 mg, 36%) as an off-white solid. LC-MS: 525.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.26 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 50 mm, 1.7 μπι); RT 1.90 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0300] Racemic compound of Example 45 was separated using a Chiralpak-ADH (250 x 20 mm, 5μιη (20 mg loading; 0.1 % DEA in rc-hexane: EtOH:MeOH (50:50); (A:B : 85 : 15) as the mobile phase) to provide the compound of Example 45 A Fraction I (+) and the compound of Example 45B Fraction II (-).
Example 45A
Synthesis of (+)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000137_0001
[0301] The compound of Example 45 A was produced as described in Example 45.
Analytical data for product Fraction I (+): 1H-NMR (OMSO-d6, 400 MHz): δ 8.21 (d, 1H), 7.45 (s, 1H), 7.20-7.18 (m, 1H), 7.17-7.10 (m, 2H), 7.01 (d, 1H), 6.33-6.30 (m, 1H), 4.74- 4.71 (m, 1H), 4.17-4.07 (m, 2H), 4.02-3.97 (m, 2H), 3.91-3.88 (m, 1H), 3.03 (t, 2H), 2.91 (s, 3H), 1.93-1.89 (m, 2H), 1.49-1.38 (m, 2H); LC-MS: 525.4 (M+1); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 3.25 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.87 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC RT = 1 1.42 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) THF (70:30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 02: +61.53 (c 0.25, DCM). Example 45B
Synthesis of (-)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000138_0001
[0302] The compound of Example 45B was produced as described in Example 45.
Analytical data for product Fraction II (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.21 (d, 1H), 7.47 (s, 1H), 7.20-7.18 (m, 1H), 7.17-7.10 (m, 2H), 7.01 (d, 1H), 6.33-6.30 (m, 1H), 4.74- 4.71 (m, lH),4.17-4.07 (m, 2H), 4.02-3.97 (m, 2H), 3.91-3.88 (m, 1H), 3.03 (t, 2H), 2.91 (s, 3H), 1.93-1.89 (m, 2H), 1.49-1.38 (m, 2H); LC-MS: 525.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.25 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.88 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 13.68 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) THF (70:30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: -52.08 (c 0.25, DCM).
Example 46
Synthesis of 7-(2, 4-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin- 4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000139_0001
Synthesis of 7-(2, 4-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)- 7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazin-2-amine
[0303] To a dry vial was added a suspension of Pd2(dba)3 (31 mg, 0.03 mmol) and (±) BINAP (63 mg, 0.15 mmol) in 1, 4-dioxane (1 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(2, 4-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.67 mmol), l-(6- methylpyrimidin-4-yl) piperidin-4-amine hydrochloride (142 mg, 0.73 mmol) and sodium tertiary butoxide (193 mg, 2.01 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 3%
MeOH:CH2Cl2 to afford 7-(2, 4-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (38 mg, 12%) as an off-white solid. LC-MS: 454.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.43 min. 5 mM NH4OAc:ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1mm, 1.7 μπι); RT 1.88 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0304] Racemic compound of Example 46 was separated using a Chiralpak-ADH (250 x 20 mm, 5μιη (30 mg loading; 0.1 % DEA in n-hexane:EtOH (A:B : 75:25) as the mobile phase) to provide the compound of Example 46A Fraction I (-) and the compound of
Example 46B Fraction II (+).
Example 46A
Synthesis of (-)-7-(2, 4-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000140_0001
[0305] The compound of Example 46A was produced as described in Example 46.
Analytical data for product Fraction I (-): 1H-NMR (OMSO-d6, 500 MHz): δ 8.35 (s, 1H), 7.43 (s, 1H), 7.33 (t, 1H), 7.10-7.00 (m, 2H), 6.71 (s, 1H), 6.18 (d, 1H), 4.98 (s, 1H), 4.33- 4.29 (m, 2H), 4.15-4.10 (m, 2H), 3.98-3.88 (m, 1H), 3.07-2.99 (m, 2H), 2.97 (s, 3H), 2.24 (s, 3H), 1.94-1.89 (m, 2H), 1.43-1.38 (m, 2H); Mass (ESI): 454.5 [M+1]; LC-MS: 454.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.50 min. 5 mM NH4OAc:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.56 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 13.28 min (CHIRALPAK- AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH (70:30); flow Rate: 1.0 mL/min); Optical rotation [<x]D 19-98: -92.88 (c 0.25, DCM).
Example 46B
Synthesis of (+)-7-(2, 4-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000140_0002
[0306] The compound of Example 46B was produced as described in Example 46.
Analytical data for product Fraction II (+): 1H-NMR (DMSO-d6, 500 MHz): δ 8.35 (s, 1H),
7.43 (s, 1H), 7.33 (t, 1H), 7.10-7.00 (m, 2H), 6.71 (s, 1H), 6.18 (d, 1H), 4.98 (s, 1H), 4.33-
4.29 (m, 2H), 4.15-4.10 (m, 2H), 3.98-3.88 (m, 1H), 3.07-2.99 (m, 2H), 2.97 (s, 3H), 2.24 (s, 3H), 1.94-1.89 (m, 2H), 1.43-1.38 (m, 2H); Mass (ESI): 454.5 [M+1]; LC-MS: 454.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.55 min. 5 mM NH4OAc:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.56 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 18.61 min (CHIRALPAK- AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH (70:30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20"01: +92.62 (c 0. 25, DCM).
Example 47
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000141_0001
Synthesis of N-( l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl- 7-(2-(trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0307] To a dry vial was added a suspension of Pd2(dba)3 (33 mg, 0.03 mmol) and (±) BINAP (67 mg, 0.10 mmol) in 1, 4-dioxane (1 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(2- (trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.72 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (190 mg, 0.79 mmol) and sodium tertiary butoxide (200 mg, 2.17 mmol) in 1, 4-dioxane (1.5 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 130 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extract was dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 3% MeOH:CH2Cl2 to afford N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (130 mg, 35%) as an off-white solid. LC-MS: 517.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.11 min. 0.05% Aq TFA:ACN; 0.8 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.76 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0308] Racemic compound of Example 47 was separated using a Chiralpak-IA (250 x 20 mm, 5μιη (40 mg loading; 0.1 % DEA in rc-hexane: EtOH:MeOH (50:50); (A:B : 90: 10) as the mobile phase) to provide the compound of Example 47A Fraction I (-) and the compound of Example 47B Fraction II (+).
Example 47A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000142_0001
[0309] The compound of Example 47A was produced as described in Example 47.
Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, 1H), 7.48- 7.40 (m, 3H), 7.32 (t, 1H), 7.13 (d, 1H), 6.53 (d, 1H), 6.14 (s, 1H), 5.02-5.00 (m, 1H), 4.23- 4.18 (m, 1H), 4.15-4.10 (m, 1H), 3.95-3.89 (m, 3H), 3.81 (s, 3H), 3.07-3.03 (nm, 2H), 3.02 (s, 3H), 2.10-2.03 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 517.5 [M+l]; LC-MS: 517.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.08 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.77 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 10.44 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 80:20; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: - 83.10 (c O. 25, DCM).
Example 47B Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000143_0001
[0310] The compound of Example 47B was produced as described in Example 47.
Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, 1H), 7.48-7.40 (m, 3H), 7.32 (t, 1H), 7.13 (d, 1H), 6.53 (d, 1H), 6.14 (s, 1H), 5.02-5.00 (m, 1H), 4.23-4.18 (m, 1H), 4.15-4.10 (m, 1H), 3.95-3.89 (m, 3H), 3.81 (s, 3H), 3.07-3.03 (m, 2H), 3.02 (s, 3H), 2.10-2.03 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 517.5 [M+1]; LC-MS: 517.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.06 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.77 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 13.94 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2Cl2:MeOH (50:50) (A:B : 80:20; flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +84.41 (c O. 25, DCM).
Example 48
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000143_0002
Synthesis of N-( l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl- 7-(2-(trifluoromethoxy) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0311] To a dry vial was added a suspension of Pd2(dba)3 (35 mg, 0.03 mmol) and (±) BINAP (71 mg, 0.11 mmol) in 1, 4-dioxane (1.25 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-8-methyl-7-(2- (trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.75 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (203 mg, 0.83 mmol) and sodium tertiary butoxide (219 mg, 2.27 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 3% MeOH:CH2Cl2 to afford N-(l -(2- methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2-(trifluoromethyl) phenyl)-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (124 mg, 32%) as an off-white solid. LC-MS: 501.5 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.04 min. 0.05% Aq TFA:ACN; 0.80 ml/min);UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.76 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0312] Racemic compound of Example 48 was separated using a Chiralpak-IA (250 x 20 mm, 5μιη (40 mg loading; 0.1 % DEA in n-hexane: EtOF MeOH (80:20); (A:B : 80:20) as the mobile phase) to provide the compound of Example 48A Fraction I (-) and the compound of Example 48B Fraction II (+).
Example 48A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000145_0001
[0313] The compound of Example 48 A was produced as described in Example 48.
Analytical data for Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.80 (d, IH), 7.75 (d, IH), 7.64-7.61 (m, IH), 7.54-7.50 (m, IH), 7.47 (s, IH), 7.30 (d, IH), 6.55 (d, IH), 6.15 (s, IH), 5.02-5.00 (m, IH), 4.25-4.20 (m, IH), 4.08-4.01 (m, IH), 3.97-3.90 (m, 3H), 3.82 (s, 3H), 3.05 (t, 2H), 2.97 (s, 3H), 2.10-2.04 (m, 2H), 2.60-2.53 (m, 2H); Mass (ESI): 501.5 [M+1]; LC-MS: 501.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.00 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.71 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 11.75 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n- hexane (B) CH2Cl2:MeOH (80:20) (A:B : 80:20; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: -28.52 (c 0. 25, DCM).
Example 48B
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2- (trifluoromethyl) phenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000145_0002
[0314] The compound of Example 48B was produced as described in Example 48.
Analytical data for Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.80 (d, IH), 7.75 (d, 1H), 7.64-7.61 (m, 1H), 7.52 (t, 1H), 7.47 (s, 1H), 7.30 (d, 1H), 6.55 (d, 1H), 6.15 (s, 1H), 5.02-5.00 (m, 1H), 4.25-4.20 (m, 1H), 4.08-4.01 (m, 1H), 3.97-3.90 (m, 3H), 3.82 (s, 3H), 3.05 (t, 2H), 2.97 (s, 3H), 2.10-2.04 (m, 2H), 2.60-2.53 (m, 2H); Mass (ESI): 501.5 [M+l]; LC-MS: 501.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.00 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.71 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 14.60 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n- hexane (B) CH2Cl2:MeOH (80:20) (A:B : 80:20; flow Rate: 1.0 mL/min); Optical rotation [a]D 20-00: +35.28 (c 0. 25, DCM).
Example 49
Synthesis of 7-(2, 4-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000146_0001
Synthesis of 7 -(2, 4-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0315] To a dry vial was added a suspension of Pd2(dba)3 (31 mg, 0.03 mmol) and (±) BINAP (63 mg, 0.10 mmol) in 1, 4-dioxane (1 mL). The suspension was degassed with argon, heated to 120 °C and stirred for 3 min. A mixture of 2-chloro-7-(2, 4-difluorophenyl)- 8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.67 mmol), l-(2- (trifluoromethyl) pyridin-4-yl) piperidin-4-amine hydrochloride (208 mg, 0.74 mmol) and sodium tertiary butoxide (194 mg, 2.01 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premixure was added. The reaction mixture was heated to 120 °C and stirred for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the mixture was concentrated in vacuo. The crude material was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by silica gel column chromatography using 4% MeOH:CH2Cl2 to afford 7-(2, 4-difluorophenyl)-8- methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (200 mg, 59%) as an off-white solid. LC-MS: 507.4 (M+1);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.14 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.70 min. ACN: 0.025% TFA (Aq); 0.50 ml/min.
[0316] Racemic compound of Example 49 was separated using a Chiralpak-ADH (250 x 20 mm, 5μιη (20 mg loading; n-hexane:EtOH (A:B : 80:20) as the mobile phase) to provide the compound of Example 49A Fraction I (-) and the compound of Example 49B Fraction II
(+)·
Example 49A
Synthesis of (-)-7-(2, 4-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000147_0001
[0317] The compound of Example 49 A was produced as described in Example 49.
Analytical data for product Fraction I (-): 1H-NMR (DMSO-d6, 400 MHz): δ 8.25 (d, 1H), 7.45 (s, 1H), 7.31 (t, 1H), 7.20-7.18 (m, 1H), 7.10-7.00 (m, 3H), 6.30 (d, 1H), 4.99-4.97 (m, 1H), 4.13-4.11 (m, 2H), 4.00-3.97 (m, 2H), 3.95-3.89 (m, 1H), 3.05 (t, 2H), 2.93 (s, 3H), 1.97-1.90 (m, 2H), 1.51-1.41 (m, 2H); LC-MS: 507.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.20 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.86 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 16.93 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) n-hexane (B) EtOH (A:B : 80:20; flow Rate: 1.0 mL/min); Optical rotation [a]D 19-97: -84.76 (c 0. 25, DCM). Example 49B
Synthesis of (+)-7-(2, 4-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000148_0001
[0318] The compound of Example 49B was produced as described in Example 49.
Analytical data for product Fraction II (+): 1H-NMR (DMSO-d6, 400 MHz): δ 8.25 (d, 1H), 7.45 (s, 1H), 7.31 (t, 1H), 7.20-7.18 (m, 1H), 7.10-7.00 (m, 3H), 6.30 (d, 1H), 4.99-4.97 (m, 1H), 4.13-4.11 (m, 2H), 4.00-3.97 (m, 2H), 3.95-3.89 (m, 1H), 3.05 (t, 2H), 2.93 (s, 3H), 1.97-1.90 (m, 2H), 1.51-1.41 (m, 2H); LC-MS: 507.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.20 min. 0.05% Aq TFA:ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μιη); RT 1.86 min. ACN: 0.025% TFA (Aq); 0.50 ml/min; Chiral HPLC: RT = 24.13 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) n-hexane (B) EtOH (A:B : 80:20; flow Rate: 1.0 mL/min); Optical rotation [a]D 20 00: +82.09 (c 0. 25, DCM).
Example 50
Synthesis of (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-amine
Figure imgf000149_0001
Synthesis of (Z)-N, N-dimethyl-2-(2-methylpyridin-4-yl) ethen-l-amine
[0319] To a stirred solution of 2, 4-dimethylpyridine (14 g, 130.65 mmol) in dry THF (150 mL) under an argon atmosphere was added 2.5 M n-butyl lithium in hexane (67.5 mL, 169.85 mmol) drop-wise for 20 min at -78 °C. The reaction mixture was stirred for 1 h at - 78°C, then diethyl amine (19.5 mL, 1.89 mmol) was added drop-wise. The reaction mixture was stirred for 1 h at -78°C then DMF (20 mL, 261.13 mmol) was added drop-wise at -78 °C and stirred for 30 min. After consumption of the starting materials (monitored by TLC), the reaction was quenched with an ammonium chloride solution (200 mL) and extracted with diethyl ether (3 x 300 mL). The combined organic extracts were washed with brine (300 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford (Z)-N, N-dimethyl-2- (2-methylpyridin-4-yl) ethen-l-amine (23 g) as orange oil. This compound was used without further purification. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.02-7.99 (m, 1H), 7.31-7.23 (m, 1H), 6.90-6.83 (m, 2H), 4.94-4.88 (m, 1H), 2.82 (s, 6H), 2.30 (s, 3H); LCMS: 163.1 (M+l);
(column; X-select C-18 (50 3.0 mm, 3.5 μπι); RT 2.32 min 5mM NH4OAc: ACN; 0.80 mL/min); HPLC (column; Acquity-UPLC-BEH-C-18 2.1 X 50 mm, 1.7 μιη); RT 1.25 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 10% MeOH/ CH2Cl2 +2 drops NH4OH solution (Rf. 0.2).
Synthesis of 2-methylisonicotinaldehyde [0320] To a stirred solution of sodium metaperiodate (91 g, 425.91 mmol) in methanol (90 mL) under an argon atmosphere was added a solution of (Z)-N, N-dimethyl-2-(2- methylpyridin-4-yl) ethen-l-amine (23 g, 141.97 mmol) in methanol (90 mL). After 5 mins an exothermic reaction was observed, then the reaction mixture was cooled to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was filtered, washed with an excess of methanol and the filtrate was concentrated in vacuo. The crude material was diluted with water (100 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were washed with brine (300 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 12% EtOAc:hexanes to afford 2-methylisonicotinaldehyde (6 g, 35%) as light brown liquid. 1H-NMR (DMSO-<¾, 500 MHz): δ 10.04 (s, 1H), 8.74-8.72 (m, 1H), 7.68 (s, 1H), 7.60-7.59 (m, 1H), 2.60 (s, 3H); LCMS: 122.7 (M+l); (column; Eclipse XDB C- 18 (150 4.6 mm, 5 μιη); RT 1.93 min 0.05% Aq TFA: ACN; 1.0 mL/min); UPLC (column; Acquity-UPLC-HSS-T3 2.1 X 100 mm, 1.8 μιη); RT 1.10 min. ACN: 0.025% Aq TFA; 0.3 mL/min; TLC: 10% MeOH/ CH2C12/NH40H solution (Rf. 0.7).
Synthesis of (S)-l-(2-methylpyridin-4-yl) but-3-en-l-ol
[0321] To a stirred solution of 1M (-) ipc2B-allyl in pentane (16.66 g (51 mL), 50.90 mmol) under an argon atmosphere was cooled to -100 °C was drop-wise added a solution of 2-methyl isonicotinaldehyde (2.8 g, 23.14 mmol) in dry ether (24 mL) at -78°C over a period of 30 min. The reaction mixture was then stirred at -100 °C for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with methanol (4 mL) and 1 N HC1 (60 mL) and warmed to RT. The reaction was extracted with EtOAc (60 mL), the aqueous layer was basified with 10%> sodium hydroxide solution (adjust to pH ~ 10) at 0 °C and extracted with EtOAc (2 x 60 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 70% EtOAc:hexanes to afford (5)-l-(2-methylpyridin-4-yl) but-3-en-l-ol (3 g, 80%) as a brown liquid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.34-8.33 (m, 1H), 7.17 (s, 1H), 7.11-7.10 (m, 1H), 5.78-5.73 (m, 1H), 5.42-5.41 (m, 1H), 5.01-4.97 (m, 2H), 4.59-4.55 (m, 1H), 2.43 (s, 3H), 2.37-2.35 (m, 2H); LCMS: 164.3 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 0.48 minO.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse-XDB-C-18 150 X 4.6 mm, 5.0 μπι); RT 7.27 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min); Chiral HPLC: 96.9% RT = 17.80 min (Chiralpak IC (250 x 4.6mm, 5μηι; mobile phase (A) 0.1 % DEA in n-hexane (B) DCM: MeOH (90: 10); flow Rate: 1.0 mL/min); TLC: EtOAc (Rf. 0.3).
Synthesis of ethyl (S)-2-(((l-(2-methylpyridin-4-yl) but-3-en-l-yl) oxy) methyl) acrylate
[0322] To a stirred solution of (S)- 1 -(2-methylpyridin-4-yl) but-3-en- 1 -ol (3.2 g, 19.63 mmol) in dry THF (38 mL) under an argon atmosphere was added 60% sodium hydride (1.4 g, 58.89 mmol) portion- wise for 15 min at 20 °C. The reaction mixture stirred for 20 mins, then warmed to RT and stirred for an additional 2 h. A solution of ethyl 2-(bromo methyl) acrylate (7.5 g, 39.26 mmol) in dry THF (38 mL) was added over a period of 20 min at 20 °C and stirred at RT for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a saturated ammonium chloride solution (120 mL) and EtOAc (150 mL). The organic layer separated and the aqueous layer was extracted with EtOAc (2 x 60 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 20% EtOAc:hexanes to afford ethyl (5)-2-(((l-(2-methylpyridin-4-yl) but-3-en-l-yl) oxy) methyl) acrylate (2.6 g, 48%) as yellow liquid. 1H-NMR (DMSO-<¾, 500 MHz): δ 8.40-8.39 (m, 1H), 7.17 (s, 1H), 7.11-7.10 (m, 1H), 6.17 (s, 1H), 5.90 (s, 1H), 5.75-5.69 (m, 1H), 5.03- 4.98 (m, 2H), 4.45-4.43 (m, 1H), 4.14-4.10 (m, 2H), 4.04-4.01 (m, 2H), 2.45 (s, 4H), 2.44- 2.35 (m, 1H), 1.21 (s, 3H); LCMS: 276.4 (M+l); (column; X-select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.90 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.70 min. ACN: 0.025% TFA (Aq); 0.50 mL/min; TLC: 75% EtOAc/hexanes (Rf. 0.7).
Synthesis of ethyl (S)-6-(2-methylpyridin-4-yl)-5, 6-dihydro-2H-pyran-3-carboxylate
[0323] To a stirred solution of ethyl (5)-2-(((l-(2-methylpyridin-4-yl) but-3-en-l-yl) oxy) methyl) acrylate (2.6 g, 9.45 mmol) in CH2CI2 (52 mL) under an argon atmosphere was added 4 N HC1 in 1, 4-dioxane (4.73 mL, 18.90 mmol) drop-wise at 0 °C. The reaction mixture was warmed to RT and stirred for 5 min. The volatile components were evaporated in vacuo. To the above residue in CH2CI2 (316 mL) under an argon atmosphere was added Grubbs (II) catalyst (401 mg, 0.47 mmol) at RT. The reaction mixture was heated to 40 °C for 6 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was cooled RT and ethyl vinyl ether (1.3 mL) was added. After consumption of the starting materials, the volatile components were concentrated in vacuo. The residue was diluted with a saturated sodium bicarbonate solution (50 mL) and extracted with EtOAc (100 mL). The combined organic extracts were washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 20- 50% EtOAc:hexanes to afford ethyl (5)-6-(2-methylpyridin-4-yl)- 5, 6-dihydro-2H-pyran-3-carboxylate (1.75 g, 75%) as a brown liquid. 1H-NMR (CDC13, 400 MHz): δ 8.49-8.48 (m, 1H), 7.23 (s, 1H), 7.15-7.08 (m, 2H), 4.69-4.64 (m, 1H), 4.50-4.41 (m, 2H), 4.26-4.21 (m, 2H), 2.61 (s, 3H), 2.57-2.48 (m, 1H), 2.42-2.32 (m, 1H), 1.29 (s, 3H); LCMS: 248.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.81 min 5 mM Aq NH4OAc: ACN; 0.80 mL/min); HPLC (column; Eclipse-XDB-C-18 150 X 4.6 mm, 5 μπι); RT 9.69 min. ACN: 5 mM NH4OAc (Aq); 1.0 mL/min; TLC: 50% EtOAc/hexanes (Rf. 0.3).
Synthesis of ethyl (6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-carboxylate
