WO2011077098A1 - Inhibitors of akt activity - Google Patents

Inhibitors of akt activity Download PDF

Info

Publication number
WO2011077098A1
WO2011077098A1 PCT/GB2010/002329 GB2010002329W WO2011077098A1 WO 2011077098 A1 WO2011077098 A1 WO 2011077098A1 GB 2010002329 W GB2010002329 W GB 2010002329W WO 2011077098 A1 WO2011077098 A1 WO 2011077098A1
Authority
WO
WIPO (PCT)
Prior art keywords
phenyl
pyrido
oxazin
aminocyclobutyl
dihydro
Prior art date
Application number
PCT/GB2010/002329
Other languages
French (fr)
Other versions
WO2011077098A9 (en
Inventor
Lixin Zhang
Graham Peter Trevitt
Hughes Miel
Frank Burkamp
Timothy Harrison
Andrew John Wilkinson
Charles-Henry Fabritius
Original Assignee
Almac Discovery Limited
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
Priority to SG2012041232A priority Critical patent/SG181517A1/en
Priority to ES10800980.4T priority patent/ES2503794T3/en
Application filed by Almac Discovery Limited filed Critical Almac Discovery Limited
Priority to MX2012007274A priority patent/MX2012007274A/en
Priority to NZ600453A priority patent/NZ600453A/en
Priority to EP10800980.4A priority patent/EP2516435B8/en
Priority to KR1020127019033A priority patent/KR20140015157A/en
Priority to AU2010334591A priority patent/AU2010334591C1/en
Priority to BRBR112012015677-2A priority patent/BR112012015677A2/en
Priority to DK10800980.4T priority patent/DK2516435T3/en
Priority to PL10800980T priority patent/PL2516435T3/en
Priority to JP2012545434A priority patent/JP5923043B2/en
Priority to CA2783340A priority patent/CA2783340C/en
Priority to RU2012131115/04A priority patent/RU2579513C2/en
Priority to CN201080059321.9A priority patent/CN102791708B/en
Priority to US13/517,219 priority patent/US9221838B2/en
Priority to SI201030774T priority patent/SI2516435T1/en
Publication of WO2011077098A1 publication Critical patent/WO2011077098A1/en
Publication of WO2011077098A9 publication Critical patent/WO2011077098A9/en
Priority to IL220153A priority patent/IL220153A/en
Priority to HK12111698.8A priority patent/HK1171013A1/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
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-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

  • the present invention relates to compounds that are useful as inhibitors of the activity of one or more isoforms of the serine / threonine kinase, AKT.
  • the present invention also relates to pharmaceutical compositions comprising these compounds and to methods of using these compounds in the treatment of cancer and methods of treating cancer.
  • AKT protein family also known as protein kinases B (PKB)
  • PKA protein kinases B
  • enzymes are members of the serine/threonine-specific protein kinase family.
  • the PKB / AKT pathway has been identified as an important regulator of cell survival signalling and apoptosis in cells. Signalling is thought to occur through a range of growth factor receptors including platelet derived growth factor, insulin growth factor and nerve growth factor, resulting in activation of phosphatidylinositol 3-OH kinase (PI-3K). This activation in turn leads to the generation of phosphatidylinositol (3,4,5) triphosphate (PIP3).
  • PIP3 binds to and in turn phosphorylates the enzyme PDK-1 , the main activator of AKT, through its pleckstrin homology domain.
  • Activated PDK-1 is responsible for a phosphorylation event at Thr308 of AKT, which induces a conformational change that facilitates further phosphorylation of AKT at Ser 473 by PDK-2.
  • PDK-1 phosphorylation of downstream kinases is not unique to AKT, as it has been reported to activate p70 S6 kinase and protein kinase C.
  • AKT AKT-like protein kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase, kinase, kinase, kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinase kinas
  • AKT 1 AKT 1
  • AKT 2 AKT 3
  • PKBpr PKB 3
  • the family members share 80% amino acid sequence homology and all retain similar regional structure. They possess a C-terminal pleckstrin homology (PH) domain, a catalytic domain, a short or helical linker region and a carboxyl terminal domain.
  • the PH domain permits binding of proteins to the cell membrane through a phospholipid interaction.
  • the catalytic domain of AKT family members contains two residues essential for kinase activation, namely Thr308 and Ser 473.
  • AKT can phosphorylate any protein containing the RXRXXS/T-B motif where X represents any amino acid and B represents bulky hydrophobic residues.
  • AKT hyper activation of AKT has been linked to the inhibition of cellular apoptosis due to phosphorylation and negative regulation of the forkhead family of transcription factors which regulate various genes responsible for instigating death processes including FKHR, FKHRL1 and AFX.
  • AKT has been reported to up-regulate genes which are known to be anti-apoptotic including IKK and CREB. It is this mixture of positive and negative regulation which highlights the importance of AKT in regulating apoptosis.
  • AKT promotes unwanted cell survival through its' phosphorylation of several key apoptotic proteins including Bad and Pro-caspase 9, thus rendering them inactive and preventing signalling through this pathway.
  • AKT activates and inhibits multiple mechanisms which have a major role in the progression of the cell cycle, ultimately leading to cell proliferation.
  • the best characterised cell cycle regulator and tumour suppressor proteins p53 can be dysregulated via AKT
  • Phosphorylated MDM2 translocates to the nucleus where it prevents p53 transcription.
  • the inhibition of p53 allows aberrant proliferation of the cell and progression towards a benign state.
  • AKT can also phosphorylate p27kip1 and p21 ; two main inhibitors of cell cycle progression, leading to loss of function, resulting in unchecked cell cycle progress and excessive proliferation.
  • AKT activation causes an increase in the rate of glycolysis by increasing the rate of glucose metabolism. It has also been reported that activated AKT stimulates the transport of amino acids and supports mTOR dependent increases in protein translation. Proangiogenic factors such as vascular endothelial growth factor (VEGF), have been reported to activate AKT, ultimately resulting in inhibition of endothelial apoptosis, as well as activating endothelial nitric oxide synthase (eNOS). The sum result of this is rapid neo-vascularisation and cell migration.
  • VEGF vascular endothelial growth factor
  • hypoxia driven angiogenesis primarily mediated by hypoxia inducible factor (HIF 1a) can lead to the induction of multiple proteins including VEGF.
  • Increased activated AKT has been reported to increase HIF-1 a expression leading to an increase in angiogenesis independent of a hypoxic environment.
  • Recent data has shown that HIF-1cr activity in invasive breast cancer is correlated with increased activated AKT-1 phosphorylation.
  • Estrogen receptor (ER) and androgen receptor (AR) inhibitors designed to inhibit cell signalling and induce apoptosis are vital tools in cancer therapies. Incidence of resistance to these drugs arises rapidly in cancers including prostate, breast and ovarian. AKT has been reported to phosphorylate androgen receptors, leading to inhibition of AR activity and blockade of normal apoptotic signalling in prostate cancer induced by androgens.
  • AKT-2 has been identified as a promoter of ERa transcription in the presence or absence of estrogen increasing the rate of proliferation of breast cancer cells.
  • Hyper-activated AKT has been reported in a range of cancers compared to normal tissues including breast, lung, prostate, gastric, ovary, pancreas, thyroid, glioblastoma and haemological cancers. Phosphorylation of AKT has also been associated with clinical characteristics including increased stage and grade of tumour and increased poor prognosis. The activation of AKT can arise from a number of different genetic mutations in the AKT/ PI-3K pathway.
  • Somatic mutations in the PI-3KCA gene have been widely reported in a large variety of tumours including breast, prostate and head and neck. A large number of these mutations will increase the copy number of the gene leading to an increase in PI-3K activity. A recent study has identified a PI-3K mutation which selectively phosphorylates AKT in colon cancer which results in increase cell proliferation and invasion.
  • AKT-2 Amplifications of AKT-2 have been reported in ovarian, pancreatic, breast and head and neck squamous cell carcinoma. No amplifications or mutations in AKT-3 have been reported to date although deletion mutations leading to hyperactivation and amplification mutations have been reported associated with AKT-1.
  • E17K One mutation; results in pathological localization of AKT-1 to the cell membrane, inducing its activation and resulting in down-stream signalling and cellular transformation. In vivo, this mutation has been shown to induce leukaemia in mice.
  • Phosphatase and tensin homolog deleted on chromosome 10 is a tumour suppressor gene known to negatively regulate AKT function.
  • loss of PTEN function results in constitutive phosphorylation of AKT and other down-stream effectors of the PI-3K pathway.
  • Loss of PTEN, due to deletion mutations or promoter methylation, has been reported in a number of different cancers including glioblastoma, endometrial, lung, breast, prostate and thyroid. This loss is commonly associated with hyperactivation of AKT.
  • LHO heterozygosity
  • AKT activation is commonly initiated at the cell surface through a signalling event at a receptor, usually one of the tyrosine kinase family.
  • a receptor usually one of the tyrosine kinase family.
  • Two tyrosine kinase receptors commonly amplified or over-expressed in cancer are HER2 and EGFR.
  • HER2 over- expressing tumours there is often a hyper-activation of AKT, this has been reported in ovarian, stomach and bladder cancer.
  • EGFR over-expressing tumours, particularly those with the EGFRvlll activating mutation selective activation of AKT has been reported in a range of cancers including non-small cell lung cancers, breast, ovarian and most commonly high grade gliomas.
  • AKT inhibitors are provided in WO 2008/070134, WO 2008/070016 and WO 2008/070041. These documents provide specific naphthyridine compounds fused to a five membered heterocycle. Other inhibitors of AKT may be found in, for example, WO 2009/1 8887 and WO 2009/148916.
  • R 1 and R 2 are each independently selected from hydrogen, aryl, C1-C10 alkyl, CN, CHO, C0 2 H, CONH 2 , CONHR 3 , CONR 3a R 3 , COR 3 , C0 2 R 3 , NH 2 , NHR 3 , NR 3a R 3b , NHCOR 3 , NHS0 2 R 3 , NR 3a COR 3 , NR 3a S0 2 R 3b , OH, OR 3 , SH, SR 3 , SOR 3 , S0 2 R 3 , S0 2 NHR 3 , S0 2 NR 3a R 3b , F, CI, Br and I, wherein each R 3 , R 3a and R 3 is independently selected from C1-C10 alkyl, including wherein R 3a and R 3b are joined to one another to form a heterocycle that includes the nitrogen to which they are attached .
  • R 1 and R 2 may be joined to one another to form an optionally substituted and optionally saturated heterocycle or carbocycle that includes the C atom to which they are attached; or R 1 and R 2 together are oxo or optionally C1-C10 alkyl O- substituted oxime;
  • W is O, S, SO, S0 2 , NR', or CR a R where R' is either hydrogen or C1-C10 alkyl, and where R a and R b are each independently selected from the members of the group from which R 1 and R 2 are selected above;
  • X is either absent or is CR 4 R 5 where R 4 and R 5 are each independently selected from the members of the group from which R 1 and R 2 are selected above; Y and Z are independently either substituted or unsubstituted nitrogen or carbon and where carbon is substituted by substituents independently selected from the members of the group from which R 1 and R 2 are selected above or when nitrogen then the substituent is selected from aryl, C1-C10 alkyl, S0 2 R 3 , CONHR 3 , CONR 3a R 3b , COR 3 and C0 2 R 3 where R 3 , R 3a and R 3b are as defined above, or where Y and Z together form an optionally substituted heterocyclyl or carbocyclic group, or where Y is S0 2 ;
  • R 7a and R 7b are independently selected from H and alkyl, including wherein R 7a and R 7b are joined to one another to form a heterocycle that includes the nitrogen to which they are attached;
  • R 63 and R 6b are independently selected from: H, (Ci-C 6 )alkyl, (C r C 6 )alkenyl, and (C r
  • C 6 )alkynyl wherein said alkyl is optionally substituted with up to three substituents selected from: OH and halo; or R 63 and R 6b can be taken together to form a monocyclic or bicyclic carbo- or heterocycle with 3-7 members in each ring, said heterocycle having one or more heteroatoms selected from N, O and S, and said carbo- or heterocycle is optionally substituted with one or more substituents selected from: (Ci-C 6 )alkyl, (C 3 -C 6 )cycloalkyl, (C C 6 )alkoxy, CO z H, halo, OH, CN and NR 3a R 3 , said alkyl, cycloalkyl and alkoxy is optionally substituted with one or more substituents selected from halo, CN, OH and NR 3a R 3b ; and ring Cy is selected from (C 3 to C 8 )cycloalkyl, alkylcycloalkyl,
  • One aspect of the invention also provides compounds selected from the following group of structures: and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
  • X is absent.
  • R 6a and R 6b together form , that is they preferably form cyclobutane.
  • the group bound to the phenyl ring in structure I is 1-aminocyclobutyl.
  • R 6a and R 6b together form where R is an alkyl group, preferably a C1 to C6 alkyl group.
  • the ring Cy is unsubstituted C 6 aryl, that is phenyl.
  • the bond between Y and Z is optionally either a single or a double bond. In preferred embodiments the bond between Y and Z is a single bond.
  • X is absent, Y is carbonyl and Z is optionally substituted amino, or X is absent, Y is optionally substituted amino and Z is carbonyl.
  • W is preferably CH 2 or O, more preferably O.
  • R 1 and R 2 are preferably hydrogen. In these embodiments when amino is substituted it is preferably methyl or acetamido substituted.
  • substituents bound to Y and Z together with Y and Z themselves form a 5 or 6 membered optionally substituted carbocyclic or herterocyclic ring.
  • the ring Cy is replaced with an iodine atom.
  • Cy is alkylcycloalkyl
  • Cy is methylcyclopropyl
  • W is either CH 2 or O. In one particularly preferred
  • W is CH 2 . In another particularly preferred embodiment W is O.
  • cyclobutane which additionally has a polar substituent at the position distal to the benzene ring to which the cyclobutane ring is bound, the polar substituent is preferably in a trans relationship to the nitrogen atom having groups R 7a and R 7 pendant therefrom.
  • polar substituent is preferably in a trans relationship to the nitrogen atom having groups R 7a and R 7 pendant therefrom.
  • alkyl group refers to an aliphatic group containing at least carbon and hydrogen and containing 1 to 15 carbon atoms, such as 1 to 10 carbon atoms.
  • Attachment to the alkyl group occurs through a carbon atom.
  • C n alkyl refers to an aliphatic group containing n carbon atoms.
  • a C1-C1 0 alkyl group contains 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.
  • An alkyl group may be straight chained or it may be branched.
  • An alkyl group may contain no ring structures or it may contain one or more rings.
  • a "cycloalkyi" group contains at least one ring. It is understood that attachment to a cycloalkyi group is via a ring of the cycloalkyi group.
  • Each ring may contain 3 to 10 atoms, such as 4 to 8 or 5 to 7 atoms.
  • Each ring may be independently selected to contain just carbon atoms or to contain both carbon atoms and from 1 to 4 heteroatoms selected from O, N and S.
  • cyclo-heteroalkyl groups i.e.
  • attachment to the cycloalkyi group may occur either through a carbon atom or, if one or more heteroatoms are contained in a ring, attachment may also occur through a heteroatom contained in a ring.
  • a cycloalkyi group may be mono-cyclic or bi-cyclic.
  • a "C n cycloalkyi” group contains n carbon atoms. All n carbon atoms may be contained in the ring(s) of the cycloalkyi group or one or more of the carbons may not be contained in the ring(s) and may instead form one or more chains branching from the ring. If a C n alkyl group is joined to a separate C m alkyl group containing m carbon atoms to form, for example, a heterocycle, the two alkyl groups contain a total number of m + n carbon atoms.
  • an alkyl group may be saturated or unsaturated.
  • the alkyl group may be an alkenyl group (i.e. contain a carbon-carbon double bond) and / or an alkynyl group (i.e. contain a carbon-carbon triple bond). If the alkyl group is unsaturated, it may contain at least 2 carbon atoms. It is understood that any unsaturated portions of an alkyl group are non-aromatic (aromatic groups fall within the scope of the definition of "aryl'). Any part of the alkyl group may be unsaturated, for example the straight, branched or cyclic portion of an alkyl group may contain a carbon-carbon double bond or a carbon-carbon triple bond. Attachment to an unsaturated alky group may occur through the unsaturated part of the alkyl group or may occur through the unsaturated part of the group.
  • an unsaturated alkyl group may contain 1 to 4 carbon-carbon double bonds or 1 to 3 carbon-carbon triple bonds or 1 to 4 of a combination of carbon-carbon double bonds and carbon-carbon triple bonds.
  • An alkyl group may be substituted with one or more heteroatoms or it may be
  • the substituents are independently selected from one another unless they form a part of a particular functional group (e.g. an amide group).
  • the heteroatom substituents may in turn be substituted with further carbon-containing groups.
  • the C n or C m prefix that defines the substituted alkyl group refers to the total number of carbons contained in the group, i.e. including the carbon atoms contained in any substituted heteroatomic groups, and the total alkyl group contains 1 to 15 carbon atoms as defined previously.
  • the alkyl group may, for example, contain one or more of CN, C0 2 H, CONH 2l CONHR, CONR a R b , C0 2 R, NH 2 , NHR, NR a R b , OH, OR, SH, SR, F, CI, Br and I, wherein each R, R a and R are independently selected groups (e.g. alkyl / aryl groups) attached to the atom to which the group joins through a carbon atom of each group, including wherein R a and R b form a heterocycle that includes the heteroatom to which they are attached.
  • a group containing two C m -C n alkyl moieties that form a cycle that includes, for example, the heteroatom to which they are attached may contain from C 2m to C 2n carbon atoms.
  • unsubstituted saturated alkyl groups containing no cyclic structures include methyl, ethyl, n-propyl, sec-propyl, n-butyl, sec-butyl, terf-butyl, pentyl (branched or unbranched), hexyl (branched or unbranched), heptyl (branched or unbranched), octyl (branched or unbranched), nonyl (branched or unbranched), and decyl (branched or unbranched).
  • unsubstitued saturated cyclic alkyl groups include cyclopropyl, cylcobutyl, cyclopentyl and cyclohexyl.
  • unsaturated alkyl groups include ethenyl, propenyl, butenyl, 2-methybutenyl and cyclohexenyl.
  • aryl group refers to a group containing at least one ring that is aromatic and containing 1 to 15 carbon atoms, such as 1 to 10 carbon atoms. Where an aryl group is stated as being substituted at a particular position, attachment of the position to the aryl group is onto the aromatic ring of the aryl group itself rather than the position being joined to the aryl group through any non-aromatic side-chain of the aryl group. For example, when R 1 is an aryl group in CR 1 , the C is attached to the aromatic part of the aryl group.
  • Each ring may be independently selected to contain only carbon atoms or to contain both carbon atoms and from 1 to 4 heteroatoms selected from O, N and S.
  • heteroaryl groups i.e. aryl groups that contain one or more heteroatoms
  • attachment to the aryl group may occur either through a carbon atom or, if one or more heteroatoms are contained in a ring, attachment may also occur through a heteroatom contained in a ring.
  • heteroatoms contained in a ring of a heteroaryl group may be substituted, for example forming an /V-oxide.
  • the aromatic group may be mono-cyclic or bi-cyclic, wherein one or both of the rings of a bi-cyclic system is aromatic.
  • aryl groups include acridinyl, phenyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl, benzotriazolyl, furanyl, naphthyl, thienyl, thiazolyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline, benzimidazolyl and melaminyl.
  • heterocycle includes within its scope both cycloalkyl groups containing one or more heteroatoms within the ring system and aryl groups containing one or more heteroatoms within the ring system.
  • Heterocyclic groups may be any of the various pyrazoles, imidazoles and triazoles and may include the oxygen and/or sulfur containing analogues of the various pyrazoles, imidazoles and triazoles, that is oxazoles, isoxazoles, thiazoles and isothiazoles and derivatives.
  • halo refers to a group selected from chlorine, fluorine, bromine and iodine.
  • the compounds of the present invention may possess some aspect of stereochemistry.
  • the compounds may possess chiral centres and / or planes and / or axes.
  • the compounds may be provided as single
  • Stereoisomers are known in the art to be molecules that have the same molecular formula and sequence of bonded atoms, but which differ in their spatial orientations of their atoms and / or groups.
  • the compounds of the present invention may possess tautomerism. Each tautomeric form is intended to fall within the scope of the invention.
  • the compounds of the present invention may be provided as a pro-drug.
  • Prodrugs are transformed, generally in vivo, from one form to the active forms of the drugs described herein.
  • a prodrug may be formed by protecting the amine appending the cyclobutane as a physiological hydrolyzable amide.
  • Y or Z or any moieties appended thereto are NH, one or more of these may be protected as a physiological hydrolyzable amide.
  • the compounds of the present invention may be provided in the form of their pharmaceutically acceptable salts or as co-crystals.
  • the compounds may be provided having protonated amine groups.
  • pharmaceutically acceptable salt refers to ionic compounds formed by the addition of an acid to a base.
  • the term refers to such salts that are considered in the art as being suitable for use in contact with a patient, for example in vivo and
  • compositions are generally chosen for their non-toxic, non-irritant characteristics.
  • co-crystal refers to a multi- component molecular crystal, which may comprise non-ionic interactions.
  • Pharmaceutically acceptable salts and co-crystals may be prepared by ion exchange chromatography or by reacting the free base or acidic form of a compound with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in one or more suitable solvents, or by mixing the compound with another pharmaceutically acceptable compound capable of forming a co-crystal.
  • Salts known in the art to be generally suitable for use in contact with a patient include salts derived from inorganic and / or organic acids, including the hydrobromide, hydrochloride, sulphate, bisulphate, nitrate, acetate, oxalate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate and tartrate. These may include cations based on the alkali and alkaline earth metals, such as sodium, potassium, calcium and magnesium, as well as ammonium,
  • the compounds of the present invention may sometimes exist as zwitterions, which are considered as part of the invention.
  • the compounds of the present invention are useful in the treatment of medical conditions associated with disordered cell growth, including, but not restricted to, cancer, in particular cancers associated with overactivity of AKT occurring either from a direct change within the kinase itself such as may occur following a mutation within any of its subunits or from increased upstream activity including but not restricted to increased PI3K or PDK activity. Increased PI3K activity may have occurred through loss of the tumor suppressor PTEN.
  • cancers include cardiac cancers, lung cancers, gastrointestinal cancers, genitourinary tract cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, hematologic cancers, skin cancers and adrenal gland cancers.
  • cancers include adrenal tumors, bile duct, bladder, blood, bone and connective tissue, brain and central nervous system, breast, cervical, colon and rectal (colorectal), endometrial, esophageal, gallbladder, head and neck, Hodgkin's Lymphoma, hypopharangeal, kidney, laryngeal, leukemias, liver, lung, lymphoma, mediastinal tumors, melanoma (malignant melanoma), mesothelioma, multiple myeloma, nasal cavity, nasopharyngeal, neuroendocrine tumors, non-Hodgkin's lymphoma, oral, oesophagus, oropharyngeal, ovarian, pancreas, paranasal sinus, parathyroid, penis, pituitary tumors, prostate, salivary gland, sarcoma, skin, spine, stomach, testicular, thyroid, urethra, uterine
  • the compounds of the present invention are also useful in preparing a medicament that is useful in treating the diseases described above, in particular cancer.
  • the compounds of the present invention may selectively inhibit one or two of the AKT protein family over the other AKT isoform(s).
  • the compounds may selectively inhibit one or two of AKT1 , AKT2 or AKT3 over the other isoform(s) of AKT.
  • the compounds of the present invention may inhibit at least AKT1 and / or AKT2.
  • the compounds may selectively inhibit AKT1 and / or AKT2 over AKT3.
  • the present invention is further directed to a method of inhibiting AKT activity which comprises administering to a mammal in need thereof a pharmaceutically effective amount of the compound of the present invention.
  • the compounds of this invention may be administered to mammals, including humans, either alone or, in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition, according to standard pharmaceutical practice.
  • the compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.
  • the present invention also includes within its scope the use of the compounds of the present invention in combination with a second drug in the treatment of cancer.
  • the second drug may be a drug that is already known in the art in the treatment of cancer.
  • the present invention also includes the use of the compounds of the invention in a regime including the step of radiotherapy.
  • cancers often become resistant to therapy.
  • the development of resistance may be delayed or overcome by the administration of a combination of drugs that includes the compounds of the present invention.
  • drugs that may be used in combination with the compounds of the present invention may target the same or a similar biological pathway to that targeted by the compounds of the present invention or may act on a different or unrelated pathway.
  • the second active ingredient may include, but is not restricted to: alkylating agents, including cyclophosphamide, ifosfamide, thiotepa, melphalan, chloroethylnitrosourea and bendamustine; platinum derivatives, including cisplatin, oxaliplatin, carboplatin and satraplatin; antimitotic agents, including vinca alkaloids (vincristine, vinorelbine and vinblastine), taxanes (paclitaxel, docetaxel), epothilones and inhibitors of mitotic kinases including aurora and polo kinases; topoisomerase inhibitors, including anthracyclines, epipodophyllotoxins, camptothecin and analogues of camptothecin; antimetabolites, including 5-fluoro uracil, capecitabine
  • alkylating agents including cyclophosphamide, ifosfamide, thiotep
  • proteosome inhibitors including bortezomib; histone deacetylase inhibitors, including valproate and SAHA; antiangiogenic drugs, including bevacizumab; monoclonal antibodies, including trastuzumab, rituximab, alemtuzumab, tositumomab, cetuximab, panitumumab; conjugates of myoclonal antibodies, including Gemtuzumab ozogamicin, Ibritumomab tiuxetan; hormonal therapies, including antiestrogens (tamoxifen, raloxifen, anastrazole, letrozole, examestane) antiandrogens (Flutamide, Biclutamide) and Luteinisng Hormone Analogues or antagonists.
  • monoclonal antibodies including trastuzumab, rituximab, alemtuzumab, tositumomab, cetuximab
  • the compounds of the present invention may be administered separately, sequentially, simultaneously, concurrently or may be chronologically staggered with one or more standard therapeutics such as any of those mentioned above.
  • the present invention also provides a pharmaceutical composition suitable for clinical use.
  • a pharmaceutical composition may comprise a pharmaceutical carrier and, dispersed therein, a therapeutically effective amount of the compounds of the invention.
  • the composition may be solid or liquid.
  • the pharmaceutical carrier is generally chosen based on the type of administration being used and the pharmaceutical carrier may for example be solid or liquid.
  • the compounds of the invention may be in the same phase or in a different phase than the pharmaceutical carrier.