[0324] To a stirred solution of ethyl (5)-6-(2-methylpyridin-4-yl)-5, 6-dihydro-2H-pyran- 3-carboxylate (1.75 g, 7.08 mmol) in MeOH (50 mL) under an argon atmosphere was added wet 10%) Pd/C (185 mg) in methanol (20 mL) at RT. The suspension was stirred under H2 atmosphere (100 Psi) for 8 h in a steel bomb. After consumption of the starting materials (monitored by LCMS), the reaction was filtered through celite and the filtrate was
concentrated in vacuo to afford ethyl (65)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- carboxylate (1.7 g, 96%) as a brown liquid [cis/trans (3: 1) isomers]. 1H-NMR (CDC13, 400 MHz): δ 8.44-8.41 (m, 1H), 7.13-7.11 (m, 1H), 7.04-7.01 (m, 1H), 4.56-4.52 (m, 1H), 4.35- 4.11 (m, 3H), 3.79-3.76 (m, 1H), 2.54 (s, 3H), 2.38-2.36 (m, 1H), 1.82-1.78 (m, 4H), 1.30 (s, 3H); LCMS: 29.6%; 250.2 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.87 min, 2.99 min 5 mM NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.5).
Synthesis of ethyl (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-carboxylate
[0325] To a stirred solution of ethyl (65)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran- 3-carboxylate (580 mg, 2.32 mmol) in EtOH (24 mL) under an argon atmosphere was added DBU (355 mg, 2.32 mmol) at RT. The reaction mixture was stirred at 90 °C for 16 h. After consumption of the starting materials (monitored by LCMS), the volatile components were evaporated in vacuo. The residue was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 50% EtOAc:hexanes to afford ethyl (3R, 65)-6-(2- methylpyridin-4-yl) tetrahydro-2H-pyran-3-carboxylate (470 mg, 81%) as a brown liquid [cis/trans (1 : 3) isomers]. 1H-NMR (CDC13, 400 MHz): δ 8.43 (d, 1H), 7.23 (s, 1H), 7.03 (d, 1H), 4.39-4.30 (m, 2H), 4.19-4.12 (m, 2H), 3.62 (t, 1H), 2.70-2.63 (m, 1H), 2.53 (s, 3H), 1.99-1.90 (m, 1H), 1.87-1.80 (m, 2H), 1.57-1.50 (m, 1H), 1.29 (s, 3H); LCMS: 250.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.99, 3.16 min 5 mM NH4OAc: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH-C-18 2.1 X 50 mm, 1.7μπι); RT 1.24 min. ACN: 0.025% TFA (Aq); 0.5 mL/min; Chiral HPLC: 24.4%, 68.2% RT = 9.14, 13.96 min (Chiralpak IA (250 x 4.6mm, 5μιη; mobile phase (A) 0.1 % TFA in n-hexane (B) DCM: MeOH (50: 50) (80: 20); flow Rate: 1.0 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.5).
[0326] Diastereomeric mixture (cis and trans-mixture) was purified by Chiralpak I A (250 x 4.6 mm: 5μιη; (20 mg loading; 0.1 % TFA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 80: 20) as mobile phase) to afford cis and trans characterization details of both fractions are as follows:
Ethyl (36',66 -6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3 -carboxylate (cis -isomer)
1H-NMR (CDC13, 400 MHz) (Cis): δ 8.41 (d, 1H), 7.11 (s, 1H), 7.01 (d, 1H), 4.57-4.51 (m, 1H), 4.39-4.32 (m, 1H), 4.28-4.21 (m, 2H), 3.75 (d, 1H), 2.56 (s, 3H), 2.55-2.54 (m, 1H), 2.39-2.31 (m, 1H), 1.89-1.75 (m, 3H), 1.31 (t, 3H); LCMS: 250.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.98 min 5 mM NH4OAc: ACN; 0.80 mL/min); HPLC (column; Acquity UPLC BEH-C-18 2.1 X 50 mm, 1.7 μπι); RT 1.34 min. ACN: 0.025% TFA (Aq); 0.5 mL/min; Chiral HPLC: 96.1% RT = 8.00 min (Chiralpak IA (250 x 4.6mm, 5μιη; mobile phase (A) 0.1 % TFA in n-hexane (B) DCM: MeOH (50: 50) (80: 20); flow Rate: 1.0 mL/min).
Ethyl (3i?,66 -6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-carboxylate (trans-isomer) 1H-NMR (CDC13, 400 MHz) (Trans): δ 8.42 (d, 1H), 7.13 (s, 1H), 7.05 (d, 1H), 4.39-4.23 (m, 2H), 4.19-4.01 (m, 2H), 3.60 (t, 1H), 2.75-2.65 (m, 1H), 2.55 (s, 3H), 2.29-2.21 (m, 1H), 2.00-1.91 (m, 1H), 1.89-1.79 (m, 1H), 1.58-1.48 (m, 1H), 1.30 (t, 3H); LCMS: 250.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.15 min 5 mM NH4OAc: ACN; 0.80 mL/min); HPLC (column; Acquity UPLC BEH-C-18 2.1 X 50 mm, 1.7 μπι); RT 1.41 min. ACN: 0.025% TFA (Aq); 0.5 mL/min; Chiral HPLC: 96.4% RT = 11.71 min (Chiralpak I A (250 x 4.6mm, 5μιη; mobile phase (A) 0.1 % TFA in n-hexane (B) DCM: MeOH (50: 50) (80: 20); flow Rate: 1.0 mL/min).
Synthesis of (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-carboxylic acid
[0327] To a stirred solution of ethyl (3R, 65)-6-(2-methylpyridin-4-yl) tetrahydro-2H- pyran-3-carboxylate (1.7 g, 6.82 mmol) in THF: EtOH (2: 1, 44 mL) under an argon atmosphere was added sodium hydroxide (2.73 g, 68.27 mmol) in water (30 mL) at RT. The reaction mixture was stirred at RT for 4 h. After consumption of the starting materials (monitored by LCMS), the volatile components were evaporated in vacuo and diluted with water (10 mL). The reaction was neutralized with a 6 N HC1 solution (10 mL) to pH ~7 and concentrated in vacuo. The residue was dissolved in MeOH (20 mL), filtered and the filtrate was concentrated in vacuo. The crude material was washed with ether (2 x 20 mL), pentane (20 mL) and dried in vacuo to afford (3R, 65)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran- 3-carboxylic acid (1.3 g, 90%) as a pale brown solid. 1H-NMR (CD3OD, 500 MHz): δ 8.35- 8.30 (m, 1H), 7.31 (s, 1H), 7.21-7.19 (m, 1H), 4.38-4.35 (m, 1H), 4.31-4.25 (m, 1H), 3.61 (t, 1H), 2.50-2.41 (m, 4H), 2.33 (s, 1H), 2.22-2.19 (m, 1H), 2.00-1.80 (m, 1H), 1.50-1.40 (m, 1H); LCMS: 222.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 0.61, 0.85 min 5 mM NH4OAc: ACN; 0.80 mL/min); Chiral HPLC: 76.5%, 19.30% RT = 7.36, 12.69 min (Chiralpak IA (250 x 4.6mm, 5μιη; mobile phase (A) 0.1 % TFA in n-hexane (B) DCM: MeOH (50: 50); flow Rate: 1.0 mL/min); TLC: 75% EtOAc/hexanes (Rf. 0.1).
Synthesis of (3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-carbonyl azide
[0328] To a stirred solution of (3R, 65)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- carboxylic acid (1.35 g, 6.10 mmol) in dry THF (63 mL) and dry DMF (10 mL) under an argon atmosphere were added triethylamine (1.36 g, 0.01 mmol) followed by isobutyl chloroformate (1.25 mL, 9.16 mmol) drop-wise at -10 °C for 10 min. The reaction mixture was stirred at -10 °C for 2 h. After consumption of the starting materials (monitored by TLC), a solution of NaN3 (600 mg, 9.16 mmol) in water (10 mL) was added and stirred at RT for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford (3R,6S)-6-(2- methylpyridin-4-yl)tetrahydro-2H-pyran-3-carbonyl azide (1.5 g,) and used without further purification. TLC: 50% EtOAc:hexanes (Rf. 0.7). Synthesis of benzyl ((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl) carbamate
[0329] To a stirred solution of (3i?,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3- carbonyl azide (1.5 g) in dry toluene (36 mL) under an argon atmosphere were added dimethylaminopyridine (150 mg, 1.21 mmol) and benzyl alcohol (987 mg, 9.28 mmol) at RT. The reaction mixture was stirred at 110 °C for 2 h. After consumption of the starting materials (monitored by TLC), the volatile components were evaporated in vacuo. The crude material was purified by silica gel column chromatography using 70% EtOAc:hexanes to afford benzyl ((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl) carbamate (500 mg, 25%) as a brown solid. 1H-NMR (CDC13, 400 MHz): δ 8.45 (d, 1H), 7.39-7.30 (m, 5H), 7.20 (s, 1H), 7.18-7.10 (m, 1H), 5.15-5.10 (m, 2H), 4.60-4.56 (m, 1H), 4.30 (d, 1H), 3.90- 3.73 (m, 2H), 3.20 (t, 1H), 2.60 (s, 3H), 2.25-2.20 (m, 1H), 2.00-1.80 (m, 1H), 1.70-1.60 (m, 1H), 1.59-1.50 (m, 1H); LCMS: 327.3 (M+l); (column; X-Select CSH-C18 (50 3.0 mm, 3.5 μπι); RT 4.84 min, 4.91 min. 5 mM NH4OAc: ACN; 0.80 ml/min); TLC: 50%
EtOAc:hexanes (Rf. 0.7).
Synthesis of (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-amine
[0330] To a stirred solution of benzyl ((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H- pyran-3-yl) carbamate (500 mg, 1.53 mmol) in methanol (30 mL) under an argon atmosphere was added 10% Pd/C (200 mg) at RT. The suspension was stirred under hydrogen
atmosphere for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was filtered through celite and the filtrate was concentrated in vacuo to afford ( R, 65)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3 -amine (280 mg) as a brown liquid. 1H- NMR (DMSO-< 5, 400 MHz): δ 8.35 (d, 1H), 7.20 (s, 1H), 7.09 (d, 1H), 4.23 (d, 1H), 3.93- 3.89 (m, 1H), 3.76-3.63 (m, 1H), 3.01 (t, 1H), 2.70-2.62 (m, 1H), 2.42 (s, 3H), 1.96-1.90 (m, 2H), 1.70-1.60 (m, 1H), 1.40-1.30 (m, 2H); LCMS: 193.3 (M+l); (column; zorbax SB-C18 (150 3.0 mm, 5 μπι); RT 6.48 min. 5 mM NH4OAc: ACN; 1.0 ml/min); TLC: 50%
EtOAc:hexanes (Rf. 0.1).
Example 51
Synthesis of 2-chloro-7-(2, 4-difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4- b] [1, 4] oxazine
Figure imgf000156_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4-dtfluorophenyl) ethan-l-one
[0331] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (500 mg, 3.14 mmol) in CH3CN (10 mL) under an argon atmosphere was added cesium carbonate (2 g, 6.28 mmol) at 0 °C. After stirring for 5 mins, 2-bromo-l-(2, 4-difluorophenyl) ethan-l-one (812 mg, 3.46 mmol) was added at 0 °C and stirred for 2 h. After consumption of the starting materials (monitored by TLC), water (20 mL) was added to the reaction and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 15-20% EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4- difluorophenyl) ethan-l-one (620 mg, 63%) as an off-white solid.1H-NMR (DMSO- 400 MHz): δ 7.85 (s, IH), 7.70-7.67 (m, IH), 7.39 (s, IH), 7.30 (t, IH), 7.19 (t, IH), 4.15 (d AB q, IH), 4.08 (d AB q, IH), 2.80 (s, 3H); LC-MS: 313.8 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.43 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.3).
Synthesis of 2-chloro-7-(2, 4-difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazine
[0332] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2,
4-difluorophenyl) ethan-l-one (600 mg, 1.91 mmol) in 1, 2-dichloroethane (12 mL) under an argon atmosphere was added sodiumtriacetoxy borohydride (812 mg, 3.83 mmol) and trifluoroacetic acid (147 mL, 1.91 mmol) at RT. The reaction mixture was stirred at RT for
16 h. After consumption of the starting materials (monitored by TLC), the reaction was basified with a IN sodium hydroxide solution (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and
concentrated in vacuo. The crude compound was purified by column chromatography using
10-15% EtOAc:hexanes to afford 2-chloro-7-(2,4-difluorophenyl)-8-methyl-7,8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (500 mg, 87%) as a pale yellow solid.1H-NMR (CDC13, 400 MHz): δ 7.70 (s, 1H), 6.99-6.81 (m, 3H), 4.92-4.90 (m, 1H), 4.24-4.20 (m, 2H), 3.10 (s, 3H); LC-MS: 298.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.20 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.6).
Example 52
Synthesis of 2-chloro-7-(2, 4-dichlorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4- b] [1, 4] oxazine
Figure imgf000157_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4-dichlorophenyl) ethan-l-one
[0333] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (500 mg, 3.14 mmol) in CH3CN (10 mL) under an argon atmosphere were added 2-bromo-l-(2, 4- dichlorophenyl) ethan-l-one (927 mg, 3.45 mmol) and cesium carbonate (2 g, 6.28 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 4 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 15-20% EtOAc :hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4-dichlorophenyl) ethan-l-one (650 mg, 60%) as an off-white solid. 1H-NMR (DMSO- 500 MHz): δ 7.92-7.88 (m, 2H), 7.69 (s, 1H), 7.60 (d, 1H), 7.49 (s, 1H), 4.31-4.29 (m, 1H), 4.06-4.01 (m, 1H), 2.77 (s, 3H); LCMS: 347 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.50 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 60% EtOAc/hexanes (Rf. 0.5).
Synthesis of 2 -chloro-7- (2, 4-dichlorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1,
4] oxazine
[0334] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4-dichlorophenyl) ethan-l-one (650 mg, 1.87 mmol) in 1, 2-dichloroethane (15 mL) under an argon atmosphere were added sodium triacetoxy borohydride (796 mg, 3.75 mmol) followed by trifluoroacetic acid (0.14 mL, 1.87 mmol) at RT. The reaction mixture was stirred at RT for 24 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with a 1 N sodium hydroxide solution (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and
concentrated in vacuo. The crude material was purified by column chromatography using 10- 20% EtOAc:hexanes to afford 2-chloro-7-(2, 4-dichlorophenyl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (350 mg, 56%) as an off-white solid. 1H-NMR (CDC13, 400 MHz): δ 7.70 (s, IH), 7.49 (s, IH), 7.25 (s, IH), 6.91 (d, IH), 5.03-5.01 (m, IH), 4.30 (d, IH), 4.21 (d, IH), 3.13 (s, 3H); LCMS: 332.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.78 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.4).
Example 53
Synthesis of 2-chloro-7-(2-chloro-4, 5-difluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b]
[1, 4] oxazine
Figure imgf000158_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethan-l-one
[0335] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (500 mg, 3.14 mmol) in acetonitrile (5 mL) under an argon atmosphere was added cesium carbonate (2.2 g, 6.91 mmol) at 0 °C. After stirring for 10 mins 2-bromo-l-(2, 4, 5-trifluorophenyl) ethan-l- one (800 mg, 3.45 mmol) was added to the reaction mixture and stirred for 30 min. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was washed with EtOAc:hexane (1 : 1, 2 x 20 mL) to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethan-l-one (550 mg, 55%) as a pale yellow solid which was used without further purification. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.88 (s, 1H), 7.70-7.63 (m, 2H), 7.53 (s, 1H), 4.17-4.10 (m, 2H), 2.81 (s, 3H); LCMS: 332.2 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.07 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc/hexanes (Rf. 0.7).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethan-l-ol
[0336] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethan-l-one (200 mg, 0.60 mmol) in MeOH (1 mL) under an argon atmosphere was added sodium borohydride (35 mg, 0.90 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with an ammonium chloride solution (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethan-l-ol (140 mg) as a pale yellow solid and used without further purification. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.69 (s, 1H), 7.68-7.53 (m, 2H), 7.41-7.39 (m, 1H), 6.00 (s, 1H), 5.20-5.18 (m, 1H), 4.20-4.17 (m, 1H), 3.99 (t, 1H), 2.83 (d, 3H);
LCMS: 334.3 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.86 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.2).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethyl methanesulfonate
[0337] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethan-l-ol (140 mg, 0.42 mmol) in CH2CI2 (2 mL) under an argon atmosphere were added triethylamine (0.12 mL, 0.84 mmol) followed by mesyl chloride (0.05 mL, 0.63 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was basified with water (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethyl methanesulfonate (200 mg) as a pale yellow solid and used without further purification. 1H-NMR (CDCI3, 500 MHz): δ 7.43 (s, 1H), 7.35-7.30 (m, 1H), 7.09-7.04 (m, 1H), 6.20 -6.18 (m, 1H), 6.13 (br s, 1H), 4.25-4.20 (m, 2H), 3.13 (s, 3H), 3.06-3.03 (m, 3H); TLC: 50% EtOAc/hexanes (Rf. 0.5).
Synthesis of 2-chloro-8-methyl-7-(2, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazine
[0338] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2, 4, 5-trifluorophenyl) ethyl methanesulfonate (200 mg, 0.48 mmol) in DMF (1 mL) under an argon atmosphere was added potassium carbonate (100 mg, 0.72 mmol) at RT. The reaction mixture was heated to 75 °C for 12 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 10% EtOAc:hexane to afford 2-chloro-8-methyl-7-(2, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (80 mg, 52%) as an off-white solid. 1H-NMR (CDC13, 500 MHz): δ 7.73 (s, 1H), 7.06-7.00 (m, 1H), 6.88-6.81 (m, 1H), 4.90 (s, 1H), 4.30-4.20 (m, 2H), 3.17 (s, 3H); LCMS: 316.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.34 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.7).
Example 54
Synthesis of 2-chloro-7-(2-chloro-4-fluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido
[5, 4-b] [1, 4] oxazine
Figure imgf000160_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2-chloro-4-fluorophenyl) ethan-l-one
[0339] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (547 mg, 3.44 mmol) in acetonitrile (5 mL) under an argon atmosphere were added cesium carbonate (2.3 g, 6.88 mmol) and 2-bromo-l-(2-chloro-4-fluorophenyl) ethan-l-one (1 g, 3.78 mmol) at 0 °C. The reaction mixture was stirred at RT for 3 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- chloro-4-fluorophenyl) ethan-l-one (1 g) as a brown solid. LCMS: 331.1 (M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.18 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 30% EtOAc/hexanes (Rf. 0.6).
Synthesis of 2-chloro-7-(2-chloro-4-fluorophenyl)-8-methyl-7 , 8-dihydro-6H-pyrimido [5, 4- b] [I, 4] oxazine
[0340] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(2- chloro-4-fluorophenyl) ethan-l-one (1 g, 3.03 mmol) in 1, 2-dichloroethane (10 mL) under an argon atmosphere were added sodium triacetoxy borohydride (1.2 g, 6.06 mmol) followed by Trifluoroacetic acid (345 mg, 3.03 mmol) at RT and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with a 1 N sodium hydroxide solution (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 15% EtOAc:hexanesto afford 2- chloro-7-(2-chloro-4-fluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (300 mg, 32%) as a pale brown solid. 1H-NMR (CDC13, 500 MHz): δ 7.74 (s, 1H), 7.21 (d, 1H), 7.00-6.93 (m, 2H), 5.03 (s, 1H), 4.33-4.29 (m, 1H), 4.25-4.20 (m, 1H), 3.15 (s, 3H); LCMS: 314.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 4.38 min. 0.05% TFA in water: ACN; 0.8 mL/min); TLC: 30% EtOAc/hexanes (Rf. 0.6).
Example 55
Synthesis of 2-chloro-7-(4-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydi
pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000162_0001
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2- (trifluoromethyl) phenyl) ethan-l-one
[0341] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (600 mg, 3.77 mmol) in acetonitrile (6 mL) under an argon atmosphere were added cesium carbonate (2.4 g, 7.54 mmol) and 2-bromo-l-(4-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (1.1 g, 4.15 mmol) at RT. The reaction mixture was stirred at RT for 3 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 20% EtOAc:hexane to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (500 mg, 36%) as an off-white solid. 1H-NMR (CDC13, 500 MHz): δ 7.56-7.50 (m, 3H), 7.40 (t, 1H), 5.75 (br s, 1H), 5.05 (s, 2H), 3.09 (d, 3H); TLC: 30% EtOAc/hexanes (R . 0.4).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2- (trifluoromethyl) phenyl) ethan-l-ol
[0342] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4- fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (500 mg, 1.37 mmol) in MeOH (2 mL) under an argon atmosphere was added sodium borohydride (76 mg, 2.06 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), volatile components were evaporated, water (50 mL) was added and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2-(trifluoromethyl) phenyl) ethan-l-ol (450 mg, 89%) as an off-white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 7.94-7.90 (m, 1H), 7.66-7.60 (m, 3H), 7.45-7.40 (m, 1H), 6.00 (s, 1H), 5.23-5.20 (m, 1H), 4.09-4.00 (m, 1H), 3.99-3.91 (m, 1H), 2.88 (d, 3H); LCMS: 366.2 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.97 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 40% EtOAc/hexanes (Rf. 0.5).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2- (trifluoromethyl) phenyl) ethyl methanesulfonate
[0343] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4- fluoro-2-(trifluoromethyl) phenyl) ethan-l-ol (450 mg, 1.23 mmol) in dichloroethane (2 mL) under an argon atmosphere were added triethylamine (0.2 mL, 1.47 mmol) and mesyl chloride (140 mg, 1.23 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was basified with water (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2- chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4-fluoro-2-(trifluoromethyl) phenyl) ethyl methanesulfonate (500 mg) as an off-white solid. 1H-NMR (CDC13, 500 MHz): δ 7.79-7.74 (m, 1H), 7.50-7.45 (m, 1H), 7.45-7.40 (m, 2H), 6.10 (br s, 1H), 4.20-4.10 (m, 2H), 3.13 (s, 1H), 3.08 (d, 3H), 2.99 (s, 3H); LCMS: 444.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.69 min 5 mM Aq NH4OAc: ACN; 0.80 mL/min); TLC: 30%
EtOAc/hexanes (Rf. 0.4).
Synthesis of 2-chloro-7 -(4-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-bJ [I, 4] oxazine
[0344] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(4- fluoro-2-(trifluoromethyl) phenyl) ethyl methanesulfonate (500 mg, 1.12 mmol) in DMF (5 mL) under an argon atmosphere was added potassium carbonate (233 mg, 1.69 mmol) at RT. The reaction mixture was heated at 80 °C for 16 h. After consumption of the starting materials (monitored by TLC), the volatile components were evaporated in vacuo. The reaction was diluted with ice cold water (50 mL) and extracted with ether (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 15% EtOAc:hexane to afford 2-chloro-7-(4-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (120 mg, 30%) as an off-white solid. 1H-NMR (CD3OD, 500 MHz): δ 7.75 (s, 1H), 7.70 (d, 1H), 7.46-7.40 (m, 1H), 7.37-7.30 (m, 1H), 5.16-5.13 (m, 1H), 4.35- 4.30 (m, 1H), 4.20-4.18 (m, 1H), 3.03 (s, 3H); LCMS: 348.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπ ; RT 4.66 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc/hexanes (R/. 0.7).
Example 56
Synthesis of 2-chloro-7-(5-chloro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydi
pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000164_0001
Synthesis of2-bromo-l-(5-chloro-2-(trifluoromethyl) phenyl) ethan-l-one
[0345] To a stirred solution of 5-chloro-2-(trifluoromethyl) benzoic acid (4 g, 17.85 mmol) in CH2CI2 (100 mL) under an argon atmosphere was added oxalyl chloride (2 mL, 19.64 mmol) and DMF (0.2 mL) at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. After consumption of acid (monitored by TLC), the volatile components were evaporated in vacuo. The obtained residue was dissolved in ether and a CH2N2 in ether solution was added at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with 48% Aq HBr solution (40 mL) at 0 °C and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with water (800 mL) and extracted with CH2CI2 (2 x 100 mL). The combined organic extracts were washed with a sodium bicarbonate solution (50 mL), water (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford 2- bromo-l-(5-chloro-2-(trifluoromethyl) phenyl) ethan-l-one (5 g, 93%) as a pale yellow liquid. 1H-NMR (CDC13, 400 MHz): δ 7.79 (d, 1H), 7.59 (d, 1H), 7.49 (s, 1H), 4.33 (s, 2H); LCMS: 301 (M-2); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.86 min. 5 mM Aq NH4OAc: ACN; 0.8 mL/min); TLC: 20% EtOAc/hexanes (Rf. 0.7).
Synthesis of l-(5-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin- 5-yl) oxy) ethan-l-one
[0346] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (2.7 g, 16.98 mmol) in CH3CN (110 mL) under an argon atmosphere was added cesium carbonate (16 g, 33.96 mmol) at 0 °C. After stirring for 5 mins 2-bromo-l-(5-chloro-2-(trifluoromethyl) phenyl) ethan-l-one (6.4 g, 21.22 mmol) was added and stirred for 30 min. After
consumption of the starting materials (monitored by TLC), the reaction was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was washed with hexane (2 x, 10 mL) to afford l-(5-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-one (5.5 g, 86%) as a pale yellow solid. LCMS: 380.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.29 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 30% EtOAc/hexanes (Rf. 0.5).
Synthesis of l-(5-chloro-2-(trifluoromethyl)phenyl)-2-((2-chloro-4-(methylamino)pyrimidin- 5-yl)oxy)ethan-l-ol)
[0347] To a stirred solution of l-(5-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-one (5.5 g, 14.43 mmol) in MeOH (110 mL) under an argon atmosphere was added sodium borohydride (1.1 g, 28.86 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with cold water. The volatile components were evaporated in vacuo, saturated ammonium chloride solution (100 mL) was added and the resulting solution was extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(5- chloro-2-(trifluoromethyl)phenyl)-2-((2-chloro-4-(methylamino)pyrimidin-5-yl) oxy) ethan- l-oi (5.5 g) as an off-white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.93 (s, 1H), 7.77 (d, 1H), 7.70-7.60 (m, 2H), 7.50-7.43 (m, 1H), 6.13 (d, 1H), 5.27-5.20 (m, 1H), 4.11-4.10 (m, 2H), 2.88 (d, 3H); LCMS: 383.1 (M+l); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 4.23 min. 0.05% Aq TFA: ACN; 0.8 mL/min); TLC: 30% EtOAc/hexanes (Rf. 0.4). Synthesis of l-(5-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin- 5-yl) oxy) ethyl methanesulfonate
[0348] To a stirred solution of l-(5-chloro-2-(trifluoromethyl)phenyl)-2-((2-chloro-4- (methylamino)pyrimidin-5-yl) oxy) ethan-l-ol) (5.3 g, 14.00 mmol) in CH2CI2 (100 mL) under an argon atmosphere was added triethylamine (6.82 mL, 49.00 mmol) followed by mesyl chloride (2.2 mL, 28.00 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 2.5 h. After consumption of the starting materials (monitored by TLC), the reaction was neutralized with a 5% sodium bicarbonate solution (100 mL) and extracted with CH2CI2 (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(5-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate (5 g, 78%) as an off-white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 7.99 (s, 1H), 7.83 (d, 1H), 7.75-7.71 (m, 2H), 7.43-7.40 (m, 1H), 6.10 (d, 1H), 4.62-4.58 (m, 1H), 4.23-4.20 (m, 1H), 3.30 (s, 3H), 2.87 (d, 3H); LCMS: 461.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.44 min. 0.05% Aq TFA: ACN; 0.8 mL/min); UPLC (column; Acquity UPLC-BEH-C 18 2.1 X 50 mm, 1.7 μπι); RT 2.49 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 30% EtOAc/hexanes (Rf. 0.5).
Synthesis of2-chloro-7-(5-chloro-2-(trifluoromethyl) phenyl) -8-methyl-l ', 8-dihydro-6H- pyrimido [5, 4-bJ [I, 4] oxazine
[0349] To a stirred solution of l-(5-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate (900 mg, 1.95 mmol) in DMF (30 mL) under an argon atmosphere was added potassium carbonate (808 mg, 5.85 mmol) at RT. The reaction mixture was stirred at 120 °C for 36 h. After consumption of the starting materials (monitored by TLC), the reaction was quenched with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 15% EtOAc:hexanes to afford 2-chloro-7-(5-chloro-2- (trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (300 mg, 42%) as a pale yellow solid. 1H-NMR (OMSO-d6, 400 MHz): δ 7.90-7.84 (m, 2H), 7.80 (d, 1H), 7.30 (s, 1H), 5.01-4.99 (m, 1H), 4.33-4.30 (m, 1H), 4.21-4.18 (m, 1H), 2.95 (s, 3H); LCMS: 365.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.75 min. 0.05% Aq TFA: ACN; 0.8 mL/min); UPLC (column; Acquity UPLC-BEH-C 18 2.1 X 50 mm, 1.7 μπι); RT 2.73 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 30% EtOAc/hexanes (Rf. 0.7).
Example 57
Synthesis of 2-chloro-7-(5-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydi pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000167_0001
Synthesis of2-bromo-l-(5-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one
[0350] To a stirred solution of copper bromide (4.1 g, 18.40 mmol) in EtOAc (25 mL) under an argon atmosphere was added l-(5-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (2 g, 9.70 mmol) in chloroform (10 mL) at 80 °C and stirred for 3 h. After consumption of the starting materials (monitored by TLC), the reaction was filtered, washed with CHCI3 and concentrated in vacuo to afford 2-bromo-l-(5-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (2 g) as a yellow liquid, used without further purification. 1H-NMR (CDC13, 500 MHz): δ 7.79-7.72 (m, IH), 7.31 (t, IH), 7.20 (d, IH), 4.32 (s, 2H); UPLC (column; Acquity UPLC BEH C- 18, 2.1 50 mm, 1.7 μιη); RT 2.45 min. ACN: 0.025% Aq TFA; 0.5 ml/min; TLC: 10% EtOAc/hexane (Rf. 0.4).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5-fluoro-2- (trifluoromethyl) phenyl) ethan-l-one
[0351] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (1.2 g, 7.50 mmol) in CH3CN (40 mL) under an argon atmosphere were added cesium carbonate (4.9 g, 15.10 mmol) and 2-bromo-l-(5-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (2.1 g, 7.50 mmol) at RT. The reaction mixture was stirred at RT for 1.5 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 20-30% EtOAc:hexanes to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5-fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (1.2 g, 47%) as an off-white solid. LCMS: 364.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.31 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc/hexane (Rf. 0.6).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5-fluoro-2- (trifluoromethyl) phenyl) ethan-l-ol
[0352] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5- fluoro-2-(trifluoromethyl) phenyl) ethan-l-one (300 mg, 0.82 mmol) in MeOH (3 mL) under an argon atmosphere was added sodium borohydride (62 mg, 1.65 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the volatile components were evaporated in vacuo. The residue was diluted with water (10 mL) and extracted with CH2CI2 (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5-fluoro-2-(trifluoromethyl) phenyl) ethan-l-ol (300 mg) as an off- white solid and used without further purification. LCMS: 366.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.03 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc/hexane (Rf. 0.6).
Synthesis of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5-fluoro-2- (trifluoromethyl) phenyl) ethyl methanesulfonate
[0353] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5- fluoro-2-(trifluoromethyl) phenyl) ethan-l-ol (300 mg, 0.82 mmol) in CH2CI2 (10 mL) under an argon atmosphere were added mesylchloride (140 mg, 1.23 mmol) and triethylamine (166 mg, 1.64 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (10 mL) and extracted with CH2CI2 (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford 2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy)-l-(5-fluoro-2-(trifluoromethyl) phenyl) ethyl methanesulfonate (405 mg) as an off- white solid and used without further purification. LCMS: 444.3 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.28 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 30% EtOAc/hexane (Rf. 0.6).
Synthesis of2-chloro-7-(5-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7 , 8-dihydro-6H- pyrimido [5, 4-b] [I, 4] oxazine
[0354] To a stirred solution of 2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy)-l-(5- fluoro-2-(trifluoromethyl) phenyl) ethyl methanesulfonate (400 mg, 0.90 mmol) in DMF (4 mL) under an argon atmosphere was added potassium carbonate (248 mg, 1.80 mmol) at RT. The reaction mixture was stirred at 80 °C for 16 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 10-20% EtOAc:hexanes to afford 2-chloro-7-(5-fluoro-2- (trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 64%) as a pale yellow solid. 1H-NMR (DMSo-< 5, 400 MHz): δ 7.80 (s, 1H), 7.79-7.74 (m, 1H), 7.25 (t, 1H), 6.90-6.86 (m, 1H), 5.01-5.00 (m, 1H), 4.27-4.22 (m, 1H), 4.15-4.10 (m, 1H), 3.06 (s, 3H); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 2.61 min. ACN: 0.025% Aq TFA; 0.5 ml/min; TLC: 30% EtOAc/hexane (Rf. 0.6).
Example 58
Synthesis of 2-chloro-7-(2-chloro-4, 5-difluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b]
[1, 4] oxazine
Figure imgf000169_0001
Synthesis of l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethan-l-one [0355] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (1.47 g, 9.24 mmol) in CH3CN (25 mL) under an argon atmosphere were added cesium carbonate (6 g, 18.49 mmol) and 2-bromo-l-(2-chloro-4, 5-difluorophenyl) ethan-l-one (2.5 g, 9.29 mmol) at RT. The reaction mixture was stirred at RT for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-one (2.5 g, 78%) as an off-white solid and used without further purification. 1H-NMR (DMSO-<¾, 500 MHz): δ 7.92-7.80 (m, 3H), 7.60 (s, 1H), 4.28 (d, 1H), 4.08 (d, 1H), 2.76 (s, 3H); TLC: 30% EtOAc/hexanes (R . 0.5).
Synthesis of l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethan-l-ol
[0356] To a stirred solution of l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-one (2.5 g, 7.18 mmol) in MeOH (25 mL) under an argon atmosphere was added sodium borohydride (531 mg, 14.36 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the volatile components were evaporated in vacuo. The residue was diluted with water (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethan-l-ol (2.2 g) as an off-white solid and used without further purification. LCMS: 350 (M+); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.39 min 5mM Aq NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc/hexanes (Rf. 0.4).
Synthesis of l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate
[0357] To a stirred solution of l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-ol (2.2 g, 6.28 mmol) in CH2CI2 (25 mL) under an argon atmosphere were added triethylamine (1.26 g, 12.57 mmol) and mesyl chloride (859 mg, 7.54 mmol) at RT. The reaction mixture was stirred at RT for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (50 mL) and extracted with CH2CI2 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(2-chloro-4, 5-difluorophenyl)-2-((2- chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate (2.5 g) as an off-white solid and used without further purification. LCMS: 428 (M+); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.39 min 5mM Aq NH4OAc: ACN; 0.80 mL/min); TLC: 50% EtOAc/hexanes (R . 0.6).
Synthesis of 2-chloro-7-(2-chloro-4, 5-difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazine
[0358] To a stirred solution of l-(2-chloro-4, 5-difluorophenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate (2.5 g, 5.84 mmol) in DMF (25 mL) under an argon atmosphere was added potassium carbonate (1.61 g, 11.68 mmol) at RT. The reaction mixture was heated at 80 °C for 6 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (200 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed with water (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 5% EtOAc:hexane to afford 2-chloro-7-(2-chloro-4, 5- difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (1.3 g, 68%) as an off-white solid. 1H-NMR (DMSO-<¾, 400 MHz): δ 7.90-7.87 (m, 1H), 7.79 (s, 1H), 7.18- 7.10 (m, 1H), 5.19-5.17 (m, 1H), 4.30 (s, 2H), 3.01 (s, 3H); LCMS: 332.1 (M+2); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.79 min 5mM Aq NH4OAc: ACN; 0.80 mL/min); TLC: 30% EtOAc/hexanes (Rf. 0.7).
Example 59
Synthesis of 2-chloro-7-(4-chloro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000171_0001
Synthesis of l-(4-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin- 5-yl) oxy) ethan-l-one
[0359] To a stirred solution of 2-chloro-4-(methylamino) pyrimidin-5-ol (250 mg, 1.57 mmol) in CH3CN (10 mL) under an argon atmosphere was added cesium carbonate (1 g, 3.14 mmol) at 0 °C. After stirring for 10 mins, 2-bromo-l-(4-chloro-2-(trifluoromethyl) phenyl) ethan-l-one (560 mg, 1.88 mmol) was added to the reaction mixture and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 5% EtOAc:hexane to afford l-(4-chloro-2- (trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethan-l-one (200 mg, 30%) as an off-white solid. 1H-NMR (CDC13, 500 MHz): δ 7.78 (s, 1H), 7.66 (d, 1H), 7.50 (s, 1H), 7.43 (d, 1H), 5.70 (s, 1H), 5.00 (s, 2H), 3.02 (d, 3H); TLC: 40%
EtOAc/hexanes (R . 0.6).
Synthesis of l-(4-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin- 5-yl) oxy) ethan-l-ol
[0360] To a stirred solution of l-(4-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-one (200 mg, 0.52 mmol) in MeOH (5 mL) under an argon atmosphere was added sodium borohydride (29 mg, 0.78 mmol) at 0 °C. The reaction mixture was warmed to RT and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the volatile components were evaporated in vacuo, water (20 mL) was added and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(4-chloro-2- (trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethan-l-ol (150 mg, 75%>) as a pale yellow solid and used without further purification. 1H-NMR (CDC13, 500 MHz): δ 7.80 (d, 1H), 7.67 (s, 1H), 7.61 (d, 1H), 7.50 (br s, 1H), 5.60 (br s, 1H), 5.55-5.49 (m, 1H), 4.10-4.07 (m, 1H), 4.01-3.97 (m, 1H), 3.05 (d, 3H), 2.67 (s, 1H); LCMS: 382.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.62 min 5mM Aq
NH4OAc: ACN; 0.80 mL/min); TLC: 40% EtOAc/hexanes (Rf. 0.5).
Synthesis of l-(4-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin- 5-yl) oxy) ethyl methanesulfonate [0361] To a stirred solution of l-(4-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethan-l-ol (150 mg, 0.39 mmol) in CH2CI2 (4 mL) under an argon atmosphere were added triethylamine (0.1 mL, 0.78 mmol) followed by mesyl chloride (0.05 mL, 0.58 mmol) at 0 °C. The reaction mixture was warmed to RT for 2 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to afford l-(4-chloro-2- (trifluoromethyl) phenyl)-2-((2-chloro-4-(methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate (180 mg) as a pale yellow solid and used without further purification. TLC: 40% EtOAc/hexanes (Rf. 0.6).
Synthesis of 2-chloro-7 -(4-chloro-2-(trifluoromethyl) phenyl) -8-methyl-l ', 8-dihydro-6H- pyrimido [5, 4-b] [I, 4] oxazine
[0362] To a stirred solution of l-(4-chloro-2-(trifluoromethyl) phenyl)-2-((2-chloro-4- (methylamino) pyrimidin-5-yl) oxy) ethyl methanesulfonate (180 mg, 0.39 mmol) in DMF (4 mL) under an argon atmosphere was added potassium carbonate (100 mg, 0.78 mmol) at RT. The reaction mixture was heated at 80 °C for 16 h. After consumption of the starting materials (monitored by TLC), the reaction was diluted with water (2 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 5% EtOAc:hexane to afford 2-chloro-7-(4-chloro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (140 mg, 36%) as a pale yellow solid. 1H-NMR (CDCI3, 400 MHz): δ 7.75 (s, 1H), 7.74 (s, 1H), 7.51 (d, 1H), 7.11 (d, 1H), 5.09-5.07 (m, 1H), 4.28-4.26 (m, 1H), 4.13-4.11 (m, 1H), 3.01 (s, 3H); TLC: 40% EtOAc/hexanes (Rf. 0.7).
Example 60
Synthesis of 7-benzyl-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8
dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000174_0001
Synthesis of 7-benzyl-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7 , 8-dihydro- 6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0363] A dry vial charged with Pd2(dba)3 (33 mg, 0.03 mmol) and (±) BINAP (67 mg, 0.10 mmol) in 1, 4-dioxane (1 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-benzyl-7-chloro-l-methyl-2, 3-dihydro-lH-pyrido [3, 4-b] [1, 4] oxazine (200 mg, 0.72 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (194 mg, 0.79 mmol) and sodium tert-butoxide (209 mg, 2.18 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst pre-mixture was added. The resultant mixture was stirred at 120 °C for a 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the volatile components were concentrated in vacuo. The residue was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 40% EtOAc:hexanes to afford 7-benzyl-N-(l-(2- methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (47 mg, 14%) as an off-white solid. LCMS: 474.5 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.07 min. 0.05% Aq TFA: ACN; 0.80 mL/min);
HPLC (column; Acquity UPLC-BEH-C18 2.1 X 50 mm, 1.7 μιη); RT 1.70 min. ACN:
0.025% Aq TFA; 0.5 mL/min; TLC: 50% EtOAc/hexanes (Rf. 0.3).
[0364] Racemic compound of Example 60 was separated using a Chiralpak IA (250 x 20 mm, 5μιη (20 mg loading; 0.1 % DEA in n-hexane: DCM: MeOH (50: 50); (85: 15) as mobile phase) to provide the compound of Example 60A Fraction I (+) and the compound of Example 60B Fraction II (-).
Example 60A
Synthesis of (+)-7-benzyl-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000175_0001
[0365] The compound of Example 60A was produced as described above. Analytical data for product Fraction (I) (+): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, IH), 7.37-7.30 (m, 3H), 7.29-7.22 (m, 3H), 6.54 (d, IH), 6.15 (s, IH), 4.01-3.97 (m, IH), 3.94-3.85 (m, 3H), 3.84 (s, 3H), 3.76-3.72 (m, IH), 3.67-3.61 (m, IH), 3.10-3.01 (m, 3H), 3.00 (s, 3H), 2.85-2.80 (m, IH), 2.04 (d, 2H), 1.59-1.47 (m, 2H); Mass (ESI): 447.4 [M+l]; LCMS: 447.5 (M+l);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.91 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC-BEH-C 18 2.1 X 50 mm, 1.7 μπι); RT 1.62 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 16.11 min
(CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH (A: B: 90: 10); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: +57.24 (c = 0. 25, DCM).
Example 60B
Synthesis of (-)-7-benzyl-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000175_0002
[0366] The compound of Example 60B was produced as described above. Analytical data for product Fraction (II) (-): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, IH), 7.37-7.30 (m, 3H), 7.29-7.22 (m, 3H), 6.54 (d, IH), 6.15 (s, IH), 4.01-3.97 (m, IH), 3.94-3.85 (m, 3H), 3.84 (s, 3H), 3.76-3.72 (m, IH), 3.67-3.61 (m, IH), 3.10-3.01 (m, 3H), 3.00 (s, 3H), 2.85-2.80 (m, IH), 2.04 (d, 2H), 1.59-1.47 (m, 2H); Mass (ESI): 447.5 [M+l]; LCMS: 447.5 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.91 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC-BEH-C 18 2.1 X 50 mm, 1.7 μπι); RT 1.62 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 95.3% RT = 18.13 min
(CHIRALPAK-IB (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH (A: B: 90: 10); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: -47.79 (c = 0. 25, DCM).
Example 61
Synthesis of 7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000176_0001
Synthesis of 7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7 , 8- dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0367] A dry vial charged with Pd2(dba)3 (88 mg, 0.09 mmol) and (±) BINAP (18 mg, 0.03 mmol) in 1, 4-dioxane (2.5 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(2-chlorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.64 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine (170 mg, 0.70 mmol) and sodium tert-butoxide (185 mg, 1.93 mmol) in 1, 4-dioxane (2.5 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the volatile components were evaporated in vacuo. The crude material was purified by column chromatography using 2% MeOH: CH2C12 to afford 7-(2-chlorophenyl)-N-(l-(2- methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (150 mg, 49%) as an off-white solid. LCMS: 467.4 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.97 min 0.05% Aq TFA: ACN; 0.80 mL/min); TLC: 5% MeOH/ CH2C12 (Rf. 0.4).
[0368] Racemic compound of Example 61 was separated using a Chiralpak-ADH (250 x
20 mm: 5μιη; (15 mg loading; 0.1% DEA in n-hexane: EtOH (A: B: 60: 40) as mobile phase) to provide the compound of Example 61 A Fraction I (-) and the compound of Example 6 IB Fraction II (+).
Example 61 A
Synthesis of (-)-7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000177_0001
[0369] The compound of Example 61 A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, IH), 7.49 (d, IH), 7.40 (s, IH), 7.37-7.30 (m, 2H), 7.05 (d, IH), 6.55 (d, IH), 6.08 (s, IH), 5.12-5.10 (m, IH), 4.24-4.19 (m, 2H), 3.99-3.90 (m, 3H), 3.84 (s, 3H), 3.08-3.01 (m, 5H), 2.11-2.09 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 467.6 [M+1]; LCMS: 467.6 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.87 min. 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column;
Eclipse XDB C-18, 150 4.6 mm, 5 μπι); RT 11.58 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; Chiral HPLC: 100% RT = 19.32 min (CHIRALPAK-ADH (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH (A: B: 60: 40); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 00: -120.38 (c = 0. 25, DCM).