  • compositions may be formulated according to their particular use and purpose by mixing, for example, excipient, binding agent, lubricant, disintegrating agent, coating material, emulsifier, suspending agent, solvent, stabilizer, absorption enhancer and / or ointment base.
  • the composition may be suitable for oral, injectable, rectal or topical administration.
  • the pharmaceutical composition may be administered orally, such as in the form of tablets, coated tablets, hard or soft gelatine capsules, solutions, emulsions, or suspensions.
  • Administration can also be carried out rectally, for example using suppositories, locally or percutaneously, for example using ointments, creams, gels or solution, or paenterally, for example using injectable solutions.
  • the compounds of the present invention may be admixed with pharmaceutically inert, inorganic or organic excipients.
  • suitable excipients include lactose, mize starch or derivatives thereof, talc or stearic acid or salts thereof.
  • suitable excipients for use with soft gelatine capsules include, for example, vegetable oils, waxes, fats and semi-solid or liquid polyols.
  • excipients include, for example, water, polyols, saccharose, invert sugar and glucose.
  • excipients include, for example, water, alcohols, polyols, glycerine and vegetable oil.
  • excipients include, for example, natural or hardened oils, waxes, fats and semi-solid or liquid polyols.
  • compositions may also contain preserving agents, solublizing agents, stabilizing agents, wetting agents, emulsifiers, sweeteners, colorants, odorants, buffers, coating agents and / or antioxidants.
  • the second drug may be provided in pharmaceutical composition with the present invention or may be provided separately.
  • a pharmaceutical formulation for oral administration may, for example, be granule, tablet, sugar coated tablet, capsule, pill, suspension or emulsion.
  • a sterile aqueous solution may be provided that may contain other substances including, for example, salts and / or glucose to make to solution isotonic.
  • the anti-cancer agent may also be administered in the form of a suppository or pessary, or may be applied topically in the form of a lotion, solution, cream, ointment or dusting powder.
  • the present invention also relates to methods of treating or preventing cancer comprising adminstering a compound according to the present invention as described herein to a subject in need thereof.
  • the method includes the use of one or more compounds of the present invention in a regime including the step of radiotherapy.
  • Examples 1 to 153 were synthesised according to the methods described subsequently. Their IC50 values were then determined as described below and are represented in the following table, in which the compound numbers correspond to the numbers in the examples.
  • Example 4 causes inhibition of GSK3
  • Method A The system consists of ThermoFinnigan LCQ Advantage Mass Spectrometer with the Surveyor LC system and 200 position autosampler.
  • the spectrometer has an electrospray source operating in positive and negative ion mode.
  • An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Luna 3micron Ci 8 50 x 2 mm; Mobile phase -A)Water 0.1 % Formic Acid B)Acetonitrile 0.1% Formic Acid
  • Method B The system consists of a Waters ZMD quadrupoie mass spectrometer linked to a Waters 1525 LC system with Waters 996 diode array detector. Sample injection is done by a Waters 2700 autosampler. The spectrometer has an electrospray source operating in positive and negative ion mode. Additional detection is achieved using a Sedex 85 evaporative light scattering detector.
  • Method C The system consists of a Waters Micromass ZQ2000 quadrupole mass spectrometer linked to a Waters Acquity UPLC system with a PDA UV detector.
  • the spectrometer has an electrospray source operating in positive and negative ion mode.
  • An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Acquity BEH C18 1.7um 100 x 2.1 mm, maintained at 40°C or Acquity BEH Shield RP18 1 Jum 100 x 2.1 mm, maintained at 40°C.
  • Method D The system consists of an Agilent Technologies 6140 single quadrupole mass spectrometer linked to an Agilent Technologies 1290 Infinity LC system with UV diode array detector and autosampler.
  • the spectrometer has a multimode ionization source (electrospray and atmospheric pressure chemical ionizations) operating in positive and negative ion mode.
  • An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Zorbax Eclipse Plus C18 RRHD 1.8micron 50 x 2.1 mm maintained at 40°C.
  • Method E The system consists of a Agilent Technologies 6120 single quadrupole mass spectrometer linked to a Agilent Technologies 1200 Preparative LC system with Multiple Wavelength detector and autosampler.
  • the spectrometer has a multimode ionization source (electrospray and atmospheric pressure chemical ionizations) operating in positive and negative ion mode. Fraction collection is mass-triggered.
  • a purification experiment is performed on each sample submitted using the following conditions: LC Column - Waters XBridgeTM Prep C18 5 ⁇ OBDTM 19 x 50mm column at room temperature. Mobile phase - A) Water 0.1 % ammonium hydroxide B) Acetonitrile/Water 95: 5 with 0.1 % ammonium hydroxide.
  • Method F The system consists of a Agilent Technologies 6120 single quadrupole mass spectrometer linked to a Agilent Technologies 1200 Preparative LC system with Multiple Wavelength detector and autosampler.
  • the spectrometer has a multimode ionization source (electrospray and atmospheric pressure chemical ionizations) operating in positive and negative ion mode. Fraction collection is mass-triggered. A purification experiment is performed on each sample submitted using the following conditions: LC Column - Waters XBridgeTMPrep C18 5pm OBDTM 30 x 100mm column at room temperature. Mobile phase - A) Water 0.1 % ammonium hydroxide B) Acetonitrile/Water 95: 5 with 0.1% ammonium hydroxide.
  • Method G The system consists of an Agilent 6110 quadrupole LC / MS mass
  • the spectrometer linked to an Agilent 1200 LC system with Agilent 1200 diode array detector. Sample injection is done by an Agilent 1200 autosampler.
  • the spectrometer has an electrospray and APCI source operating in positive and negative ion mode.
  • Step 1 tert-butyl (1-(4-(3-(dimethylamino)-2-phenylacryloyl)phenyl)cyclobutyl)carbamate
  • a solution of tert-butyl (1-(4-(2-phenylacetyl)phenyl)cyclobutyl)carbamate (25.0 g, 68 mmol) in ⁇ , ⁇ -dimethylformamide dimethylacetal (150 mL) was heated to 80°C under a nitrogen atmosphere for 18 hours. The resulting brown solution was allowed to cool to room temperature and concentrated to dryness under reduced pressure to give the desired product as a yellow/brown solid (30.6 g, quantitative yield).
  • Step 2 tert-butyl (1-(4-(5-oxo-3-phenyl-5.6.7.8-tetrahvdroquinolin-2- vQphenvQcyclobutvDcarbamate
  • Step 3 2-(4-(1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihvdroquinolin-5(6H)-one
  • tert-butyl (1 -(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (11 mg, 0.023 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 Terf-butyl (1-(4-(5-(hvdroxyirnino)-3-phenyl-5,6,7,8-tetrahvclroauinoliri-2- vDphenvDcyclobutyQcarbamate
  • Step 2 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin-5(6H)-one oxime
  • Tert-butyl (1-(4-(5-(hydroxyimino)-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (7 mg, 0.017 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times.
  • Step 1 tert-butyl (1-(4-(5-(methoxyimino)-3-phenyl-5,6,7,8-tetrahvdroquinolin-2- yl)phenyl)cyclobutyl)carbannate
  • Step 2 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7.8-dihydroquinolin-5(6H)-one O- methyl oxime
  • Step 1 Tert-butyl 1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-5,6,7 l 8- tetrahydronaphthalen-2-yl)phenyl)cvclobutyl)carbamate
  • Step 2 terf-butyl (1-(4-(8-phenyl-4.5-dihvdro-2H-pyrazolor3.4-flquinolin-7- yl)phenyl)cyclobutyl)carbamate
  • Step 3 1-(4-(8-phenyl-4,5-dihvdro-2H-pyrazolof3,4-f1auinolin-7- vDphenyQcyclobutanamine
  • Step 1 tert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3,4-f1auinolin-7- yl)phenyl)cyclobutyl)carbamate and te/ -butyl (1-(4-(1-methyl-8-phenyl-4 1 5-dihydro-1 H- pyrazolor3,4-f1quinolin-7-yl)phenyl)cvclobutyl)carbamate
  • reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 99:1 to 70:30) to give the desired product fert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihydro-2H- pyrazolo[3,4-f]quinolin-7-yl)phenyl)cyclobutyl)carbamate as a white solid (14 mg, 15% yield).
  • Step 2A 1 -(4-(2-methyl-8-phenyl-4.5-dihvdro-2H-pyrazolof3.4-f1quinolin-7- yl)phenyl)cyclobutanamine
  • Step 1 tert-butyl (1-(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl-5,6,7.8- tetrahydroquinolin-2-yl)phenyl)cvclobutyl)carbamate
  • reaction mixture was diluted with saturated sodium bicarbonate solution (40 mL) and extracted with ethyl acetate (3 x 40 mL). The combined organic phases were washed with 50:50 waterbrine (40 mL), then brine (40 mL), dried over Na 2 S0 , filtered and concentrated to dryness under reduced pressure to give the desired product as a bright yellow solid (1.21 g, 99% yield).
  • Step 2 methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-5-oxo-3-phenyl- 5,6,7,8-tetrahydroquinoline-6-carboxylate
  • Step 3 tert-butyl (1-(4-(3-hvdroxy-8-phenyl-4.5-dihvdro-2H-pyrazolor3,4-flquinolin-7- vDphenvDcyclobutyDcarbamate
  • Step 4 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4,5-dihvdro-2H-pyrazolof3,4-flquinolin- 3-ol
  • tert-butyl (1-(4-(3-hydroxy-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (19 mg, 0.037 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Example 8 7-(4-(1-aminocvclobutyl)phenvH-3a-methyl-8-phenyl-4,5-dihvdro-2H- pyrazolor3,4-f
  • Step 1 methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cvclobutyl)phenyl)-6-methyl-5-oxo-3- phenyl-5,6,7,8-tetrahydroquinoline-6-carboxylate
  • Step 2 tert-butyl (1-(4-(3a-methyl-3-oxo-8-phenyl-3.3a,4.5-tetrahvdro-2H-pyrazolor3.4- f1quinolin-7-yl)phenyl)cvclobutyl)carbamate
  • Step 3 7-(4-(1-aminocvclobutyl)phenyl)-3a-methyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3,4- fiquinolin-3(3aH)-one
  • tert-butyl (1-(4-(3a-methyl-3-oxo-8-phenyl-3,3a,4,5-tetrahydro-2H-pyrazolo[3,4-f]quinolin- 7-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.023 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1-(4-(3-(methylthioV8-phenyl-4,5-dihvdro-2H-pyrazolo[3.4-f1quinolin-7- vnphenvOcvclobutvOcarbamate
  • Step 2 1 -(4-(3-(methylthio)-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4-f1quinolin-7- yl)phenyl)cvclobutanamine
  • tert-butyl (1-(4-(3-(methylthio)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate 53 mg, 0.098 mmol was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4- f
  • Step 2 1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihvdro-2H-pyrazolo[3,4-flquinolin-7- vOphenyQcvclobutanamine
  • tert-butyl (1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (40 mg, 0.070 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1-(4-(3-(methylamino)-8-phenyl-4,5-dihvdro-2H-pyrazolof3.4-f1quinolin- 7-yl)phenyl)cvclobutyl)carbamate
  • tert-butyl (1-(4-(3-(rnethylamino)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (47 mg, 0.090 mmol) was dissolved in TFA (2 ml.) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1 -(4-(3-((2-hvdroxyethyl)amino)-8-phenyl-4,5-dihvdro-2H-pyrazolof3.4- f1quinolin-7-yl)phenyl)cvclobutyl)carbamate
  • Step 2 2-((7-(4-(1 -aminocvclobutyl)phenyl)-8-phenyl-4,5-dihvdro-2H-pyrazolo[3,4- f1quinolin-3-yl)amino)ethanol
  • tert-butyl (1-(4-(3-((2-hydroxyethyl)amino)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-7-yl)phenyl)cyclobutyl)carbamate 25 mg, 0.045 mmol was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1 -(4-(3-morpholino-8-phenyl-4,5-dihvdro-2H-pyrazolof3,4-f]quinolin-7- vDphenyQcyclobutvDcarbamate
  • reaction mixture was concentrated to dryness under reduced pressure and purified via Biotage chromatography (silica, cyclohexane:ethyl acetate, gradient elution from 92:2 to 100:0) to give a white solid (30 mg, 30% yield).
  • Step 2 1-(4-(3-morpholino-8-phenyl-4,5-dihydro-2H-pyrazolof3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
  • tert-butyl (1-(4-(3-morpholino-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (30 mg, 0.052 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1-(4-(6-(hvdroxyimino)-5-oxo-3-phenyl-5,6,7,8-tetrahvdroquinolin-2- yQphenvOcyclobutvQcarbamate
  • the reaction mixture was stirred at 50°C under a nitrogen atmosphere for 1 hour then allowed to cool to room temperature and concentrated to dryness under reduced pressure.
  • the residue was redissolved in water (5 mL), acidified with 1 M HCI solution (5 mL) and extracted into ethyl acetate (3 x 10 mL).
  • the combined organic phases were washed with water (20 mL), brine (20 mL), dried over Na 2 S0 4 , filtered and concentrated to dryness under reduced pressure.
  • the residue was purified via Biotage chromatography (silica, hexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a brown solid (436 mg, 41% yield)).
  • Step 2 tert-butyl (1-(4-(3-hvdroxy-2-methyl-8-phenyl-4,5-dihvdro-3H-imidazof4.5- f1quinolin-7-yl)phenyl)cyclobutyl)carbamate
  • Step 3 7-(4-(1 -aminocvclobutyl)phenyl)-2-methyl-8-phenyl-4,5-dihydro-3H-imidazof4.5- flquinolin-3-ol
  • tert-butyl (1-(4-(3-hydroxy-2-methyl-8-phenyl-4,5-dihydro-3H-imidazo[4,5-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (10 mg, 0.019 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihvdro-3H-imidazof4,5-f1quinolin-7- vDphenvDcvclobutvDcarbamate
  • tert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihydro-3H-irnidazo[4,5-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate 25 mg, 0.049 mmol was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Example 16 1 -(4-(9-phenyl-5,6-dihvdropyridor2.3-h1quinazolin-8-yl)phenyl)- cvclobutanamine
  • Step 1 terf-butyl (1-(4-(9-phenyl-5.6-dihydropyridor2,3-hlauinazolin-8- yl)phenyl)cvclobutyl)carbamate
  • Step 2 1 -(4-(9-phenyl-5,6-dihvdropyridof2,3-hlquinazolin-8-yl)phenyl)cvclobutanamine
  • 1-(4-(9-phenyl-5,6-dihydropyrido[2,3-h]quinazolin-8- yl)phenyl)cyclobutyl)carbamate (12 mg, 0.023 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times.
  • Example 17 8-(4-(9-phenyl- -dihvdropyridor2,3-h1quinazolin-2-amine
  • Step 1 teff-putyl (1-(4-(2-amino-9-phenyl-5,6-dihvdropyridoi2,3-hlquinazolin-8- yl)phenyl)cyclobutyl)carbamate
  • Step 2 8-(4-(9-phenyl-5.6-dihvdropyridor2,3-h1quinazolin-2-amine
  • Step 1 tert-butyl (1-(4-(7,7-dimethyl-5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cvclobutyl)carbamate
  • Step 2 2-(4-(1 -aminocvclobutyl)phenyl)-7,7-dimethyl-3-phenyl-7.8-dihvdroquinolin-5(6H)- one
  • Step 1 tert-butyl (1-(4-(6-((dimethylamino)methylene)-7,7-dimethyl-5-oxo-3-phenyl- 5,6,7,8-tetrahydroquinolin-2-yl)phenyl)cvclobutyl)carbamate
  • Step 2 tert-butyl (1-(4-(4.4-dimethyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4-flquinolin-7- yl)phenyl)cyclobutyl)carbamate
  • Step 3 1-(4-(4,4-dimethyl-8-phenyl-4,5-dihydro-2H-pyrazolor3,4-f
  • tert-butyl (1-(4-(4,4-dimethyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (8 mg, 0.015 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (1 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times.
  • Step 1 tert-butyl (1-(4-(5-oxo-3-phenyl-6,7.8,9-tetrahvdro-5H-cvcloheptarblpyridin-2- yOphenvOcyclobutvQcarbamate
  • Step 2 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-6.7,8,9-tetrahvdro-5H- cvclohepta[b]pyridin-5-one
  • Example 21 1-(4-(9-phenyl-2,4,5.6-tetrahvdropyrazolor3'.4':3,4lcvcloheptari.2- b1pyridin-8-yl)phenyl)cvclobutanamine
  • Step 1 tert-butyl (1-(4-(6-((dimethylamino)rnethylene)-5-oxo-3-phenyl-6,7,8,9-tetrahvdro- 5H-cvcloheptafblpyridin-2-yl)phenyl)cyclobutyl)carbamate
  • Step 2 tert-butyl f1-(4-f9-phenyl-2,4.5,6-tetrahvdropyrazolof3',4':3.4lcvcloheptaf1.2- b1pyridin-8-yl)phenyl)cyclobutyl)carbamate
  • reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 99:1 to 60:40) and preparative HPLC (Method G, gradient 5 to 95% 0.1 %FA/ACN in 0.1 %FA/H 2 O, 20 minutes run) to give the desired product as a white solid (50 mg, 58% yield).
  • Step 3 1 -(4-(9-phenyl-2.4.5.6-tetrahvdropyrazolof3'.4':3.41cvcloheptaf 1 ,2-blPyridin-8- vQphenyQcvclobutanamine
  • Example 22 2-(4-(1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihvdro-1 ,6- naphthyridin-5(6H)-one
  • Step 1 tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahvdro-1.6-naphthyridin-2- vDphenvDcvclobutvDcarbamate
  • Step 2 2-(4-( 1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihydro-1 ,6-naphthyridin-5(6H)- one
  • Step 1 tert-butyl (1-(4-(6-methyl-5-oxo-3-phenyl-5,6.7.8-tetrahvdro-1 ,6-naphthyridin-2- vQphenvOcvclobutvOcarbamate
  • Step 2 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-7,8-dihvdro-1 ,6-naphthyridin-5(6H)- one
  • Step 1 tert-butyl (1-(4-(6-(cvanomethyl)-5-oxo-3-phenyl-5.6.7.8-tetrahvdro-1.6- naphthyridin-2-yl)phenyl)cvclobutyl)carbarnate
  • Step 2 2-(2-(4-(1 -aminocvclobutvnphenyl)-5-oxo-3-phenyl-7,8-dihydro-1 ,6-naphthyridin- 6(5H)-yl)acetonitrile
  • Step 1 tert-butyl (1-(4-(6-(2-(dimethylamino)ethyl)-5-oxo-3-phenyl-5.6.7.8-tetrahvdro-1.6- naphthyridin-2-yl)phenyl)cvclobutyl)carbamate
  • Step 2 2-(4-(1-aminocvclobutyl)phenyl)-6-(2-dimethylamino)ethyl)-3-phenyl-7,8-di ,6-naphthyridin-5(6H)-one
  • Example 26 8-(4-(1 -aminocvclobutyl)phenyl)-9-phenyl-5,6-dihvdro- ⁇ 1.2.41triazolor3,4-fin .61naphthyridin-3(2H)-one
  • Stepl tert-butyl (1-(4-(3-phenyl-5-thioxo-5,6,7,8-tetrahvdro-1.6-naphthyridin-2- vQphenvOcyclobutvOcarbamate
  • Step 2 tert-butyl (1 -(4-(3-oxo-9-phenyl-2.3.5.6-tetrahvdro-ri.2.41triazolor3.4- firi,61naphthyridin-8-yl)phenyl)cvclobutyl)carbamate
  • Step 3 8-(4-(1-aminocvclobutyl)phenyl)-9-phenyl-5,6-dihvdro-n.2.41triazolor3.4- flM .61naphthyridin-3(2H)-one
  • Step 1 tert-butyl (1-(4-(6-ethyl-3-phenyl-5.6,7,8-tetrahvdro-1.6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate
  • Step 2 1-(4-(6-ethyl-3-phenyl-5,6,7,8-tetrahvdro-1 ,6-naphthyridin-2- vQphenvDcvclobutanamine
  • Step 1 tert-butyl (1-(4-(5-hvdrazono-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate
  • Step 2 tert-butyl (1 -(4-(9-phenyl-5.6-dihvdro-f1 ,2.41triazolor3.4-fin ,6lnaphthyridin-8- yl)phenyl)cvclobutyl)carbamate
  • Step 3 1-(4-(9-phenyl-5.6-dihvdro-ri .2.41triazolor3,4-flM .61naphthyridin-8- yDphenvDcvclobutanamine
  • Example 29 1-(4-(3-(1-methyl-1H-imidazol-4-yl)-9-phenyl-5.6-dihydro- ri.2,41triazolof3.4-firi.61naphthyridin-8-yl)phenvhcvclobutanamine
  • Step 1 teit-butyl (1-(4-(3-(1-methyl-1 H-imidazol-4-vn-9-phenyl-5,6-dihvdro- f1 ,2,4ltriazolor3,4-f1[1 ,6lnaphthyridin-8-yl)phenyl)cvclobutyl)carbamate
  • Step 2 1 -(4-f 3-( 1 -methyl-1 H-imidazol-4-yl)-9-phenyl-5.6-dihvdro-M ,2,41triazolo[3.4- firi ,61naphthyridin-8-yl)phenyl)cyclobutanamine
  • Step 1 tert-butyl (1-(4-(5-cyano-6-hydroxy-3-phenylpyridin-2- Yl)phenyl)cyclobutyl)carbamate
  • Step 2 tert-butyl (1-(4-(5-carbamoyl-6-hydroxy-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate
  • Step 3 tert-butyl (1-(4-(5-amino-6-hvdroxy-3-phenylpyridin-2- vDphenvQcvclobutvDcarbamate
  • Step 4 tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2.3-biri,4loxazin-6- vQphenvDcyclobutvDcarbamate
  • Step 5 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1H-pyridor2.3-biri,41oxazin-2(3H)-one
  • tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (8 mg, 0.017 mmol) was reacted to afford the title compound (6 mg, 73%).
  • Step 1 tert-butyl (1-(4-(1-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2.3-biri,4loxazin- 6-yl)phenyl)cvclobutyl)carbamate
  • Example 32 6-(4-(1 -aminocvclobutyl)phenyl)-7-phenyM -(prop-2-vn-1 -yl)-1 H- PVridor2,3-bin,41oxazin-2(3H)-one
  • Step 1 tert-butyl (1-(4-(2-oxo-7-Dhenyl-1 -(prop-2-vn-1-yl)-2,3-dihvdro-1 H-pyridor2,3- bl[1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 1 tert-butyl (1 -(4-n-(cvanomethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2.3- blf 1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyridof2,3- blf 1 ,41oxazin-1 -vDacetonitrile
  • Example 34 6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-(dimethylamino)ethyl)-7-phenyl- 1 H-pyridor2.3-blM .41oxazin- -one
  • Step 1 tert-butyl (1-(4-(1-(2-(dimethylamino)ethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- pyridor2,3-biri,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-(dimethylamino)ethyl)-7-phenyl-1 H- pyridor2,3-bir .41oxazin-2(3H)-one
  • Step 1 tert-butyl (1 -(4-(1-(2-amino-2-oxoethyl)-2-oxo-7-phenyl-2,3-dihvdro- H- pyridof2.3-b1[1.41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 2-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridoi2.3- b1f1.41oxazin-1-yl)acetamide
  • Step 1 tert-butyl (1-(4-(1-(2-oxo -1-(2-oxopropyl)-7-phenyl-2.3-dihvdro-1 H-pyridor2.3- bli1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-oxopropyl)-7-phenyl-1 H-pyrido[2,3- b1M.41oxazin-2(3H)-one
  • Step 1 tert-butyl (1-(4-(7-phenyl-2.3-dihvdro-1 H-pyridof2.3-blf1.4loxazin-6- yl)phenyl)cvclobutyl)carbamate
  • the aqueous phase was extracted with dichloromethane (3x) using a phase separator (Isolute ® SPE).
  • the combined organic phases were concentrated to dryness under reduced pressure.
  • the resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 2% MeOH in DCM) to give the title compound as a white solid (66 mg, 48%).
  • Step 2 1-(4-(7-phenyl-2.3-dihvdro- H-pyridor2,3-bl[1.41oxazin-6- yl)phenyl)cyclobutanamine
  • Step 1 tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihvdro-1H-pyridor2,3-biri,41oxazin-6- yl)phenyl)cyclobutyl)carbamate
  • Step 2 Ethyl 2-(6-(4-(1-(tert-butoxycarbonylamino)cvclobutyl)phenyl)-7-phenyl-1 H- pyridof2,3-bl[1 ,4loxazin-2(3H)-ylidene)hvdrazinecarboxylate
  • Step 3 tert-butyl (1-(4-(1-oxo-8-phenyl-2.4-dihvdro-1 H-pyridof2.3-biri.2.4ltriazolor4,3- d1H ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
  • Step 4 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-2.4-dihydro-1 H-pyridor2,3- blf 1.2,41triazolof4,3-din .41oxazin-1 -one
  • Step 1 tert-butyl (1-(4-(2-hvdrazono-7-phenyl-2.3-dihvdro-1 H-pyrido[2,3-b1f1.4loxazin-6- yl)phenyl)cvclobutyl)carbamate
  • Step 2 tert-butyl (1-(4-(8-phenyl-4H-pyridor2.3-biri.2.41triazolor4.3-dlf1.41oxazin-7- vDphenyOcyclobutyQcarbamate
  • Step 3 1-(4-(8-phenyl-4H-pyridof2.3-biri.2.41triazolo[4.3-dir .41oxazin-7- vDphenvDcvclobutanamine
  • Example 40 1-(4-(1-methyl-7-phenyl-2,3-dihvdro-1H-pyridor2.3-b1M.41oxazin-6- vnphenvDcyclobutanamine
  • Step 1 tert-butyl (1-(4-(1-methyl-7-phenyl-2.3-dihvdro-1 H-pyridor2.3-biri.4loxazin-6- vDphenvDcvclobutvDcarbamate
  • Step 2 1-(4-(1-methyl-7-phenyl-2.3-dihvdro-1 H-pyrido[2.3-b1f1.4loxazin-6- yQphenyQcvclobutanamine
  • Example 41 1-(6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-2.3-dihvdro-1H- pyridor2,3-blH ,4loxazin-1 -vDethanone
  • Step 1 tert-butyl (1-(4-(1-acetyl-7-phenyl-2.3-dihvdro-1 H-pyridor2.3-b1f1.41oxazin-6- vQphenvDcvclobutyDcarbamate
  • Step 2 1-(6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- b][1 ,4loxazin-1-yl)ethanone
  • Example 42 1-((1H-imidazol-2-yl)rnethyl)-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl- 1 H-pyridor2,3-biri .41oxazin -one
  • Step 1 tert-butyl (1 -(4-(1-((1 H-imidazol-2-yl)methyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- Pyridor2,3-b1f1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 1-(( H-imidazol-2-yl)methyl)-6-(4-(1-aminocvclobutyl)Dhenyl)-7-phenyl-1 H- pyridor2.3-blf1.4loxazin-2(3H)-one
  • Step 1 tert-butyl (1 -(4-(1-(cvanomethyl)-7-phenyl-2.3-dihvdro-1 H-pyrido[2.3- bl[1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 2-(6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-2.3-dihydro-1 H-pyridof2.3- blf 1 ,4loxazin-1 -vDacetonitrile
  • Example 45 6-(4-(1 -aminocyclobutyl)phenv0-7-phenyl-1 -(pyridin-4-ylmethylH H- pyridor2,3-biri,41oxazin-2(3H)-one
  • Step 1 tert-butyl n-(4-(2-oxo-7-phenyl-1-(pyridin-4-ylmethyl)-2,3-dihvdro-1H-pyridor2,3- b)n,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-4-ylmethyl)-1 H-pyridor2,3- bin .41oxazin-2(3H)-one
  • Example 46 6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 -(pyridin-3-Ylmethyl)-1 H- pyridor2.3-bU1.41oxazin-2(3H)-one
  • Step 1 tert-butyl n-(4-(2-oxo-7-phenyl-1-(pyridin-3-ylmethyl)-2.3-dihvdro-1 H-Pyridof2,3- b1f1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-3-ylmethyl)-1 H-pyhdof2,3- bin.41oxazin-2(3H)-one
  • Step 1 tert-butyl (1-(4-(7-phenyl-2-thioxo-2.3-dihvdro-1 H-pyridof2,3-b]ri.4loxazin-6- yl)phenyl)cvclobutyl)carbamate
  • Step 2 (E)-tert-butyl (1-(4-(2-hvdrazono-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- bl[1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 3 tert-butyl (1-(4-(9-phenyl-5.6-dihvdro-ri.2,4ltriazolof3.4-firi ,6lnaphthyridin-8- vDphenvQcvclobutvDcarbamate
  • Step 4 1-(4-(1-ethyl-8-phenyl-4H-pyridof2.3-b1f1.2,4ltriazolof4.3-dlf1.4loxazin-7- vDphenvQcyclobutanamine
  • Example 48 6-(4-(1-aminocvclobutyl)phenyl)-1 -(cyclobutylmethyl)-7-phenyl-1 H- pyridor2,3-b1M,41oxazin-2(3 -one
  • Step 1 tert-butyl n-(4-(1-(cvclobutylmethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-Pyridor2.3- blf1.4loxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Example 49 ethyl 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyridor2,3-b1[1 ,41oxazin-1 -vDacetate
  • Step 1 ethyl 2-(6-(4-(1-(te/ -butoxycarbonylamino)cvclobutyl)phenyl)-2-oxo-7-phenyl-2 l 3- dihvdro-1 H-pyridof2,3-biri ,41oxazin-1 -vDacetate
  • Step 2 ethyl 2-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- Pyridof2,3-b1f ,4loxazin-1-yl)acetate
  • Example 50 7-(4-(1 -aminocyclobutyl)phenyl)-8-phenyl-4H-pyridor2 1 3- biri.2.41triazolor4,3-diri.4loxazin-1-amine
  • Step 1 tert-butyl (1-(4-(1-amino-8-phenyl-4H-pyridof2.3-blf1.2.41triazolor3,4- d1f1,4]oxazin-7-yl)phenyl)cvclobutyl)carbamate
  • Step 2 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4H-pyhdor2,3-biri .2,41triazolor4,3- dlf1 ,4loxazin-1 -amine
  • Example 51 6-(4-(1 -aminocvclobutyl)phenvn-1 -(2-methoxyethyl)-7-phenyl-1 H- pyridor2,3-biri,41oxazin-2(3 -one
  • Step 1 tert-butyl (1-(4-(1-(2-methoxyethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- b1f1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-methoxyethyl)-7-phenyl-1 H-pyrido[2,3- bin .41oxazin-2(3H)-one
  • Step 1 terf-butyl 1-(4-(1-ethyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-Pyridof2,3-blf1 ,4loxazin-6- vDphenvDcyclobutylcarbamate
  • reaction mixture was quenched by the addition of saturated sodium bicarbonate solution (5 ml_) and extracted into ethyl acetate (3 x 5 ml_).
  • the organic phase was washed with 50:50 water:brine (2 x 10 mL) then brine (10 mL), dried over Na 2 S0 4 , filtered and concentrated to dryness under reduced pressure.
  • the residue was purified by Biotage chromatography
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-1-ethyl-7-phenyl-1 H-pyrido[2,3-blf1 ,41oxazin- 2(3H)-one
  • Example 53 1-((1 H-imidazol-5-yl)methvn-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl- 1 H-pyridf>r2.3-b]ri .41oxazin- -one
  • Step 1 tert-butyl (1-(4-(1-((1 H-imidazol-5-yl)methyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof2,3-biri,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 1-((1 H-imidazol-5-yl)methyl)-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1 H- pyridor2.3-biri.41oxazin-2(3H)-one
  • tert-butyl (1-(4-(1-((1 H-imidazol-5-yl)methyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.022 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times.
  • Example 54 2-(e-(4-f1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H- pyridor2,3-biri,41oxazin-1-yl)propanenitrile
  • Step 1 tert-butyl (1-(4-(1-(1-cyanoethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- blf1 ,4loxazin-6-vnphenyl)cyclobutyl)carbamate
  • Step 2 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- biri,4loxazin-1-yl)propanenitrile
  • Step 1 te/t-butyl 1-(4-(1-(cvclopropylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyridor2,3- biri,41oxazin-6-yl)phenyl)cyclobutylcarbamate
  • reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na 2 S0 4 , filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (40 mg, 72% yield).
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-1-(cyclopropylmethyl)-7-phenyl-1 H-pyrido[2,3- blH ,41oxazin-2(3H)-one
  • Step 1 tert-butyl (1 -(4-(2-oxo-7-phenyl-1-(pyridin-2-ylmethyl)-2,3-dihvdro-1H-pyridof2.3- blH ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-2-ylmethyl)-1 H-pyridor2,3- blH .41oxazin-2(3H)-one
  • Step 1 fert-butyl 1 -(4-(1-isopropyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2.3- blfl ⁇ loxazin-e-vDphenvDcyclobutylcarbamate
  • reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 ml) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na 2 S0 4 , filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexaneiethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (13 mg, 48% yield).
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-1 -isopropyl-7-phenyl-1 H-pyridof2.3- blH ,41oxazin-2(3H)-one
  • fert-Butyl 1 -(4-(1-isopropyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (7 mg, 0.014 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette.
  • Step 1 tert-butyl 1-(4-(1-cvclopentyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- blf1 ,4loxazin-6-yl)phenyl)cvclobutylcarbamate
  • reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 3 mL). The combined organic phases were dried over Na 2 S0 4 , filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (12 mg, 21% yield).
  • tert-Butyl 1 -(4-(1 -cyclopentyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (12 mg, 0.022 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether ( ⁇ 2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette.
  • Step 1 ferf-butyl 1-(4-(2-oxo-7-phenyl-1-(thiazol-4-ylmethvn-2.3-dihvdro-1 H-pyridor2,3- biri,41oxazin-6-yl)phenyl)cvclobutylcarbamate
  • reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na 2 S0 4 , filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (54 mg, 90% yield).
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(thiazol-4-ylmethyl)-1 H-pyridof2,3- bin .41oxazin-2(3H)-one
  • Example 60 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(2,2,2-trifluoroethyl)-1H- pyridor2.3-b 1.41oxazin-2(3H)-one
  • Step 1 tert-butyl 1-(4-(2-oxo-7-phenyl-1-(2,2.2-trifluoroethyl)-2.3-dihvdro-1 H-pyridor2.3- biri,41oxazin-6-yl)phenyl)cyclobutylcarbamate
  • reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na 2 S0 4) filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (31 mg, 53% yield).
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1 -(2.2,2-trifluoroethyl)-1 H-pyridor2,3- biri.41oxazin-2(3H)-one
  • Step V tert-butyl 1-(4-(1-cvclobutyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyrido[2,3- b1f1 ,4loxazin-6-yl)phenyl)cyclobutylcarbamate
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-1-cvclobutyl-7-phenyl-1 H-pyridoi2,3- bin ,41oxazin-2(3H)-one
  • Example 62 3-(6-(4-(1 -aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- pyrido[2,3-b1[1 ,41oxazin-1 -vDpropanenitrile
  • Step 1 tert-butyl (1-(4-(1-(2-cvanoethvn-2-oxo-7-prienyl-2.3-dihvdro-1 H-pyridor2.3- b1H ,4]oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 2 3-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- blf 1 ,4]oxazin-1 -vDpropanenitrile
  • Step 1 Tert-butyl (1-(4-(1 -methyl-8-phenyl-4H-pyridor2.3-biri,2.4ltriazolor4.3- d]f ,4loxazin-7-yl)phenyl)cvclobutyl)carbamate
  • Step 2 1-(4-(1-methyl-8-phenyl-4H-pyridor2,3-biri,2.4ltriazolor4.3-diri,4loxazin-7- vDphenvQcvclobutanamine
  • Example 64 4-(6-(4-(1 -aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof2,3-b1M .41oxazin-1 -vDbutanenitrile
  • Step 1 tert-butyl (1 -(4-(1-(3-cvanopropyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- blf1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 4-(6-(4-(1 -aminocvclobutvnphenyl)-2-oxo-7-phenyl-2,3-dihvdro-1H-pyrido[2.3- blf 1 ,4loxazin-1 -vDbutanenitrile
  • Example 65 e-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(tetrahvdro-2H-pyran-4- v))-1H-Pyridor2.3-b1i1.41oxazin-2(3H)-one
  • Step 1 tert-butyl (1-(4-(6-hvdroxy-3-phenyl-5-((tetrahvdro-2H-pyran-4-yl)amino)pyridin-2- vDphenvDcyclobutvDcarbamate
  • Step 2 tert-butyl (1-(4-(2-oxo-7-phenyl-1-(tetrahvdro-2H-pyran-4-yl)-2,3-dihvdro-1 H- Pyrido[2,3-bl[1 ,4loxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Step 3 6-(4-(1-arninocvclobutyl)phenyl)-7-phenyl-1-(tetrahvdro-2H-pyran-4-yl)-1 H- pyridof2.3-bin ,41oxazin-2(3H)-one
  • Step 1 tert-butyl (1-(4-(1-(4-cvanobutyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2,3- blf1.41oxazin-6-yl)phenyl)cvclobutyl)carbamate
  • Example 67 1-allyl-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1H-pyridor2,3- bin.41oxazin-2(3H)-one
  • Step 1 tert-butyl 1-(4-(1-allyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3-biri,4loxazin-6- vDphenyQcyclobutylcarbamate
  • Step 1 tert-butyl (1-(4-(1-(2-hvdroxyethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyrido[2,3- bl[1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Step 2 6-(4-(1-aminocyclobutyl)phenyl)-1-(2-hvdroxyethyl)-7-phenyl-1 H-pyrido[2,3- blM ,41oxazin-2(3H)-one
  • Step 1 Tert-butyl (1-(4-(8-phenyl-1-propyl-4H-pyridor2.3-biri .2,41triazolor4.3- dlf1 ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
  • Step 2 1-(4-(8-phenyl-1-propy
  • Example 70 1-(4-(1-(methylsulfonyl)-7-phenyl-2,3-dihydro-1 H-pyridor2,3- biri.41oxazin-6-yl)phenyl)cvclobutanamine
  • Step 1 tert-butyl f1-(4-(1-(methylsulfonyl)-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- b1f1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • Example 71 6-(4-(1-aminocyclobutyl)phenyl)-3-(2-hvdroxyethyl)-7-phenyl-1 H- pyridor2.3-biri.41oxazin- -one
  • Step 1 tert-butyl (1 -(4-(3-(2-hvdroxyethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- biri,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
  • the reaction mixture was transferred to a 250ml round bottom flask and sodium bicarbonate saturated solution (40 ml) and sodium bicarbonate (1g) was added. The resulted mixture was heated to reflux for 24 h. The reaction mixture was cooled down to room temperature. The organic phase was separated. The aqueous phase was extracted with ethyl acetate (45ml). The organic phases were combined and washed with water (30mlX2), brine (25ml) then concentrated at reduced pressure. The residue was suspended in DCM (10ml).
  • Step 2 6-(4-(1-aminocvclobutyl)phenyl)-3-(2-hvdroxyethyl)-7-phenyl-1 H-pyridof2,3- blf .4loxazin-2(3H)-one
  • Example 72 6-(4-(1 -aminocvclobutyl)phenyl)-4-methyl-7-phenyl-3,4- dihvdropyridor2.3-blpyrazin-2(1H)-one
  • Step 1 methyl 2-((6-(4-(1-((fer -butoxycarbonyl)amino)cvclobutyl)phenyl)-3-nitro-5- phenylpyridin-2-yl)(methyl)amino)acetate
  • Step 2 tert-butyl (1-(4-(4-methyl-2-oxo-7-phenyl-1 ,2,3,4-tetrahvdropyridor2,3-blpyrazin-6- yl)phenyl)cyclobutyl)carbamate
  • Step 3 6-(4-( 1 -aminocvclobutyl)phenyl)-4-methyl-7-phenyl-3,4-dihvdropyridor2,3- b]pyrazin-2(1 H)-one
  • Step 1 fe/f-butyl (1 -(4-(2,2-dioxido-7-phenyl-1 H-pyridof3,2-e1f1.3,4loxathiazin-6- vQphenvQcvclobutvOcarbamate
  • reaction mixture was concentrated to dryness under reduced pressure and redissolved in anhydrous methanol (5 mL), followed by the addition of potassium carbonate (641 mg, 4.63 mmol) and heating to 60°C overnight.
  • the reaction mixture was allowed to cool to room temperature and concentrated to dryness under reduced pressure.
  • the residue was redissolved in ethyl acetate (5 mL) and washed with 50:50 waterbrine (2 x 5 mL), dried over Na 2 S0 , filtered and concentrated to dryness under reduced pressure to give a brown oil. This was purified by Biotage chromatography (cyclohexane:ethyl

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Oncology (AREA)
  • Hematology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)
  • Quinoline Compounds (AREA)
  • Other In-Based Heterocyclic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention relates to a series of compounds with particular activity as inhibitors of the serine-threonine kinase AKT. Also provided are pharmaceutical compositions comprising same as well as methods for treating cancer.

Description

INHIBITORS OF AKT ACTIVITY
The present invention relates to compounds that are useful as inhibitors of the activity of one or more isoforms of the serine / threonine kinase, AKT. The present invention also relates to pharmaceutical compositions comprising these compounds and to methods of using these compounds in the treatment of cancer and methods of treating cancer.
BACKGROUND TO THE INVENTION
The AKT protein family, also known as protein kinases B (PKB), are known to be involved in a wide variety of biological processes including cell proliferation,
differentiation, apoptosis, tumorigenesis, as well as glycogen synthesis and glucose uptake. These enzymes are members of the serine/threonine-specific protein kinase family.
The PKB / AKT pathway has been identified as an important regulator of cell survival signalling and apoptosis in cells. Signalling is thought to occur through a range of growth factor receptors including platelet derived growth factor, insulin growth factor and nerve growth factor, resulting in activation of phosphatidylinositol 3-OH kinase (PI-3K). This activation in turn leads to the generation of phosphatidylinositol (3,4,5) triphosphate (PIP3). Activated PIP3 binds to and in turn phosphorylates the enzyme PDK-1 , the main activator of AKT, through its pleckstrin homology domain. Activated PDK-1 is responsible for a phosphorylation event at Thr308 of AKT, which induces a conformational change that facilitates further phosphorylation of AKT at Ser 473 by PDK-2.
PDK-1 phosphorylation of downstream kinases is not unique to AKT, as it has been reported to activate p70 S6 kinase and protein kinase C.
The activation of AKT influences multiple events within the cell including the inhibition of apoptosis, the progression of the cell cycle, cellular survival, metabolism, angiogenesis and hormone resistance.
Presently three family members of AKT have been identified, AKT 1 , AKT 2 and AKT 3 (also known as PKBpr, PKB/3 and PKBy). The family members share 80% amino acid sequence homology and all retain similar regional structure. They possess a C-terminal pleckstrin homology (PH) domain, a catalytic domain, a short or helical linker region and a carboxyl terminal domain. The PH domain permits binding of proteins to the cell membrane through a phospholipid interaction. The catalytic domain of AKT family members contains two residues essential for kinase activation, namely Thr308 and Ser 473. In turn AKT can phosphorylate any protein containing the RXRXXS/T-B motif where X represents any amino acid and B represents bulky hydrophobic residues.
Turning to the cellular function of AKT, hyper activation of AKT has been linked to the inhibition of cellular apoptosis due to phosphorylation and negative regulation of the forkhead family of transcription factors which regulate various genes responsible for instigating death processes including FKHR, FKHRL1 and AFX. Conversely AKT has been reported to up-regulate genes which are known to be anti-apoptotic including IKK and CREB. It is this mixture of positive and negative regulation which highlights the importance of AKT in regulating apoptosis. AKT promotes unwanted cell survival through its' phosphorylation of several key apoptotic proteins including Bad and Pro-caspase 9, thus rendering them inactive and preventing signalling through this pathway. AKT activates and inhibits multiple mechanisms which have a major role in the progression of the cell cycle, ultimately leading to cell proliferation. The best characterised cell cycle regulator and tumour suppressor proteins p53 can be dysregulated via AKT
phosphorylation and activation of the main p53 negative regulator MDM2 .
Phosphorylated MDM2 translocates to the nucleus where it prevents p53 transcription. The inhibition of p53 allows aberrant proliferation of the cell and progression towards a benign state.
In a similar fashion, AKT can also phosphorylate p27kip1 and p21 ; two main inhibitors of cell cycle progression, leading to loss of function, resulting in unchecked cell cycle progress and excessive proliferation.
AKT activation causes an increase in the rate of glycolysis by increasing the rate of glucose metabolism. It has also been reported that activated AKT stimulates the transport of amino acids and supports mTOR dependent increases in protein translation. Proangiogenic factors such as vascular endothelial growth factor (VEGF), have been reported to activate AKT, ultimately resulting in inhibition of endothelial apoptosis, as well as activating endothelial nitric oxide synthase (eNOS). The sum result of this is rapid neo-vascularisation and cell migration.
Hypoxia driven angiogenesis, primarily mediated by hypoxia inducible factor (HIF 1a) can lead to the induction of multiple proteins including VEGF. Increased activated AKT has been reported to increase HIF-1 a expression leading to an increase in angiogenesis independent of a hypoxic environment. Recent data has shown that HIF-1cr activity in invasive breast cancer is correlated with increased activated AKT-1 phosphorylation.
Estrogen receptor (ER) and androgen receptor (AR) inhibitors designed to inhibit cell signalling and induce apoptosis, are vital tools in cancer therapies. Incidence of resistance to these drugs arises rapidly in cancers including prostate, breast and ovarian. AKT has been reported to phosphorylate androgen receptors, leading to inhibition of AR activity and blockade of normal apoptotic signalling in prostate cancer induced by androgens.
In a similar manner, activation of AKT leads to phosphorylation of ER resulting in an inhibition of tamoxifen mediated apoptosis or tumour regression, coupled with the creation of an estrogen independent signalling pathway. Activated AKT-2 has been identified as a promoter of ERa transcription in the presence or absence of estrogen increasing the rate of proliferation of breast cancer cells.
Hyper-activated AKT has been reported in a range of cancers compared to normal tissues including breast, lung, prostate, gastric, ovary, pancreas, thyroid, glioblastoma and haemological cancers. Phosphorylation of AKT has also been associated with clinical characteristics including increased stage and grade of tumour and increased poor prognosis. The activation of AKT can arise from a number of different genetic mutations in the AKT/ PI-3K pathway.
Somatic mutations in the PI-3KCA gene have been widely reported in a large variety of tumours including breast, prostate and head and neck. A large number of these mutations will increase the copy number of the gene leading to an increase in PI-3K activity. A recent study has identified a PI-3K mutation which selectively phosphorylates AKT in colon cancer which results in increase cell proliferation and invasion.
Any mutation which increases the activity of the PI-3K pathway will ultimately result in an increased activation of AKT. Gene amplifications are common occurrences in cancer. Amplifications of AKT-2 have been reported in ovarian, pancreatic, breast and head and neck squamous cell carcinoma. No amplifications or mutations in AKT-3 have been reported to date although deletion mutations leading to hyperactivation and amplification mutations have been reported associated with AKT-1. One mutation; E17K, results in pathological localization of AKT-1 to the cell membrane, inducing its activation and resulting in down-stream signalling and cellular transformation. In vivo, this mutation has been shown to induce leukaemia in mice.
Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a tumour suppressor gene known to negatively regulate AKT function. In cancer, loss of PTEN function results in constitutive phosphorylation of AKT and other down-stream effectors of the PI-3K pathway. Loss of PTEN, due to deletion mutations or promoter methylation, has been reported in a number of different cancers including glioblastoma, endometrial, lung, breast, prostate and thyroid. This loss is commonly associated with hyperactivation of AKT. Recent studies have shown that loss of heterozygosity (LOH) at the PTEN gene was directly correlated to increased AKT activation and chemoresistance in gastric carcinomas and decreased progesterone receptor expression in breast carcinomas.
AKT activation is commonly initiated at the cell surface through a signalling event at a receptor, usually one of the tyrosine kinase family. Two tyrosine kinase receptors commonly amplified or over-expressed in cancer are HER2 and EGFR. In HER2 over- expressing tumours there is often a hyper-activation of AKT, this has been reported in ovarian, stomach and bladder cancer. Similarly in EGFR over-expressing tumours, particularly those with the EGFRvlll activating mutation, selective activation of AKT has been reported in a range of cancers including non-small cell lung cancers, breast, ovarian and most commonly high grade gliomas.
Examples of AKT inhibitors are provided in WO 2008/070134, WO 2008/070016 and WO 2008/070041. These documents provide specific naphthyridine compounds fused to a five membered heterocycle. Other inhibitors of AKT may be found in, for example, WO 2009/1 8887 and WO 2009/148916.
SUMMARY OF THE INVENTION
In a first aspect the present invention provides a compound according to Formula (I):
Figure imgf000006_0001
(I)
wherein:
R1 and R2 are each independently selected from hydrogen, aryl, C1-C10 alkyl, CN, CHO, C02H, CONH2, CONHR3, CONR3aR3 , COR3, C02R3, NH2, NHR3, NR3aR3b, NHCOR3, NHS02R3, NR3aCOR3 , NR3aS02R3b, OH, OR3, SH, SR3, SOR3, S02R3, S02NHR3, S02NR3aR3b, F, CI, Br and I, wherein each R3, R3a and R3 is independently selected from C1-C10 alkyl, including wherein R3a and R3b are joined to one another to form a heterocycle that includes the nitrogen to which they are attached .
Wherein separate R1 and R2 may be joined to one another to form an optionally substituted and optionally saturated heterocycle or carbocycle that includes the C atom to which they are attached; or R1 and R2 together are oxo or optionally C1-C10 alkyl O- substituted oxime;
W is O, S, SO, S02, NR', or CRaR where R' is either hydrogen or C1-C10 alkyl, and where Ra and Rb are each independently selected from the members of the group from which R1 and R2 are selected above;
X is either absent or is CR4R5 where R4 and R5 are each independently selected from the members of the group from which R1 and R2 are selected above; Y and Z are independently either substituted or unsubstituted nitrogen or carbon and where carbon is substituted by substituents independently selected from the members of the group from which R1 and R2 are selected above or when nitrogen then the substituent is selected from aryl, C1-C10 alkyl, S02R3, CONHR3, CONR3aR3b, COR3 and C02R3 where R3, R3a and R3b are as defined above, or where Y and Z together form an optionally substituted heterocyclyl or carbocyclic group, or where Y is S02;
R7a and R7b are independently selected from H and alkyl, including wherein R7a and R7b are joined to one another to form a heterocycle that includes the nitrogen to which they are attached; and
R63 and R6b are independently selected from: H, (Ci-C6)alkyl, (CrC6)alkenyl, and (Cr
C6)alkynyl, wherein said alkyl is optionally substituted with up to three substituents selected from: OH and halo; or R63 and R6b can be taken together to form a monocyclic or bicyclic carbo- or heterocycle with 3-7 members in each ring, said heterocycle having one or more heteroatoms selected from N, O and S, and said carbo- or heterocycle is optionally substituted with one or more substituents selected from: (Ci-C6)alkyl, (C3-C6)cycloalkyl, (C C6)alkoxy, COzH, halo, OH, CN and NR3aR3 , said alkyl, cycloalkyl and alkoxy is optionally substituted with one or more substituents selected from halo, CN, OH and NR3aR3b; and ring Cy is selected from (C3 to C8)cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaryl and aryl, wherein m is 0, 1 , 2, 3, 4 or 5, and each R8 is independently selected from alkyl, CN, CHO, C02H, CONH2, CONHR9, CONHR9aR9b, COR9, C02R9, NH2, NHR9, NR9aR9 , NHCOR9, NHS02R9, NR9aCOR9b, NR9aS02R9b, OH, OR9, SH, SR9, F, CI, Br and I, wherein each R9, R9a and R9 is independently selected from alkyl, including wherein R9a and R9b form a heterocycle that includes the nitrogen to which they are attached or Cy may be iodine; and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
One aspect of the invention also provides compounds selected from the following group of structures:
Figure imgf000008_0001
and pharmaceutically acceptable salts, stereoisomers and tautomers thereof. In preferred embodiments X is absent.
In preferred embodiments R6a and R6b together form
Figure imgf000008_0002
, that is they preferably form cyclobutane. In further preferred embodiments the group bound to the phenyl ring in structure I is 1-aminocyclobutyl.
In preferred embodiments R6a and R6b together form
Figure imgf000008_0003
where R is an alkyl group, preferably a C1 to C6 alkyl group.
In preferred embodiments the ring Cy is unsubstituted C6 aryl, that is phenyl.
In a further embodiment of the compound of the invention the phenyl ring shown in Formula (I) is replaced with a pyridine ring.