Example 61B
Synthesis of (+)-7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000177_0002
[0370] The compound of Example 6 IB was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, 1H), 7.49 (d, 1H), 7.40 (s, 1H), 7.37-7.30 (m, 2H), 7.05 (d, 1H), 6.55 (d, 1H), 6.08 (s, 1H), 5.12-5.10 (m, 1H), 4.24- 4.19 (m, 2H), 3.99-3.90 (m, 3H), 3.84 (s, 3H), 3.08-3.01 (m, 5H), 2.11-2.09 (m, 2H), 1.60- 1.50 (m, 2H); Mass (ESI): 467.5 [M+1]; LCMS: 467.5 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.87 min. 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB C-18, 150 4.6 mm, 5 μπι); RT 11.58 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; Chiral HPLC: 99.5% RT = 32.89 min (CHIRALPAK-ADH (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH (A: B: 60: 40); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: +116.32 (c = 0. 25, DCM).
Example 62
Synthesis of 7-(2, 4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000178_0001
Synthesis of 7-(2, 4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7 , 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0371] A dry vial charged with Pd2(dba)3 (28 mg, 0.03 mmol) and (±) BINAP (56 mg, 0.09 mmol) in 1, 4-dioxane (2 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(2, 4-dichlorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.60 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine (138 mg, 0.66 mmol) and sodium tert-butoxide (175 mg, 1.81 mmol) in 1, 4-dioxane (2 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (50 mL) and extracted with 20% MeOH/ CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 2% MeOH: CH2CI2 to afford 7-(2, 4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4- yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (110 mg, 33%) as an off-white solid. 1H-NMR (CDC13, 400 MHz): δ 7.88 (d, 1H), 7.52 (s, 1H), 7.43 (s, 1H), 7.22- 7.20 (m, 1H), 7.00-6.98 (m, 1H), 6.55 (d, 1H), 6.08 (s, 1H), 4.94-4.92 (m, 1H), 4.79-4.71 (m, 1H), 4.20-4.10 (m, 2H), 4.00-3.90 (m, 1H), 3.89 (s, 3H), 3.80-3.75 (m, 2H), 3.06-3.00 (m, 5H), 2.16-2.10 (m, 2H), 1.60-1.58 (m, 2H); LCMS: 501.7 (M+1); (column; X-Select CSH C- 18 (50 3.0 mm, 3.5 μιη); RT 3.09 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μιη); RT 1.84 min. ACN: 0.025% Aq TFA; 0.5 ml/min; TLC: 10% MeOH/ CH2C12 (R . 0.5).
[0372] Racemic compound of Example 62 was separated using a Chiralpak IA (250 x 20 mm: 5μιη; (50 mg loading; 0.1 % DEA in rc-hexane: CH2C12: MeOH (50: 50); (A: B: 55: 45) as mobile phase) to provide the compound of Example 62A Fraction I (-) and the compound of Example 62B Fraction II (+).
Example 62A
Synthesis of (-)-7-(2, 4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000179_0001
[0373] The compound of Example 62A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.73-7.71 (m, 1H), 7.56 (s, 1H), 7.40 (s, 1H), 7.37-7.31 (m, 1H), 7.04-7.00 (m, 1H), 6.55 (d, 1H), 6.17 (s, 1H), 5.10-5.08 (m, 1H), 4.25-4.16 (m, 2H), 3.97-3.80 (m, 3H), 3.83 (s, 3H), 3.10-3.06 (m, 1H), 3.05 (s, 3H), 3.04-3.01 (m, 1H), 2.10-2.07 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 501.9 [M+1]; LCMS: 501 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.61 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.2% RT = 11.84 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 55: 45); flow Rate: 1.0 mL/min); Optical rotation [<x]D 135.24 (c = 0. 25, DCM).
Example 62B
Synthesis of (+)-7-(2, 4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000180_0001
[0374] The compound of Example 62B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.73-7.71 (m, 1H), 7.56 (s, 1H), 7.40 (s, 1H), 7.37-7.31 (m, 1H), 7.04-7.00 (m, 1H), 6.55 (d, 1H), 6.17 (s, 1H), 5.10-5.08 (m, 1H), 4.25-4.16 (m, 2H), 3.97-3.80 (m, 3H), 3.83 (s, 3H), 3.10-3.06 (m, 1H), 3.05 (s, 3H), 3.04-3.01 (m, 1H), 2.10-2.07 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 501.8 [M+l]; LCMS: 501 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.61 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.1% RT = 19.64 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 55: 45); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +95.85 (c = 0. 25, DCM).
Example 63
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine .0.
Figure imgf000181_0001
F
F
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazin-2 -amine
[0375] A dry vial charged with Pd2(dba)3 (36 mg, 0.04 mmol) and (±) BINAP (74 mg, 0.11 mmol) in 1, 4-dioxane (1.25 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-8-methyl-7-(2, 4, 5-trifluorophenyl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.79 mmol), l-(2-methoxypyridin-4-yl) piperidin- 4-amine hydrochloride (385 mg, 1.58 mmol) and sodium tert-butoxide (152 mg, 1.58 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the volatile components were evaporated in vacuo. The residue was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 3% MeOH: CH2C12 to afford N-(l -(2 -methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (100 mg, 26%) as an off-white solid. LCMS: 487.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.40 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.65 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 5% MeOH/ CH2C12 (Rf. 0.4).
[0376] Racemic compound of Example 63 was separated using a Chiralpak IA (250 x 20 mm: 5μιη; (30 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 70: 30) as mobile phase) to provide the compound of Example 63 A Fraction I (-) and the compound of Example 63B Fraction II (+).
Example 63A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000182_0001
[0377] The compound of Example 63 A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.76-7.72 (m, 1H), 7.44 (s, 1H), 7.30-7.23 (m, 1H), 7.00-6.92 (m, 1H), 6.57 (d, 1H), 6.16 (s, 1H), 5.00-4.97 (m, 1H), 4.20- 4.16 (m, 2H), 3.98-3.90 (m, 3H), 3.82 (s, 3H), 3.10 (s, 3H), 3.09-3.05 (m, 2H), 2.10-2.04 (m, 2H), 1.61-1.50 (m, 2H); Mass (ESI): 487.5 [M+1]; LCMS: 487 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.36 min. 0.05% Aq TFA: ACN; 0.80 mL/min);
UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.61 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.5% RT = 9.20 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 02: -78.51 (c = 0. 25, CH2C12).
Example 63B
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000182_0002
[0378] The compound of Example 63B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.78-7.72 (m, 1H), 7.48 (s, 1H), 7.32-7.25 (m, 1H), 7.00-6.92 (m, 1H), 6.57 (d, 1H), 6.16 (s, 1H), 5.00-4.97 (m, 1H), 4.20- 4.16 (m, 2H), 3.98-3.90 (m, 3H), 3.82 (s, 3H), 3.10 (s, 3H), 3.09-3.05 (m, 2H), 2.10-2.03 (m, 2H), 1.61-1.50 (m, 2H); Mass (ESI): 487.6 [M+l]; LCMS: 487 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.36 min. 0.05% Aq TFA: ACN; 0.80 mL/min);
HPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.62 min. ACN:
0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.7% RT = 19.10 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [<x]D 19-97: +83.61 (c = 0. 25, CH2C12).
Example 64
Synthesis of 7-(2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin
methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000183_0001
Synthesis of 7-(2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0379] A dry vial charged with Pd2(dba)3 (37 mg, 0.04 mmol) and Xantphos (70 mg, 0.12 mmol) in 1, 4-dioxane (1.25 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(2-chloro-4-fluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.80 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (389 mg, 1.60 mmol) and cesium carbonate (366 mg, 1.12 mmol) in 1, 4- dioxane (1.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the volatile components were evaporated in vacuo. The residue was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 3% MeOH: CH2C12 to afford 7-(2-chloro-4-fluorophenyl)-N-( 1 -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (100 mg, 25.8%) as an off-white solid. LCMS: 485 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.45 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.70 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 5% MeOH/ CH2C12 (Rf. 0.3).
[0380] Racemic compound of Example 64 was separated using a Chiralpak IA (250 x 4.6 mm: 5μιη; (50 mg loading; 0.1 % DEA in rc-hexane: CH2C12: MeOH (50: 50) (A: B: 70: 30) as mobile phase) to provide the compound of Example 64A Fraction I (-) and the compound of Example 64B Fraction II (+).
Example 64A
Synthesis of (-)-7-(2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4- yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000184_0001
[0381] The compound of Example 64A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.72-7.70 (m, 1H), 7.39 (s, 1H), 7.31 (d, 1H), 7.06 (d, 2H), 6.53 (d, 1H), 6.14 (s, 1H), 5.08-5.06 (m, 1H), 4.20-4.12 (m, 2H), 3.94-3.87 (m, 3H), 3.80 (s, 3H), 3.08-3.06 (m, 2H), 3.05 (s, 3H), 2.10-2.02 (m, 2H), 1.60- 1.50 (m, 2H); Mass (ESI): 485 [M+l]; LCMS: 485.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.67 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.74 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.8% RT = 8.42 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20"01 : -122.20 (c = 0. 25, CH2C12).
Example 64B
Synthesis of (+)-7-(2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4- yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000185_0001
[0382] The compound of Example 64B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.72-7.70 (m, 1H), 7.39 (s, 1H), 7.31 (d, 1H), 7.05 (d, 2H), 6.53 (d, 1H), 6.14 (s, 1H), 5.08-5.06 (m, 1H), 4.20-4.12 (m, 2H), 3.94-3.87 (m, 3H), 3.80 (s, 3H), 3.08-3.06 (m, 2H), 3.05 (s, 3H), 2.10-2.02 (m, 2H), 1.60- 1.50 (m, 2H); Mass (ESI): 485.1 [M+1]; LCMS: 485.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.66 min. 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μιη); RT 1.74 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.4% RT = 14.00 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": +120.27 (c = 0. 25, CH2C12).
Example 65
Synthesis of 7-(4-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000185_0002
Synthesis of 7-(4-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin- 4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2 -amine
[0383] A dry vial charged with Pd2(dba)3 (33 mg, 0.03 mmol) and (±) BINAP (67 mg, 0.10 mmol) in 1, 4-dioxane (1.25 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(4-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.72 mmol), l-(2-methoxypyridin-4- yl) piperidin-4-amine hydrochloride (192 mg, 0.79 mmol) and sodium tert-butoxide (207 mg, 2.16 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by preparative HPLC (Column YMC polymer C-18 (300 x 20 mm, ΙΟμιη (70 mg loading;
CH3CN: 0.05% TFA (0.1/90, 2/80, 15/70, 25/20, 35/10)) to afford 7-(4-fiuoro-2- (trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (115 mg, 31 >) as an off-white solid. LCMS: 519.6 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.12 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.77 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 10% MeOH/ CH2C12
(Rf. 0.3).
[0384] Racemic compound of Example 65 was separated using a Chiralpak IA (250 x 20 mm: 5μιη; (20 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 80: 20) as mobile phase) to provide the compound of Example 65 A Fraction I (-) and the compound of Example 65B Fraction II (+).
Example 65A
Synthesis of (-)-7-(4-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000186_0001
[0385] The compound of Example 65 A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.74-7.71 (m, 1H), 7.57 (d, 1H), 7.47 (s, 1H), 7.42-7.31 (m, 2H), 6.54 (d, 1H), 6.15 (s, 1H), 5.01-4.99 (m, 1H), 4.22-4.20 (m, 1H), 4.09-4.01 (m, 1H), 3.99-3.90 (m, 3H), 3.82 (s, 3H), 3.02 (t, 2H), 2.98 (s, 3H), 2.10-2.02 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 519.1 [M+1]; LCMS: 519.1 (M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.77 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.83 min. ACN:
0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 17.68 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) CH2C12: MeOH (50: 50) (A: B: 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 03: -44.67 (c = 0. 25, DCM); TLC: 10% MeOH/ CH2C12 (Rf. 0.3).
Example 65B
Synthesis of (+)-7-(4-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000187_0001
[0386] The compound of Example 65B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.74-7.71 (m, 1H), 7.57 (d, 1H), 7.47 (s, 1H), 7.42-7.31 (m, 2H), 6.54 (d, 1H), 6.15 (s, 1H), 5.01-4.99 (m, 1H), 4.22-4.20 (m, 1H), 4.09-4.01 (m, 1H), 3.99-3.90 (m, 3H), 3.82 (s, 3H), 3.02 (t, 2H), 2.98 (s, 3H), 2.10-2.02 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 519.1 [M+1]; LCMS: 519.1 (M+1); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.77 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.83 min. ACN:
0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 98.5% RT = 21.87 min (CHIRALPAK-IA (250 x 4.6mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +45.28 (c = 0. 25, DCM).
Example 66
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(tetrahydi
pyran-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000188_0001
Synthesis ofN-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(tetrahydro-2H-pyran- 4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazin-2 -amine
[0387] A dry vial charged with Pd2(dba)3 (42 mg, 0.04 mmol) and (±) BINAP (86 mg, 0.13 mmol) in 1, 4-dioxane (1.25 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-8-methyl-7-(tetrahydro-2H-pyran-4-yl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.92 mmol), l-(2-methoxypyridin-4-yl) piperidin- 4-amine hydrochloride (247 mg, 1.01 mmol) and sodium tert-butoxide (267 mg, 2.78 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 3% MeOH: CH2C12 to afford N-(l -(2 -methoxypyridin-4-yl) piperidin- 4-yl)-8-methyl-7-(tetrahydro-2H-pyran-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (150 mg, 37%) as an off-white solid. LCMS: 441.1 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 1.91 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.27 min. ACN: 0.025% Aq TFA; 0.5 ml/min.
[0388] Racemic compound of Example 66 was separated using a Chiralpak IA (250 x 20 mm: 5μιη; (50 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50) (A: B: 70: 30) as mobile phase) to provide the compound of Example 66A Fraction I (-) and the compound of Example 66B Fraction II (+).
Example 66A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(tetrahydro- 2H-pyran-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000189_0001
[0389] The compound of Example 66A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 500 MHz): δ 7.73 (d, 1H), 7.33 (s, 1H), 6.56 (d, 1H), 6.18 (s, 1H), 4.40-4.36 (m, 1H), 4.02-3.88 (m, 5H), 3.86 (s, 3H), 3.80-3.76 (m, 1H), 3.44-3.36 (m, 2H), 3.32-3.30 (m, 1H), 3.20 (s, 3H), 3.06 (t, 2H), 2.13-2.00 (m, 3H), 1.62-1.45 (m, 6H); Mass (ESI): 441.2 [M+1]; LCMS: 441.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 1.97 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column;
Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.30 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.3% RT = 10.94 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: -25.16 (c = 0. 25, CH2C12).
Example 66B
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(tetrahydro- 2H-pyran-4-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000189_0002
[0390] The compound of Example 66B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 500 MHz): δ 7.73 (d, 1H), 7.33 (s, 1H), 6.56 (d, 1H), 6.18 (s, 1H), 4.40-4.36 (m, 1H), 4.02-3.88 (m, 5H), 3.86 (s, 3H), 3.80-3.76 (m, 1H), 3.44-3.36 (m, 2H), 3.32-3.30 (m, 1H), 3.20 (s, 3H), 3.06 (t, 2H), 2.13-2.00 (m, 3H), 1.62-1.45 (m, 6H); Mass (ESI): 441.1 [M+1]; LCMS: 441 (M+1); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 1.88 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column;
Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.30 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 22.65 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [<x]D 19-98: +28.01 (c = 0. 25, CH2C12).
Example 67
Synthesis of 7-(5-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000190_0001
Synthesis of 7-(5-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin- 4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0391] A dry vial charged with Pd2(dba)3 (31 mg, 0.03 mmol) and (±) BINAP (64 mg, 0.10 mmol) in 1, 4-dioxane (0.5 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(5-chloro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.68 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (200 mg, 0.82 mmol) and sodium tert-butoxide (197 mg, 2.06 mmol) in 1, 4-dioxane (0.5 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the volatile components were concentrated in vacuo. The crude material was purified by column chromatography using 4% MeOH: CH2C12 to afford 7-(5-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (140 mg, 38%) as an off-white solid. LCMS: 535.8 (M+l); (column; Eclipse XDB C-18 (150 4.6 mm, 5 μπι); RT 11.96 min. 5mMAq NH4OAc: ACN; 1.0 mL/min); TLC: 5% MeOH/ CH2C12 (Rf. 0.3). [0392] Racemic compound of Example 67 was separated using a Chiralpak IA (250 x 4.6 mm: 5μιη; (30 mg loading; 0.1 % DEA in rc-hexane: CH2C12: MeOH (50: 50); (A: B: 80: 20) as mobile phase) to provide the compound of Example 67A Fraction I (-) and the compound of Example 67B Fraction II (+).
Example 67A
Synthesis of (-)-7-(5-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000191_0001
[0393] The compound of Example 67A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.80 (d, IH), 7.75 (d, IH), 7.58 (d, IH), 7.50 (s, IH), 7.28-7.25 (m, IH), 6.56 (d, IH), 6.18-6.16 (m, IH), 5.02-5.00 (m, IH), 4.24-4.20 (m, IH), 4.09-4.04 (m, IH), 3.98-3.90 (m, 3H), 3.83 (s, 3H), 3.10-3.02 (m, 2H), 3.00 (s, 3H), 2.10 (d, 2H), 1.60-1.51 (m, 2H); Mass (ESI): 535.1 [M+1]; LCMS: 535 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.80 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.6% RT= 9.55 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": - 68.51 (c = 0. 25, CH2C12).
Example 67B
Synthesis of (+)-7-(5-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000192_0001
[0394] The compound of Example 67B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.80 (d, IH), 7.75 (d, IH), 7.58 (d, IH), 7.50 (s, IH), 7.28-7.25 (m, IH), 6.56 (d, IH), 6.18-6.16 (m, IH), 5.02-5.00 (m, IH), 4.24-4.20 (m, IH), 4.09-4.04 (m, IH), 3.98-3.90 (m, 3H), 3.83 (s, 3H), 3.10-3.02 (m, 2H), 3.00 (s, 3H), 2.10 (d, 2H), 1.60-1.51 (m, 2H); Mass (ESI): 535.1 [M+l]; LCMS: 535.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.80 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.85 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT= 18.01 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: +66.65 (c = 0. 25, CH2C12).
Example 68
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 8-dimethyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000192_0002
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 8-dimethyl-l ', 8-dihydi pyrimido [5, 4-b] [I, 4] oxazin-2 -amine [0395] A dry vial charged with Pd2(dba)3 (57 mg, 0.06 mmol) and (±) BINAP (117 mg, 0.18 mmol) in 1, 4-dioxane (1 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7, 8-dimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 1.25 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine (286 mg, 1.38 mmol) and sodium tert-butoxide (362 mg, 3.76 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (20 mL) and extracted EtOAc (2 x 50 mL), 20% MeOH/ CH2C12 (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 2-5% MeOH: CH2C12 to afford N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 8-dimethyl- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (300 mg, 64%>) as a yellow solid. 1H-NMR (CDC13, 500 MHz): δ 7.88 (d, 1H), 7.41 (s, 1H), 6.40-6.38 (m, 1H), 6.08-6.05 (m, 2H), 4.00-3.90 (m, 3H), 3.87 (s, 3H), 3.79-3.72 (m, 2H), 3.57-3.50 (m, 2H), 3.10 (s, 3H), 3.02 (t, 2H), 2.11-2.02 (m, 3H), 1.60-1.50 (m, 4H), 1.30 (d, 6H); LCMS: 371 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 1.93 min. 0.05% Aq TFA: ACN; 0.80 ml/min); HPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.28 min. ACN: 0.025% Aq TFA; 0.5 ml/min; TLC: 5% MeOH/ CH2C12 (R . 0.2).
[0396] Racemic compound of Example 68 was separated using a Chiralpak IA (250 x 20 mm: 5μιη; (50 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 70: 30) as mobile phase) to provide the compound of Example 68 A Fraction I (-) and the compound of Example 68B Fraction II (+).
Example 68A
Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 8-dimethyl-7, 8-dihydro-
6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000193_0001
[0397] The compound of Example 68 A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (DMSO-<¾, 500 MHz): δ 7.78 (d, 1H), 7.39 (s, 1H), 6.53- 6.50 (m, 1H), 6.10-6.07 (m, 2H), 3.95-3.78 (m, 5H), 3.75 (s, 3H), 3.59-3.53 (m, 1H), 2.99 (s, 3H), 2.90 (t, 2H), 1.89-1.81 (m, 2H), 1.45-1.35 (m, 2H), 1.17 (d, 3H); Mass (ESI): 371
[M+1]; LCMS: 371 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 1.94 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.26 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 10.46 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n- hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20-00: -19.08 (c = 0. 25, DCM).
Example 68B
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 8-dimethyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000194_0001
[0398] The compound of Example 68B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (DMSO-<¾, 500 MHz): δ 7.78 (d, 1H), 7.39 (s, 1H), 6.53-6.50 (m, 1H), 6.10-6.07 (m, 2H), 3.95-3.78 (m, 5H), 3.75 (s, 3H), 3.59-3.53 (m, 1H), 2.99 (s, 3H), 2.90 (t, 2H), 1.89-1.81 (m, 2H), 1.45-1.35 (m, 2H), 1.17 (d, 3H); Mass (ESI): 371 [M+1]; LCMS: 371 (M+1); (column; X-Select CSH C-18 (50 x 3.0 mm, 3.5 μπι); RT 1.94 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.27 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 19.24 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min);