The bond between Y and Z is optionally either a single or a double bond. In preferred embodiments the bond between Y and Z is a single bond.
In preferred embodiments X is absent, Y is carbonyl and Z is optionally substituted amino, or X is absent, Y is optionally substituted amino and Z is carbonyl. In these embodiments W is preferably CH2 or O, more preferably O. In these same embodiments R1 and R2 are preferably hydrogen. In these embodiments when amino is substituted it is preferably methyl or acetamido substituted.
In preferred embodiments the substituents bound to Y and Z together with Y and Z themselves form a 5 or 6 membered optionally substituted carbocyclic or herterocyclic ring.
In particularly preferred embodiments the substituents bound to Y and Z together with Y and Z themselves form a moiety selected from the group:
Figure imgf000009_0001
In one particularly preferred embodiment of the compound of the invention the ring Cy is replaced with an iodine atom.
In preferred embodiments where Cy is alkylcycloalkyl, Cy is methylcyclopropyl.
In preferred embodiments W is either CH2 or O. In one particularly preferred
embodiment W is CH2. In another particularly preferred embodiment W is O.
Where the compound of the invention has R63 and R6b groups cooperating so as to
Figure imgf000009_0002
together form that is they preferably form cyclobutane, which additionally has a polar substituent at the position distal to the benzene ring to which the cyclobutane ring is bound, the polar substituent is preferably in a trans relationship to the nitrogen atom having groups R7a and R7 pendant therefrom. Other preferred embodiments of the compounds according to the invention appear throughout the specification and in particular in the examples. Particularly preferred are those named compounds having greater activity as tested. Compounds having higher activity are more preferred over those having lower activity.
DETAILED DESCRIPTION OF THE INVENTION
The term "alkyl group" refers to an aliphatic group containing at least carbon and hydrogen and containing 1 to 15 carbon atoms, such as 1 to 10 carbon atoms.
Attachment to the alkyl group occurs through a carbon atom.
A "Cn alkyl" group refers to an aliphatic group containing n carbon atoms. For example, a C1-C10 alkyl group contains 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.
An alkyl group may be straight chained or it may be branched.
An alkyl group may contain no ring structures or it may contain one or more rings.
For example, a "cycloalkyi" group contains at least one ring. It is understood that attachment to a cycloalkyi group is via a ring of the cycloalkyi group. Each ring may contain 3 to 10 atoms, such as 4 to 8 or 5 to 7 atoms. Each ring may be independently selected to contain just carbon atoms or to contain both carbon atoms and from 1 to 4 heteroatoms selected from O, N and S. For cyclo-heteroalkyl groups (i.e. cycloalkyi groups that contain one or more heteroatoms), attachment to the cycloalkyi group may occur either through a carbon atom or, if one or more heteroatoms are contained in a ring, attachment may also occur through a heteroatom contained in a ring.
For example, a cycloalkyi group may be mono-cyclic or bi-cyclic.
Thus, a "Cn cycloalkyi" group contains n carbon atoms. All n carbon atoms may be contained in the ring(s) of the cycloalkyi group or one or more of the carbons may not be contained in the ring(s) and may instead form one or more chains branching from the ring. If a Cn alkyl group is joined to a separate Cm alkyl group containing m carbon atoms to form, for example, a heterocycle, the two alkyl groups contain a total number of m + n carbon atoms.
An alkyl group may be saturated or unsaturated. Thus, the alkyl group may be an alkenyl group (i.e. contain a carbon-carbon double bond) and / or an alkynyl group (i.e. contain a carbon-carbon triple bond). If the alkyl group is unsaturated, it may contain at least 2 carbon atoms. It is understood that any unsaturated portions of an alkyl group are non-aromatic (aromatic groups fall within the scope of the definition of "aryl'). Any part of the alkyl group may be unsaturated, for example the straight, branched or cyclic portion of an alkyl group may contain a carbon-carbon double bond or a carbon-carbon triple bond. Attachment to an unsaturated alky group may occur through the unsaturated part of the alkyl group or may occur through the unsaturated part of the group.
For example, an unsaturated alkyl group may contain 1 to 4 carbon-carbon double bonds or 1 to 3 carbon-carbon triple bonds or 1 to 4 of a combination of carbon-carbon double bonds and carbon-carbon triple bonds.
An alkyl group may be substituted with one or more heteroatoms or it may be
unsubstituted (i.e. not contain any heteroatoms). If more than one hetero-substituent is present, the substituents are independently selected from one another unless they form a part of a particular functional group (e.g. an amide group).
The heteroatom substituents may in turn be substituted with further carbon-containing groups. In this case, the Cn or Cm prefix that defines the substituted alkyl group refers to the total number of carbons contained in the group, i.e. including the carbon atoms contained in any substituted heteroatomic groups, and the total alkyl group contains 1 to 15 carbon atoms as defined previously.
Accordingly, if the alkyl group is substituted, it may, for example, contain one or more of CN, C02H, CONH2l CONHR, CONRaRb, C02R, NH2, NHR, NRaRb, OH, OR, SH, SR, F, CI, Br and I, wherein each R, Ra and R are independently selected groups (e.g. alkyl / aryl groups) attached to the atom to which the group joins through a carbon atom of each group, including wherein Ra and Rb form a heterocycle that includes the heteroatom to which they are attached. A group containing two Cm-Cn alkyl moieties that form a cycle that includes, for example, the heteroatom to which they are attached may contain from C2m to C2n carbon atoms.
Examples of unsubstituted saturated alkyl groups containing no cyclic structures include methyl, ethyl, n-propyl, sec-propyl, n-butyl, sec-butyl, terf-butyl, pentyl (branched or unbranched), hexyl (branched or unbranched), heptyl (branched or unbranched), octyl (branched or unbranched), nonyl (branched or unbranched), and decyl (branched or unbranched).
Examples of unsubstitued saturated cyclic alkyl groups include cyclopropyl, cylcobutyl, cyclopentyl and cyclohexyl.
Examples of unsaturated alkyl groups include ethenyl, propenyl, butenyl, 2-methybutenyl and cyclohexenyl.
The term "aryl group" refers to a group containing at least one ring that is aromatic and containing 1 to 15 carbon atoms, such as 1 to 10 carbon atoms. Where an aryl group is stated as being substituted at a particular position, attachment of the position to the aryl group is onto the aromatic ring of the aryl group itself rather than the position being joined to the aryl group through any non-aromatic side-chain of the aryl group. For example, when R1 is an aryl group in CR1, the C is attached to the aromatic part of the aryl group.
Each ring may be independently selected to contain only carbon atoms or to contain both carbon atoms and from 1 to 4 heteroatoms selected from O, N and S. For heteroaryl groups (i.e. aryl groups that contain one or more heteroatoms), attachment to the aryl group may occur either through a carbon atom or, if one or more heteroatoms are contained in a ring, attachment may also occur through a heteroatom contained in a ring.
It is noted that the heteroatoms contained in a ring of a heteroaryl group may be substituted, for example forming an /V-oxide.
For example, the aromatic group may be mono-cyclic or bi-cyclic, wherein one or both of the rings of a bi-cyclic system is aromatic. Examples of aryl groups include acridinyl, phenyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl, benzotriazolyl, furanyl, naphthyl, thienyl, thiazolyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline, benzimidazolyl and melaminyl.
It is noted that the term "heterocycle" includes within its scope both cycloalkyl groups containing one or more heteroatoms within the ring system and aryl groups containing one or more heteroatoms within the ring system.
Heterocyclic groups may be any of the various pyrazoles, imidazoles and triazoles and may include the oxygen and/or sulfur containing analogues of the various pyrazoles, imidazoles and triazoles, that is oxazoles, isoxazoles, thiazoles and isothiazoles and derivatives.
The term "halo" refers to a group selected from chlorine, fluorine, bromine and iodine.
The present invention will now be further described. In the following passages different aspects of the invention are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.
For completeness, it is also noted that certain chemical formulae used herein define delocalized systems. This definition is known in the art as a definition of aromaticity and may indicate the presence of, for example, a mono-, di- or tri-cyclic system that contains (4n+2) electrons where n is an integer. In other words, these systems may display Hiickel aromaticity.
In whatever aspect, the compounds of the present invention may possess some aspect of stereochemistry. For example, the compounds may possess chiral centres and / or planes and / or axes. As such, the compounds may be provided as single
stereoisomers, single diastereomers, mixtures of stereoisomers or as racemic mixtures. Stereoisomers are known in the art to be molecules that have the same molecular formula and sequence of bonded atoms, but which differ in their spatial orientations of their atoms and / or groups. In addition, the compounds of the present invention may possess tautomerism. Each tautomeric form is intended to fall within the scope of the invention.
In addition, the compounds of the present invention may be provided as a pro-drug. Prodrugs are transformed, generally in vivo, from one form to the active forms of the drugs described herein. For example, a prodrug may be formed by protecting the amine appending the cyclobutane as a physiological hydrolyzable amide. Alternatively or additionally, Y or Z or any moieties appended thereto are NH, one or more of these may be protected as a physiological hydrolyzable amide.
In those compounds of the invention where X is 'absent' there is instead a single covalent bond such that W and Y are bound to the same carbon atom, that atom with R1 and R2 substitution.
In addition, the compounds of the present invention may be provided in the form of their pharmaceutically acceptable salts or as co-crystals. For example, the compounds may be provided having protonated amine groups.
The term "pharmaceutically acceptable salt" refers to ionic compounds formed by the addition of an acid to a base. The term refers to such salts that are considered in the art as being suitable for use in contact with a patient, for example in vivo and
pharmaceutically acceptable salts are generally chosen for their non-toxic, non-irritant characteristics.
The term "co-crystal" refers to a multi- component molecular crystal, which may comprise non-ionic interactions.
Pharmaceutically acceptable salts and co-crystals may be prepared by ion exchange chromatography or by reacting the free base or acidic form of a compound with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid or base in one or more suitable solvents, or by mixing the compound with another pharmaceutically acceptable compound capable of forming a co-crystal.
Salts known in the art to be generally suitable for use in contact with a patient include salts derived from inorganic and / or organic acids, including the hydrobromide, hydrochloride, sulphate, bisulphate, nitrate, acetate, oxalate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate and tartrate. These may include cations based on the alkali and alkaline earth metals, such as sodium, potassium, calcium and magnesium, as well as ammonium,
tetramethylammonium, tetraethylammonium. Further reference is made to the number of literature sources that survey suitable pharmaceutically acceptable salts, for example the Handbook of pharmaceutical salts published by lUPAC.
In addition, the compounds of the present invention may sometimes exist as zwitterions, which are considered as part of the invention.
The compounds of the present invention are useful in the treatment of medical conditions associated with disordered cell growth, including, but not restricted to, cancer, in particular cancers associated with overactivity of AKT occurring either from a direct change within the kinase itself such as may occur following a mutation within any of its subunits or from increased upstream activity including but not restricted to increased PI3K or PDK activity. Increased PI3K activity may have occurred through loss of the tumor suppressor PTEN.
For example, cancers include cardiac cancers, lung cancers, gastrointestinal cancers, genitourinary tract cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, hematologic cancers, skin cancers and adrenal gland cancers.
For example, cancers include adrenal tumors, bile duct, bladder, blood, bone and connective tissue, brain and central nervous system, breast, cervical, colon and rectal (colorectal), endometrial, esophageal, gallbladder, head and neck, Hodgkin's Lymphoma, hypopharangeal, kidney, laryngeal, leukemias, liver, lung, lymphoma, mediastinal tumors, melanoma (malignant melanoma), mesothelioma, multiple myeloma, nasal cavity, nasopharyngeal, neuroendocrine tumors, non-Hodgkin's lymphoma, oral, oesophagus, oropharyngeal, ovarian, pancreas, paranasal sinus, parathyroid, penis, pituitary tumors, prostate, salivary gland, sarcoma, skin, spine, stomach, testicular, thyroid, urethra, uterine, vaginal and vulvar.
The compounds of the present invention are also useful in preparing a medicament that is useful in treating the diseases described above, in particular cancer. The compounds of the present invention may selectively inhibit one or two of the AKT protein family over the other AKT isoform(s). For example, the compounds may selectively inhibit one or two of AKT1 , AKT2 or AKT3 over the other isoform(s) of AKT.
For example, the compounds of the present invention may inhibit at least AKT1 and / or AKT2. For example, the compounds may selectively inhibit AKT1 and / or AKT2 over AKT3.
The present invention is further directed to a method of inhibiting AKT activity which comprises administering to a mammal in need thereof a pharmaceutically effective amount of the compound of the present invention.
The compounds of this invention may be administered to mammals, including humans, either alone or, in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition, according to standard pharmaceutical practice. The compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.
The present invention also includes within its scope the use of the compounds of the present invention in combination with a second drug in the treatment of cancer. The second drug may be a drug that is already known in the art in the treatment of cancer. The present invention also includes the use of the compounds of the invention in a regime including the step of radiotherapy.
In particular, cancers often become resistant to therapy. The development of resistance may be delayed or overcome by the administration of a combination of drugs that includes the compounds of the present invention.
For example, drugs that may be used in combination with the compounds of the present invention may target the same or a similar biological pathway to that targeted by the compounds of the present invention or may act on a different or unrelated pathway.
Depending on the disease to be treated, a variety of combination partners may be coadministered with the compounds of the present invention. The second active ingredient may include, but is not restricted to: alkylating agents, including cyclophosphamide, ifosfamide, thiotepa, melphalan, chloroethylnitrosourea and bendamustine; platinum derivatives, including cisplatin, oxaliplatin, carboplatin and satraplatin; antimitotic agents, including vinca alkaloids (vincristine, vinorelbine and vinblastine), taxanes (paclitaxel, docetaxel), epothilones and inhibitors of mitotic kinases including aurora and polo kinases; topoisomerase inhibitors, including anthracyclines, epipodophyllotoxins, camptothecin and analogues of camptothecin; antimetabolites, including 5-fluoro uracil, capecitabine, cytarabine, gemcitabine, 6-mercaptopurine, 6- thioguanine, fludarabine, methotrexate and premetrexed; protein kinase inhibitors, including imatinib, gefitinib, sorafenib, sunitinib, erlotinib, dasatinib, and lapatinib;
proteosome inhibitors, including bortezomib; histone deacetylase inhibitors, including valproate and SAHA; antiangiogenic drugs, including bevacizumab; monoclonal antibodies, including trastuzumab, rituximab, alemtuzumab, tositumomab, cetuximab, panitumumab; conjugates of myoclonal antibodies, including Gemtuzumab ozogamicin, Ibritumomab tiuxetan; hormonal therapies, including antiestrogens (tamoxifen, raloxifen, anastrazole, letrozole, examestane) antiandrogens (Flutamide, Biclutamide) and Luteinisng Hormone Analogues or antagonists.
With regard to combination therapy the compounds of the present invention may be administered separately, sequentially, simultaneously, concurrently or may be chronologically staggered with one or more standard therapeutics such as any of those mentioned above.
The present invention also provides a pharmaceutical composition suitable for clinical use.
In particular, a pharmaceutical composition may comprise a pharmaceutical carrier and, dispersed therein, a therapeutically effective amount of the compounds of the invention. The composition may be solid or liquid. The pharmaceutical carrier is generally chosen based on the type of administration being used and the pharmaceutical carrier may for example be solid or liquid. The compounds of the invention may be in the same phase or in a different phase than the pharmaceutical carrier.
Pharmaceutical compositions may be formulated according to their particular use and purpose by mixing, for example, excipient, binding agent, lubricant, disintegrating agent, coating material, emulsifier, suspending agent, solvent, stabilizer, absorption enhancer and / or ointment base. The composition may be suitable for oral, injectable, rectal or topical administration.
For example, the pharmaceutical composition may be administered orally, such as in the form of tablets, coated tablets, hard or soft gelatine capsules, solutions, emulsions, or suspensions. Administration can also be carried out rectally, for example using suppositories, locally or percutaneously, for example using ointments, creams, gels or solution, or paenterally, for example using injectable solutions.
For the preparation of tablets, coated tablets or hard gelatine capsules, the compounds of the present invention may be admixed with pharmaceutically inert, inorganic or organic excipients. Examples of suitable excipients include lactose, mize starch or derivatives thereof, talc or stearic acid or salts thereof. Suitable excipients for use with soft gelatine capsules include, for example, vegetable oils, waxes, fats and semi-solid or liquid polyols.
For the preparation of solutions and syrups, excipients include, for example, water, polyols, saccharose, invert sugar and glucose.
For injectable solutions, excipients include, for example, water, alcohols, polyols, glycerine and vegetable oil.
For suppositories and for local and percutaneous application, excipients include, for example, natural or hardened oils, waxes, fats and semi-solid or liquid polyols.
The pharmaceutical compositions may also contain preserving agents, solublizing agents, stabilizing agents, wetting agents, emulsifiers, sweeteners, colorants, odorants, buffers, coating agents and / or antioxidants.
For combination therapies, the second drug may be provided in pharmaceutical composition with the present invention or may be provided separately.
Thus, a pharmaceutical formulation for oral administration may, for example, be granule, tablet, sugar coated tablet, capsule, pill, suspension or emulsion. For parenteral injection for, for example, intravenous, intramuscular or subcutaneous use, a sterile aqueous solution may be provided that may contain other substances including, for example, salts and / or glucose to make to solution isotonic. The anti-cancer agent may also be administered in the form of a suppository or pessary, or may be applied topically in the form of a lotion, solution, cream, ointment or dusting powder.
The present invention also relates to methods of treating or preventing cancer comprising adminstering a compound according to the present invention as described herein to a subject in need thereof. In preferred embodiments the method includes the use of one or more compounds of the present invention in a regime including the step of radiotherapy.
EXAMPLES
The present invention will now be described in relation to several examples.
Examples 1 to 153 were synthesised according to the methods described subsequently. Their IC50 values were then determined as described below and are represented in the following table, in which the compound numbers correspond to the numbers in the examples.
Example IC50 in IMAP IC50 in IMAP IC50 in IMAP assay
Number assay vs AKT1 assay vs AKT2 vs AKT3
1 ++ ++ +
2 +++ ++ +
3 ++ + +
4 ++++ +++
5 +++ ++ +
6 +++ ++ ++
7 ++++ ++ +
8 +++ +++ +
9 +++ ++ +
10 +++ ++
11 +++ +++ +
12 ++++ ++ +
13 +++ +++ ++ Example IC50 in IMAP IC50 in IMAP IC50 in ID
Number assay vs AKT1 assay vs AKT2 vs AKT3
14 +++ ++ ++
15 +++ ++ ++
16 ++++ +++ ++
17 +++ ++ ++
18 ++ ++ +
19 +++ +++ +
20 +++ ++ +
21 +++ ++ +
22 ++++ ++ +
23 +++ ++ +
24 ++ +++ +
25 +++ ++ +
26 ++++ ++ +
27 ++ + +
28 ++ +++ +
29 ++++ +++ +
30 ++ ++ ++
31 ++++ +++ ++
32 +++ +++ +
33 ++++ ++++ +++
34 ++ ++ +
35 ++ +++ +
36 +++ ++ ++
37 +++ ++ +
38 +++ +++ ++
39 +++ ++ +
40 +++ ++ +
41 ++ +++ +
42 ++ +++ ++
43 +++ ++++ + Example IC50 in IMAP IC50 in IMAP IC50 in I AP assay
Number assay vs AKT1 assay vs AKT2 vs AKT3
44 ++ ++ +
45 ++ +++ ++
46 +++ +++ +++
47 ++++ +++ ++
48 +++ ++++ +++
49 +++ +++ +++
50 ++++ +++ ++
51 +++ +++ ++
52 +++ ++++ ++
53 ++ ++ ++
54 +++ +++ ++
55 +++ ++++ +++
56 +++ +++ +++
57 ++ +++ ++
58 ++ +++ ++
59 ++ +++ ++
60 +++ ++++ ++
61 ++ +++ ++
62 ++ +++ ++
63 +++ ++ +
64 +++ +++ ++
65 ++ ++ +
66 +++ +++ ++
67 +++ ++++ ++
68 +++ +++ +
69 ++++ +++ ++
70 +++ +++ +
71 +++ ++ ++
72 +++ ++ +
73 ++ +++ + Example IC50 in IMAP IC50 in IMAP IC50 in IMAP assay
Number assay vs AKT1 assay vs AKT2 vs AKT3
74 ++ ++ +
75 ++ ++ +
76 +++ ++++ +
77 +++ ++ +
78 +++ +++ +
79 ++++ +++ ++
80 +++ +++ +
81 ++++ +++ ++
82 ++ ++++ ++
83 ++ ++ ++
84 +++ ++ ++
85 +++ ++++ +++
86 +++ ++++ ++
87 +++ ++ +
88 + + +
89 +++ +++ ++
90 ++ +++ +
91 +++ ++++ ++
92 +++ ++ ++
93 ++++ ++++ ++
94 +++ ++ +
95 +++ +++ +
96 +++ ++ +
97 +++ ++ ++
98 ++++ +++ +++
99 +++ +++ ++
100 +++ ++ ++
101 +++ +++ ++
102 +++ ++++ ++
103 ++ +++ ++ Example IC50 in IMAP IC50 in IMAP IC50 in IMAP assay
Number assay vs AKT1 assay vs AKT2 vs AKT3
104 +++ ++ +
105 ++++ ++++ ++
106 +++ +++ ++
107 ++++ ++ ++
108 +++ +++ ++
109 ++++ ++++ +++
110 +++ +++ +++
111 ++ ++ +
112 +++ ++ ++
113 ++++ +++ ++
114 ++ ++ +
115 +++ +++ ++
116 ++ ++ ++
117 ++ ++ ++
118 +++ ++ ++
119 +++ +++ ++
120 ++++ +++ +++
121 +++ ++ ++
122 +++ ++ ++
123 ++++ ++ ++
124 ++ ++ ++
125 +++ ++ ++
126 +++ +++ ++
127 ++ + +
128 ++++ +++ +++
129 +++ ++ ++
130 +++ + +
131 ++ +++ ++
132 +++ ++ +
133 ++ + + Example IC50 in IMAP IC50 in IMAP IC50 in IMAP assay
Number assay vs AKT1 assay vs AKT2 vs AKT3
134 +++ ++ +
135 +++ ++++ ++
136 +++ ++ ++
137 ++++ +++ ++
138 ++++ +++ ++
139 +++ +++ ++
140 +++ +++ ++
141 +++ +++ +++
142 ++ ++ ++
143 ++++ ++++ +++
144 +++ ++++ +++
145 ++++ ++++ +++
146 ++ +++ +
147 +++ +++ +
148 +++ +++ +
149 +++ ++ +
150 ++++ ++++ ++
151 ++ +++ +
152 +++ ++++ ++
153 ++ ++ +
Key
+ IC50 > 10μΜ
++ 1 Μ < IC50 < 10μΜ
+++ 0.1 μΜ < ΙΟ50≤1μΜ
++++ IC50 <0.1μΜ
In addition, the phosphorylation status of various members of the AKT/PI3K pathway was investigated via ELISA. For example, Example 4 causes inhibition of GSK3
phosphorylation with an Ιθ50<0.1μΜ measured at 48h. It can be seen from the Table that several Examples exhibit selectivity for one or more AKT isoforms over the other isoform(s). For example, a greater activity for AKT1 and / or AKT2 is observed compared to AKT3.
In addition, the activity of compounds in in vitro cell proliferation assays was investigated. Representative examples (eg.4, 28 and 31 ) show inhibition of proliferation of PC3 and/or LnCaP cell lines with an IC50 < 10 μΜ.
Abbreviations
AcOH: Acetic acid; nBuLi: n-Butyllithium; BINAP: 2,2'-bis(diphenylphosphino)-1 , 1 '- binaphthyl; DBU: 1 ,8-Diazabicyclo[5.4.0]undec-7-ene; DCM: Dichloromethane; DIPEA: Diisopropylethylamine; DMA: A/./V-Dimethyl acetamide; DMAP: 4-Dimethylaminopyridine; DME: Dimethoxy ethane; DMF: A/.W-Dimethylformamide; DMSO: Dimethylsulfoxide; EDCI: 1-Ethyl-3-(3'-dimethylaminopropyl)carbodiimide; EtOAc: Ethyl acetate; h:Hour: HATU: 0-(7-Azabenzotriazol-1 -y\)-N,N,N', Λ/'-tetramethyluronium hexafluorophosphate; HCI: Hydrochloric acid; HOBt: 1-Hydroxybenzotriazole; HPLC: High Pressure Liquid Chromatography; IMS: Industrial methylated spirits: M: Molar; MeOH: Methanol; NMP: N- Methyl-2-pyrrolidone; NMR: Nuclear Magnetic Resonance; Min: Minutes; RT: Room temperature; SCX: SCX- strong cation exchange; TBAF: Tetra-n-butylammonium fluoride; TEA: Triethylamine; TFA: Trifluoroacetic acid; THF: Tetrahydrofuran; TMSCI: Trimethylsilyl chloride
General Experimental Conditions
1H NMR spectra were recorded at ambient temperature using a Varian Unity Inova (400MHz) spectrometer with a triple resonance 5mm probe, a Bruker Avance DRX (400MHz) with a 5mm inverse detection triple resonance TXI probe, a Bruker Avance (500MHz) spectrometer with a 5mm QNP probe or a Bruker Avance DPX (300MHz) spectrometer with a standard 5mm dual frequency probe. Chemical shifts are expressed in ppm relative to tetramethylsilane.
High Pressure Liquid Chromatography - Mass Spectrometry (LCMS) experiments to determine retention times (RT) and associated mass ions were performed using one of the following methods.
Method A: The system consists of ThermoFinnigan LCQ Advantage Mass Spectrometer with the Surveyor LC system and 200 position autosampler. The spectrometer has an electrospray source operating in positive and negative ion mode. An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Luna 3micron Ci8 50 x 2 mm; Mobile phase -A)Water 0.1 % Formic Acid B)Acetonitrile 0.1% Formic Acid
Gradient - Time flow %A %B
0.00 0.6 95 5
7.00 0.6 5 95
8.00 0.6 5 95
8.20 0.6 95 5
1 1.00 0.6 95 5
Split - Ι ΟΟμΙ/min split to the ESI source with inline Surveyor DAD detection.
Detection - MS, UV
MS ionisation method - Electrospray (positive and negative ion)
Total experiment time - 11 mins (approx)
Method B: The system consists of a Waters ZMD quadrupoie mass spectrometer linked to a Waters 1525 LC system with Waters 996 diode array detector. Sample injection is done by a Waters 2700 autosampler. The spectrometer has an electrospray source operating in positive and negative ion mode. Additional detection is achieved using a Sedex 85 evaporative light scattering detector.