Optical rotation [a]D 19": +12.94 (c = 0. 25, DCM).
Example 69 Synthesis of 7-(5-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000195_0001
Synthesis of 7-(5-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin- 4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0399] A dry vial charged with Pd2(dba)3 (33 mg, 0.03 mmol) and (±) BINAP (67 mg, 0.10 mmol) in 1, 4-dioxane (1.5 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(5-fluoro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.72 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine (164 mg, 0.79 mmol) and sodium tert-butoxide (207 mg, 2.16 mmol) in 1, 4-dioxane (1.5 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 4% MeOH: CH2C12 to afford 7-(5-fluoro-2-(trifluoromethyl) phenyl)- N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (100 mg, 27%) as an off-white solid. LCMS: 519.1 (M+l); (column; X- Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.54 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μπι); RT 1.75 min. ACN:
0.025% Aq TFA; 0.5 mL/min; TLC: 5% MeOH/ CH2C12 (R . 0.4).
[0400] Racemic compound of Example 69 was separated using a Chiralpak IA (250 x 4.6 mm: 5μιη; (25 mg loading; 0.1 % DEA in n-hexane: THF: MeOH (50: 50); (A: B: 70: 30) as mobile phase) to provide the compound of Example 69A Fraction I (-) and the compound of Example 69B Fraction II (+).
Example 69A Synthesis of (-)-7-(5-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000196_0001
[0401] The compound of Example 69A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.88-7.83 (m, IH), 7.74 (d, IH), 7.49 (s, IH), 7.29 (t, IH), 6.99 (d, IH), 6.55-6.51 (m, IH), 6.16 (s, IH), 5.02-5.00 (m, IH), 4.22 (d, IH), 4.08 (d, IH), 3.97-3.89 (m, 3H), 3.81 (s, 3H), 3.08-3.01 (m, 2H), 3.00 (s, 3H), 2.10-2.06 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 519 [M+l]; LCMS: 519.1 (M+l);
(column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.45 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.75 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 6.62 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B; 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 00: - 26.80 (c = 0. 25, CH2C12).
Example 69B
Synthesis of (+)-7-(5-fluoro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000196_0002
[0402] The compound of Example 69B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.88-7.83 (m, 1H), 7.74 (d, 1H), 7.49 (s, 1H), 7.29 (t, 1H), 6.99 (d, 1H), 6.55-6.51 (m, 1H), 6.16 (s, 1H), 5.02-5.00 (m, 1H), 4.22 (d, 1H), 4.08 (d, 1H), 3.97-3.89 (m, 3H), 3.81 (s, 3H), 3.08-3.01 (m, 2H), 3.00 (s, 3H), 2.10-2.06 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 519 [M+1]; LCMS: 519 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.42 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.74 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 100% RT = 9.84 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A:B; 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +38.64 (c = 0. 25, CH2C12).
Example 70
Synthesis of 7-(2-chloro-4, 5-difluorophenyl)-N-(3-fluoro-4-(4-methyl-lH-imidazol-l-yl) phenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000197_0001
Synthesis of 7-(2-chloro-4, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8- methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0403] A dry vial charged with Pd2(dba)3 (34 mg, 0.03 mmol) and (±) BINAP (70 mg, 0.11 mmol) in 1, 4-dioxane (1.25 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(2-chloro-4, 5-difluorophenyl)-8-methyl-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.75 mmol), l-(2-methoxypyridin-4-yl) piperidin- 4-amine (201 mg, 0.83 mmol) and sodium tert-butoxide (217 mg, 0.22 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (20 mL) and extracted with CH2C12 (2 x 25 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 3% MeOH:CH2Cl2 to afford 7-(2-chloro-4, 5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (100 mg, 26%) as an off-white solid. 1H-NMR (CD3OD, 400 MHz): δ 7.72 (d, 1H), 7.55-7.50 (m, 1H), 7.41 (s, 1H), 6.92-6.89 (m, 1H), 6.55 (d, 1H), 6.16 (s, 1H), 5.09-5.07 (m, 1H), 4.24-4.11 (m, 2H), 3.99-3.90 (m, 3H), 3.83 (s, 3H), 3.10-3.03 (m, 5H), 2.10-2.05 (m, 2H), 1.60-1.50 (m, 2H); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μιη); RT 1.74 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 5% MeOH/ CH2C12 (Rf. 0.4).
[0404] Racemic compound of Example 70 was separated using a Chiralpak IA (250 x 4.6 mm: 5μιη; (25 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 70: 30) as mobile phase) to provide the compound of Example 70A Fraction I (-) and the compound of Example 70B Fraction II (+).
Example 70A
Synthesis of (-)-7-(2-chloro-4, 5-difluorophenyl)-N-(3-fluoro-4-(4-methyl-lH-imidazol-l- yl) phenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000198_0001
[0405] The compound of Example 70A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, 1H), 7.54-7.55 (m, 1H), 7.44 (s, 1H), 6.91 (t, 1H), 6.55 (d, 1H), 6.15 (s, 1H), 5.10-5.06 (m, 1H), 4.21-4.11 (m, 2H), 3.99-3.90 (m, 3H), 3.81 (s, 3H), 3.10-3.01 (m, 5H), 2.10-2.05 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 502.9 [M+1]; LCMS: 502.9 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.47 min. 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB C-18, 150 x 4.6 mm, 5.0 μπι); RT 12.51 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; Chiral HPLC: 97.1% RT = 7.78 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) CH2C12: MeOH (50: 50) (A: B; 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 02: -88.64 (c = 0. 25, CH2C12).
Example 70B
Synthesis of (+)-7-(2-chloro-4, 5-difluorophenyl)-N-(3-fluoro-4-(4-methyl-lH-imidazol- 1-yl) phenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000199_0001
[0406] The compound of Example 70B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.73 (d, IH), 7.53 (t, IH), 7.44 (s, IH), 6.91 (t, IH), 6.55 (d, IH), 6.15 (s, IH), 5.10-5.06 (m, IH), 4.21-4.11 (m, 2H), 3.99- 3.90 (m, 3H), 3.81 (s, 3H), 3.10-3.01 (m, 5H), 2.10-2.05 (m, 2H), 1.60-1.50 (m, 2H); Mass (ESI): 502.9 [M+l]; LCMS: 502.9 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.46 min. 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB C- 18, 150 4.6 mm, 5.0 μπι); RT 12.49 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; Chiral HPLC: 96.9% RT = 18.34 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B; 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +95.05 (c = 0. 25, CH2C12).
Example 71
Synthesis of 7-(4-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000200_0001
Synthesis of 7-(4-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin- 4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-bJ [I, 4] oxazin-2- amine
[0407] A dry vial charged with Pd2(dba)3 (25 mg, 0.02 mmol) and (±) BINAP (47 mg, 0.08 mmol) in 1, 4-dioxane (1 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(4-chloro-2-(trifluoromethyl) phenyl)-8-methyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.55 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (148 mg, 0.60 mmol) and sodium tert-butoxide (158 mg, 1.60 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 2 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (50 mL) and extracted with CH2C12 (2 x 25 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 10-50% EtOAc:hexanes to afford 7-(4-chloro-2- (trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (150 mg, 51%) as a pale yellow solid.
LCMS: 535.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.69 min 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB-C18 (150 X 4.6 mm, 5.0 μπι); RT 12.59 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; TLC: 5% MeOH/ CH2C12 (R/.
0.3).
[0408] Racemic compound of Example 71 was separated using a Chiralpak IA (250 x 4.6 mm: 5μιη; (25 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 85: 15) as mobile phase) to provide the compound of Example 71 A Fraction I (-) and the compound of Example 7 IB Fraction II (+).
Example 71A Synthesis of (-)-7-(4-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000201_0001
[0409] The compound of Example 71 A was produced as described above. Analytical data for product Fraction I (-): 1H-NMR (CD3OD, 400 MHz): δ 7.80 (s, 1H), 7.73 (d, 1H), 7.65 (d, 1H), 7.46 (s, 1H), 7.30 (d, 1H), 6.53 (d, 1H), 6.13 (s, 1H), 5.00-4.98 (m, 1H), 4.21 (d, 1H), 4.04 (d, 1H), 3.92-3.87 (m, 3H), 3.81 (s, 3H), 3.02 (t, 2H), 2.99 (s, 3H), 2.08 (d, 2H), 1.60- 1.50 (m, 2H); Mass (ESI): 535.7 [M+1]; LCMS: 535.6 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.20 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μιη); RT 1.90 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.5% RT = 11.49 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B; 75: 25); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 00: -61.95 (c = 0. 25, CH2C12).
Example 71B
Synthesis of (+)-7-(4-chloro-2-(trifluoromethyl) phenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000201_0002
[0410] The compound of Example 71B was produced as described above. Analytical data for product Fraction II (+): 1H-NMR (CD3OD, 400 MHz): δ 7.80 (s, 1H), 7.73 (d, 1H), 7.65 (d, 1H), 7.46 (s, 1H), 7.30 (d, 1H), 6.53 (d, 1H), 6.13 (s, 1H), 5.00-4.98 (m, 1H), 4.21 (d, 1H), 4.04 (d, 1H), 3.92-3.87 (m, 3H), 3.81 (s, 3H), 3.02 (t, 2H), 2.99 (s, 3H), 2.08 (d, 2H), 1.60- 1.50 (m, 2H); Mass (ESI): 535.7 [M+1]; LCMS: 535.6 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.20 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μπι); RT 1.89 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 98.9% RT = 14.08 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in rc-hexane (B) CH2C12: MeOH (50: 50) (A: B; 75: 25); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 01: +56.36 (c = 0. 25, CH2C12).
Example 72
Synthesis of 8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)- 7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000202_0001
Synthesis of 8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7-(3 , 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0411] A dry vial charged with Pd2(dba)3 (33 mg, 0.72 mmol) and (±) BINAP (66 mg, 0.10 mmol) in 1, 4-dioxane (1 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-8-methyl-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (230 mg, 0.73 mmol), (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro- 2H-pyran-3 -amine (154 mg, 0.80 mmol) and sodium tert-butoxide (210 mg, 2.19 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), The reaction was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 3% MeOH: CH2C12 to afford 8-methyl-N-((3i?, 6S)-6-(2- methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazin-2-amine (225 mg, 60%) as a brown solid. 1H-NMR (DMSO- d6, 500 MHz): δ 8.36-8.34 (m, 1H), 7.48 (s, 1H), 7.22-7.20 (m, 1H), 7.20-7.13 (m, 3H), 6.30- 3.627 (m, 1H), 4.78-4.76 (m, 1H), 4.30 (d, 1H), 4.20-4.14 (m, 3H), 3.90-3.83 (m, 1H), 3.20 (t, 1H), 2.99 (s, 3H), 2.42 (s, 3H), 2.07-1.98 (m, 2H), 1.70-1.63 (m, 1H), 1.50-1.43 (m, 1H); LCMS: 472.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.36 min 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB C-18 150 X 4.6 mm, 5.0 μπι); RT 10.88 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; TLC: 5% MeOH/ CH2C12 (R/. 0.4).
[0412] Racemic compound of Example 72 was separated using a (Chiralpak-IA (250 x 4.6 mm, 5μιη (25 mg loading; 0.1 % DEA in rc-hexane: CH2C12: MeOH (50: 50); (A: B: 85: 15) as mobile phase) to provide the compound of Example 72A Fraction I (+) and the compound of Example 72B Fraction II (-).
Example 72A
Synthesis of (+)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000203_0001
[0413] The compound of Example 72A was produced as described above. Analytical data for product Fraction I (+): 1H-NMR (DMSO-<¾, 400 MHz): δ 8.37-8.35 (m, 1H), 7.48 (s, 1H), 7.21-7.19 (m, 1H), 7.17-7.10 (m, 3H), 6.29 (d, 1H), 4.78-4.76 (m, 1H), 4.30 (d, 1H), 4.18-4.16 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 2.99 (s, 3H), 2.41 (s, 3H), 2.08-1.99 (m, 2H), 1.71-1.61 (m, 1H), 1.50-1.41 (m, 1H); Mass (ESI): 472.1 [M+l]; LCMS: 472.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.34 min. 0.05% Aq TFA: ACN; 0.80 ml/min); HPLC (column; Eclipse XDB-C-18, 150 x 4.6 mm, 5.0 μπι); RT 10.94 min. ACN: 5mM Aq NH4OAc; 0.5 mL/min; Chiral HPLC: 99.2% RT = 16.10 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D +32.17 (c = 0. 25, CH2C12).
Example 72B
Synthesis of (-)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-7-(3, 4, 5-trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000204_0001
[0414] The compound of Example 72B was produced as described above. Analytical data for product Fraction II (-): 1H-NMR (DMSO-< 5, 400 MHz): δ 8.37-8.35 (m, 1H), 7.48 (s, 1H), 7.21-7.19 (m, 1H), 7.17-7.10 (m, 3H), 6.29 (d, 1H), 4.78-4.76 (m, 1H), 4.30 (d, 1H), 4.18-4.16 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 2.99 (s, 3H), 2.41 (s, 3H), 2.08-1.99 (m, 2H), 1.71-1.61 (m, 1H), 1.50-1.41 (m, 1H); Mass (ESI): 472.1 [M+l]; LCMS: 472.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.38 min. 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB-C-18, 150 4.6 mm, 5.0 μπι); RT 11.00 min. ACN: 5mM Aq NH4OAc; 1.0 mL/min; Chiral HPLC: 99.6% RT = 15.15 min
(CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 02: - 42.24 (c = 0. 25, CH2C12).
Example 73
Synthesis of 8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)- 7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000205_0001
Synthesis of 8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7-(o- tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0415] A dry vial charged with Pd2(dba)3 (38 mg, 0.04 mmol) and (±) BINAP (78 mg, 0.12 mmol) in 1, 4-dioxane (1 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-8-methyl-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (230 mg, 0.83 mmol), (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- amine (176 mg, 0.91 mmol) and sodium tert-butoxide (240 mg, 2.51 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 2% MeOH: CH2C12 to afford 8-methyl-N-((3i?, 65)-6-(2-methylpyridin-4-yl) tetrahydro-2H- pyran-3-yl)-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (190 mg, 49%) as a brown solid. 1H-NMR (DMSO-<¾, 500 MHz): δ 8.39-8.37 (m, 1H), 7.42 (s, 1H), 7.22-7.10 (m, 5H), 6.83 (d, 1H), 6.39 (br s, 1H), 4.97-4.95 (m, 1H), 4.30 (d, 1H), 4.18-4.03 (m, 2H), 3.90-3.81 (m, 1H), 3.20 (t, 1H), 2.96 (s, 3H), 2.42 (s, 3H), 2.35 (s, 3H), 2.07-1.97 (m, 2H), 1.70-1.64 (m, 1H), 1.50-1.42 (m, 1H), 1.22-1.20 (m, 1H); LCMS: 432.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.30 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.68 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 5% MeOH/ CH2C12 (Rf. 0.4).
[0416] Racemic compound of Example 73 was purified by (Chiral pack-ADH (250 x 4.6 mm, 5μιη (30 mg loading; 0.1 % DEA in n-hexane: EtOH: MeOH (50: 50); (A: B: 75: 25) as mobile phase) to provide the compound of Example 73 A Fraction I (+) and the compound of Example 73B Fraction II (-)
Example 73A Synthesis of (+)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000206_0001
[0417] The compound of Example 73 A was produced as described above. Analytical data for product Fraction I (+) 1H-NMR (DMSO-< 5, 500 MHz): δ 8.37-8.35 (m, 1H), 7.42 (s, 1H), 7.24-7.10 (m, 5H), 6.86 (d, 1H), 6.20 (d, 1H), 4.92-4.89 (m, 1H), 4.30 (d, 1H), 4.18-4.10 (m, 2H), 4.08-4.03 (m, 1H), 3.90-3.81 (m, 1H), 3.20 (t, 1H), 2.94 (s, 3H), 2.41 (s, 3H), 2.36 (s, 3H), 2.08-1.98 (m, 2H), 1.70-1.62 (m, 1H), 1.52-1.42 (m, 1H); Mass (ESI): 432.1 [M+l]; LCMS: 432.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.35 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH-C-18, 50 2.1 mm, 1.7 μπι); RT 1.63 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.6% RT = 10.86 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +75.15 (c = 0. 25, CH2C12).
Example 73B
Synthesis of (-)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-7-(o-tolyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000206_0002
[0418] The compound of Example 73B was produced as described above. Analytical data for product Fraction II (-): 1H-NMR (DMSO-< 5, 500 MHz): δ 8.37-8.35 (m, 1H), 7.42 (s, 1H), 7.24-7.10 (m, 5H), 6.86 (d, 1H), 6.20 (d, 1H), 4.92-4.89 (m, 1H), 4.29 (d, 1H), 4.18-4.10 (m, 2H), 4.08-4.03 (m, 1H), 3.90-3.81 (m, 1H), 3.20 (t, 1H), 2.92 (s, 3H), 2.41 (s, 3H), 2.36 (s, 3H), 2.08-1.98 (m, 2H), 1.70-1.62 (m, 1H), 1.52-1.42 (m, 1H); Mass (ESI): 432.2 [M+l]; LCMS: 432.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.33 min. 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity BEH-C-18, 50 2.1 mm, 1.7 μπι); RT 1.60 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.3% RT = 10.30 min (CHIRALPAK-IA (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 85: 15); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": - 87.04 (c = 0. 25, CH2C12).
Example 74
Synthesis of 7-(3, 5-difluorophenyl)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000207_0001
Synthesis of 7-(3, 5-dtfluorophenyl)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2 -amine
[0419] A dry vial charged with Pd2(dba)3 (38 mg, 0.04 mmol) and (±) BINAP (78 mg, 0.12 mmol) in 1, 4-dioxane (1 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 2-chloro-7-(3, 5-difluorophenyl)-8-methyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (250 mg, 0.83 mmol), (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H- pyran-3-amine (177 mg, 0.88 mmol) and sodium tert-butoxide (242 mg, 2.51 mmol) in 1, 4- dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (30 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 2% MeOH: CH2C12 to afford 7-(3, 5-difluorophenyl)-8-methyl-N- ((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1 , 4] oxazin-2-amine (230 mg, 60%) as a pale yellow solid. 1H-NMR (DMSO-<¾, 500 MHz): δ 8.37-8.35 (m, 1H), 7.47 (s, 1H), 7.21-7.17 (m, 2H), 7.16-7.10 (m, 1H), 6.92-6.89 (m, 2H), 6.30-6.28 (m, 1H), 4.81-4.79 (m, 1H), 4.30 (d, 1H), 4.20-4.10 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 3.00 (s, 3H), 2.41 (s, 3H), 2.01-1.90 (m, 2H), 1.71-1.61 (m, 1H), 1.50-1.40 (m, 1H); LCMS: 454.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.32 min 0.05% Aq TFA: ACN; 0.80 mL/min); HPLC (column; Eclipse XDB C-18 150 X 4.6 mm, 5.0 μπι); RT 10.70 min. ACN: 5 mM Aq NH4OAc; 1.0 mL/min; TLC: 5% MeOH/ CH2C12 (Rf. 0.4).
[0420] Racemic compound of Example 74 was separated using a (Chiralpak-ADH (250 x 4.6 mm, 5μιη (40 mg loading; 0.1 % DEA in rc-hexane: CH2C12: MeOH (50: 50); (A: B: 70: 30) as mobile phase to provide the compound of Example 74 A Fraction I (+) and the compound of Example 74B Fraction II (-).
Example 74A
Synthesis of (+)-7-(3, 5-difluorophenyl)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000208_0001
[0421] The compound of Example 74 was produced as described above. Analytical data for product Fraction I (+): 1H-NMR (OMSO-d6, 400 MHz): δ 8.37-8.35 (m, 1H), 7.47 (s,
1H), 7.22-7.17 (m, 2H), 7.16-7.10 (m, 1H), 6.92-6.88 (m, 2H), 6.28 (d, 1H), 4.80-4.78 (m,
1H), 4.30 (d, 1H), 4.20-4.10 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 2.99 (s, 3H), 2.44 (s,
3H), 2.06-1.95 (m, 2H), 1.70-1.60 (m, 1H), 1.50-1.43 (m, 1H); Mass (ESI): 454.4 [M+l];
LCMS: 454.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.30 min.
0.05% Aq TFA: ACN; 0.80 ml/min); HPLC (column; Eclipse XDB C-18, 150 x 4.6 mm, 5.0 μπι); RT 10.89 min. ACN: 5 mM aq NH4OAc; 1.0 mL/min; Chiral HPLC: 98.3% RT = 8.94 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) EtOH: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D +41.63 (c = 0. 25, CH2C12).
Example 74B
Synthesis of (-)-7-(3, 5-difluorophenyl)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000209_0001
[0422] The compound of Example 74B was produced as described above. Analytical data for product Fraction II (-): 1H-NMR (DMSO-<¾, 400 MHz): δ 8.37-8.35 (m, 1H), 7.47 (s, 1H), 7.22-7.17 (m, 2H), 7.16-7.10 (m, 1H), 6.92-6.88 (m, 2H), 6.28 (d, 1H), 4.80-4.78 (m, 1H), 4.30 (d, 1H), 4.20-4.10 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 2.99 (s, 3H), 2.44 (s, 3H), 2.06-1.95 (m, 2H), 1.70-1.60 (m, 1H), 1.50-1.43 (m, 1H); Mass (ESI): 454.4 [M+l]; LCMS: 454.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.28 min. 0.05% Aq TFA: ACN; 0.80 ml/min); HPLC (column; Eclipse XDB C-18, 150 4.6 mm, 5.0 μπι); RT 10.76 min. ACN: 5 mM aq NH4OAc; 1.0 mL/min; Chiral HPLC: 99.6% RT = 17.14 min (CHIRALPAK-AD-H (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n- hexane (B) EtOH: MeOH (50: 50) (A: B: 70: 30); flow Rate: 1.0 mL/min); Optical rotation [a]D 19-97: -36.17 (c = 0. 25, CH2C12).
Example 75
Synthesis of l-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)- 2-(2, 4, 5-trifluorophenyl)-2, 3-dihydro-lH-pyrido [3, 4-b] [1, 4] oxazin-7-amine
Synthesis of l-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-2- ( 2, 4, 5-trifluorophenyl)-2, 3-dihydro- lH-pyrido [ 3, 4-b ][ 1, 4]oxazin- 7 -amine
[0423] A dry vial charged with Pd2(dba)3 (36 mg, 0.04 mmol) and (±) BINAP (74 mg, 0.12 mmol) in 1, 4-dioxane (1.25 mL) at RT was degassed with argon and stirred at 120 °C for 3 min. A mixture of 7-chloro-l-methyl-2-(2, 4, 5-trifluorophenyl)-2, 3-dihydro-lH-pyrido [3, 4-b] [1, 4] oxazine (250 mg, 0.79 mmol), (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro- 2H-pyran-3 -amine (167 mg, 0.87 mmol) and sodium tert-butoxide (228 mg, 2.38 mmol) in 1, 4-dioxane (1.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction was diluted with water (50 mL) and extracted with CH2C12 (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column
chromatography using 3% MeOH: CH2C12 to afford l-methyl-N-((3i?, 6S)-6-(2- methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-2-(2, 4, 5-trifluorophenyl)-2, 3-dihydro- \H- pyrido [3, 4-b] [1, 4] oxazin-7-amine (105 mg, 28%) as an off-white solid. LCMS: 472.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.31 min 0.05% Aq TFA: ACN; 0.80 mL/min); UPLC (column; Acquity UPLC BEH C-18 50 X 2.1 mm, 1.7 μπι); RT 1.64 min. ACN: 0.025% Aq TFA; 0.5 mL/min; TLC: 5% MeOH/ CH2C12 (Rf. 0.3).
[0424] Racemic compound of Example 75 was separated using a Chiralpak IC (250 x 4.6 mm: 5μιη; (15 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (50: 50); (A: B: 80: 20) as mobile phase) to provide the compound of Example 75 A Fraction I (+) and the compound of Example 75B Fraction II (-).
Example 75A
Synthesis of (+)-l-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-2-(2, 4, 5-trifluorophenyl)-2, 3-dihydro-lH-pyrido [3, 4-b] [1, 4] oxazin-7-amine
Figure imgf000211_0001
[0425] The compound of Example 75 was produced as described above. Analytical data for product Fraction I (+): 1H-NMR (DMSO-< 5, 400 MHz): δ 8.36-8.34 (m, 1H), 7.69-7.61 (m, 1H), 7.50 (s, 1H), 7.21 (s, 1H), 7.16-7.14 (m, 1H), 7.09-7.00 (m, 1H), 6.28 (d, 1H), 5.00- 4.98 (m, 1H), 4.29 (d, 1H), 4.18-4.10 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 3.00 (s, 3H), 2.41 (s, 3H), 2.05-1.92 (m, 2H), 1.70-1.61(m, 1H), 1.50-1.40 (m, 1H); Mass (ESI): 472.1 [M+1]; LCMS: 472.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.45 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.62 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.4% RT = 16.05 min (CHIRALPAK-IC (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n- hexane (B) CH2C12: MeOH (50: 50) (A: B; 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 19-95: +42.00 (c = 0. 25, CH2C12).
Example 75B
Synthesis of (-)-l-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-2-(2, 4, 5-trifluorophenyl)-2, 3-dihydro-lH-pyrido [3, 4-b] [1, 4] oxazin-7-amine
Figure imgf000211_0002
[0426] The compound of Example 75B was produced as described above. Analytical data for product Fraction II (-): 1H-NMR (DMSO-< 5, 400 MHz): δ 8.36-8.34 (m, 1H), 7.69-7.61 (m, 1H), 7.50 (s, 1H), 7.21 (s, 1H), 7.16-7.14 (m, 1H), 7.09-7.00 (m, 1H), 6.28 (d, 1H), 5.00- 4.98 (m, 1H), 4.29 (d, 1H), 4.18-4.10 (m, 3H), 3.90-3.80 (m, 1H), 3.20 (t, 1H), 3.00 (s, 3H), 2.41 (s, 3H), 2.05-1.92 (m, 2H), 1.70-1.61(m, 1H), 1.50-1.40 (m, 1H); Mass (ESI): 472.1 [M+1]; LCMS: 472.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.43 min. 0.05% Aq TFA: ACN; 0.80 ml/min); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μπι); RT 1.59 min. ACN: 0.025% Aq TFA; 0.5 mL/min; Chiral HPLC: 99.3% RT = 17.81 min (CHIRALPAK-IC (250 x 4.6 mm, 5 μιη; mobile phase (A) 0.1 % DEA in n- hexane (B) CH2C12: MeOH (50: 50) (A: B; 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 19-96: -49.53 (c = 0. 25, CH2C12).
Example 76
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7- (trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000212_0001
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl- 7-(trifluoromethyl)- 7, 8- dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2-amine
[0427] To a dry vial was added a suspension of Pd2(dba)3 (21 mg, 0.02 mmol) and (±) BINAP (44 mg, 0.07 mmol) in 1, 4-dioxane (0.25 mL) at room temperature. The suspension was degassed with argon, heated to 120 °C, and stirred at 120 °C for 3 min. A mixture of 2- chloro-8-methyl-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (120 mg, 0.47 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (126 mg, 0.52 mmol), sodium tert-butoxide (136 mg, 1.42 mmol) in 1, 4-dioxane (0.25 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by preparative HPLC (X-select CSH C-18 (250 x 19 mm, 5μιη (65 mg loading; CH3CN: 0.05% TFA (0.1/90, 1/90, 18/70, 25/30, 30/10, 35/10)) to afford N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(trifluoromethyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (30 mg, 15%) as an off-white solid. LCMS: 425 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.11 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 ml/min) (Gradient); TLC: 5% MeOH/ CH2C12 (R 0.3).
[0428] Racemic compound of Example 76 was separated using a Chiralpak-IB column (250 x 20 mm, 5μιη) (15 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (80: 20) (A: B: 85: 15) as mobile phase) to provide the compound of Example 76A (Fraction I) and the compound of Example 76B (Fraction II).
Example 76A
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(trifluoromethyl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000213_0001
[0429] The compound of Example 76A was produced as described in Example 76.
Analytical data for product Fraction I: 1H NMR (CD3OD, 400 MHz): δ 7.72 (d, 1H), 7.43 (d, 1H), 6.51 (d, 1H), 6.15 (s, 1H), 4.49 (d, 1H), 4.29-4.21 (m, 1H), 3.97-3.90 (m, 4H), 3.80 (s, 3H), 3.11 (s, 3H), 3.05 (t, 2H), 2.09-2.01 (m, 2H), 1.58-1.50 (m, 2H); Mass (ESI): 425.4 [M+1]; LCMS: 425 (M+1); (column; X-select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.01 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 8/90; flow rate 0.8 ml/min (Gradient); UPLC (column; Acquity BEH C-18, 50 x 2.1 mm, 1.7 μ); RT 1.43 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 mL/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10: diluent: CH3CN: Water; Chiral HPLC: 98.4% RT = 12.33 min (Chiralpak-IB (250 x 4.6 mm, 5μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (80: 20) (A: B; 85: 15); flow Rate: 1.0 mL/min).
Example 76B Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(trifluoromethyl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000214_0001
[0430] The compound of Example 76B was produced as described in Example 76.
Analytical data for product Fraction II: 1H NMR (CD3OD, 400MHz): δ 7.72 (d, 1H), 7.43 (d, 1H), 6.51 (d, 1H), 6.15 (s, 1H), 4.49 (d, 1H), 4.29-4.21 (m, 1H), 3.97-3.90 (m, 4H), 3.80 (s, 3H), 3.11 (s, 3H), 3.05 (t, 2H), 2.09-2.01 (m, 2H), 1.58-1.50 (m, 2H); Mass (ESI): 425.4 [M+l]; LCMS: 425 (M+l); (column; X-select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.00 min. mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 8/90; flow rate 0.8 ml/min (Gradient); UPLC (column; Acquity BEH C-18, 50 2.1 mm, 1.7 μ); RT 1.45 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 mL/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10: diluent: CH3CN: Water; Chiral HPLC: 96.6% RT = 15.17 min (Chiralpak-IB (250 x 4.6 mm, 5μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (80: 20) (A: B; 85: 15); flow Rate: 1.0 mL/min).
Example 77
Synthesis of 8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)- 7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000214_0002
Synthesis of 8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7- (trifluoromethyl)-l ', 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazin-2- amine
[0431] To a dry vial was added a suspension of Pd2(dba)3 (36 mg, 0.04 mmol) and (±) BINAP (73 mg, 0.12 mmol) in 1, 4-dioxane (1 mL) at room temperature. The suspension was degassed with argon, heated to 120 °C, and stirred at 120 °C for 3 min. A mixture of 2- chloro-8-methyl-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (200 mg, 0.79 mmol), (3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3 -amine (166 mg, 0.87 mmol) and sodium tert-butoxide (227 mg, 2.37 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction mixture was filtered and the filtrate was concentrated in vacuo. The crude material was purified by column chromatography using 3% MeOH/ CH2CI2 to afford (8-methyl-N-((3i?, 65)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3-yl)-7- (trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine (85 mg, 26%) as colorless oil. 1H NMR (CD3OD, 400 MHz): δ 8.34 (d, 1H), 7.46 (s, 1H), 7.32 (s, 1H), 7.25 (d, 1H), 4.50 (d, 1H), 4.39 (d, 1H), 4.31-4.26 (m, 2H), 4.00-3.91 (m, 2H), 3.30-3.27 (m, 4H), 2.53 (s, 3H), 2.26- 2.20 (m, 1H), 2.10-2.04 (m, 1H), 1.76-1.60 (m, 2H); TLC: 5%MeOH/ CH2C12 (R 0.3).
[0432] Racemic compound of Example 77 was separated using a Chiralpak-IC column (250 x 20 mm, 5μιη) (15 mg loading; 0.1 % DEA in n-hexane: CH2C12: MeOH (80: 20) (A: B: 80: 20) as mobile phase) to provide the compound of Example 77A (Fraction I (-)) and the compound of Example 77B (Fraction II (+)).
Example 77A
Synthesis of (-)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
[0433] The compound of Example 77A was produced as described in Example 77.
Analytical data for product Fraction I (-): 1H NMR (CD3OD, 400 MHz): δ 8.34 (d, 1H), 7.46 (s, 1H), 7.32 (s, 1H), 7.25 (d, 1H), 4.50 (d, 1H), 4.39 (d, 1H), 4.31-4.26 (m, 2H), 4.00-3.91 (m, 2H), 3.30-3.27 (m, 4H), 2.53 (s, 3H), 2.26- 2.20 (m, 1H), 2.10-2.04 (m, 1H), 1.76-1.60 (m, 2H); Mass (ESI): 410.4 [M+1]; LCMS: 410 (M+1); (column; Ascentis Express C-18 (50 3.0 mm, 2.7 μπι); RT 1.52 min; mobile phase: 0.025% Aq TFA+5% ACN:
5%ACN+0.025% Aq TFA; T/B%: 0.01/5, 0.5/5, 3/100, 5/100; flow rate: 1.2 mL/min (Gradient); HPLC (column; Eclipse XDB-C-18, 150 4.6 mm, 5.0 μπι); RT 9.43 min.
mobile phase: ACN: 5mM Aq NH4OAc; flow rate: 1.0 mL/min; Gradient program: T/B% 0.01/80, 3/80, 10/10, 20/10: diluent: CH3CN: Water; Chiral HPLC: 99.7% RT = 21.03 min (Chiralpak-IC (250 x 4.6 mm, 5μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (80: 20) (A: B; 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: -83.21 (c = 0. 25, CH2C12).
Example 77B
Synthesis of (+)-8-methyl-N-((3R, 6S)-6-(2-methylpyridin-4-yl) tetrahydro-2H-pyran-3- yl)-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000216_0001
[0434] The compound of Example 77B was produced as described in Example 77.
Analytical data for product Fraction II (+): 1H NMR (CD3OD, 400 MHz): δ 8.34 (d, 1H), 7.46 (s, 1H), 7.32 (s, 1H), 7.25 (d, 1H), 4.50 (d, 1H), 4.39 (d, 1H), 4.31-4.26 (m, 2H), 4.00- 3.91 (m, 2H), 3.30-3.27 (m, 4H), 2.53 (s, 3H), 2.26- 2.20 (m, 1H), 2.10-2.04 (m, 1H), 1.76- 1.60 (m, 2H); Mass (ESI): 410.4 [M+1]; LCMS: 410 (M+1); (column; Ascentis Express C-18 (50 x 3.0 mm, 2.7 μπι); RT 1.49 min; mobile phase: 0.025% Aq TFA+5% ACN:
5%ACN+0.025% Aq TFA; T/B%: 0.01/5, 0.5/5, 3/100, 5/100; flow rate: 1.2 mL/min (Gradient); HPLC (column; Eclipse XDB-C-18, 150 4.6 mm, 5.0 μπι); RT 9.41 min.
mobile phase: ACN: 5mM Aq NH4OAc; flow rate: 1.0 mL/min; Gradient program: T/B% 0.01/80, 3/80, 10/10, 20/10: diluent: CH3CN: Water; Chiral HPLC: 98.3% RT = 23.06 min (Chiralpak-IC (250 x 4.6 mm, 5μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (80: 20) (A: B; 80: 20); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 0°: +44.00 (c = 0. 25, CH2C12). Example 78
Synthesis of 2-chloro-8-methyl-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1,
4] oxazine
Figure imgf000217_0001
Synthesis of ethyl 2-((tert-butoxycarbonyl) amino)-3, 3, 3-trifluoro-2-hydroxypropanoate
[0435] To a stirred solution of ethyl 3, 3, 3-trifluoro-2-oxopropanoate (25 g, 147.05 mmol) in CH2CI2 (250 mL) under an argon atmosphere was added Boc-amine (19 g, 161.76 mmol) at room temperature. The reaction mixture was stirred for 16 h at room temperature. After consumption of the starting materials (monitored by TLC), the volatiles were evaporated in vacuo to obtain ethyl 2-((fert-butoxycarbonyl) amino)-3, 3, 3-trifluoro-2- hydroxypropanoate (40 g, 95%) as pale yellow syrup used in the next step without further purification. TLC: 20% EtOAc/ hexane (R 0.5).
Synthesis of ethyl (Z)-2-((tert-butoxycarbonyl) imino)-3, 3, 3-trifluoropropanoate
[0436] To a stirred solution of ethyl 2-((tert-butoxycarbonyl) amino)-3, 3, 3-trifluoro-2- hydroxypropanoate (20 g, 69.56 mmol) in ether (300 mL) under an argon atmosphere were added trifluoroacetic anhydride (14.6 g, 69.56 mmol) and pyridine (11.2 mL, 139.12 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was filtered. The filtrate was washed with water (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo to obtain ethyl (Z)-2-((tert-butoxycarbonyl) imino)-3, 3, 3- trifluoropropanoate (11 g, 59%>) as an off- white solid used in the next step without further purification. 1H NMR (DMSO-<¾, 400 MHz): δ 4.26-4.11 (m, 2H), 1.38 (s, 9H), 1.19 (t, 3H); TLC: 20% EtOAc/ hexane (R/. 0.5).
Synthesis of tert-butyl (Z)-(l, 1, l-trifluoro-3-hydroxypropan-2-ylidene) carbamate [0437] To a stirred solution of ethyl (Z)-2-((tert-butoxycarbonyl) imino)-3, 3, 3- trifluoropropanoate (11 g, 40.81 mmol) in THF (2.75 mL) under an argon atmosphere was added lithium aluminum hydride (4.65 g, 122.44 mmol) portion wise at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (5 mL) and basified with a 15% sodium hydroxide solution (5 mL) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h. The reaction mixture was filtered and washed with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain tert-butyl (Z)-(l, 1, l-trifluoro-3- hydroxypropan-2-ylidene) carbamate (6 g, 64%) as an off-white solid. 1H NMR (DMSO 400 MHz): δ 6.11 (br s, 1H), 5.04 (br s, 1H), 4.21-4.07 (m, 1H), 3.69-3.45 (m, 2H), 1.37 (d, 9H); TLC: 30% EtOAc/ hexane (R 0.3).
Synthesis of tert-butyl (Z)-(3-((2, 4-dichloropyrimidin-5-yl) oxy)-l, 1, l-trifluoropropan-2- ylidene) carbamate
[0438] To a stirred solution of tert-butyl (Z)-(l, 1, l-trifluoro-3-hydroxypropan-2- ylidene) carbamate (65 g, 26.20 mmol) in THF (180 mL) under an argon atmosphere were added diisopropyl azodicarboxylate (7.9 g, 39.30 mmol), triphenylphosphine (10 g, 39.30 mmol) and 2, 4-dichloropyrimidin-5-ol (4.3 g, 26.20 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 4 h. After consumption of the starting materials (monitored by TLC), the volatiles were concentrated in vacuo. The crude material was purified by column chromatography using 20-30% EtOAc: hexane to afford tert-butyl (Z)-(3- ((2, 4-dichloropyrimidin-5-yl) oxy)-l, 1 , l-trifluoropropan-2-ylidene) carbamate (3.5 g, 36%>) as an off-white solid. 1H NMR (CDC13, 400 MHz): δ 8.21 (s, 1H), 5.21-5.18 (m, 1H), 4.74 (br s, 1H), 4.43-4.18 (m, 2H), 1.48 (s, 9H); TLC: 50% EtOAc/ hexane (R 0.5).
Synthesis of 3 -((2, 4-dichloropyrimidin-5-yl) oxy)-l, 1, l-trifluoropropan-2-amine hydrochloride
[0439] To a stirred solution of tert-butyl (Z)-(3-((2, 4-dichloropyrimidin-5-yl) oxy)-l, 1, l-trifluoropropan-2-ylidene) carbamate (3.5 g, 9.30 mmol) in CH2CI2 (30 mL) under an argon atmosphere was added 4M HC1 in 1, 4-dioxane (10 mL) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the volatiles were concentrated in vacuo. The crude material was washed with n-pentane (2 x 10 mL) to obtain 3-((2, 4-dichloropyrimidin-5-yl) oxy)-l, 1, 1- trifluoropropan-2-amine hydrochloride (2.2 g, as HC1 salt) as an off-white solid. TLC: 30% EtOAc/ hexane (R 0.3).
Synthesis of 2-chloro-7-(trifluoromethyl)-7 , 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazine
[0440] To a stirred solution of 3-((2, 4-dichloropyrimidin-5-yl) oxy)-l, 1, 1- trifluoropropan-2-amine hydrochloride (2.2 g, 7.97 mmol) in 1, 4-dioxane (5.5 mL) under an argon atmosphere was added diisopropylethylamine (5.5 mL, 31.88 mmol) at room temperature. The reaction mixture was stirred at 120 °C for 16 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain 2-chloro-7-(trifluoromethyl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (1.6 g, 84%) as a pale yellow solid. 1H NMR (CDC13, 400 MHz): δ 7.90 (s, 1H), 7.43 (br s, 1H), 5.01-4.97 (m, 1H), 4.61-4.58 (m, 1H), 4.17-4.09 (m, 1H); LCMS: 239.8 (M+l); (column; Ascentis Express C-18 (50 3.0 mm, 2.7 μιη); RT 1.82 min; mobile phase: 0.025% Aq TFA+5% ACN: ACN+5% 0.025% Aq TFA; flow rate: T/B%: 0.01/5, 0.5/5, 3/100, 5/100; 1.2 mL/min) (Gradient); TLC: 30% EtOAc/ hexane (R 0.4).
Synthesis of 2-chloro-8-methyl-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
[0441] To a stirred solution of 2-chloro-7-(trifluoromethyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (1.6 g, 6.69 mmol) in DMF (20 mL) under an argon atmosphere was added sodium hydride (193 mg, 8.03 mmol) at 0 °C. The reaction mixture was stirred for 5 min at 0 °C. Then methyl iodide (1.14 g, 8.03 mmol) was added to the reaction mixture at 0 °C and stirred for 5 min at 0 °C. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and
concentrated in vacuo. The crude material was purified by column chromatography using 20% EtOAc: hexane to afford 2-chloro-8-methyl-7-(trifluoromethyl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazine (1.2 g, 71%) as a pale yellow syrup. 1H NMR (400 MHz, CDC13): δ 7.82 (s, 1H), 4.62 (d, 1H), 4.03-4.00 (m, 1H), 3.99-3.95 (m, 1H), 3.31 (s, 3H); LCMS: 253.8 (M+l); (column; Ascentis Express C-18 (50 3.0 mm, 2.7 μπι); RT 2.18 min; mobile phase: 0.025% Aq TFA+5% ACN: ACN+5% 0.025% Aq TFA; T/B%: 0.01/5, 0.5/5, 3/100, 5/100; flow rate: 1.2 mL/min) (Gradient);TLC: 30% EtOAc/ hexane (R 0.3).
Example 79
Synthesis of 2-chloro-6a, 7, 8, 9-tetrahydro-6H-pyrimido [5, 4-b] pyrrolo [1, 2-d] [1, 4] oxazine
CH2CI2 0 °C-RT, 1 h
Figure imgf000220_0001
Synthesis of 2, 4-dichloro-5-((4-methoxybenzyl) oxy) pyrimidine
[0442] To a stirred solution of 2, 4-dichloropyrimidin-5-ol (7 g, 42.68 mmol) in acetone (100 mL) under an argon atmosphere were added l-(chloromethyl)-4-methoxybenzene (10 g, 64.02 mmol), potassium carbonate (14.7 g, 106.70 mmol) and sodium iodide (600 mg, 4.26 mmol) at room temperature. The reaction mixture was stirred for 16 h at room temperature. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 10% EtOAc: hexanes to afford 2, 4- dichloro-5-((4-methoxybenzyl) oxy) pyrimidine (6.2 g, 51%) as a pale yellow solid. 1H-NMR (CDCI3, 500 MHz): δ 8.19 (s, 1H), 7.34 (d, 2H), 6.93 (d, 2H), 5.19 (s, 2H), 3.80 (s, 3H);
TLC: 30% EtOAc/ hexane (R/. 0.6).
Synthesis of (l-(2-chloro-5-((4-methoxybenzyl) oxy) pyrimidin-4-yl) pyrrolidin-2-yl) methanol
[0443] To a stirred solution of 2, 4-dichloro-5-((4-methoxybenzyl) oxy) pyrimidine (100 mg, 0.35 mmol) in 2-methyl THF (1 mL) under an argon atmosphere were added pyrrolidin- 2-ylmethanol (40 mg, 0.39 mmol) and triethylamine (0.12 mL, 0.87 mmol) at room
temperature. The reaction mixture was stirred at 70 °C for 8 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 50% EtOAc: hexanes to afford (l-(2-chloro-5-((4- methoxybenzyl) oxy) pyrimidin-4-yl) pyrrolidin-2-yl) methanol (120 mg, 98%) as colorless syrup. 1H-NMR (CDC13, 500 MHz): δ 7.67 (s, 1H), 7.41 (d, 2H), 6.91 (d, 2H), 4.90 (s, 2H), 4.48-4.41 (m, 1H), 3.88-3.78 (m, 5H), 3.69-3.60 (m, 2H), 3.21 (br s, 1H), 2.00-1.80 (m, 4H); LCMS: 350.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.91 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 50% EtOAc/ hexane (R 0.3).
Synthesis of 2-chloro-4-(2-(hydroxymethyl) pyrrolidin-l-yl) pyrimidin-5-ol
[0444] To a stirred solution of (l-(2-chloro-5-((4-methoxybenzyl) oxy) pyrimidin-4-yl) pyrrolidin-2-yl) methanol (120 mg, 0.34 mmol) in CH2CI2 (2 mL) under an argon atmosphere was added trifluoroacetic acid (1 mL) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with a saturated sodium bicarbonate solution (20 mL) and extracted with 5% MeOH/ CH2CI2 (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain 2-chloro-4-(2- (hydroxymethyl) pyrrolidin-l-yl) pyrimidin-5-ol (50 mg 64%) as a white solid. 1H-NMR (DMSO-< 5, 400 MHz): δ 9.80 (s, 1H), 7.50 (s, 1H), 4.79 (br s, 1H), 4.50-4.40 (m, 1H), 3.75- 3.69 (m, 1H), 3.67-3.60 (m, 1H), 3.59-3.50 (m, 1H), 2.00-1.71 (m, 4H); LCMS: 230.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.06 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 50% EtOAc/ hexane (R 0.3).
Synthesis of 2-chloro-6a, 7, 8, 9-tetrahydro-6H-pyrimido [5, 4-bJ pyrrolo [I, 2-dJ [I, 4] oxazine
[0445] To a stirred solution of 2-chloro-4-(2-(hydroxymethyl) pyrrolidin-l-yl) pyrimidin- 5-ol (1.8 g, 0.56 mmol) in THF (20 mL) under an argon atmosphere were added
triphenylphosphine (2.2 g, 0.85 mmol) and diisopropyl azodicarboxylate (1.7 mL, 0.85 mmol) at room temperature. The reaction mixture was stirred at 70 °C for 16 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 30% EtOAc: hexanes to afford 2-chloro-6a, 7, 8, 9-tetrahydro-6H-pyrimido [5, 4-b] pyrrolo [1, 2-d [1, 4] oxazine (800 mg, 66%>) as an off- white solid. 1H-NMR (CDC13, 500 MHz): δ 7.68 (d, 1H), 4.52 (d, 1H), 3.78-3.70 (m, 2H), 3.60-3.56 (m, 1H), 3.39 (t, 1H), 2.20-2.10 (m, 2H), 2.08-1.99 (m, 1H), 1.53-1.40 (m, 1H); LCMS: 212.2 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.73 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 50% EtOAc/ hexanes (R 0.5).
Example 80
Synthesis of 2-chloro-7, 7-dimethyl-8-phenyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
Figure imgf000222_0001
Synthesis of 2-((2-chloro-5-methoxypyrimidin-4-yl) amino)-2-methylpropan-l-ol:
[0446] To a stirred solution of 2, 4-dichloro-5-methoxypyrimidine (5 g, 27.93 mmol) in 1, 4-dioxane (50 mL) under an argon atmosphere were added diisopropylethylamine (7.2 g, 55.86 mmol) and 2-amino-2-methylpropan-l-ol (2.4 g, 27.93 mmol) at room temperature. The reaction mixture was stirred at 130 °C for 48 h. After consumption of the starting materials (monitored by TLC), the volatiles were evaporated in vacuo. The residue was diluted with a saturated sodium bicarbonate solution (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 10% EtOAc: hexanes to afford 2-((2-chloro-5-methoxypyrimidin-4-yl) amino)-2- methylpropan-l-ol (5.5 g, 84%) as a white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 7.69 (s, 1H), 6.19 (s, 1H), 6.11 (t, 1H), 3.82 (s, 3H), 3.45 (d, 2H), 1.33 (s, 6H); TLC: 50% EtOAc: hexanes (R 0.3).
Synthesis of 2-chloro-4-((l -hydroxy-2-methylpropan-2-yl) amino) pyrimidin-5-ol:
[0447] To a stirred solution of 2-((2-chloro-5-methoxypyrimidin-4-yl) amino)-2- methylpropan-l-ol (5.5 g, 23.80 mmol) in CH2CI2 (100 mL) was added boron tribromide (34 mL, 357.14 mmol) at 0 °C under an argon atmosphere. The reaction mixture was warmed to room temperature and stirred for 24 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was quenched with methanol (50 mL) at 0 °C and concentrated in vacuo. The residue was dissolved in EtOAc (2 x 100 mL) and washed with a sodium bicarbonate solution (2 x 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain 2-chloro-4-((l-hydroxy-2-methylpropan- 2-yl) amino) pyrimidin-5-ol (4 g, 81%) as a white solid used in the next step without further purification. 1H-NMR (DMSO-< 5, 500 MHz): δ 10.38 (br s, 1H), 7.48 (s, 1H), 6.03 (s, 1H), 5.11 (br s, 1H), 3.45 (s, 2H), 1.37 (s, 6H); TLC: 80% EtOAc: hexanes (R 0.3).