An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Luna 3micron C18(2) 30 x 4.6mm or equivalent. Mobile phase - A)Water 0.1 % Formic Acid B)Methanol 0.1 % Formic Acid
Gradient - Time flow %A %B
0.00 2.0 95 5
0.50 2.0 95 5
4.50 2.0 5 95
5.50 2.0 5 95
6.00 2.0 95 5
Split - 200pl/min split to the ESI source with inline Waters 996 DAD detection.
Detection - MS, ELS, UV MS ionisation method - Electrospray (positive and negative ion)
Total experiment time - 6 mins (approx)
Method C: The system consists of a Waters Micromass ZQ2000 quadrupole mass spectrometer linked to a Waters Acquity UPLC system with a PDA UV detector. The spectrometer has an electrospray source operating in positive and negative ion mode. An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Acquity BEH C18 1.7um 100 x 2.1 mm, maintained at 40°C or Acquity BEH Shield RP18 1 Jum 100 x 2.1 mm, maintained at 40°C. Mobile phase - A)Water 0.1 % Formic Acid B)Acetonitrile 0.1 % Formic Acid
Gradient - Time flow ml/min %A %B
0.00 0.4 95 5
0.40 0.4 95 5
6.00 0.4 5 95
6.80 0.4 5 95
7.00 0.4 95 5
8.00 0.4 95 5
Detection - MS, UV PDA
MS ionisation method - Electrospray (positive/negative ion)
Method D: The system consists of an Agilent Technologies 6140 single quadrupole mass spectrometer linked to an Agilent Technologies 1290 Infinity LC system with UV diode array detector and autosampler. The spectrometer has a multimode ionization source (electrospray and atmospheric pressure chemical ionizations) operating in positive and negative ion mode. An LC-MS experiment is performed on each sample submitted using the following conditions: LC Column - Zorbax Eclipse Plus C18 RRHD 1.8micron 50 x 2.1 mm maintained at 40°C. Mobile phase - A) Water 0.1 % Formic Acid B) Acetonitrile 0.1 % Formic Acid. Gradient - Time Flow mL/min %A %B
0.00 1.0 95 5
1.80 1.0 0 100
2.20 1.0 0 100
2.21 1.0 95 5
2.50 1.0 95 5
Detection - MS, UV
MS ionisation method - Multimode (positive and negative ion)
Total experiment time - 2.50 mins (approx)
Method E: The system consists of a Agilent Technologies 6120 single quadrupole mass spectrometer linked to a Agilent Technologies 1200 Preparative LC system with Multiple Wavelength detector and autosampler. The spectrometer has a multimode ionization source (electrospray and atmospheric pressure chemical ionizations) operating in positive and negative ion mode. Fraction collection is mass-triggered. A purification experiment is performed on each sample submitted using the following conditions: LC Column - Waters XBridge™ Prep C18 5μιη OBD™ 19 x 50mm column at room temperature. Mobile phase - A) Water 0.1 % ammonium hydroxide B) Acetonitrile/Water 95: 5 with 0.1 % ammonium hydroxide.
Gradient - Time Flow mL/min %A %B,
0.00 20.0 95 5
7.00 20.0 0 100
8.00 20.0 0 100
8.20 20.0 95 5
Detection - MS, UV
MS ionisation method - Multimode (positive and negative ion)
Total experiment time - 10 mins (approx)
Method F: The system consists of a Agilent Technologies 6120 single quadrupole mass spectrometer linked to a Agilent Technologies 1200 Preparative LC system with Multiple Wavelength detector and autosampler. The spectrometer has a multimode ionization source (electrospray and atmospheric pressure chemical ionizations) operating in positive and negative ion mode. Fraction collection is mass-triggered. A purification experiment is performed on each sample submitted using the following conditions: LC Column - Waters XBridge™Prep C18 5pm OBD™ 30 x 100mm column at room temperature. Mobile phase - A) Water 0.1 % ammonium hydroxide B) Acetonitrile/Water 95: 5 with 0.1% ammonium hydroxide.
Gradient - Time Flow ml_/min %A %B
0.00 60.0 95 5
14.00 60.0 0 100
16.00 60.0 0 100
16.50 60.0 95 5
Detection - MS, UV
MS ionisation method - Multimode (positive and negative ion)
Total experiment time - 18 mins (approx)
Method G: The system consists of an Agilent 6110 quadrupole LC / MS mass
spectrometer linked to an Agilent 1200 LC system with Agilent 1200 diode array detector. Sample injection is done by an Agilent 1200 autosampler. The spectrometer has an electrospray and APCI source operating in positive and negative ion mode.
A purification experiment is performed on each sample submitted using the following conditions: LC Column - Sunfire 5micron C18(2) 15x 4.6mm or equivalent. Mobile phase -A)Water 0.1 % Formic Acid B)ACN 0.1 % Formic Acid
Split - 40/min split to the ESI / APCI source
Detection - MS, UV
MS ionisation method - Electrospray and APCI (positive and negative ion)
Total experiment time - 6 or 20 mins (approx)
Microwave experiments were carried out using a CEM Explorer™ or Biotage Initator™ instruments. Temperatures from 60-300°C can be achieved, and pressures of up to 20bar can be reached. Unless otherwise indicated, the nomenclature of structures was using "structure=name" from ChemBioDraw 11 (CambridgeSoft).. Some compounds named using the software mentioned above have stereocentres marked 'S' and 'R' where they would normally be designated 'cis' or 'trans'. For these compounds the structures presented are representative of the compounds made.
Unless otherwise indicated, starting materials and intermediates were obtained from commercial suppliers, prepared according to literature procedures (for example WO 2008/070016, WO 2009/148887 and WO 2009/148916) or by standard transformations obvious to one skilled in the art.
Example 1 : 2-(4-(1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin-5(6H)- one
Figure imgf000030_0001
Step 1 : tert-butyl (1-(4-(3-(dimethylamino)-2-phenylacryloyl)phenyl)cyclobutyl)carbamate A solution of tert-butyl (1-(4-(2-phenylacetyl)phenyl)cyclobutyl)carbamate (25.0 g, 68 mmol) in Ν,Ν-dimethylformamide dimethylacetal (150 mL) was heated to 80°C under a nitrogen atmosphere for 18 hours. The resulting brown solution was allowed to cool to room temperature and concentrated to dryness under reduced pressure to give the desired product as a yellow/brown solid (30.6 g, quantitative yield). 1H-NMR (500 MHz, CDCI3) δ 7.14-7.48 (10H, m), 5.09 (1 H, br s), 2.74 (6H, s), 2.45-2.60 (4H, m), 2.04-2.16 (1 H, m), 1.80-1.90 (1 H, m), 1.10-1.52 (9H, br m).
Step 2: tert-butyl (1-(4-(5-oxo-3-phenyl-5.6.7.8-tetrahvdroquinolin-2- vQphenvQcyclobutvDcarbamate
To acetic acid (120 mL) was added molecular sieves (5 A, 1.50 g), tert-butyl (1-(4-(3- (dimethylamino)-2-phenylacryloyl)phenyl)cyclobutyl)carbamate (8.00 g, 19.0 mmol) and 3-amino-2-cyclohexene-1-one (3.20 g, 28.8 mmol). The reaction mixture was heated to 100°C under a nitrogen atmosphere for 2 hours. The mixture was allowed to cool to room temperature and concentrated under reduced pressure. The residue was partitioned between water (160 mL) and dichloromethane (160 mL) and decanted from the molecular sieves. The layers were separated and the aqueous phase extracted into dichloromethane (2 x 160 mL). The combined organic phases were washed with saturated NaHC03 solution (2 x 160 mL), brine (160 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow/brown solid. This was purified by Biotage chromatograpy (silica, hexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as a yellow solid (3.30 g, 37% yield). 1H-NMR (500 MHz, CDCI3) δ 8.30 (1 H, s), 7.37 (2H, d), 7.30 (2H, d), 7.24-7.27 (3H, m), 7.15-7.20 (2H, m), 5.01 (1H, br s), 3.26 (2H, t), 2.75 (2H, t), 2.42-2.55 (4H, m), 2.26 (2H, p), 2.00- 2.10 (1 H, m), 1.73-1.85 (1 H, m), 1.12-1.44 (9H, br m). LCMS (Method A) RT = 7.54 min, M+H+ = 469.15.
Step 3: 2-(4-(1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihvdroquinolin-5(6H)-one
tert-butyl (1 -(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (11 mg, 0.023 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (10 mg, 91% yield). 1H-NMR (500 MHz, CD3OD) δ 8.29 (1H, s), 7.51 (2H, d), 7.45 (2H, d), 7.31 (3H, t), 7.21 (2H, dd), 3.26 (2H, t), 2.80 (2H, t), 2.72-2.81 (2H, m), 2.53-2.62 (2H, m), 2.30 (2H, t), 2.20-2.30 (1 H, m), 1.92-2.01 (1 H, m). LCMS (Method A) RT = 4.09 min, M+H+ = 368.91.
Example 2: 2-(4-(1-aminocvclobutyl)phenvH-3-phenyl-7,8-dihvdroquinolin-5(6H)- one oxime
Figure imgf000031_0001
Step 1 : Terf-butyl (1-(4-(5-(hvdroxyirnino)-3-phenyl-5,6,7,8-tetrahvclroauinoliri-2- vDphenvDcyclobutyQcarbamate
To a solution of ie/i-butyl-(1-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin- 5(6H)-one (80 mg, 0.17 mmol) in anhydrous ethanol (2.0 mL) was added hydroxylamine hydrochloride (65 mg, 0.9 mmol) and pyridine (75 μΙ_, 0.9 mmol). The resulting yellow solution was heated to 90°C for 2 hours under a nitrogen atmosphere. The reaction mixture was allowed to cool to room temperature and concentrated to dryness under reduced pressure. The residue was mixed with water (2.0 mL) and extracted into dichloromethane (3 x 2.0 mL). The combined organic phases were washed with brine (2 x 4.0 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. The residue was purified by Biotage chromatography (silica, hexane:ethyl acetate, gradient elution from 95:5 to 60:40) to give the desired product as a white solid (47 mg, 56% yield). 1H NMR (500 MHz, CH3OD) 8.30 (s, 1H), 7.30 (d, 2H), 7.26 (m, 5H), 7.19 (d, 2H), 3.19 (m, 2H), 2.87 (m, 2H), 2.45-2.50 (m, 4H), 2.0-2.1 (m, 1H), 1.90-1.99 (m, 2H), 1.80-1.86 (m, 1H), 1.2-1.4 (br s, 9H).
Step 2: 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin-5(6H)-one oxime Tert-butyl (1-(4-(5-(hydroxyimino)-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (7 mg, 0.017 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (3 mg, 35% yield). 1H NMR (500 MHz, CH3OD) 8.45 (s, 1H), 7.46 (m, 5H), 7.34 (d, 2H), 7.21 (d, 2H), 3.10 (t, 2H), 2.90 (t, 2H), 2.74-2.78 (m, 2H), 2.56-2.6 (m, 2H), 2.20-2.28 (m, 1 H), 2.02-2.10 (m, 2H), 1.93-1.98 (m, 1 H). Example 3: 2-(4-(1 -aminocvclobutyl)phenyl)-3-phenyl-7.8-dihvdroquinolin-5(6H)- one O-methyl oxime
Figure imgf000033_0001
Step 1: tert-butyl (1-(4-(5-(methoxyimino)-3-phenyl-5,6,7,8-tetrahvdroquinolin-2- yl)phenyl)cyclobutyl)carbannate
To a solution of tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (90 mg, 0.19 mmol) in anhydrous ethanol (2.0 ml.) was added O-methylhydroxylamine hydrochloride (80 mg, 0.96 mmol) and pyridine (80 μΙ_, 0.99 mmol). The resulting yellow solution was heated to 90°C under a nitrogen atmosphere. The reaction mixture was allowed to cool to room temperature and concentrated to dryness under reduced pressure. The residue was mixed with water (2.0 ml_) and extracted into ethyl acetate (3 x 2.0 ml_). The combined organic phases were washed with brine (2 x 4.0 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. The residue was purified by Biotage chromatography (silica, hexane:ethyl acetate, gradient elution from 95:5 to 60:40) to give the desired product as a white solid (44 mg, 46% yield). 1H-NMR (500 MHz, CD3OD) δ 8.29 (1 H, s), 7.35 (2H, d), 7.26-7.32 (5H, m), 7.16-7.21 (2H, m), 4.01 (3H, s), 3.02 (2H, t), 2.83 (2H. t), 2.36-2.52 (4H, m), 2.04-2.16 (1 H, m), 2.01 (2H, p), 1.81-1.92 (1 H, m), 1.15- 1.50 (9H, br m). LCMS (Method A) RT = 8.28 min, M+H+ = 498.10.
Step 2: 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7.8-dihydroquinolin-5(6H)-one O- methyl oxime
tert-butyl (1-(4-(5-(methoxyimino)-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (44 mg, 0.088 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (32 mg, 73% yield). 1H-NMR (500 MHz, CD3OD) δ 8.35 (1 H, s), 7.48 (2H, d), 7.43 (2H, d), 7.30 (3H, t), 7.18-7.23 (2H, m), 4.02 (3H, s), 3.05 (2H, t), 2.85 (2H, t), 2.73-2.81 (2H, m), 2.54-2.63 (2H, m), 2.19-2.30 (1 H, m), 2.03 (2H, p), 1.91-2.03 (1 H, m). LCMS (Method A) RT = 4.57 min, M+H+ = 398.10.
Example 4: 1 -(4-(8-phenyl- ,5-dihydro-2H-pyrazolo[3.4- 1quinolin-7- vPphenvQcyclobutanamine
Figure imgf000034_0001
Step 1 : Tert-butyl 1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-5,6,7l8- tetrahydronaphthalen-2-yl)phenyl)cvclobutyl)carbamate
Tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (500 mg, 1.06 mmol) was dissolved in 5 ml of anhydrous Ν,Ν-dimethylfomamide dimethyl acetale. The resulting mixture was heated to 100°C under a nitrogen atmosphere for 2.5 hours. TLC and LC-MS analysis showed complete consumption of the starting material. The reaction mixture was concentrated to dryness under reduced pressure. The crude residue was slurried in n-hexane (10 ml) for 1 hour. The suspension was filtered and the solid was dried until constant weigh to give the desired product as a bright yellow solid (420 mg, 76% yield). LCMS (Method A): RT = 6.89 min, M+H+ = 524; RT = 8.37 min, M+H+ = 497
Step 2: terf-butyl (1-(4-(8-phenyl-4.5-dihvdro-2H-pyrazolor3.4-flquinolin-7- yl)phenyl)cyclobutyl)carbamate
To a solution of terf-butyl 1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydronaphthalen-2-yl)phenyl)cyclobutyl)carbamate (40 mg, 0.07 mmol) in anhydrous ethanol (1500 pL) was added hydrazine monohydrate (15 μΙ_, 0.23 mmol). The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 2.5 hours. The reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 95:5 to 60:40) and slurried in diethyl ether (1 mL) to give the desired product as a white solid (6 mg, 16% yield). 'H NMR (500 MHz, CH3OD) 8.15 (s, 1 H), 7.22-7.37 (m, 9H), 3.22 (t, 2H), 3.00 (t, 2H), 2.45-2.51 (m, 4H), 2.03-2.09 (m, 1 H), 1.86-1.89 (m, 1 H), 1.3-1.5 (br s, 9H).
Step 3: 1-(4-(8-phenyl-4,5-dihvdro-2H-pyrazolof3,4-f1auinolin-7- vDphenyQcyclobutanamine
Terf-butyl (1 -(4-(8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (6 mg, 0.012 mmol) was dissolved in TFA (0.8 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (3 mg, 50% yield). 1H NMR (500 MHz, CH3OD) 8.16 (s, 1 H), 7.58 (s, 1 H), 7.49 (d, 2H), 7.43 (d, 2H), 7.30 (m, 3H), 3.24 (t, 2H), 3.02 (t, 2H), 2.78 (m, 2H), 2.59 (m, 2H), 2.23 (m, 1 H), 1.98 (m, 1 H).
Example 5: 1 -(4-(2-methyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3,4-f1quinolin-7- vQphenvDcvclobutanamine
Example 6: 1-(4-(1-methyl-8-phenyl-4,5-dihvdro-1 H-pyrazolor3,4-f|quinolin-7- vQphenvDcvclobutanamine
Figure imgf000035_0001
Step 1 : tert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3,4-f1auinolin-7- yl)phenyl)cyclobutyl)carbamate and te/ -butyl (1-(4-(1-methyl-8-phenyl-415-dihydro-1 H- pyrazolor3,4-f1quinolin-7-yl)phenyl)cvclobutyl)carbamate
To a solution of fert-butyl 1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydronaphthalen-2-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.19 mmol) in anhydrous ethanol (2 mL) was added methyl hydrazine (30 μί, 0.57 mmol). The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 99:1 to 70:30) to give the desired product fert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihydro-2H- pyrazolo[3,4-f]quinolin-7-yl)phenyl)cyclobutyl)carbamate as a white solid (14 mg, 15% yield). 1H NMR (500 MHz, CH3OD) 8.10 (s, 1 H), 7.36 (d, 2H), 7.29 (d, 2H), 7.20-7.22 (m, 6H), 4.99 (s, 1H), 3.94 (s, 3H), 3.20-3.36 (br s, 2H), 2.86-3.01 (m, 4H), 2.43-2.55 (m, 2H), 1.99-2.10 (m, 1H), 1.74-1.85 (m, 1 H), 1.49-1.68 (br s, 9H). H-1H NOESY (500 MHz, CH3OD) positive correlation between 7.22 (s, 1 H) and 3.94 (s, 3H).
Mixted fractions from the column (45mg) were purified by preparative HPLC (method G, gradient 5 to 95% 0.1%FA/ACN in 0.1%FA/H2O, 20 minutes run) to give the desired product fe/ -butyl (1-(4-(1-methyl-8-phenyl-4,5-dihydro-1 H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate as a white solid (9 mg, 10% yield). 1H NMR (500 MHz, CH3OD) 7.83 (s, 1H), 7.37 (m, 3H), 7.29 (m, 5H), 7.22 (m, 2H), 5.00 (s, 1 H), 4.16 (s, 3H), 3.22-3.33 (br m, 2H), 2.87-2.94 (m, 2H), 2.41-2.55 (m, 4H), 2.04 (m, 1 H), 1.57 (m, 1 H), 1.50-1.62 (br s, 9H). LCMS (Method A): RT = 7.36 min, M+H+ = 507.
Step 2A: 1 -(4-(2-methyl-8-phenyl-4.5-dihvdro-2H-pyrazolof3.4-f1quinolin-7- yl)phenyl)cyclobutanamine
Figure imgf000036_0001
Terf-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (14 mg, 0.027 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (-1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (1 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (7 mg, 50% yield). 1H NMR (500 MHz, CH3OD) 8.20 (s, 1H), 7.50 (s, 1 H), 7.47 (d, 2H), 7.43 (d, 2H), 7.28 (m, 3H), 7.22 (m, 2H), 3.94 (s, 1 H), 3.24 (t, 2H), 2.97 (t, 2H), 2.70-2.80 (br m, 2H), 2.52-2.61 (br m, 2H), 2.15-2.29 (br m, 1 H), 1.88-2.00 (br m, 1 H). Step 2B. 1-(4-(1-methyl-8-phenyl- .5-dihvdro-1 H-pyrazolo[3.4-flauinolin-7- vDphenvDcyclobutanamine
Figure imgf000037_0001
Tert-butyl (1 -(4-(1-methyl-8-phenyl-4,5-dihydro-1 H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (3 mg, 0.006 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (1 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (0.65 mg, 22% yield). 1H NMR (500 MHz, CH3OD) 8.06 (s, 1 H), 7.50 (d, 2H), 7.43 (d, 3H), 7.27-7.36 (br m, 5H), 4.19 (s, 1 H), 3.22 (t, 2H), 2.93 (t, 2H), 2.73-2.83 (br m, 2H), 2.54-2.66 (br m, 2H), 2.18-2.33 (br m, 1 H), 1.92-2.03 (br m, 1 H). LCMS (Method A): RT = 4.21 min, M+H+ = 407
Example 7: 7-(4-(1 -aminocvclobutyl)phenyl)-8-phenyl-4,5-dihvdro-2H-pyrazolof3.4- flquinolin-3-ol
Figure imgf000037_0002
Step 1 : tert-butyl (1-(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl-5,6,7.8- tetrahydroquinolin-2-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (1.00 g, 2.13 mmol) in anhydrous tetrahydrofuran
(10 mL) was added dropwise potassium tert-butoxide, 1M in THF (4.69 mL, 4.69 mmol) over a period of 10 min to give a deep red suspension. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 15 min. To this was added dropwise carbon disulfide (142 μΙ_, 2.35 mmol) over a period of 5 min to give a deep red solution. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 30 min. Methyl iodide (294 μΙ, 4.69 mmol) was added and the mixture stirred at room temperature for 90 min. The reaction mixture was diluted with saturated sodium bicarbonate solution (40 mL) and extracted with ethyl acetate (3 x 40 mL). The combined organic phases were washed with 50:50 waterbrine (40 mL), then brine (40 mL), dried over Na2S0 , filtered and concentrated to dryness under reduced pressure to give the desired product as a bright yellow solid (1.21 g, 99% yield). H-NMR (500 MHz, CDCI3) δ 8.40 (1 H, s), 7.37 (2H, d), 7.31 (2H, d), 7.23-7.28 (3H, m), 7.17-7.21 (2H, m), 3.40 (2H, t), 3.31 (2H, br s), 2.50 (3H, s), 2.46 (3H, s), 2.42-2.57 (4H, m), 2.00- 2.12 (1 H, m), 1.75-1.87 (1 H, m), 1.12-1.47 (9H, br m). LCMS (Method A) RT = 8.64 min, M+H+ = 573.13.
Step 2: methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-5-oxo-3-phenyl- 5,6,7,8-tetrahydroquinoline-6-carboxylate
To a stirred solution of tert-butyl (1-(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl- 5,6,7,8-tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (140 mg, 0.24 mmol) in anhydrous tetrahydrofuran (2.8 mL) was added a solution of sodium hydroxide (96 mg, 2.4 mmol) in methanol (2.2 mL). The reaction mixture was heated to 50°C under a nitrogen atmosphere for 2 hours. After allowing to cool to room temperature the mixture was diluted with 1 M HCI solution (3 mL) and extracted into ethyl acetate (3 x 6 mL). The combined organic phases were washed with brine (3 x 6 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give the desired product as a brown solid (126 mg, quantitative yield). LCMS (Method A) RT = 7.56 min, M+H+ = 527.14.
Step 3: tert-butyl (1-(4-(3-hvdroxy-8-phenyl-4.5-dihvdro-2H-pyrazolor3,4-flquinolin-7- vDphenvDcyclobutyDcarbamate
To a solution of methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-5-oxo-3- phenyl-5,6,7,8-tetrahydroquinoline-6-carboxylate (60 mg, 0.1 1 mmol) in anhydrous ethanol (1500 pL) was added hydrazine monohydrate (9 μί, 0. 9 mmol). The reaction mixture was heated to reflux under a nitrogen atmosphere for 18 hours. After cooling to room temperature the reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, dichloromethane:methanol, gradient elution from 99: 1 to 90:10) and slurried in diethyl ether (1 mL) to give the desired product as a white solid (19 mg, 33% yield). 1H-NMR (500 MHz, CD3OD) δ 7.94 (1 H, s), 7.32 (2H, d), 7.21-7.29 (5H, m), 7.14-7.21 (2H, m), 3.18 (2H, t), 2.81 (2H, t), 2.31-2.50 (4H, m), 2.00-2.12 (1 , br m), 1.76-1.91 (1 H, br m), 1.09-1.48 (br m). LCMS (Method A) RT = 5.57 min, M+H+ = 509.17.
Step 4: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4,5-dihvdro-2H-pyrazolof3,4-flquinolin- 3-ol
tert-butyl (1-(4-(3-hydroxy-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (19 mg, 0.037 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (3 mg, 17% yield). 1H-NMR (500 MHz, CD3OD) δ 8.05 (1 H, s), 7.49 (2H, d), 7.44 (2H, d), 7.27-7.34 (3H, m), 7.20-7.27 (2H, m), 3.25 (2H, t), 2.87 (2H, t), 2.72-2.82 (2H, m), 2.54-2.65 (2H, m), 2.18-2.30 (1 H, m), 1.90-2.03 (1 H, m). LCMS (Method A) RT = 3.40 min, M+H+ = 409.19.
Example 8: 7-(4-(1-aminocvclobutyl)phenvH-3a-methyl-8-phenyl-4,5-dihvdro-2H- pyrazolor3,4-f|quinolin-3(3 -one
Figure imgf000039_0001
7-(4-(1-aminocyclobutyl)phenyl)-3a-methyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-3(3a/-/)-one
Step 1 : methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cvclobutyl)phenyl)-6-methyl-5-oxo-3- phenyl-5,6,7,8-tetrahydroquinoline-6-carboxylate
A solution of methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-5-oxo-3- phenyl-5,6,7,8-tetrahydroquinoline-6-carboxylate (65 mg, 0.12 mmol) in anhydrous THF (1.0 mL) was cooled to 0°C under a nitrogen atmosphere, followed by the dropwise addition of potassium tert-butoxide solution (1 M in THF, 150 μί, 0.15 mmol). The mixture was stirred at 0°C for 15 minutes followed by the addition of methyl iodide (10 μΐ_, 0.16 mmol). The reaction mixture was allowed to warm to room temperature and stirred for a further 4 hours. The mixture was diluted with ethyl acetate (2.0 mL), washed with 1M HCI solution (2.0 mL), brine (2 x 2.0 mL), dried over Na2S04, filtered and
concentrated to dryness under reduced pressure to give the desired product as a brown solid (54 mg, 83% yield). LCMS (Method A) RT = 7.92 min, M+H+ = 541.18.