Synthesis of tert-butyl (2-chloro-5-hydroxypyrimidin-4-yl) (l-hydroxy-2-methylpropan-2-yl) carbamate:
[0448] To a stirred solution of 2-chloro-4-((l-hydroxy-2-methylpropan-2-yl) amino) pyrimidin-5-ol (4.3 g, 19.90 mmol) in CH2CI2 (50 mL) under an argon atmosphere were added triethylamine (4.3 mL, 29.80 mmol) and Boc anhydride (4.3 g, 19.90 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 h. After
consumption of the starting materials (monitored by TLC), the volailes were evaporated in vacuo. The crude material was purified by column chromatography using 40% EtOAc:
hexanes to afford tert-butyl (2-chloro-5-hydroxypyrimidin-4-yl) (l-hydroxy-2-methylpropan- 2-yl) carbamate (3 g, 48%) as a white solid. 1H-NMR (DMSO-< 5, 500 MHz): δ 8.00 (s, 1H), 6.61 (s, 1H), 5.07 (t, 1H), 3.49 (d, 2H), 1.50 (s, 9H), 1.37 (s, 6H); TLC: 50% EtOAc: hexanes (R 0.5).
Synthesis of tert-butyl 2 -chloro-7, 7-dimethyl-6, 7-dihydro-8H-pyrimido [5, 4-b] [1, 4] oxazine-8-carboxylate: [0449] To a stirred solution of tert-butyl (2-chloro-5-hydroxypyrimidin-4-yl) (1-hydroxy- 2-methylpropan-2-yl) carbamate (3 g, 9.46 mmol) in THF (30 mL) under an argon atmosphere were added triphenylphosphine (2.9 g, 11.36 mmol) and diisopropyl
azodicarboxylate (2.3 g, 11.35 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 1 h. After consumption of the starting materials (monitored by TLC), the volailes were evaporated in vacuo. The crude material was purified by column chromatography using 20% EtOAc: hexanes to afford tert-butyl 2-chloro-7, 7-dimethyl-6, 7- dihydro-8H-pyrimido [5, 4-b] [1, 4] oxazine-8-carboxylate (1.5 g, 53%>) as a white solid. 1H- NMR (DMSO-<¾, 400 MHz): δ 8.11 (s, 1H), 4.01 (s, 2H), 1.52 (s, 9H), 1.38 (s, 6H); LCMS: 300.3 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.69 min; mobile phase: 5 mM Aq NH4OAc: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 30% EtOAc: hexanes (R 0.3).
Synthesis of 2-chloro-7 , / 1 -dimethyl-'/ ', 8-dihydro-6H-pyrimido [5, 4-b] [I, 4] oxazine:
[0450] To a stirred solution of tert-butyl 2-chloro-7, 7-dimethyl-6, 7-dihydro-8H- pyrimido [5, 4-b] [1, 4] oxazine-8-carboxylate (500 mg, 1.67 mmol) in CH2C12 (5 mL) under an argon atmosphere was added 4M HC1 in 1, 4-dioxane (5 mL) at 0 °C. The reaction mixture was stirred for 16 h at room temperature. After consumption of the starting materials
(monitored by TLC), the volatiles were evaporated in vacuo. The residue was diluted with a saturated sodium bicarbonate solution (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain the 2-chloro-7, 7-dimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (260 mg, 78%) as a white solid. 1H-NMR (DMSO-<¾, 400 MHz): δ 8.50 (s, 1H), 7.71 (s, 2H), 1.20 (s, 6H); TLC: 50% EtOAc: hexanes (R/. 0.3).
Synthesis of 2 -chloro-7 , 7, 8-trimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine
[0451] To a stirred solution of 2-chloro-7, 7-dimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (50 mg, 0.25 mmol) in DMF (0.5 mL) under an argon atmosphere were added sodium hydride (8.8 mg, 0.37 mmol) and methyl iodide (35.5 mg, 0.25 mmol) at 0 °C. The reaction mixture was stirred for 30 min at 0 °C. After consumption of the starting materials (monitored by TLC), the volatiles were evaporated in vacuo. The residue was diluted with ice cold water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 20% EtOAc: hexanes to afford 2-chloro-7, 7, 8- trimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (30 mg, 56%) as a white solid. 1H NMR (DMSO-< 5, 400 MHz): δ 7.68 (s, 1H), 3.93 (s, 2H), 3.00 (s, 3H), 1.24 (s, 6H); TLC: 50% EtOAc: hexanes (R/. 0.4).
Example 81
Synthesis of 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine]
Figure imgf000225_0001
Synthesis of (l-((2-chloro-5-((4-methoxybenzyl) oxy) pyrimidin-4-yl) amino) cyclopentyl) methanol
[0452] To a stirred solution of 2, 4-dichloro-5-((4-methoxybenzyl) oxy) pyrimidine (1.5 g, 5.26 mmol) in 1, 4-dioxane (4 mL) under an argon atmosphere were added
diisopropylethylamine (1.3 g, 21.05 mmol) and (1-aminocyclopentyl) methanol
hydrochloride (1 g, 6.84 mmol) at room temperature. The reaction mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC), the volatiles were evaporated in vacuo to obtain (l-((2-chloro-5-((4-methoxybenzyl) oxy) pyrimidin-4-yl) amino) cyclopentyl) methanol (700 mg, 37%) as a brown liquid. LCMS: 364.4 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 4.29 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 20% EtOAc/ hexane (R 0.4). Synthesis of2-chloro-4-((l-(hydroxymethyl) cyclopentyl) amino) pyrimidin-5-ol
[0453] To a stirred solution of (l-((2-chloro-5-((4-methoxybenzyl) oxy) pyrimidin-4-yl) amino) cyclopentyl) methanol (120 mg, 0.33 mmol) in CH2CI2 (1.2 mL) under an argon atmosphere was added trifluoroacetic acid (150 mg, 1.32 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 2 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was quenched with a saturated sodium bicarbonate solution (10 mL) and extracted with CH2CI2 (2 x 10 mL). The aqueous layer was acidified with a citric acid solution up to pfT5 and extracted with EtOAc (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain 2-chloro-4-((l-(hydroxymethyl) cyclopentyl) amino) pyrimidin-5-ol (80 mg, 99%) as an off-white solid used in the next step without further purification. 1H-NMR (DMSO-<¾, 400 MHz): δ 10.30 (br s, 1H), 8.13-8.11 (m, 1H), 7.46 (s, 1H), 6.13 (s, 1H), 3.70 (s, 2H), 2.00-1.93 (m, 2H), 1.90-1.88 (m, 2H), 1.80-1.61 (m, 4H); TLC: 50% EtOAc/ hexane (R/. 0.1).
Synthesis of tert-butyl (2-chloro-5-hydroxypyrimidin-4-yl) (l-(hydroxymethyl) cyclopentyl) carbamate
[0454] To a stirred solution of 2-chloro-4-((l-(hydroxymethyl) cyclopentyl) amino) pyrimidin-5-ol (500 mg, 2.05 mmol) in 1, 4-dioxane: water (1 : 1, 10 mL) under an argon atmosphere were added sodium hydroxide (98 mg, 2.46 mmol) and Boc anhydride (538 mg, 2.46 mmol) at 0 °C. The reaction mixture was stirred for 3 h at 0 °C. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (2 x 10 mL). The aqueous layer was acidified with a citric acid solution up to pH 5 and extracted with EtOAc (2 x 10 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain tert-butyl (2- chloro-5-hydroxypyrimidin-4-yl) (l-(hydroxymethyl) cyclopentyl) carbamate (700 mg, 99%) as a brown liquid. LCMS: 344.1 (M+l); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.70 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 50% EtOAc/ hexane (R 0.6).
Synthesis of tert-butyl 2 '-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-bJ
[ 1, 4]oxazine]-8 '-carboxylate [0455] To a stirred solution of tert-butyl (2-chloro-5-hydroxypyrimidin-4-yl) (1- (hydroxymethyl) cyclopentyl) carbamate (150 mg, 0.43 mmol) in THF (2 mL) under an argon atmosphere were added diisopropyl azodicarboxylate (132 g, 0.65 mmol) and triphenylphosphine (171 mg, 0.65 mmol) at 0 °C. The reaction mixture was stirred for 30 min at 0 °C. After consumption of the starting materials (monitored by TLC), the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 10% EtOAc; hexanes to afford tert-butyl 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine]-8'- carboxylate (60 mg, 42%) as colorless liquid. 1H-NMR (CD3OD, 400 MHz): δ 7.93 (s, 1H), 3.99 (s, 2H), 2.08-1.94 (m, 4H), 1.80-1.65 (m, 4H), 1.95 (s, 9H); TLC: 20% EtOAc/ hexane (R 0.7).
Synthesis of 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazinej
[0456] To a stirred solution of tert-butyl 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'- pyrimido [5, 4-b] [1, 4] oxazine]-8'-carboxylate (400 mg, 1.23 mmol) in CH2CI2 (4 mL) under an argon atmosphere was added trifluoroacetic acid (561 mg, 4.92 mmol) at 0 °C. The reaction mixture was warmed to room temperature and stirred for 16 h. After consumption of the starting materials (monitored by TLC), the reaction mixture was quenched with a saturated sodium bicarbonate solution (20 mL) and extracted with CH2CI2 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to obtain 2'-chloro-6'H, 8'H-spiro [ cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine] (270 mg, 97%) as a brown solid. 1H-NMR (CDC13, 400 MHz): δ 7.75 (s, 1H), 5.50-5.43 (m, 1H), 3.90 (s, 2H), 1.89-1.80 (m, 4H), 1.79-1.70 (m, 4H); TLC: 20% EtOAc/ hexane (R 0.3).
Synthesis of 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine]
[0457] To a stirred solution of 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine] (100 mg, 0.44 mmol) in DMF (1 mL) under an argon atmosphere was added sodium hydride (26 mg, 0.66 mmol) and methyl iodide (94 mg, 0.66 mmol) at 0 °C. The reaction mixture was stirred for 5 min at 0 °C. After consumption of the starting materials (monitored by TLC), the reaction mixture was quenched with ice water (10 mL) and extracted with EtOAc (2 x 10 mL). The combined organic extracts were washed with water, dried over sodium sulfate, filtered and concentrated in vacuo to obtain 2'-chloro-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine] (100 mg, 95%) as a brown liquid. 1H-NMR (CDC13, 400 MHz): δ 7.61 (s, 1H), 3.82 (s, 2H), 3.09 (s, 3H), 1.87-1.80 (m, 4H), 1.73-1.67 (m, 4H); LCMS: 240.1 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 3.32 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min) (Gradient); TLC: 20% EtOAc/ hexane (R 0.3).
Example 82
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-6a, 7, 8, 9-tetrahydi
pyrimido [5, 4-b] pyrrolo [1, 2-d] [1, 4] oxazin-2-amine
Figure imgf000228_0001
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-6a, 7, 8, 9-tetrahydro-6H-pyrimido [5, 4-b] pyrrolo [I, 2-d] [I, 4] oxazin-2-amine
[0458] To a dry vial was added a suspension of Pd2(dba)3 (54 mg, 0.05 mmol) and (±) BINAP (110 mg, 0.19 mmol) in 1, 4-dioxane (2.5 mL) at room temperature. The suspension was degassed with an argon atmosphere, heated to 120 °C, and stirred at 120 °C for 3 min. A mixture of 2-chloro-6a, 7, 8, 9-tetrahydro-6H-pyrimido [5, 4-b] pyrrolo [1, 2-d] [1, 4] oxazine (250 mg, 0.12 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (316 mg, 0.13 mmol) and sodium tert-butoxide (350 mg, 3.55 mmol) in 1, 4-dioxane (2.5 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction mixture was filtered and the filtrate was concentrated in vacuo. The crude material was purified by column chromatography using 8% MeOH: CH2C12 to afford N-(l -(2 -methoxypyridin-4-yl) piperidin-4-yl)-6a, 7, 8, 9-tetrahydro-6H-pyrimido [5, 4- b] pyrrolo [1, 2-d] [1, 4] oxazin-2-amine (130 mg, 28%) as an off-white solid. LCMS: 383.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 1.21 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 9/90; flow rate: 0.80 mL/min)
(Gradient); UPLC (column; Acquity UPLC BEH C-18 2.1 X 50 mm, 1.7 μ); RT 1.27 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 mL/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10; diluent: CH3CN: Water; TLC: 10% MeOH/ CH2C12 (R 0.3).
[0459] Racemic compound of Example 82 was separated using a Chiralpak-AD-H column (250 x 20 mm, 5μιη (20 mg loading; 0.1 % DEA in n-hexane: EtOH (A: B: 85: 15) as mobile phase) to provide the compound of Example 82A (Fraction I (+)) and the compound of Example 82B (Fraction II (-)).
Example 82A
Synthesis of (+)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-6a, 7, 8, 9-tetrahydi
pyrimido [5, 4-b] pyrrolo [1, 2-d] [1, 4] oxazin-2-amine
Figure imgf000229_0001
[0460] The compound of Example 82A was produced as described in Example 82.
Analytical data for product Fraction I (+): 1H-NMR (CD3OD, 400 MHz): δ 7.74 (d, 1H), 7.33 (s, 1H), 6.54 (d, 1H), 6.14 (s, 1H), 4.47-4.40 (m, 1H), 3.91-3.84 (m, 3H), 3.81 (s, 3H), 3.77- 3.70 (m, 1H), 3.62-3.49 (m, 2H), 3.32-3.30 (m, 1H), 3.02 (t, 2H), 2.20-1.90 (m, 5H), 1.59- 1.45 (m, 3H); Mass (ESI): 383.4 [M+1]; LCMS: 383.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.77 min; mobile phase: 5 mM Aq NH4OAc: ACN; T/B%:
0.01/10, 0.5/10, 4/90, 9/90; flow rate 0.8 ml/min (Gradient); UPLC (column; Acquity UPLC BEH C-18, 2.1 50 mm, 1.7 μ); RT 1.31 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 mL/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10: Diluent: CH3CN: Water: Chiral HPLC: 99.5% RT = 6.37 min (Chiralpak-IA (250 x 4.6 mm, 5μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 50: 50); flow Rate: 1.0 mL/min); Optical rotation [a]D 20 02: +29.12 (c = 0. 25, CH2C12).
Example 82B Synthesis of (-)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-6a, 7, 8, 9-tetrahydi pyrimido [5, 4-b] pyrrolo [1, 2-d] [1, 4] oxazin-2-amine
Figure imgf000230_0001
[0461] The compound of Example 82B was produced as described in Example 82.
Analytical data for product Fraction II (-): 1H-NMR (CD3OD, 400 MHz): δ 7.74 (d, 1H), 7.33 (s, 1H), 6.54 (d, 1H), 6.14 (s, 1H), 4.47-4.40 (m, 1H), 3.91-3.84 (m, 3H), 3.81 (s, 3H), 3.77- 3.70 (m, 1H), 3.62-3.49 (m, 2H), 3.32-3.30 (m, 1H), 3.02 (t, 2H), 2.20-1.90 (m, 5H), 1.59- 1.45 (m, 3H); Mass (ESI): 383.4 [M+1]; LCMS: 383.4 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 3.78 min; mobile phase: 5 mM Aq NH4OAc: ACN; T/B%:
0.01/10, 0.5/10, 4/90, 9/90 flow rate 0.8 ml/min (Gradient); UPLC (column; Acquity UPLC BEH C-18, 2.1 50 mm, 1.7 μ); RT 1.31 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 ml/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10: diluent: CH3CN: Water; Chiral HPLC: 99.3% RT = 9.02 min (Chiralpak-IA (250 x 4.6 mm, 5μιη; mobile phase (A) 0.1 % DEA in n-hexane (B) CH2C12: MeOH (50: 50) (A: B: 50: 50); flow Rate: 1.0 mL/min); Optical rotation [a]D 19": -24.80 (c = 0. 25, CH2C12).
Example 83
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 7, 8-trimethyl-7, 8-dihydro-
6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine
Figure imgf000230_0002
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-7, 7, 8-trimethyl-7 , 8-dihydro-6H- pyrimido [5, 4-b] [I, 4] oxazin-2 -amine
[0462] To a dry vial was added a suspension of Pd2(dba)3 (32 mg, 0.03 mmol) and (±) BINAP (65 mg, 0.10 mmol) in 1, 4-dioxane (1.5 mL) at room temperature. The suspension was degassed with argon, heated to 120 °C, and stirred at 120 °C for 3 min. A mixture of 2- chloro-7, 7, 8-trimethyl-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazine (150 mg, 0.70 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (187 mg, 0.77 mmol) and sodium tert-butoxide (202 mg, 2.10 mmol) in 1, 4-dioxane (1.5 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction mixture was diluted with water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 4% MeOH: CH2C12 to afford N-(l -(2 -methoxypyridin-4-yl) piperidin-4-yl)-7, 7, 8-trimethyl-7, 8-dihydro-6H- pyrimido [5 , 4-b] [1, 4] oxazin-2-amine (60 mg, 22%) as an off-white solid. 1H-NMR
(CD3OD, 500 MHz): δ 7.76 (d, 1H), 7.35 (s, 1H), 6.55 (d, 1H), 6.18 (s, 1H), 3.95-3.90 (m, 3H), 3.82 (s, 3H), 3.81 (s, 2H), 3.09 (s, 3H), 3.08-3.03 (m, 2H), 2.07 (d, 2H), 1.60-1.50 (m, 2H), 1.30 (s, 6H); Mass (ESI): 385 [M+1]; LCMS: 385 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μιη); RT 2.05 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 8/90; flow rate 0.8 ml/min (Gradient); UPLC (column; Acquity UPLC BEH C- 18 2.1 X 50 mm, 1.7 μ); RT 1.37 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 mL/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10: Diluent: CH3CN: Water; TLC: 5% MeOH/ CH2C12 (R 0.3).
Example 84
Synthesis of N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8'-methyl-6'H, 8'H-spiro
[cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazin]-2'-amine
Figure imgf000232_0001
Synthesis ofN-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8'-methyl-6'H, 8'H-spiro
fcyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazin] -2'-amine
[0463] To a dry vial was added a suspension of Pd2(dba)3 (25 mg, 0.02 mmol) and (±) BINAP (51 mg, 0.08 mmol) in 1, 4-dioxane (1 mL) at room temperature. The suspension was degassed swith argon, heated to 120 °C, and stirred at 120 °C for 3 min. A mixture of 2'- chloro-8'-methyl-6'H, 8'H-spiro [cyclopentane-1, 7'-pyrimido [5, 4-b] [1, 4] oxazine] (130 mg, 0.54 mmol), l-(2-methoxypyridin-4-yl) piperidin-4-amine hydrochloride (124 mg, 0.59 mmol) and sodium tert-butoxide (156 mg, 1.63 mmol) in 1, 4-dioxane (1 mL) was degassed and the catalyst premix was added. The resultant mixture was stirred at 120 °C for 16 h in a sealed tube. After consumption of the starting materials (monitored by TLC and LCMS), the reaction mixture was diluted with water (20 mL) and extracted with CH2C12 (2 x 20 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by column chromatography using 3% MeOH: CH2C12 to afford N-(l -(2 -methoxypyridin-4-yl) piperidin-4-yl)-8'-methyl-6'H, 8'H-spiro [cyclopentane- 1, 7'-pyrimido [5, 4-b] [1, 4] oxazin] -2'-amine (50 mg, 22%) as an off-white solid. 1H-NMR
(CD3OD, 400 MHz): δ 7.72-7.70 (m, 1H), 7.30 (s, 1H), 6.52 (d, 1H), 6.13 (s, 1H), 3.90-3.83 (m, 3H), 3.81 (s, 3H), 3.78 (s, 2H), 3.04-3.02 (m, 2H), 3.01 (s, 3H), 2.10-2.00 (m, 2H), 1.90- 1.80 (m, 4H), 1.70-1.63 (m, 4H), 1.57-1.46 (m, 2H); Mass (ESI): 411.5 [M+1]; LCMS: 41 1.2 (M+1); (column; X-Select CSH C-18 (50 3.0 mm, 3.5 μπι); RT 2.21 min; mobile phase: 0.05% Aq TFA: ACN; T/B%: 0.01/10, 0.5/10, 4/90, 8/90; flow rate: 0.8 ml/min (Gradient); UPLC (column; Acquity UPLC BEH C-18, 2.1 x 50 mm, 1.7 μ); RT 1.54 min; mobile phase: ACN: 0.025% Aq TFA; flow rate: 0.5 rnL/min; Gradient program: T/B% 0.01/90, 0.5/90, 3/10, 6/10: Diluent: CH3CN: Water; TLC: 5% MeOH/ CH2C12 (R 0.3).
Example 85
In Vitro Cell screening Assay and Quantification of Αβ(ι-χ and Αβ(ι-42) Peptides [0464] Human neuroglioma H4 cells were transfected with a pcDNA3.1 plasmid expressing human wild type APP751 cDNA and a stable cell line was generated using G418 selection. Cells were plated at 15,000 cells/well in Costar 96-well plates and placed at 37°C and 5% C02. Six hours after plating, cells were washed three times with Pro293™
chemically defined medium, followed by addition of compounds (0.003-10 μΜ, final DMSO concentration of 0.33%). Plates were incubated overnight (16-18 h) and supernatant was removed for quantification of Αβ peptides by sandwich ELISA. Cytotoxicity was evaluated using Cell-Titer 96W AQueous One Solution Cell Proliferation Assay according to the manufacturer's protocol.
ELISA measurements of Αβ peptides
[0465] Αβ peptide levels were quantified by sandwich ELISA. 96-well plates were coated with C-terminal specific Αβ antibodies recognizing either Αβ37, Αβ38, Αβ40, Αβ42, Αβ43 or a N-terminal specific Αβ antibody to detect Αβ 1-x. Plates were then blocked overnight at 4°C with 1% bovine serum albumin (BSA) in PBS-T. Plates were washed and 100 μΐ of cultured cell supernatant or synthetic Αβ peptide standards and a detection antibody (4G8-HRP) were applied to the blocked plate and incubated overnight at 4°C. The next day, wells were washed before the addition of detection substrate (TMB peroxidase). Plates were then read for absorbance at 450 nm on a Molecular Devices SpectraMax M5e Microplate Reader.
[0466] Compound-treated samples were normalized to samples treated with DMSO alone (no inhibition) and to samples treated with DAPT. IC50 values were calculated from values reported as percent of DMSO controls using nonlinear regression, based on a sigmoidal dose- response (variable slope) model. GraphPAD software from Prism used for calculation.
Table III. Biological Assay
Figure imgf000233_0001
Figure imgf000234_0001
1 u amine
Figure imgf000235_0001
Figure imgf000236_0001
Figure imgf000237_0001
Figure imgf000238_0001
1 XF 2-amine (-)-7-(4-fluorophenyl)-N-(l-(2- methoxypyridin-4-yl) piperidin-B 4-yl)-8-methyl-7, 8-dihydro-6H- 1.2097 pyrimido [5, 4-b] [1, 4] oxazin-
Figure imgf000239_0001
IF 2-amine
(+)-8-methyl-7-(o-tolyl)-N-(l- (2-(trifluoromethyl) pyridin-4-A yl) piperidin-4-yl)-7, 8-dihydro- 0.2051
6H-pyrimido [5, 4-b] [1, 4]
Figure imgf000239_0002
oxazin-2-amine
(-)-8-methyl-7-(o-tolyl)-N-(l-(2-
(trifluoromethyl) pyridin-4-yl)
B piperidin-4-yl)-7, 8-dihydro-6H- 1.7502 pyrimido [5, 4-b] [1, 4] oxazin- 2-amine
(+)-7-(4-fluorophenyl)-8- methyl-N-( 1 -(2-(trifluoromethyl)
A pyridin-4-yl) piperidin-4-yl)-7, 0.122
8-dihydro-6H-pyrimido [5, 4-b]
[1, 4] oxazin-2-amine
(-)-7-(4-fluorophenyl)-8-methyl-
N-( 1 -(2-(trifluoromethyl)
B pyridin-4-yl) piperidin-4-yl)-7, 5.7283
8-dihydro-6H-pyrimido [5, 4-b]
[1, 4] oxazin-2-amine
Figure imgf000239_0003
(-)-7-(2, 4-difluorophenyl)-N-(l- (2-methoxypyridin-4-yl)
A piperidin-4-yl)-8-methyl-7, 8- 0.9716 dihydro-6H-pyrimido [5, 4-b] [1,
4] oxazin-2-amine
(+)-7-(2, 4-difluorophenyl)-N-
( 1 -(2-methoxypyridin-4-yl)
B piperidin-4-yl)-8-methyl-7, 8- 0.0775 dihydro-6H-pyrimido [5, 4-b] [1,
4] oxazin-2-amine
Figure imgf000240_0001
[1,4] oxazin-2-amine
Figure imgf000241_0001
amine
Figure imgf000242_0001
b] [ 1 ,4]oxazin-2-amine (+)-7-(4-fluoro-2- (trifluoromethyl)phenyl)-N-(l -
(2-methoxypyridin-4- 0.1913B
yl)piperidin-4-yl)- 8 -methyl- 7, 8 -
H 1 IA dihydro-6H-pyrimido[5,4- b] [ 1 ,4]oxazin-2-amine
(-)-N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7-
3.1057A (tetrahydro-2H-pyran-4-yl)-7,8-
1 U> dihydro-6H-pyrimido[5,4- b] [ 1 ,4]oxazin-2-amine
(+)-N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-8-methyl-7-
0.9774B (tetrahydro-2H-pyran-4-yl)-7,8-
1 U> dihydro-6H-pyrimido[5,4- b] [ 1 ,4]oxazin-2-amine
(-)-7-(5-chloro-2- (trifluoromethyl)phenyl)-N-(l -
(2-methoxypyridin-4- 0.4001A
yl)piperidin-4-yl)- 8 -methyl- 7, 8 -
1 dihydro-6H-pyrimido[5,4-
CI b] [ 1 ,4]oxazin-2-amine
(+)-7-(5-chloro-2- (trifluoromethyl)phenyl)-N-(l -
(2-methoxypyridin-4- 0.0916B
yl)piperidin-4-yl)- 8 -methyl- 7, 8 -
1 dihydro-6H-pyrimido[5,4-
CI b] [ 1 ,4]oxazin-2-amine
(-)-N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-7,8-dimethyl- 0.677A
7,8-dihydro-6H-pyrimido[5,4-
H 1 b ] [ 1 ,4 ] oxazin-2- amine
(+)-N-(l -(2-methoxypyridin-4- yl)piperidin-4-yl)-7,8-dimethyl- 1.144B
7,8-dihydro-6H-pyrimido[5,4-
H 1 b] [ 1 ,4]oxazin-2-amine
Figure imgf000244_0001
F 2-amine
Figure imgf000245_0001
2-amine
Figure imgf000246_0001
Figure imgf000247_0001