Step 2: tert-butyl (1-(4-(3a-methyl-3-oxo-8-phenyl-3.3a,4.5-tetrahvdro-2H-pyrazolor3.4- f1quinolin-7-yl)phenyl)cvclobutyl)carbamate
To a solution of methyl 2-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-6-methyl- 5-oxo-3-phenyl-5,6,7,8-tetrahydroquinoline-6-carboxylate (54 mg, 0.10 mmol) in anhydrous ethanol (1300 pL) was added hydrazine monohydrate (8 pL, 0.16 mmol). The reaction mixture was heated to reflux under a nitrogen atmosphere for 5 hours, additional hydrazine monohydrate (40 pL, 0.82 mmol) added, and the mixture heated at reflux for 18 hours. After cooling to room temperature the reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica,
hexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a yellow solid (12 mg, 23% yield). 1H-NMR (500 MHz, CDCI3) δ 8.72 (1 H, s), 8.09 (1 H, s), 7.34 (2H, d), 7.30 (2H, d), 7.23-7.28 (3H, m), 7.16-7.22 (2H, m), 5.05 (1H, br s), 3.32- 3.44 (1 H, m), 3.19-3.32 (1 H, br m), 2.41-2.60 (4H, br m), 2.26-2.35 (1 H, dd), 1.97-2.11 (2H, m), 1.74-1.86 (1 H, m), 1.45 (3H, s), 1.14-1.44 (9H, br m). LCMS (Method A) RT = 6.60 min, M+H+ = 523.17.
Step 3: 7-(4-(1-aminocvclobutyl)phenyl)-3a-methyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3,4- fiquinolin-3(3aH)-one
tert-butyl (1-(4-(3a-methyl-3-oxo-8-phenyl-3,3a,4,5-tetrahydro-2H-pyrazolo[3,4-f]quinolin- 7-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.023 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (9 mg, 60% yield). 1H-NMR (500 MHz, CD3OD) δ 8.15 (1H, s), 7.49 (2H, d), 7.44 (2H, d), 7.29-7.36 (3H, m), 7.20-7.26 (2H, m), 3.35-3.45 (1 H, m), 3.16-3.26 (1H, dd), 2.72-2.84 (2H, m), 2.54-2.66 (2H, m), 2. 9-2.39 (2H, m), 1.92-2.09 (2H, m), 1.43 (3H, s). LCMS (Method A) RT = 3.76 min, M+H+ = 423.16.
Example 9: 1 -(4-(3-(methylthio)-8-phenyl-4,5-dihydro-2H-pyrazolor3,4-flquinolin-7- yl)phenyl)cyclobutanami
Figure imgf000041_0001
Step 1 : tert-butyl (1-(4-(3-(methylthioV8-phenyl-4,5-dihvdro-2H-pyrazolo[3.4-f1quinolin-7- vnphenvOcvclobutvOcarbamate
To a solution of tert-butyl (1-(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (200 mg, 0.35 mmol) in anhydrous ethanol (1.7 mL) was added hydrazine monohydrate (25 μΙ_, 0.52 mmol). The reaction mixture was heated to 80°C under a nitrogen atmosphere for 2 hours. After cooling to room temperature the reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane:ethyl acetate, gradient elution from 90:10 to 80:20) to give the desired product as a yellow solid (53 mg, 28% yield). 1H-NMR (500 MHz, CDCI3) δ 7.94 (1H, s), 7.16-7.24 (4H, m), 7.11-7.16 (3H, m), 7.06-7.10 (2H, m), 3.16 (2H, t), 2.83 (2H, t), 2.35 (3H, s), 2.15-2.45 (4H, br m), 1.88-2.01 (1 H, m), 1.64-1.77 (1 H, m), 1.00-1.35 (9H, br m). LCMS (Method A) RT = 6.94 min, M+H+ = 539.15.
Step 2: 1 -(4-(3-(methylthio)-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4-f1quinolin-7- yl)phenyl)cvclobutanamine
tert-butyl (1-(4-(3-(methylthio)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (53 mg, 0.098 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as an off-white solid (24 mg, 37% yield). 1H-NMR (500 MHz, CD3OD) δ 8.14 (1 H, br s), 7.48 (2H, d), 7.43 (2H, d), 7.27-7.34 (3H, m), 7.21- 7.27 (2H, m), 3.26 (2H, t), 2.97 (2H, br t), 2.72-2.82 (2H, m), 2.53-2.63 (2H, m), 2.49 s), 2.19-2.30 (1 H, m), 1.91-2.02 (1 H, m). LCMS (Method A) RT = 4.06 min, M+H+ = 439.14.
Example 10: 1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4- flquinolin-7-yl)phenyl)cyclobutanamine
Figure imgf000042_0001
Step 1 : tert-butyl (1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4- f|quinolin-7-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(3-(methylthio)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-7-yl)phenyl)cyclobutyl)carbamate (158 mg, 0.29 mmol) in a mixture of THF (2 mL) and methanol (2 mL) was added a solution of oxone® (monopersulfate compound, 1.08 g, 1.76 mmol) in water (4 mL) dropwise. The reaction mixture was stirred at room temperature for 4 hours, then quenched by the dropwise addition of saturated NaHC03 solution (4 mL) and extracted into ethyl acetate (3 x 8 mL). The combined organic phases were washed with saturated NaHC03 solution (8 mL), brine (8mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (silica, hexane:ethyl acetate, gradient elution from 92:12 to 100:0) to give the desired product as a white solid (44mg, 26% yield). 1H-NMR (500 MHz, CDCI3) δ 7.85 (1 H, s), 7.27 (2H, d), 7.17-7.25 (5H, m), 7.11-7.16 (2H, m), 3.24 (2H, t), 3.14 (2H, t), 3.15 (3H, s), 2.18-2.51 (4H, br m), 1.96-2.10 (1 H, br m), 1.68-1.80 (1 H, br m), 1.09-1.38 (9H, br m). LCMS (Method A) RT = 6.59 min, M+H+ = 571.11.
Step 2: 1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihvdro-2H-pyrazolo[3,4-flquinolin-7- vOphenyQcvclobutanamine
tert-butyl (1-(4-(3-(methylsulfonyl)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (40 mg, 0.070 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as an off-white solid (18 mg, 37% yield). 1H-NMR (500 MHz, CD3OD) δ 8.10 (1H, s), 7.50 (2H, d), 7.44 (2H, d), 7.31-7.35 (3H, m), 7.24- 7.29 (2H, m), 3.32 (2H, t), 3.28 (3H, s), 3.24 (2H, t), 2.74-2.82 (2H, m), 2.54-2.64 (2H, m), 2.19-2.30 (1 H, m), 1.92-2.03 (1H, m). LCMS (Method A) RT = 3.81 min, M+H+ = 471.16.
Example 11 : 7-(4-(1 -aminocvclobutyl)phenyl)-N-methyl-8-phenyl-4.5-dihydro-2H- pyrazolof3.4-f1quinolin-3-amine
Figure imgf000043_0001
Step 1 : tert-butyl (1-(4-(3-(methylamino)-8-phenyl-4,5-dihvdro-2H-pyrazolof3.4-f1quinolin- 7-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.18 mmol) in ethanol (873 μΙ) was added methylamine (2M in THF, 87 μΙ, 0.18 mmol) and the mixture heated to 80°C under a nitrogen atmosphere for 2 hours. Further methylamine (2M in THF, 873 μΙ, 1.746 mmol) was added and the mixture heated to 80°C under a nitrogen atmosphere for 2 hours. Hydrazine monohydrate (25.4 μΙ, 0.52 mmol) was added and the mixture heated to 80°C for 18 hours. The reaction mixture was concentrated to dryness under reduced pressure and purified via Biotage chromatography (silica, DCM:MeOH, gradient elution from 100:0 to 90:10) to give a colourless solid (47 mg, 52% yield). 1H- NMR (500 MHz, CDCI3) δ 7.63 (1 H, s), 7.32 (2H, d), 7.27 (2H, d), 7.16-7.22 (3H, m), 7.09-7.16 (2H, m), 5.06 (1 H, br s), 3.10 (2H, t), 2.68 (3H, s), 2.22-2.60 (6H, br m), 1.97- 2.09 (1 H, m), 1.72-1.84 (1 H, m), 1.03-1.53 (9H, br m). LCMS (Method A) RT = 5.34 min, M+H+ = 522.19. Step 2: 7-(4-(1 -aminocvclobutyl)phenyl)-N-methyl-8-phenyl-4.5-dihvdro-2H-pyrazolor3,4- f quinolin-3-amine
tert-butyl (1-(4-(3-(rnethylamino)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (47 mg, 0.090 mmol) was dissolved in TFA (2 ml.) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as an off-white solid (31 mg, 53% yield). 1H-N R (500 MHz, CD3OD) δ 8.08 (1 H, s), 7.50 (2H, d), 7.44 (2H, d), 7.30-7.50 (3H, m), 7.23- 7.27 (2H, m), 3.30 (2H, t), 2.98 (3H, s), 2.87 (2H, t), 2.73-2.81 (2H, m), 2.54-2.64 (2H, m), 2.19-2.31 (1 H, m), 1.92-2.04 (1 H, m). LCMS (Method A) RT = 3.28 min, M+H+ = 422.18.
Example 12: 2-((7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4,5-dihydro-2H- Pyrazolor3,4-f|quinolin-3-yl)amino)ethanol
Figure imgf000044_0001
Step 1 : tert-butyl (1 -(4-(3-((2-hvdroxyethyl)amino)-8-phenyl-4,5-dihvdro-2H-pyrazolof3.4- f1quinolin-7-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl-5, 6,7,8- tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.18 mmol) in ethanol (873 μΙ) was added ethanolamine (10.6 μΙ, 0.18 mmol) and the mixture heated to 80 °C under a nitrogen atmosphere for 2 hours. Further ethanolamine (106 μΙ, 1.75 mmol) was added and the mixture heated to 80 °C under a nitrogen atmosphere for 18 hours.
Hydrazine monohydrate (25.4 μΙ, 0.52 mmol) was added and the mixture heated to 80 °C for 3 hours. The reaction mixture was concentrated to dryness under reduced pressure and purified via Biotage chromatography (silica, DCM:MeOH, gradient elution from 100:0 to 90:10) to give a colourless solid (51 mg, 53% yield). 1H-NMR (500 MHz, CDCI3) δ 7.67 (1 H, s), 7.18-7.28 (4H, br m), 7.10-7.16 (3H, m), 7.01-7.08 (2H, m), 5.1 (1 H, br s), 3.74 (2H, t), 3.35 (2H, t), 3.15 (2H, t), 2.66 (2H, t), 2.19-2.58 (4H, br m), 1.95-2.08 (1 H, m), 1.69-1.81 (1 H, m), 1.03-1.49 (9H, br m). LCMS (Method A) RT = 5.20 min, M+H+ = 552.24.
Step 2: 2-((7-(4-(1 -aminocvclobutyl)phenyl)-8-phenyl-4,5-dihvdro-2H-pyrazolo[3,4- f1quinolin-3-yl)amino)ethanol
tert-butyl (1-(4-(3-((2-hydroxyethyl)amino)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-7-yl)phenyl)cyclobutyl)carbamate (25 mg, 0.045 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as an off-white solid (22 mg, 71% yield). 1H-NMR (500 MHz, CD3OD) δ 8.06 (1 H, s), 7.50 (2H, d), 7.44 (2H, d), 7.29-7.34 (3H, m), 7.22-7.28 (2H, m), 3.78 (2H, t), 3.41 (2H, t), 3.30 (2H, t), 2.90 (2H, t), 2.71-2.82 (2H, m), 2.54-2.66 (2H, m), 2.19-2.30 (1 H, m), 1.91-2.03 (1 H, m). LCMS (Method A) RT = 3.30 min, M+H+ = 452.17.
Example 13: 1-(4-(3-morpholino-8-phenyl-4,5-dihvdro-2H-pyrazolor3,4-flquinolin-7- vDphenvDcyclobutanamine
Figure imgf000045_0001
Step 1 : tert-butyl (1 -(4-(3-morpholino-8-phenyl-4,5-dihvdro-2H-pyrazolof3,4-f]quinolin-7- vDphenyQcyclobutvDcarbamate
To a solution of tert-butyl (1-(4-(6-(bis(methylthio)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.18 mmol) in ethanol (873 μΙ) was added morpholine (76 μΙ, 0.87 mmol) and the mixture heated to 80 °C under a nitrogen atmosphere for 2 hours. Hydrazine monohydrate (12.7 μΙ, 0.26 mmol) was added and the mixture heated to 80 °C for 1 hour. Further hydrazine monohydrate (72 μΙ, 1.48 mmol) was added and the mixture heated to 80 "C for 18 hours. The reaction mixture was concentrated to dryness under reduced pressure and purified via Biotage chromatography (silica, cyclohexane:ethyl acetate, gradient elution from 92:2 to 100:0) to give a white solid (30 mg, 30% yield). 1H-NMR (500 MHz, CDCI3) δ 7.67 (1H, s), 7.31 (2H, d), 7.23-7.28 (2H, m), 7.18-7.24 (3H, m), 7.07-7.14 (2H, m), 4.95-5.44 (1 H, br m), 3.82 (4H, t), 3.24 (2H, t), 3.21 (4H, t), 2.91 (2H, t), 2.19-2.64 (4H, br m), 1.97-2.09 (1H, m), 1.73-1.84 (1 H, m), 1.10-1.46 (9H, br m). LCMS (Method A) RT = 6.18 min, M+H+ = 578.24.
Step 2: 1-(4-(3-morpholino-8-phenyl-4,5-dihydro-2H-pyrazolof3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
tert-butyl (1-(4-(3-morpholino-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (30 mg, 0.052 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as an off-white solid (23 mg, 63% yield). H-NMR (500 MHz, CD3OD) δ 8.08 (1 H, s), 7.49 (2H, d), 7.44 (2H, d), 7.29-7.33 (3H, m), 7.22- 7.27 (2H, m), 3.86 (4H, t), 3.26 (2H, t), 3.22 (4H, t), 2.98 (2H, t), 2.73-2.82 (2H, m), 2.54- 2.64 (2H, m), 2.19-2.30 (1 H, m), 1.92-2.03 (1 H, m). LCMS (Method A) RT = 3.74 min, M+H+ = 478.23.
Example 14: 7-(4-(1-aminocvclobutyl)phenyl)-2-methyl-8-phenyl-4,5-dihvdro-3H- imidazor4,5-flquinotin-3-o
Figure imgf000046_0001
Step 1 : tert-butyl (1-(4-(6-(hvdroxyimino)-5-oxo-3-phenyl-5,6,7,8-tetrahvdroquinolin-2- yQphenvOcyclobutvQcarbamate
To a solution of tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (1.00 g, 2.134 mmol) in anhydrous THF (30 mL) was added dropwise potassium tert-butoxide (1 in THF, 2.56 ml, 2.56 mmol) over a period of 10 min to give a deep red solution. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 30 min. To this was added dropwise isopentyl nitrite (0.286 ml, 2.134 mmol) over a period of 5 min to give a red solution. The reaction mixture was stirred at 50°C under a nitrogen atmosphere for 1 hour then allowed to cool to room temperature and concentrated to dryness under reduced pressure. The residue was redissolved in water (5 mL), acidified with 1 M HCI solution (5 mL) and extracted into ethyl acetate (3 x 10 mL). The combined organic phases were washed with water (20 mL), brine (20 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified via Biotage chromatography (silica, hexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a brown solid (436 mg, 41% yield)). 1H-N R (500 MHz, CDCI3) δ 8.40 (1 H, s), 7.40 (2H, d), 7.32 (2H, d), 7.25-7.29 (3H, m), 7.18-7.22 (2H, m), 5.04 (1 H, br s), 3.38 (2H, t), 3.29 (2H, t), 2.26-2.59 (4H, br m), 2.01-2.12 (1 H, m), 1.76-1.86 (1 H, m), 1.15-1.45 (9H, br m). LCMS (Method A) RT = 6.74 min, M+H+ = 498.13.
Step 2: tert-butyl (1-(4-(3-hvdroxy-2-methyl-8-phenyl-4,5-dihvdro-3H-imidazof4.5- f1quinolin-7-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(6-(hydroxyimino)-5-oxo-3-phenyl-5,6,7,8- tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (300 mg, 0.603 mmol) in ethanol (3.0 mL) was added acetaldehyde (68.1 μΙ, 1.206 mmol) followed by ammonium hydroxide (329 pi, 2.53 mmol). The reaction mixture was heated at 80°C under a nitrogen atmosphere for 2 hours, then allowed to cool to room temperature and concentrated to dryness under reduced pressure and dried to give the crude product as a brown solid (306 mg, 97% yield).
A 50 mg portion of this was further purified by Biotage chromatography (silica,
DCM:MeOH, gradient elution from 100:0 to 50:50) to give the desired product as a brown solid ( 0 mg). H-NMR (500 MHz, CDCI3) δ 7.70 (1 H, s), 7.15-7.24 (7H, m), 7.09-7.14 (2H, br m), 3.19 (2H, t), 2.91 (2H, t), 2.20-2.52 (4H, br m), 2.30 (3H, s), 1.93-2.06 (1 H, m), 1.69-1.79 (1 H, m), 1.06-1 .45 (9H, br m). LCMS (Method A) RT = 5.00 min, M+H+ = 523.19.
Step 3: 7-(4-(1 -aminocvclobutyl)phenyl)-2-methyl-8-phenyl-4,5-dihydro-3H-imidazof4.5- flquinolin-3-ol
tert-butyl (1-(4-(3-hydroxy-2-methyl-8-phenyl-4,5-dihydro-3H-imidazo[4,5-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (10 mg, 0.019 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as a pale brown solid (10 mg, 80% yield). 1H- NMR (500 MHz, CD3OD) δ 7.92 (1 H, s), 7.48 (2H, d), 7.43 (2H, d), 7.30-7.34 (3H, m), 7.22-7.26 (2H, m), 3.42 (2H, t), 3.18 (2H, t), 2.72-2.81 (2H, m), 2.63 (3H, s), 2.54-2.64 (2H, m), 2.18-2.29 (1 H, m), 1.91-2.02 (1 H, m). LCMS (Method A) RT = 3.14 min, M+H+ = 423.12.
Example 15: 1-(4-(2-methyl-8-phenyl-4,5-dihydro-3H-imidazor4lS-flquinolin-7- vDphenvDcyclobutanamine
Figure imgf000048_0001
Step 1 : tert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihvdro-3H-imidazof4,5-f1quinolin-7- vDphenvDcvclobutvDcarbamate
To a solution of tert-butyl (1-(4-(3-hydroxy-2-methyl-8-phenyl-4,5-dihydro-3H- imidazo[4,5-f]quinolin-7-yl)phenyl)cyclobutyl)carbamate (276 mg, 0.528 mmol) in anhydrous DMF (1.4 mL) was added triethyl phosphite (181 μΙ, 1.056 mmol) and heated to 100 °C under a nitrogen atmosphere overnight. The reaction mixture was allowed to cool to room temperature, diluted with 50:50 watenbrine (10 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were washed with 50:50 water.brine (2 x 10 mL), brine (10 mL) dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified via Biotage chromatography (silica,
DCM:MeOH, gradient elution from 100:0 to 90:10) to give the desired product as a pale brown solid (100 mg, 37% yield). 1H-NMR (500 MHz, CDCI3) δ 7.74 (1 H, s), 7.28 (2H, d), 7.23 (2H, d), 7.13-7.17 (3H, m), 7.06-7.11 (2H, m), 5.09 (1 H, br s), 3.25 (2H, t), 2.92 (2H, t), 2.16-2.59 (4H, br m), 2.41 (3H, s), 1.95-2.08 (1H, m), 1.70-1.82 (1 H, m), 1.05-1.47 (9H, br m). LCMS (Method A) RT = 4.66 min, M+H+ = 507.20. Step 2: 1-(4-(2-methyl-8-phenyl-4,5-dihvdro-3H-imidazof4,5-f1quinolin-7- vDphenvQcvclobutanamine
tert-butyl (1-(4-(2-methyl-8-phenyl-4,5-dihydro-3H-irnidazo[4,5-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (25 mg, 0.049 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried to give the desired product as an off-white solid (23 mg, 74% yield). 1H-NMR (500 MHz, CD3OD) δ 7.90 (1 H, s), 7.49 (2H, d), 7.44 (2H, d), 7.31 -7.35 (3H, m), 7.23- 7.27 (2H, m), 3.44 (2H, t), 3.20 (2H, t), 2.73-2.81 (2H, m), 2.72 (3H, s), 2.55-2.65 (2H, m), 2.19-2.30 (1H, m), 1.92-2.03 (1 H, m). LCMS (Method A) RT = 3.02 min, M+H+ = 407.14.
Example 16: 1 -(4-(9-phenyl-5,6-dihvdropyridor2.3-h1quinazolin-8-yl)phenyl)- cvclobutanamine
Figure imgf000049_0001
Step 1 : terf-butyl (1-(4-(9-phenyl-5.6-dihydropyridor2,3-hlauinazolin-8- yl)phenyl)cvclobutyl)carbamate
To a solution of ferf-butyl 1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydronaphthalen-2-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.09 mmol) in anhydrous ethanol (1000 μί) was added sodium ethoxide solution (21 % in ethanol, 150 μί, 0.38 mmol) and formamidine acetate (20 mg, 0.19 mmol). The reaction mixture was heated to 80°C under a nitrogen atmosphere for 2 hours. It was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 95:5 to 60:40) and slurried in diethyl ether (1 mL) to give the desired product as an off-white solid (12 mg, 26% yield). 1H NMR (500 MHz, CH3OD) 9.03 (s, 1 H), 8.66 (s, 1 H), 8.59 (s, 1 H), 7.31 (bs, 4H), 7.25 (dd, 3H), 7.21 (m, 2H), 3.23 (m, 2H), 3.16 (m, 2H), 2.31-2.45 (m, 4H), 1.92-1.97 (m, 1 H), 1.72- 1.74 (m, 1 H), 1.2-1.5 (br s, 9H). LCMS (Method A): RT = 7.64 min, M+H+ = 505.
Step 2: 1 -(4-(9-phenyl-5,6-dihvdropyridof2,3-hlquinazolin-8-yl)phenyl)cvclobutanamine Tert-butyl (1-(4-(9-phenyl-5,6-dihydropyrido[2,3-h]quinazolin-8- yl)phenyl)cyclobutyl)carbamate (12 mg, 0.023 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (8 mg, 70% yield). H NMR (500 MHz, CH3OD) 9.09 (s, 1 H), 8.74 (s, 1H), 8.67 (s, 1 H), 7.52 (d, 2H), 7.43 (d, 2H), 7.32 (m, 3H), 7.26 (m, 2H), 3.22 (m, 2H), 2.75 (m, 2H), 2.59 (m, 2H), 2.23 (m, 1 H), 1.98 (m, 1 H). LCMS (Method A): RT = 4.26 min, M+2H+ = 406.
Example 17: 8-(4-(9-phenyl- -dihvdropyridor2,3-h1quinazolin-2-amine
Figure imgf000050_0001
Step 1 : teff-putyl (1-(4-(2-amino-9-phenyl-5,6-dihvdropyridoi2,3-hlquinazolin-8- yl)phenyl)cyclobutyl)carbamate
To a solution of terf-butyl 1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-5,6,7,8- tetrahydronaphthalen-2-yl)phenyl)cyclobutyl)carbamate (45 mg, 0.085 mmol) in anhydrous ethanol (1500 pL) was added sodium ethoxide solution (21 % in ethanol, 130 μΐ, 0.34 mmol) and guanidine nitrate (22 mg, 0.17 mmol). The reaction mixture was heated to 80°C under a nitrogen atmosphere for one hour. It was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica,
dichloromethane: methanol, gradient elution from 99:1 to 90:10) and slurried in diethyl ether (1 mL) to give the desired product as an off-white solid (10 mg, 23% yield). 1H NMR (500 MHz, ds-DMSO) 8.36 (s, 1 H), 8.25 (s, 1 H), 7.31 (m, 5H), 7.25 (m 4H), 6.56 (bs, 1 H), 3.11 (t, 2H), 2.91 (t, 2H), 2.33 (br m, 4H), 1.93-2.04 (m, 1 H), 1.70- 1.82 (m, 1 H), 1.1- 1.41 (br s, 9H). LCMS (Method A): RT = 6.23 min, M+H+ = 520.
Step 2: 8-(4-(9-phenyl-5.6-dihvdropyridor2,3-h1quinazolin-2-amine
Tert-butyl (1-(4-(2-amino-9-phenyl-5,6-dihydropyrido[2,3-h]quinazolin-8- yl)phenyl)cyclobutyl)carbamate (10 mg, 0.019 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (2 mg, 20% yield). 1H NMR (500 MHz, d6-DMSO) 8.04 (br s, 2H), 8.31 (s, 1 H), 8.23 (s, 1H), 7.40 (d, 2H), 7.35 (d, 2H), 7.29 (m, 3H), 7.20 (m, 2H), 3.13 (t, 2H), 2.94 (t, 2H), 2.56 (m, 4H), 2.07-2.19 (br m, 1 H), 1.74-1.85 (br m, 1H). LCMS (Method A): RT = 3.58 min, M+H+ = 420.
Example 18: 2-(4-(1-aminocyclobutyl)phenyl)-7,7-dimethyl-3-phenyl-7,8- dihvdroquinolin-5(6H)-one
Figure imgf000051_0001
Step 1 : tert-butyl (1-(4-(7,7-dimethyl-5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(3-(dimethylamino)-2- phenylacryloyl)phenyl)cyclobutyl)carbamate (166 mg, 0.40 mmol) in acetic acid (1 mL) was added molecular sieves (10 mg) and 3-amino-5,5-dimethylcyclohex-2-enone (82 mg, 0.59 mmol) under nitrogen. The resulting mixture was heated at 100 °C for 2 h. After cooled down to room temperature, the mixture was partitioned between water (10 mL) and dichloromethane (10 mL). The layers were separated and the aqueous phase extracted into dichloromethane (2 x 10 mL). The combined organic phases were washed with saturated NaHC03 solution (20 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 20% EtOAc in hexane) to give the title compound (80 mg, 41%). 1H NMR (500 MHz, CDCI3): 8.27 (1 H, s), 7.37 (2H, d), 7.30 (2H,d), 7.25 -7.24 (3H, m), 7.19 -7.17 (2H, m), 5.06 (1 H, br s), 3.13 (2H, s), 2.59 (2H, s), 2.55-2.25 (4H, m), 2.09 -2.01 (1 H, m), 1.82 -1.77 (1 H, m), 1.44-1.15 (9H, br), 1.17 (6H, s).