Claims

We claim:
1. A compound of Formula
Figure imgf000248_0001
or a pharmaceutically acceptable salt thereof,
wherein:
R is phenyl, C1-C4 alkylene-phenyl, -Ci-C6 alkyl or 3- to 7-membered monocyclic heterocycle, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH, -CN, -NH2, -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci- C4 alkyl, -N(Ci-C4 alkyl)(C(0)(Ci-C4 alkyl)), -C(0)NH-Ci-C4 alkyl, -C(0)N(Ci-C4 alkyl)2, hydroxy-substituted C1-C4 alkyl, -S(0)2-Ci-C4 alkyl, -S(0)2-halo-substituted C1-C4 alkyl, - S(0)2-NH-Ci-C4 alkyl,
-S(0)2-N(Ci-C4 alkyl)2, -NH-S(0)2-C C4 alkyl, -N(C C4 alkyl)-S(0)2-C C4 alkyl, -C C4 alkoxy, halo-substituted C1-C4 alkoxy, 3- to 7-membered monocyclic heterocycle, C3-C8 monocyclic cycloalkyl and -C(0)NH2;
Y is 4- to 6- membered nonaromatic heterocycle, which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C1-C4 alkoxy, halo-substituted C1-C4 alkoxy, -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino- substituted C1-C4 alkoxy, (C1-C4 alkyl)2N-Ci-C4 alkoxy, -NH-C1-C4 alkyl, -CN, -OH and -NH2; and
Z is nitrogen-containing 5- to 6-membered aromatic monocyclic heterocycle which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -NH2, -OH, -C1-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkyl, C1-C4 alkoxy, halo-substituted C1-C4 alkoxy and 3- to 7-membered monocyclic heterocycle.
2. A compound of claim 1, wherein R is phenyl or C1-C4 alkylene -phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -OH, -CN, -NH2, -C1-C4 alkyl, halo-substituted C1-C4 alkyl, amino-substituted C1-C4 alkyl, -NH-C1-C4 alkyl, -NHC(0)-Ci-C4 alkyl, -N(Ci-C4
alkyl)(C(0)(Ci-C4 alkyl)), -C(0) H-Ci-C4 alkyl, -C(0)N(Ci-C4 alkyl)2, hydroxy-substituted C1-C4 alkyl, -S(0)2-Ci-C4 alkyl, -S(0)2-halo-substituted C1-C4 alkyl, -S(0)2-NH-Ci-C4 alkyl, -S(0)2-N(Ci-C4 alkyl)2, - H-S(0)2-Ci-C4 alkyl, -N(C C4 alkyl)-S(0)2-CrC4 alkyl, -C C4 alkoxy, halo-substituted C1-C4 alkoxy, 3- to 7-membered monocyclic heterocycle, C3-C8 monocyclic cycloalkyl and -C(0)NH2; or a pharmaceutically acceptable salt thereof.
3. The compound of any one of claims 1-2, wherein R is phenyl or C1-C4 alkylene- phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, -C1-C4 alkyl, -C1-C4 alkoxy, halo-substituted C1-C4 alkyl and halo-substituted C1-C4 alkoxy; or a pharmaceutically acceptable salt thereof.
4. The compound of any one of claims 1-3, wherein Y is piperidinyl or azetidinyl, each of which is unsubstituted; or a pharmaceutically acceptable salt thereof.
5. The compound of any one of claims 1-4, wherein Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, halo-substituted C1-C4 alkyl and -C1-C4 alkoxy; or a pharmaceutically acceptable salt thereof.
6. The compound of any one of claims 1-5, wherein:
R is phenyl or C1-C4 alkylene-phenyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen or -C1-C4 alkyl;
Y is piperidinyl; and Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, halo- substituted C1-C4 alkyl and -C1-C4 alkoxy; or a pharmaceutically acceptable salt thereof.
7. The compound of any one of claims 1-6, wherein:
R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C1-C4 alkyl; and
Z is pyridinyl which is unsubstituted or substituted with -C1-C4 alkoxy; or a pharmaceutically acceptable salt thereof.
8. The compound of any one of claims 1-7, wherein R is phenyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen and -C1-C4 alkyl; or a pharmaceutically acceptable salt thereof.
9. The compound of any one of claims 1-8, wherein R is phenyl which is unsubstituted; or a pharmaceutically acceptable salt thereof.
10. The compound of any one of claims 1-8, wherein R is phenyl substituted with two or three -F; or a pharmaceutically acceptable salt thereof.
11. The compound of any one of claims 1-10, wherein each -C1-C4 alkyl is methyl and each C1-C4 alkylene is methylene; or a pharmaceutically acceptable salt thereof.
12. The compound of any one of claims 1-8 and 10-11, wherein Ris phenyl substituted with one methyl; or a pharmaceutically acceptable salt thereof.
13. The compound of any one of claims 1-12, wherein Y is piperidinyl; or a
pharmaceutically acceptable salt thereof.
14. The compound of any one of claims 1-12, wherein Y is azetidinyl; or a pharmaceutically acceptable salt thereof.
15. The compound of any one of claims 1-14, wherein Z is pyridinyl or pyrimidinyl, each of which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of -C1-C4 alkyl, -CF3 and -C1-C4 alkoxy; or a pharmaceutically acceptable salt thereof.
16. The compound of any one of claims 1-15, wherein Z is pyridinyl substituted with one -CF3; or a pharmaceutically acceptable salt thereof.
17. The compound of any one of claims 1-15, wherein Z is pyridinyl substituted with one methyl; or a pharmaceutically acceptable salt thereof.
18. The compound of any one of claims 1-15, wherein Z is pyridinyl substituted with one methoxy; or a pharmaceutically acceptable salt thereof.
19. The compound of any one of claims 1-15, wherein Z is pyrimidinyl which is unsubstituted or substituted with one to three -C1-C4 alkyl; or a pharmaceutically acceptable salt thereof.
20. The compound of any one of claims 1-15 and 19, wherein Z is pyrimidinyl substituted with one methyl; or a pharmaceutically acceptable salt thereof.
21. A compound selected from the group consisting of:
N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-phenyl-7,8-dihydro-6H- pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-phenyl-7,8-dihydro-6H- pyrimido[5,4-b][l,4]oxazin-2-amine; N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(o-tolyl)-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(o-tolyl)-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine;
8-methyl-N-(l-(6-methylpyrimidm^
dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-8-methyl-N-(l-(6-methylpyrimidin-4-yl)piperidin-4-yl)-7-(3,4,5-trifl^
dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
7- (3,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dih 6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-7-(3 ,5 -difluorophenyl)-N-( 1 -(2-meth^
dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8-dih 6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-N-(l-(2-methoxypyridin-4-yl)pip^
dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(-)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluoroph^ dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
8- methyl-7-phenyl-N-(l-(2-(trifluoromethyl)pyridin-4-yl)piperidin-4-yl)-7,8-dihyd pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-8-methyl-7-phenyl-N-(l-(2-(trifluoromethyl)pyridin-4-yl)piperidin-4-yl)-7,8-dih pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine; (+)-7-benzyl-8-methyl-N-( 1 -(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro-6H- pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(3, 5-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(4-fluorophenyl)-8-methyl-N-(l -(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-benzyl-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(4-fluorophenyl)-N-(l -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-8-methyl-7-(o-tolyl)-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(4-fluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(2, 4-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(2, 4-difluorophenyl)-8-methyl-N-(l-(6-methylpyrimidin-4-yl) piperidin-4-yl)-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(-)-N-( 1 -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2-(trifluoromethoxy) phenyl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; (+)-N-( 1 -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7-(2-(trifluoromethoxy) phenyl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; and
(+)-7-(2, 4-difluorophenyl)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)- 7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-benzyl-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine;
(-)-7-benzyl-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro- 6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-7-(2,4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(2,4,5-trifluorophenyl)-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(4-fluoro-2-(trifluoromethyl)phenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(5-chloro-2-(trifluoromethyl)phenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(5-fluoro-2-(trifluoromethyl)phenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(2-chloro-4,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl- 7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(3,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l ,4]oxazin-2-amine;
(+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(o-tolyl)-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(3,5-difluorophenyl)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H- pyran-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(2,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l ,4]oxazin-2-amine;
(-)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(2,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l ,4]oxazin-2-amine; or a pharmaceutically acceptable salt thereof.
22. The compound of claim 21 selected from the group consisting of:
7-(3,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro- 6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-7-(3,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8-dihydro- 6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(3,4,5-trifluorophenyl)-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine; (+)-7-(4-fluorophenyl)-N-(l -(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8-dihydro- 6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-(2, 4-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl) piperidin-4-yl)-8-methyl-7, 8- dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine; and
(+)-8-methyl-N-(l-(2-(trifluoromethyl) pyridin-4-yl) piperidin-4-yl)-7-(3, 4, 5- trifluorophenyl)-7, 8-dihydro-6H-pyrimido [5, 4-b] [1, 4] oxazin-2-amine;
(+)-7-benzyl-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro-6H- pyrimido[5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-7-(2-chlorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8-dihydro- 6H-pyrimido [5 ,4-b] [ 1 ,4]oxazin-2-amine;
(+)-7-(2,4-dichlorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7-(2,4,5-trifluorophenyl)-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(2-chloro-4-fluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl-7,8- dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(5-fluoro-2-(trifluoromethyl)phenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8- methyl-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-7-(2-chloro-4,5-difluorophenyl)-N-(l-(2-methoxypyridin-4-yl)piperidin-4-yl)-8-methyl- 7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(3,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l ,4]oxazin-2-amine; (+)-7-(3,5-difluorophenyl)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H- pyran-3-yl)-7,8-dihydro-6H-pyrimido[5,4-b][l,4]oxazin-2-amine;
(+)-8-methyl-N-((3R,6S)-6-(2-methylpyridin-4-yl)tetrahydro-2H-pyran-3-yl)-7-(2,4,5- trifluorophenyl)-7,8-dihydro-6H-pyrimido[5,4-b][l ,4]oxazin-2-amine; or a pharmaceutically acceptable salt thereof.
23. A pharmaceutical composition comprising a pharmaceutically acceptable carrier or vehicle and an effective amount of a compound or a pharmaceutically acceptable salt of a compound of any one of claims 1-22.
24. A method for treating a neurodegenerative disease, comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable salt of a compound of any one of claims 1-22.
25. The method of claim 24, wherein the neurodegenerative disease is panic disorder, obsessive compulsive disorder, delusional disorder, drug-induced psychosis, post-traumatic stress disorder, age-related cognitive decline, attention deficit/hyperactivity disorder, personality disorder of the paranoid type, personality disorder of the schizoid type, dyskinesia, choreiform condition, psychosis associated with Parkinson's disease, psychotic symptoms associated with Alzheimer's disease, mood disorder, or dementia.
26. A method for treating Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable salt of a compound of any one of claims 1-22.
27. A method for improving an impaired cognitive function, comprising administering to a subject having impaired cognitive function an effective amount of a compound or a pharmaceutically acceptable salt of a compound of any one of claims 1-22.
28. A method for ameliorating a symptom of Alzheimer's disease, comprising administering to a subject in need thereof an effective amount of a compound or a pharmaceutically acceptable salt of a compound of any one of claims 1-22.
29. The method of any one of claims 26 and 28, wherein Alzheimer's disease is early onset Alzheimer's disease.
30. The method of any one of claims 24-29, wherein the subject is a human.
31. The method of claim 28, wherein the symptom is progressive loss of memory, progressive loss of cognition, progressive loss of reasoning and/or progressive loss of judgment.
32. The method of claim 27, wherein the cognitive function impaired is one or more of attention, learning, delayed memory, working memory, visual learning, speed of processing, vigilance, verbal learning, visual motor function, social cognition, long term memory or executive function.
PCT/US2015/020114 2014-03-13 2015-03-12 Fused morpholinopyrimidines and methods of use thereof WO2015138689A1 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201461952704P 2014-03-13 2014-03-13
US61/952,704 2014-03-13
US201462054673P 2014-09-24 2014-09-24
US62/054,673 2014-09-24

Publications (1)

Publication Number Publication Date
WO2015138689A1 true WO2015138689A1 (en) 2015-09-17

Family

ID=54072390

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2015/020114 WO2015138689A1 (en) 2014-03-13 2015-03-12 Fused morpholinopyrimidines and methods of use thereof

Country Status (1)

Country Link
WO (1) WO2015138689A1 (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100298359A1 (en) * 2007-12-06 2010-11-25 Xianhai Huang Gamma secretase modulators
US20110201605A1 (en) * 2010-02-17 2011-08-18 Karlheinz Baumann Heteroaryl substituted piperidines

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100298359A1 (en) * 2007-12-06 2010-11-25 Xianhai Huang Gamma secretase modulators
US20110201605A1 (en) * 2010-02-17 2011-08-18 Karlheinz Baumann Heteroaryl substituted piperidines

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
RICHTER, L ET AL.: "Amyloid beta 42 peptide (Abeta42)-lowering compounds directly bind to Abeta and interfere with amyloid precursor protein (APP) transmembrane dimerization", PNAS, vol. 107, no. 33, 2010, pages 14597 - 14602, XP055017429, Retrieved from the Internet <URL:http://www.pnas.org/content/107/33/14597.full.pdf> *

Similar Documents

Publication Publication Date Title
US9771378B2 (en) Fused morpholinopyrimidines and methods of use thereof
US9139594B2 (en) Fused aminodihydropyrimidone derivatives
EP2285800B1 (en) Quinoline or isoquinoline substituted p2x7 antagonists
AU2020203035A1 (en) CDK Inhibitors
EP2379553B1 (en) Bicyclic compounds for the reduction of beta-amyloid production
TWI468402B (en) Compounds for the reduction of β-amyloid production
US7935815B2 (en) Imidazoyl pyridine compounds and salts thereof
KR20230163575A (en) Inhibitors of (alpha-v)(beta-6) integrin
IL299415A (en) Pyrimidin-2-ylamino-1h-pyrazols as lrrk2 inhibitors for use in the treatment of neurodegenerative disorders
WO2012009309A1 (en) Compounds for the reduction of beta-amyloid production
KR20180095833A (en) Fused tricyclic heterocyclic compounds as HIV integrase inhibitors
US20170050980A1 (en) Oxadiazine compounds and methods of use thereof
EP3722298A1 (en) Pyrrolo(pyrazolo)pyrimidine derivative as lrrk2 inhibitor
EP4149937A1 (en) Htt modulators for treating huntington&#39;s disease
EP3097103B1 (en) Diazabicyclo[4.3.1]decane derivatives for treatment of psychiatric disorders
EP3255993B1 (en) Inhibitors of the renal outer medullary potassium channel
WO2015066697A1 (en) Fused morpholinopyrimidines and methods of use thereof
WO2015066696A1 (en) Fused morphlinopyrimidines and methods of use thereof
WO2023224894A1 (en) Macrocycles as lrrk2 inhibitors, pharmaceutical compositions, and uses thereof
WO2016201168A1 (en) Oxadiazine compounds and methods of use thereof
WO2015138689A1 (en) Fused morpholinopyrimidines and methods of use thereof
US11970493B2 (en) Autotaxin inhibitor compounds
CN116322692A (en) Autotaxin inhibitor compounds
WO2023177563A2 (en) Fused pyrazole amide analogs as glucosylceramide synthase inhibitors

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15761707

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC , EPO FORM 1205A DATED 09.01.2017.

122 Ep: pct application non-entry in european phase

Ref document number: 15761707

Country of ref document: EP

Kind code of ref document: A1