Step 2: 2-(4-(1 -aminocvclobutyl)phenyl)-7,7-dimethyl-3-phenyl-7.8-dihvdroquinolin-5(6H)- one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1 -(4-(7,7-dimethyl-5-oxo-3-phenyl-5,6,7,8- tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate (30 mg, 0.06 mmol) was reacted to afford the title compound as a white solid (27 mg, 87%). LCMS (Method A): RT = 4.48 min, M-NH2 = 380. 1H NMR (500 MHz, MeOD): 8.26 (1H, s), 7.50 (2H, d), 7.43 (2H, d), 7.31-7.29 (3H, m), 7.21-7.19 (2H, m), 3.15 (2H, s), 2.78-2.72 (2H, m), 2.66 (2H, s), 2.60- 2.54 (2H, m), 2.27-2.18 (1 H, m), 2.00 -1.91 (1 H, m), 1.17 (6H, s).
Example 19: 1 -(4-(4.4-dimethyl-8-phenyl-4,5-dihvdro-2H-pyrazolo[3,4-f]quinolin-7- vDphenvDcyclobutanamine
Figure imgf000052_0001
Step 1 : tert-butyl (1-(4-(6-((dimethylamino)methylene)-7,7-dimethyl-5-oxo-3-phenyl- 5,6,7,8-tetrahydroquinolin-2-yl)phenyl)cvclobutyl)carbamate
Tert-butyl (1-(4-(7,7-dimethyl-5-oxo-3-phenyl-5,6,7,8-tetrahydroquinolin-2- yl)phenyl)cyclobutyl)carbamate (1g, 2 mmol) was dissolved in 10 ml of anhydrous N,N- dimethylfomamide dimethyl acetale. The resulting mixture was heated to 100°C under a nitrogen atmosphere for 48 hours. TLC and LC-MS analysis showed complete consumption of the starting material. The reaction mixture was concentrated to dryness under reduced pressure. The crude residue was slurried in n-hexane (10 ml) for 1 hour. The suspension was filtered and the solid was dried until constant weigh to give the desired product as a bright yellow solid (420 mg, 76% yield). LCMS (Method A): RT = 8.51 min, M+H+ = 552; RT = 8.81 min, M+H+ = 525 (rerf-butyl(1-(4-(6-
(hydroxymethylene)-717-dirnethyl-5-oxo-3-phenyl-5,6,718-tetrahydroquinolin-2- yl)cyclobutyl)carbamate.
Step 2: tert-butyl (1-(4-(4.4-dimethyl-8-phenyl-4,5-dihvdro-2H-pyrazolor3.4-flquinolin-7- yl)phenyl)cyclobutyl)carbamate
To a solution of reri-butyl (1-(4-(6-((dimethylamino)methylene)-7,7-dimethyl-5-oxo-3- phenyl-5,6>7,8-tetrahydroquinolin-2-yl)phenyl)cyclobutyl)carbamate
(180 mg, 0.33 mmol) in anhydrous ethanol (4 mL) was added hydrazine hydrate (48 μί, 1 mmol). The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 99:1 to 60:40) and PREP HPLC (Method A) to give the desired product as a white solid (8 mg, 5% yield). 1H NMR (500 MHz, CDCI3) 8.12 (s, 1 H), 7.27- 7.48 (br m, 5H), 7.25-7.18 (br m, 5H), 5.03 (br s, 1 H), 3.16 (s, 2H), 2.23-2.68 (br m, 4H), 1.97-2.12 (m, 1 H), 1.75-1.88 (m, 1H), 1.11-1.57 (br s, 9H + 6H). LCMS (Method A): RT = 6.62 min, M+H+ = 521.
Step 3: 1-(4-(4,4-dimethyl-8-phenyl-4,5-dihydro-2H-pyrazolor3,4-f|quinolin-7- vDphenvDcvclobutanamine
tert-butyl (1-(4-(4,4-dimethyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutyl)carbamate (8 mg, 0.015 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (1 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (4 mg, 50% yield). 1H NMR (500 MHz, CH3OD) 8.18 (s, 1H), 7.63 (s, 1 H), 7.47 (d, 2H), 7.41 (d, 2H), 7.19-7.33 (br m, 5H), 3.08 (s, 2H), 2.68-2.79 (br m, 2H), 2.51-2.64 (br m, 2H), 2.15-2.30 (br m, 1H), 1.87-2.01 (br m, 1 H), 1.35 (s, 6H). LCMS (Method A): RT = 3.03 min, M+H+ = 421. Example 20: 2-(4-(1 -aminocvclobutyl)phenyl)-3-prtenyl-6.7.8.9-tetrahydro-5H- cvcloheptafblPyridin-5-one
Figure imgf000054_0001
Step 1 : tert-butyl (1-(4-(5-oxo-3-phenyl-6,7.8,9-tetrahvdro-5H-cvcloheptarblpyridin-2- yOphenvOcyclobutvQcarbamate
To a solution of tert-butyl (1-(4-(3-(dimethylamino)-2- phenylacryloyl)phenyl)cyclobutyl)carbamate (1 19 mg, 0.28 mmol) in acetic acid (1 mL) was added molecular sieves (10 mg) and cycloheptane-1 ,3-dione (36 μΙ_, 0.42 mmol) under nitrogen. The resulting mixture was heated at 100 °C for 2 h. After cooled down to room temperature, the mixture was partitioned between water (10 mL) and
dichloromethane (10 mL). The layers were separated and the aqueous phase extracted into dichloromethane (2 x 10 mL). The combined organic phases were washed with saturated NaHC03 solution (20 mL), brine (20 mL), dried over Na2S04l filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 20% EtOAc in hexane) to give the title compound (32 mg, 24%). H NMR (500 MHz, CDCI3): 8.09 (1 H, s), 7.38 (2H, d), 7.30 (2H,d), 7.25 -7.24 (3H, m), 7.19 -7.17 (2H, m), 5.04 (1 H, br s), 3.29 (2H,t), 2.87 (2H, t), 2.60 -2.25 (4H, m), 2.07 -2.01 (3H, m), 1.99-1.94 (2H, m), 1.84 -1.75 (1 H, m), 1.42-1.15 (9H, br).
Step 2: 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-6.7,8,9-tetrahvdro-5H- cvclohepta[b]pyridin-5-one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(5-oxo-3-phenyl-6,7,8,9-tetrahydro-5H- cyclohepta[b]pyridin-2-yl)phenyl)cyclobutyl)carbamate (14 mg, 0.03 mmol) was reacted to afford the title compound as a white solid (14 mg, 100%). LCMS (Method A): RT = 4.26 min, M-NH2 = 366. Ή NMR (500 MHz, MeOD): 8.06 (1 H, s), 7.50 (2H, d), 7.42 (2H, d), 7.29-7.28 (3H, m), 7.20-7.18 (2H, m), 3.30 (2H, t), 2.89 (2H, t), 2.78-2.72 (2H, m), 2.59 - 2.54 (2H, m), 2.25-2.19 (1 H, m), 2.06-2.01 (2H, m), 1.98 -1.92 (3H, m). Example 21 : 1-(4-(9-phenyl-2,4,5.6-tetrahvdropyrazolor3'.4':3,4lcvcloheptari.2- b1pyridin-8-yl)phenyl)cvclobutanamine
Figure imgf000055_0001
Step 1 : tert-butyl (1-(4-(6-((dimethylamino)rnethylene)-5-oxo-3-phenyl-6,7,8,9-tetrahvdro- 5H-cvcloheptafblpyridin-2-yl)phenyl)cyclobutyl)carbamate
Tert-butyl (1-(4-(5-oxo-3-phenyl-6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridine-2- yl)phenyl)cyclobutyl)carbamate (270 mg, 0.56 mmol) was dissolved in 3 ml of anhydrous Ν,Ν-dimethylfomamide dimethyl acetale. The resulting mixture was heated to 100°C under a nitrogen atmosphere for 18 hours. TLC and LC-MS analysis showed complete consumption of the starting material. The reaction mixture was concentrated to dryness under reduced pressure. The crude residue was slurried in n-hexane (5 ml) for 1 hour. The suspension was filtered and the solid was dried until constant weigh to give the desired product as a bright yellow solid (190 mg, 63% yield). LCMS (Method A): RT = 6.55 min, M+H+ = 538; RT = 8.43 min, M+H+ = 511 (rert-butyl(1-(4-(6- (hydroxymethylene)-5-oxo-3-phenyl-6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-2- yl)cyclobutyl)carbamate.
Step 2: tert-butyl f1-(4-f9-phenyl-2,4.5,6-tetrahvdropyrazolof3',4':3.4lcvcloheptaf1.2- b1pyridin-8-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(6-((dimethylamino)methylene)-5-oxo-3-phenyl-6,7,8,9- tetrahydro-5H-cyclohepta[b]pyridin-2-yl)phenyl)cyclobutyl)carbamate (90 mg, 0.17 mmol) in anhydrous ethanol (2 mL) was added hydrazine hydrate (25 μΙ_, 0.5 mmol). The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. The reaction mixture was concentrated to dryness under reduced pressure, purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 99:1 to 60:40) and preparative HPLC (Method G, gradient 5 to 95% 0.1 %FA/ACN in 0.1 %FA/H2O, 20 minutes run) to give the desired product as a white solid (50 mg, 58% yield). 1H NMR (500 MHz, CDCI3) 7.49 (s, 1 H), 7.38 (d, 2H), 7.29 (d, 2H), 7.21-7.25 (br m, 5H + 1 H), 3.25 (m, 2H), 2.94 (t, 2H), 2.53-2.61 (br m, 1 H), 2.42-2.52 (m, 2H), 2.31- 2.41 (br m, 1 H), 2.14-2.21 (m, 2H), 1.98-2.10 (m, 1H), 1.74-1.85 (m, 1H), 1.30-1.43 (br s, 9H). LCMS (Method A): RT = 6.09 min, M+H+ = 507.
Step 3: 1 -(4-(9-phenyl-2.4.5.6-tetrahvdropyrazolof3'.4':3.41cvcloheptaf 1 ,2-blPyridin-8- vQphenyQcvclobutanamine
Tert-butyl (1-(4-(9-phenyl-2,4,5,6-tetrahydropyra2olo[3',4':3,4]cyclohepta[1 ,2-b]pyridin-8- yl)phenyl)cyclobutyl)carbamate (10 mg, 0.019 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (1 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (2 mg, 20% yield). 1H NMR (500 MHz, CH3OD) 8.69 (s, 1 H), 7.66 (s, 1 H), 7.56 (d, 2H), 7.2 (d, 2H), 7.34 (m, 3H), 7.29 (m, 2H), 3.47-3.53 (m, 1 H), 3.03 (m, 2H), 2.74-2.82 (br m, 2H), 2.56-2.67 (br m, 2H), 2.19- 2.34 (br m, 3H), 1.91- 2.03 (m, 1 H). LCMS (Method A): RT = 3.03 min, M+H+ = 407.
Example 22: 2-(4-(1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihvdro-1 ,6- naphthyridin-5(6H)-one
Figure imgf000056_0001
Step 1 : tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahvdro-1.6-naphthyridin-2- vDphenvDcvclobutvDcarbamate
To a solution of tert-butyl (1-(4-(3-(dimethylamino)-2- phenylacryloyl)phenyl)cyclobutyl)carbamate (952 mg, 2.27 mmol) in acetic acid (10 mL) was added molecular sieves (100 mg), ammonium acetate (524 mg, 6.80 mmol) and piperidine-2,4-dione (385 mg, 3.40 mmol) under nitrogen. The resulting mixture was heated at 100 °C for 2 h. After cooled down to room temperature, the mixture was partitioned between water (100 mL) and dichloromethane (100 mL). The layers were separated and the aqueous phase extracted into dichloromethane (2 x 50 mL). The combined organic phases were washed with saturated NaHC03 solution (100 ml_), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 100% EtOAc in hexane) to give the title compound as a yellow solid (353 mg, 33%). 1H NMR (500 MHz, MeOD): 8.28 (1 H, s), 7.38-7.34 (4H, m), 7.32-7.29 (3H, m), 7.23-7.21 (2H, m), 3.69 (2H, t), 3.25 (2H, t), 2.51-2.40 (4H, m), 2.13-2.05 (1 H, m), 1.91-1.84 (1 H, m), 1.44-1.18 (9H, br).
Step 2: 2-(4-( 1 -aminocvclobutyl)phenyl)-3-phenyl-7,8-dihydro-1 ,6-naphthyridin-5(6H)- one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydro-1 ,6- naphthyridin-2-yl)phenyl)cyclobutyl)carbamate (10 mg, 0.021 mmol) was reacted to afford the title compound (7 mg, 69%). LCMS (Method A): RT = 3.62 min, Μ+Ι-Γ = 370. H NMR (500 MHz, MeOD) 8.30 (1 H, s), 7.51 (2H, d), 7.45 (2H, d), 7.31 (3H, m), 7.22 (2H, m), 3.70 (2H, t), 3.27(2H, t), 2.85-2.70 (2H, m), 2.65-2.50 (2H, m), 2.30-2.20 (1 H, m), 2.05-1.90 (1 H,m).
Example 23: 2-(4-(1 -aminocvclobutyl)phenyl)-6-methyl-3-phenyl-7,8-dihvdro-1 ,6- naphthyridin-5(6H)-one
Figure imgf000057_0001
Step 1 : tert-butyl (1-(4-(6-methyl-5-oxo-3-phenyl-5,6.7.8-tetrahvdro-1 ,6-naphthyridin-2- vQphenvOcvclobutvOcarbamate
To a solution of tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (30 mg, 0.06 mmol) in dry DMF (1 mL) was added sodium hydride (3 mg, 0.07 mmol) at 0 °C under nitrogen. After 1 h at 0 °C, iodomethane (17 μΙ_, 0.19 mmol) was added and the resulting mixture was stirred for an additional 10 min at 0 °C. A saturated solution of NH4CI was added and the mixture was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 50% EtOAc in hexane) to give the title compound (22 mg, 71 %). 1H NMR (500 MHz,CDCI3): 8.35 (1 H, s), 7.34 (2H, d), 7.29 (2H,d), 7.24-7.22 (3H, m), 7.19-7.17 (2H, m), 5.06 (1 H, br s), 3.70 (2H, t), 3.29 (2H, t), 3.19 (3H, s), 2.54-2.30 (4H, m), 2.08-2.00 (1 H, m), 1.83-1.74 ( H, m), 1.44-1.18 (9H, br).
Step 2: 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-7,8-dihvdro-1 ,6-naphthyridin-5(6H)- one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(6-methyl-5-oxo-3-phenyl-5,6,7,8-tetrahydro- 1 ,6-naphthyridin-2-yl)phenyl)cyclobutyl)carbamate (29 mg, 0.06 mmol) was reacted to afford the title compound (32 mg, 87%). LCMS (Method A): RT = 3.76 min, M-NH2 = 367. 1H NMR (500 MHz, MeOD): 8.29 (1 H,s), 7.49 (2H, d), 7.42 (2H, d), 7.30-7.29 (3H, m), 7.21 -7.19 (m, 2H), 3.80 (2H, t), 3.30 (2H, t), 3.20 (3H, s), 2.78 -2.72 (m, 2H), 2.60-2.54 (m, 2H), 2.27-2.18 (m, 1 H), 2.00-1.91 (m, 1 H).
Example 24: 2-(2-(4-(1-aminocvclobutyl)phenyl)-5-oxo-3-phenyl-7,8-dihvdro-1,6-
Figure imgf000058_0001
Step 1 : tert-butyl (1-(4-(6-(cvanomethyl)-5-oxo-3-phenyl-5.6.7.8-tetrahvdro-1.6- naphthyridin-2-yl)phenyl)cvclobutyl)carbarnate
To a solution of tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (31 mg, 0.07 mmol) in dry DMF (1 mL) was added sodium hydride (3 mg, 0.07 mmol) at 0 °C under nitrogen. After 1 h at 0 °C, 2- bromoacetonitrile (14 μί., 0.20 mmol) was added and the resulting mixture was stirred for an additional 30 min at 0 °C. A saturated solution of NH4CI was added and the mixture was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 40% EtOAc in hexane) to give the title compound (10 mg, 29%). Ή NMR (500 MHz,CDCI3): 8.36 (1 H, s), 7.35 (2H, d), 7.30 (2H,d), 7.27-7.26 (3H, m), 7:20-7.18 (2H, m), 5.05 (1 H, br s), 4.59 (2H, s), 3.86 (2H, t), 3.40 (2H, t), 2.55-2.25 (4H, m), 2.10-2.02 (1 H, m), 1.85-1.76 (1 H, m), 1.44-1.18 (9H, br).
Step 2: 2-(2-(4-(1 -aminocvclobutvnphenyl)-5-oxo-3-phenyl-7,8-dihydro-1 ,6-naphthyridin- 6(5H)-yl)acetonitrile
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1 -(4-(6-(cyanomethyl)-5-oxo-3-phenyl-5, 6,7,8- tetrahydro-1 ,6-naphthyridin-2-yl)phenyl)cyclobutyl)carbamate (17 mg, 0.03 mmol) was reacted to afford the title compound (15 mg, 71%). LCMS (Method A): RT = 3.80 min, M- NH2 = 392. Ή NMR (500 MHz, MeOD): 8.33 (1 H,s), 7.50 (2H, d), 7.42 (2H, d), 7.31 - 7.29 (3H, m), 7.22-7.20 (m, 2H), 4.66 (2H, s), 3.92 (2H, t), 3.38 (2H, t), 2.78 -2.72 (m, 2H), 2.60-2.54 (m, 2H), 2.27-2.18 (m, 1 H), 2.00-1.91 (m, 1 H).
Example 25: 2-(4-(1-aminocvclobutyl)phenyl)-6-(2-dimethylamino)ethyl)-3-phenyl-
7.8-dihvdro-1.6-naphthyridin-5(6H)-one
Figure imgf000059_0001
Step 1 : tert-butyl (1-(4-(6-(2-(dimethylamino)ethyl)-5-oxo-3-phenyl-5.6.7.8-tetrahvdro-1.6- naphthyridin-2-yl)phenyl)cvclobutyl)carbamate
To a solution of terf-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (25 mg, 0.05 mmol) in dry DMF (1 mL) was added sodium hydride (5 mg, 0.106 mmol) at 0 °C under nitrogen. After 1h at 0 °C, a solution of 2-chloro-N,N-dimethylethanamine hydrochloride (9 mg, 0.064 mmol) and potassium carbonate (9 mg, 0.064) in 1 ml_ of DMF was added and the resulting mixture was stirred for one hour at 40°C. The reaction mixture was cooled to0°C and a saturated solution of NH CI was added while stirring, keeping the temperature below 10°C. The resulting mixture was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by PREP HPLC (method G) to give the desired product as a white solid (5 mg, 17% yield). Ή NMR (500 MHz,CDCI3): 8.35 (1 H, s), 7.34 (2H, d), 7.29 (2H,d), 7.23-7.25 (3H, m), 7.17-7.20 (2H, m), 5.01 (1H, s), 3.77 (4H, m), 3.28 (2H, t), 2.63 (2H, t), 2.47 (2H + 1 H, m), 2.35 (6H, s), 2.24-2.32 (2H, m), 1.98-2.12 (1H, m), 1.67-1.89 (9H, br s).
Step 2: 2-(4-(1-aminocvclobutyl)phenyl)-6-(2-dimethylamino)ethyl)-3-phenyl-7,8-di ,6-naphthyridin-5(6H)-one
Tert-butyl (1-(4-(6-(2-(dimethylamino)ethyl)-5-oxo-3-phenyl-5,6,7,8-tetrahydro-1 ,6- naphthyridin-2-yl)phenyl)cyclobutyl)carbamate (5 mg, 0.01 mmol) dissolved in TFA (0.5 ml.) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (1 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (1.66 mg, 35% yield). 1H NMR (500 MHz, CH3OD) 8.22 (s, 1 H), 7.50 (d, 2H), 7.39 (d, 2H), 7.33 (d, 2H), 7.18-7.22 (m, 3H), 7.08-7.13 (m, 2H), 3.90 (t, 2H), 3.76 (t, 2H), 3.39 (t, 2H), 3.24 (t, 2H), 2.94 (s, 6H), 2.60-2.70 (br m, 2H), 2.43-2.52 (br m, 2H), 2.09-2.19 (br m, 1 H), 1.80- 1.95 (br m, 1 H). LCMS (Method A): RT = 3.03 min, M+H+ = 441.
Example 26: 8-(4-(1 -aminocvclobutyl)phenyl)-9-phenyl-5,6-dihvdro- Γ1.2.41triazolor3,4-fin .61naphthyridin-3(2H)-one
Figure imgf000060_0001
Stepl : tert-butyl (1-(4-(3-phenyl-5-thioxo-5,6,7,8-tetrahvdro-1.6-naphthyridin-2- vQphenvOcyclobutvOcarbamate
To a solution of tert-butyl (1-(4-(5-oxo-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (223 mg, 0.47 mmol) in toluene (15 mL) was added lawesson's reagent (192 mg, 0.47 mmol) under nitrogen. The resulting mixture was heated under reflux for 2 h. After cooled down to room temperature, the mixture was concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 1% MeOH in dichloromethane) to give the title compound as a yellow solid (231 mg, quantitative). Ή NMR (500 MHz, MeOD): 8.76 (1 H, s), 7.37 -7.33 (4H, m), 7.31-7.28 (3H, m), 7.22-7.20 (2H, m), 3.69 (2H, t), 3.25 (2H, t), 2.49 -2.39 (4H, m), 2.15-2.05 (1 H, m), 1.91-1.80 (1 H, m), 1.45-1.15 (9H, br).
Step 2: tert-butyl (1 -(4-(3-oxo-9-phenyl-2.3.5.6-tetrahvdro-ri.2.41triazolor3.4- firi,61naphthyridin-8-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(3-phenyl-5-thioxo-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (55 mg, 0.1 13 mmol) in THF (7 mL) was added ethyl carbozate (23.6 mg, 0.226 mmol) and mercury acetate (54.1 mg, 0.17 mmol) under nitrogen. The resulting mixture was stirred at r.t. for 2 h. The mixture was concentrated to dryness under reduced pressure and partitioned between DCM (15 ml) and water ( 5 ml). The organic phase was separated and concentrated to dryness under reduced pressure. The resulting residue was suspended in toluene (7 ml). The resulting mixture was heated under reflux for 3 h. After cooled down to room temperature, the mixture was concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 4% MeOH in dichloromethane) to give tert-butyl (1 -(4-(3-oxo-9-phenyl-2,3,5,6-tetrahydro-[1 ,2,4]triazolo[3,4-f][1 ,6]naphthyridin-8- yl)phenyl)cyclobutyl)carbamate (32 mg,56%). LCMS (Method A): RT = 6.29 min, M+1 = 510. Ή NMR (500 MHz, CDCI3): 9.83 (1 H,s), 8.10 (1 H, s), 7.29 (2H, d), 7.22 (2H, d), 7.19-7.18 (3H, m), 7.13-7.11 (2H, m), 5.1 (1 H, br), 4.01 (2H, t), 3.36 (2H, t), 2.43-2.28 (4H, m), 2.02-1.97 (1 H, m), 1.76-1.70 (1 H, m), 1.30-1.14 (9H, br).
Step 3: 8-(4-(1-aminocvclobutyl)phenyl)-9-phenyl-5,6-dihvdro-n.2.41triazolor3.4- flM .61naphthyridin-3(2H)-one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl ( -(4-(3-oxo-9-phenyl-2, 3, 5, 6-tetrahydro- [1 ,2,4]triazolo[3,4-f][1 ,6]naphthyridin-8-yl)phenyl)cyclobutyl)carbamate (10 mg, 0.019 mmol) was reacted to afford the title compound (6 mg, 58%). LCMS (Method A): RT = 3.51 min, M+2 = 41 1.2. 1H NMR (500 MHz, MeOD): 8.23 (1 H,s), 7.52 (2H, d), 7.43 (2H, d), 7.34-7.32 (3H, m), 7.26 -7.24 (2H, m), 4.09 ( 2H, t), 3.46 (2H, t), 2.81 -2.75 (2H, m), 2.62-2.56 (2H, m), 2.28-2.22 (1 H, m), 2.01-1.95 (1 H, m). Example 27: 1-(4-(6-ethyl-3-phenyl-5,6.7.8-tetrahvdro-1.6-naphthyridin-2- vDphenvDcvclobutanamin
Figure imgf000062_0001
Step 1 : tert-butyl (1-(4-(6-ethyl-3-phenyl-5.6,7,8-tetrahvdro-1.6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(3-phenyl-5-thioxo-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (41 mg, 0.08 mmol) in dry ethanol (1 mL) was added Raney Nickel (1 mL) under nitrogen. The resulting mixture was heated under reflux for 4 h. After cooled down to room temperature, the mixture was filtered on celite (rinse few times with ethanol) and concentrated to dryness under reduced pressure to give the title compound (28 mg, 74%). Ή NMR (500 Hz,CDCI3): 7.29 (1 H, s), 7.22 -7. 8 (4H, m), 7.15-7.14 (3H, m), 7.07 -7.06 (2H, m), 4.98 (1H, br s), 3.64 (2H,s), 3.09 (2H, t), 2.83 (2H, t), 2.58 (2H, q), 2.50-2.20 (4H, m), 2.00 -1.91 (1 H, m), 1.74 -1.66 (1 H, m), 1.40-1.10 (9H, br), 1.15 (3H, t).
Step 2: 1-(4-(6-ethyl-3-phenyl-5,6,7,8-tetrahvdro-1 ,6-naphthyridin-2- vQphenvDcvclobutanamine
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1 -(4-(6-ethyl-3-phenyl-5,6,7,8-tetrahydro-1 ,6- naphthyridin-2-yl)phenyl)cyclobutyl)carbamate (14 mg, 0.03 mmol) was reacted to afford the title compound (18 mg, quantitative). LCMS (Method A): RT = 0.44 min, M-NH2 = 367. 1H NMR (500 MHz.MeOD) 7.76 (1 H, s), 7.45 (2H, d), 7.42 (2H, d), 7.30-7.29 (3H, m), 7.19-7.18 (2H, m), 3.47 (2H, q), 3.46 (2H, t), 3.39 (2H, br s), 3.32 (2H, m), 2.77 -2.71 (2H, m), 2.60-2.54 (2H, m), 2.27-2.18 (1 H, m), 1.99-1.90 (1 H, m), 1.50 (3H, t). Example 28: 1-(4-(9-phenyl-5,6-dihvdro-ri,2,41triazolor3,4-firi .61naphthyridin-8- vQphenyDcvclobutanamine
Figure imgf000063_0001
Step 1 : tert-butyl (1-(4-(5-hvdrazono-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(3-phenyl-5-thioxo-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (127 mg, 0.26 mmol) in THF (2 ml_) was added hydrazine. H20 (63 μΙ_, 1.31 mmol) and mercuric acetate (125 mg, 0.39 mmol) to give a black solution under nitrogen. The resulting mixture was stirred at room temperature for 2 h. After cooled down to room temperature, the mixture was filtered on celite (rinse with dichloromethane) and concentrated to dryness under reduced pressure. The resulting black yellow oil was used directly in the following step without any further purification. LCMS (Method A): RT = 4.77 min, M+H+ = 484.
Step 2: tert-butyl (1 -(4-(9-phenyl-5.6-dihvdro-f1 ,2.41triazolor3.4-fin ,6lnaphthyridin-8- yl)phenyl)cvclobutyl)carbamate
A solution of tert-butyl (1-(4-(5-hydrazono-3-phenyl-5,6,7,8-tetrahydro-1 ,6-naphthyridin-2- yl)phenyl)cyclobutyl)carbamate (40 mg, 0.08 mmol) in triethyl orhtoformate (1 mL) was heated at 150 °C for 1.5h under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by preparative HPLC (method G, gradient 5 to 95% 0.1 %FA/ACN in 0.1 %FA/H2O) to give the title compound (9 mg, 22%). Ή NMR (500 MHz, CDCI3): 8.47 (1 H, s), 8.29 (1 H, s), 7.37 (2H, d), 7.31 (2H, d), 7.28-7.26 (3H, m), 7.23-7.21 (2H, m), 5.07 (1 H, br s), 4.42 (2H, t), 3.50 (2H, t), 2.55-2.25 (4H, m), 2.10-2.02 (1 H, m), 1.85-1.76 (1 H, m), 1.45-1.20 (9H, br).
Step 3: 1-(4-(9-phenyl-5.6-dihvdro-ri .2.41triazolor3,4-flM .61naphthyridin-8- yDphenvDcvclobutanamine
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4- f][1 ,6]naphthyridin-8-yl)phenyl)cyclobutyl)carbamate (9 mg, 0.02 mmol) was reacted to afford the title compound (10 mg, quantitative). LCMS (Method A): RT = 3.53 min, M-NH2 = 377. H NMR (500 MHz, MeOD) 8.70 (1 H, s), 8.41 (1H, s), 7.52 (2H, d), 7.43 (2H, d), 7.33-7.31 (3H, m), 7.29-7.27 (2H, m), 4.57 (2H, t), 3.51 (2H, t), 2.79-2.73 (2H, m), 2.60 - 2.54 (2H, m), 2.27-2.20 (1H, m), 1.99-1.92 (1 H, m).
Example 29: 1-(4-(3-(1-methyl-1H-imidazol-4-yl)-9-phenyl-5.6-dihydro- ri.2,41triazolof3.4-firi.61naphthyridin-8-yl)phenvhcvclobutanamine
Figure imgf000064_0001
Step 1 : teit-butyl (1-(4-(3-(1-methyl-1 H-imidazol-4-vn-9-phenyl-5,6-dihvdro- f1 ,2,4ltriazolor3,4-f1[1 ,6lnaphthyridin-8-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(5-hydrazono-3-phenyl-5,6,7,8-tetrahydro-1,6- naphthyridin-2-yl)phenyl)cyclobutyl)carbamate (22mg, 0.04 mmol) in dry DMF (1 ml_) was added EDCI (11 mg, 0.06 mmol), HOBt.H20(8 mg, 0.06 mmol) and 1-methyl-1H- imidazole-4-carboxylic acid (8 mg, 0.06 mmol) under nitrogen. The reaction mixture was heated at 60 °C overnight. After cooled down to room temperature, the mixture was partitioned between saturated NaHC03 solution and ethyl acetate. The layers were separated and the organic phase washed with water and brine, dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by preparative HPLC (method G, gradient 5 to 95% 0.1%FA/ACN in
0.1%FA/H2O) to give the title compound (1 mg, 5%). Ή NMR (500 MHz, CDCI3): 8.49 (1 H, s), 7.74 (1 H, s), 7.54 (1H, s), 7.38 (2H, d), 7.31 (2H, d), 7.28-7.23 (5H, m), 5.01 (1 H, br s), 4.94 (2H, t), 3.81 (3H, s), 3.49 (2H, t), 2.60-2.30 (4H, m), 2.10-2.01 (1 H, m), 1.85- 1.76 (1 H, m), 1.45-1.20 (9H, br).
Step 2: 1 -(4-f 3-( 1 -methyl-1 H-imidazol-4-yl)-9-phenyl-5.6-dihvdro-M ,2,41triazolo[3.4- firi ,61naphthyridin-8-yl)phenyl)cyclobutanamine
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(3-(1-methyl-1 H-imidazol-4-yl)-9-phenyl-5,6- dihydro-[1 ,2,4]triazolo[3,4-f][1 ,6]naphthyridin-8-yl)phenyl)cyclobutyl)carbamate (9 mg, 0.02 mmol) was reacted to afford the title compound (10 mg, quantitative). LCMS (Method A): RT = 3.53 min, M-NH2 = 377. Ή NMR (500 MHz, MeOD): 8.43 (1 H, s), 7.99 (1 H, br s), 7.91 (1 H, br s), 7.52 (2H, d), 7.44 (2H, d), 7.33-7.27 (5H, m), 3.89 (3H, s), 3.54 (2H, t), 3.32 (2H, t), 2.79-2.73 (2H, m), 2.61-2.55 (2H, m), 2.27-2.20 (1 H, m), 2.00-1.92 (1 H, m).
Example 30: 6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1H-pyridor2,3-b1f1,4loxazin 2(3H)-one
Figure imgf000065_0001
Step 1 : tert-butyl (1-(4-(5-cyano-6-hydroxy-3-phenylpyridin-2- Yl)phenyl)cyclobutyl)carbamate
To a suspension of sodium hydride (3.58g, 0.089 mol) in DMF (150 ml) at 0°C was added a solution of 2-cyanoacetamide (3.48 g, 0.041mol) and (E)-tert-butyl (1-(4-(3- (dimethylamino)-2-phenylacryloyl)phenyl)cyclobutyl)carbamate (15 g, 0.0357 mol) in methanol (5.5 ml) and DMF (150 ml). The resulting mixture was heated at 40 °C for 16 h. After cooled down to room temperature, the mixture was added to a solution of acetic (50 ml) in water (300 ml) slowly. The resulting precipitate was filtered and the filter cake was partitioned between water (250 ml) and dichloromethane ( 50 ml). The layers were separated and the aqueous phase extracted into dichloromethane (150 ml). The combined organic phases were washed with water (200 ml) and brine (150 ml), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give the title compound as a yellow solid (14.6 g, 93%). LCMS (Method A): RT = 6.59 min, M+1 = 442. 1H NMR (500 MHz, CDCI3): 7.89 (1 H, s), 7.37-7.32 (2H, m), 7.22-7.16 (5H, m), 7.03-6.98 (2H, m), 2.50-2.37 (4H, m), 2.06-1.97 (1 H, m), 1.82-1.77 (1 H, m), 1.45-1.05 (9H, br).
Step 2: tert-butyl (1-(4-(5-carbamoyl-6-hydroxy-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate
To a suspension of tert-butyl (1-(4-(5-cyano-6-hydroxy-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (14.6g, 3.58g, 0.033 mol) in ethanol (112 ml) and water (112 ml) at room temperature was added sodium hydroxide solution (6M, 54.4 ml) and hydrogenperoxide (30%, 21.9 ml). The resulting mixture was heated at 50 °C for 16 h. After cooled down to 0 °C, the mixture was added acetic acid to adjust pH==4. The resulting precipitate was filtered and the filter cake was dried to give the title compound as white solid (10.8g, 72%). LCMS (Method A): RT = 5.88 min, M+1 = 460. Ή NMR (500 MHz, DMSO-d6): 9.65 (1H, br), 8.10 (1 H, s), 7.25-7.05 (7H, m), 6.98 (2H, m), 2.45-2.25 (4H, m), 2.0-1.95 (1 H, m), 1.79-1.75 (1 H, m), 1.3-1.0 (9H, br).
Step 3: tert-butyl (1-(4-(5-amino-6-hvdroxy-3-phenylpyridin-2- vDphenvQcvclobutvDcarbamate
To a solution of bromine (5.99g, 0.037 mol) in water (350 ml) was added a solution of sodium hydroxide (15.02g, 0.37 mol) in water (350 ml). The resulting solution was stirred at 0 °C for 10 min. To this solution was added tert-butyl (1-(4-(5-carbamoyl-6-hydroxy-3- phenylpyridin-2-yl)phenyl)cyclobutyl)carbamate (10.8g, 0.0235 mol). The resulting mixture was heated at 65 °C for 24 h. After cooled down to room temperature, the mixture was extracted with ethyl acetate (200 ml). The layers were separated and the aqueous phase extracted into ethyl acetate (2 x 200 ml). The combined organic phases were washed with water (400 ml) and brine (300ml), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 5% methanol in DCM) to give the title compound (1.8 g, 18%). LCMS (Method A): RT = 6.13 min, M+1 = 432. H NMR (500 MHz, CDCI3): 7.45-6.9 (10H, m), 7.37 (2H, d), 5.0 (1 H, br), 2.4-2.1 (4H, m), 2.09 -1.95 (1 H, m), 1.85 -1.7 (1 H, m), 1.4-1.0 (9H, br).
Step 4: tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2.3-biri,4loxazin-6- vQphenvDcyclobutvDcarbamate
To a solution of tert-butyl (1-(4-(5-amino-6-hydroxy-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (1.8g, 4.17 mmol), DIPEA (3.63 ml, 20 mmol) in DCM (100 ml) at 0 °C was added a solution of chloroacetyl chloride (0.465 ml) in DCM (20 ml). The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure. To the residue was added MIBK (120 ml) and saturated sodium bicarbonate (120 ml). The resulting mixture was heated at 115 °C for 16 h. After cooled down to room temperature, the MIBK was removed under reduced pressure. The residue was extracted with ethyl acetate (75 ml). The layers were separated and the aqueous phase extracted into ethyl acetate (2 x 50 ml). The combined organic phases were washed with water (150 ml) and brine (150ml), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 2% methanol in DCM) to give the title compound (0.82 g, 42%). LCMS (Method A): RT = 6.55 min, M+1 = 472. 1H NMR (500 MHz, CDCI3): 7.3-7.0 (10H, m), 4.96 (1H, br ), 4.82 (2H, s), 2.55-2.25 (4H, m), 2.1 -1.9 (1 H, m), 1.8 -1.65 (1 H, m), 1.35-1.1 (9H, br).
Step 5: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1H-pyridor2.3-biri,41oxazin-2(3H)-one Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (8 mg, 0.017 mmol) was reacted to afford the title compound (6 mg, 73%). LCMS (Method A): RT = 3.59 min, M+1 = 373. H NMR (500 MHz, MeOD): 7.40 (2H, d), 7.37 (2H, d), 7.33 (1 H, s), 7.32-7.28 (3H, m), 7.19 -7.17 (2H, m), 4.93 (2H, s), 2.78 -2.72 (2H, m), 2.60-2.54 (2H, m), 2.27-2.19 (1 H, m), 2.00-1.91 (1 H, m).
Example 31: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-phenyl-1H-pyrido[2,3 b]H,41oxazin-2(3H)-one
Figure imgf000067_0001
Step 1 : tert-butyl (1-(4-(1-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2.3-biri,4loxazin- 6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (60 mg, 0.13 mmol) in dry DMF (1 mL) was added sodium hydride (6 mg, 0.14 mmol) at 0 °C under nitrogen. After 1 h at 0 °C, iodomethane (9 μΐ_, 0.14 mmol) was added and the resulting mixture was stirred for an additional 10 min at 0 °C. A saturated solution of NH4CI was added and the mixture was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 30% EtOAc in cyclohexane) to give the title compound as a white solid (38 mg, 60%). Ή NMR (500 MHz,CDCI3): 7.31-7.24 (8H, m), 7.21-7.19 (2H,m), 5.01 (1 H, br s), 4.90 (2H, s), 3.39 (3H, s),2.55-2.25 (4H, m), 2.09-2.01 (1H, m), 1.84-1.76 (1H, m), 1.44-1.15 (9H, br). Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1 -methyl-7-phenyl-1 H-pyridof2,3-b1f1 ,41oxa2in- 2(3H)-one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1 -(4-(1-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxa2in-6-yl)phenyl)cyclobutyl)carbamate (37 mg, 0.08 mmol) was reacted to afford the title compound (25 mg, 78%). LCMS (Method A): RT = 3.90 min, M+2H+ = 387. H NMR (500 MHz, MeOD): 7.54 (1 H,s), 7.40 (2H, d), 7.37 (2H, d), 7.29- 7.27 (3H, m), 7.23 -7.21 (m, 2H), 4.95 (2H, s), 3.42 (3H, s), 2.77 -2.71 (m, 2H), 2.59-2.53 (m, 2H), 2.26-2.17 (m, 1 H), 1.99-1.90 (m, 1 H).
Example 32: 6-(4-(1 -aminocvclobutyl)phenyl)-7-phenyM -(prop-2-vn-1 -yl)-1 H- PVridor2,3-bin,41oxazin-2(3H)-one
Figure imgf000068_0001
Step 1 : tert-butyl (1-(4-(2-oxo-7-Dhenyl-1 -(prop-2-vn-1-yl)-2,3-dihvdro-1 H-pyridor2,3- bl[1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (23 mg, 0.05 mmol) in dry DMF (1 mL) was added sodium hydride (3 mg, 0.05 mmol) at 0 °C under nitrogen. After 1 h at 0 °C, propargyl bromide (16 μί, 0.15 mmol) was added and the resulting mixture was stirred for an additional 1 h at 0 °C. A saturated solution of NH4CI was added and the mixture was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 30% EtOAc in cyclohexane) to give the title compound (17 mg, 68%). Ή NMR (500 MHz,CDCI3): 7.50 (1 H, s), 7.31-7.24 (7H, m), 7.22-7.20 (2H,m), 5.01 (1 H, br s), 4.92 (2H, s), 4.73 (2H, d), 2.55-2.25 (4H, m), 2.32 (1 H, t), 2.09-2.01 (1 H, m), 1.85-1.76 (1 H, m), 1.44-1.15 (9H, br). Step 2: 6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(prop-2-vn-1-yl)-1 H-pyridof2,3- bin ,41oxazin-2(3H)-one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one tert-butyl (1 -(4-(2-oxo-7-phenyl-1 -(prop-2-yn-1 -yl)-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (6 mg, 0.01 mmol) was reacted to afford the title compound (3 mg, 40%). LCMS (Method A): RT = 4.12 min, M+2H+ =41 1. 1H NMR (500 MHz, MeOD): 7.69 (1 H,s), 7.41 (2H, d), 7.38 (2H, d), 7.30- 7.29 (3H, m), 7.23 -7.21 (m, 2H), 4.98 (2H, s), 4.85 (2H, s), 2.80 (1 H, t), 2.77 -2.71 (m, 2H), 2.59-2.53 (m, 2H), 2.26-2.17 (m, 1 H), 1.97-1.92 (m, 1 H).
Example 33: 2-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro- H- pyridor2,3-biri.41oxazin-1-yl)acetonitrile
Figure imgf000069_0001
Step 1 : tert-butyl (1 -(4-n-(cvanomethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2.3- blf 1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (14 mg, 0.03 mmol) in dry DMF (1 mL) was added sodium hydride (2 mg, 0.05 mmol) at 0 °C under nitrogen. After 1 h at 0 °C, 2- bromoacetonitrile (7 μΙ_, 0.09 mmol) was added and the resulting mixture was stirred for an additional 4h at room temperature. Water was added and the mixture was extracted with ethyl acetate (3x). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 40% EtOAc in hexane) to give the title compound (6 mg, 40%). 1H NMR (500 MHz,CDCI3): 7.36 (1 H, s), 7.30-7.26 (7H, m), 7.22-7.20 (2H,m), 5.01 (1 H, br s), 4.96 (2H, s), 4.88 (2H, s), 2.55-2.25 (4H, m), 2.10-2.02 (1 H, m), 1.85-1.76 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyridof2,3- blf 1 ,41oxazin-1 -vDacetonitrile
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1 -(4-(1 -(cyanomethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (6 mg, 0.02 mmol) was reacted to afford the title compound (6 mg, quantitative). LC S (Method A): RT = 3.95 min, Μ+2ΗΓ =412. Ή NMR (500 MHz, MeOD): 7.69 (1 H,s), 7.43 (2H, d), 7.38 (2H, d), 7.31-7.30 (3H, m), 7.25-7.24 (m, 2H), 5.13 (2H, s), 5.03 (2H, s), 2.77 -2.72 (m, 2H), 2.59- 2.53 (m, 2H), 2.26-2.17 (m, 1 H), 1.99-1.92 (m, 1 H).
Example 34: 6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-(dimethylamino)ethyl)-7-phenyl- 1 H-pyridor2.3-blM .41oxazin- -one
Figure imgf000070_0001
Step 1 : tert-butyl (1-(4-(1-(2-(dimethylamino)ethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- pyridor2,3-biri,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (19 mg, 0.04 mmol) in dry DMF (1 ml.) was added sodium hydride (8 mg, 0.18 mmol) at 0 °C under nitrogen. After 1 h at 0 °C, 2-chloro-N,N- dimethylethanamine. hydrochloride (18 mg, 0.12 mmol) was added and the resulting mixture was stirred for an additional 5h at 40 °C. Water was added and the mixture was extracted with ethyl acetate (3x). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 3% MeOH in dichloromethane) to give the title compound (7 mg, 32%). Ή NMR (500 MHz,CDCI3): 7.38 (1 H, s), 7.31-7.24 (7H,m), 7.20-7.18 (2H,m), 5.01 (1 H, br s), 4.88 (2H, s), 4.08 (2H,t), 2.60 (2H, t), 2.55- 2.25 (4H, m), 2.33 (6H, s), 2.09-2.01 (1 H, m), 1.85-1.76 (1H, m), 1.44-1.15 (9H, br).
Step 2: 6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-(dimethylamino)ethyl)-7-phenyl-1 H- pyridor2,3-bir .41oxazin-2(3H)-one
Following the procedure 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin- 5(6H)-one, tert-butyl (1-(4-(1-(2-(dimethylamino)ethyl)-2-oxo-7-phenyl-2,3-dihydro-1H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (7mg, 0.01 mmol) was reacted to afford the title compound (7 mg, quantitative). LCMS (Method A): RT = 2.92 min, M+H+ =443. Ή NMR (500 MHz, MeOD): 7.63 (1 H,s), 7.41 (2H, d), 7.38 (2H, d), 7.31-7.29 (3H, m), 7.25-7.23 (m, 2H), 5.00 (2H, s), 4.45 (2H, t), 3.48 (2H, t), 3.02 (6H, s), 2.77 -2.72 (m, 2H), 2.59-2.53 (m, 2H), 2.25-2.19 (m, 1 H), 1.99-1.92 (m, 1 H).
Example 35: 2-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dirivdro-1H- pyridor2,3-biri ,41oxazin-1 -vQacetamide
Figure imgf000071_0001
Step 1 : tert-butyl (1 -(4-(1-(2-amino-2-oxoethyl)-2-oxo-7-phenyl-2,3-dihvdro- H- pyridof2.3-b1[1.41oxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (19 mg, 0.04 mmol) in dry DMF (1 mL) was added potassium carbonate (12 mg, 0.08 mmol) and 2-chloroacetamide (12 mg, 0.12 mmol) at room temperature under nitrogen. The reaction mixture was stirred overnight at room temperature and concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 80% AcOEt in hexane) to give the title compound (14 mg, 67%). 1H NMR (500 MHz,CDCI3): 7.32 (1 H, s), 7.28-7.23 (7H,m), 7.16-7.14 (2H,m), 6.22 (1 H, br s), 5.65 (1 H, br s), 5.09 (1 H, br s), 4.94 (2H, s), 4.52 (2H,s), 2.55-2.25 (4H, m), 2.09-2.01 (1 H, m), 1.85-1.76 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 2-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridoi2.3- b1f1.41oxazin-1-yl)acetamide
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(1-(2-amino-2-oxoethyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (10 mg, 0.02 mmol) was reacted to afford the title compound (10 mg, quantitative). LCMS (Method A): RT = 3.43 min, M-NH2 =412. Ή NMR (500 MHz, MeOD): 7.39 (2H, d), 7.36 (2H, d), 7.34 (1 H,s), 7.28-7.26 (3H, m), 7.18-7.17 (m, 2H), 5.01 (2H, s), 4.72 (2H, s), 2.76 -2.70 (m, 2H), 2.57-2.51 (m, 2H), 2.25-2.16 (m, 1 H), 1.98-1.89 (m, 1 H). Example 36: 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-oxopropyl)-7-phenyl-1H- pyridor2.3-blH,41oxazin-2(3H)-one
Figure imgf000072_0001
Step 1 : tert-butyl (1-(4-(1-(2-oxo -1-(2-oxopropyl)-7-phenyl-2.3-dihvdro-1 H-pyridor2.3- bli1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of re -butyl (1-(4-(2-oxo-1-(2-oxopropyl)-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (60 mg, 0.13 mmol) in dry DMF (2.5 mL) was added sodium hydride (5 mg, 0.14 mmol) at 0 °C under nitrogen. After 1h at 0 °C, chloroacetone (11 DL, 0.14 mmol) was added and the resulting mixture was stirred for two hours at 0°C. A saturated solution of NH4CI was added while stirring, keeping the temperature below 10°C. The resulting mixture was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage chromatography (silica, hexane: ethyl acetate, gradient elution from 100:0 to 55:45) to give the desired product as a white solid (41 mg, 60% yield). Ή NMR (500 MHz,CDCI3): 7.12-7.28 (9H, m), 6.82 (1H,s), 4.95 (2H, s), 4.54 (2H, s), 2.36-2.52 (4H, m), 2.23 (3H, s), 1.98-2.12 (1 H, m), 1.69-1.75 (1 H, m), 1.47 (9H, br s). LC S (Method A): RT = 6.87 min, M+H+ = 528.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-oxopropyl)-7-phenyl-1 H-pyrido[2,3- b1M.41oxazin-2(3H)-one
Tert-butyl (1-(4-(1-(2-oxo -1-(2-oxopropyl)-7-phenyl-2,3-dihydro- H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (5 mg, 0.009 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (0.5 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (2.2 mg, 46% yield). 1H NMR (500 MHz, CH3OD) 7.15-7.50 (m, 10H), 5.01 (s, 2H), 4.98 (s, 2H), 2.71-2.81 (m, 2H), 2.53-2.63 (m, 2H), 2.30 (s, 3H), 2.18-2.27 (br m, 1 H), 1.92- 2.03 (br m, 1 H). LCMS (Method A): RT = 3.03 min, M+H+ = 428.
Example 37: 1-(4-(7-phenyl-2,3-dihydro-1H-pyrido[2,3-bin.41oxazin-6- yQphenyQcyclobutanamine
Figure imgf000073_0001
Step 1 : tert-butyl (1-(4-(7-phenyl-2.3-dihvdro-1 H-pyridof2.3-blf1.4loxazin-6- yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (143 mg, 0.42 mmol) in dry THF (4 mL) was added boron trifluoride diethyl etherate (91 μί, 0.73 mmol) and sodium borohydride (27 mg, 0.73 mmol) at 0 °C under nitrogen. The resulting mixture was stirred for 4 h at 0 °C. AcOEt (5 mL) and a saturated solution of NaHC03 (10 mL) were added. The aqueous phase was extracted with dichloromethane (3x) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 2% MeOH in DCM) to give the title compound as a white solid (66 mg, 48%). Ή NMR (500 MHz,CDCI3): 7.21-7.26 (7H, m), 7.14-7.12 (2H,m), 6.97 (1 H, s), 4.48 (2H, t), 3.48 (2H, br s),2.55-2.25 (4H, m), 2.06-1.98 (1 H, m), 1.82-1.73 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 1-(4-(7-phenyl-2.3-dihvdro- H-pyridor2,3-bl[1.41oxazin-6- yl)phenyl)cyclobutanamine
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (15 mg, 0.03 mmol) was reacted to afford the title compound (15 mg.quantitative). LCMS (Method A): RT = 3.80 min, M-NH2=341. 1H NMR (500 MHz, MeOD):7.31 (4H, s), 7.22-7.21 (3H, m), 7.12-7.11 (2H, m), 7.01 (1 H, s), 4.45 (2H, t), 3.45 (2H, t), 2.75 -2.70 (m, 2H), 2.56-2.50 (m, 2H), 2.24-2.15 (m, 1 H), 1.97-1.88 (m, 1 H). Example 38: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-2,4-dihydro-1 H-pyridor2,3- bin.2,41triazolor4,3-diri,41oxazin-1-one
Figure imgf000074_0001
Step 1 : tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihvdro-1H-pyridor2,3-biri,41oxazin-6- yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (211 mg, 0.45 mmol) in toluene (15 mL) was added lawesson's reagent (181 mg, 0.45 mmol) under nitrogen. The resulting mixture was heated under reflux for 2 h. After cooled down to room temperature, the mixture was concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 5% MeOH in dichloromethane) to give the title compound as a yellow solid (124 mg, 57%). 1H NMR (500 MHz,CDCI3): 9.47 (1H, br s), 7.30-7.24 (7H, m), 7.18 (1 H, s), 7.16-7.14 (2H, m), 5.17 (2H, s), 5.04 (1 H, br s), 2.55-2.25 (4H, m), 2.10-2.01 (1 H, m), 1.85-1.76 (1 H, m), 1.44-1.15 (9H, br).
Step 2: Ethyl 2-(6-(4-(1-(tert-butoxycarbonylamino)cvclobutyl)phenyl)-7-phenyl-1 H- pyridof2,3-bl[1 ,4loxazin-2(3H)-ylidene)hvdrazinecarboxylate
To a solution of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (20 mg, 0.04 mmol) in dry THF (1 mL) was added ethyl hydrazinecarboxylate (21 mg, 0.20 mmol) and mercuric acetate (21 mg, 0.06 mmol) under nitrogen. The reaction mixture was stirred for 2h at room temperature, filtered on celite and concentrated to dryness under reduced pressure to give the title compound which was used without purification in the next step.
Step 3: tert-butyl (1-(4-(1-oxo-8-phenyl-2.4-dihvdro-1 H-pyridof2.3-biri.2.4ltriazolor4,3- d1H ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A solution of Ethyl 2-(6-(4-(1-(tert-butoxycarbonylamino)cyclobutyl)phenyl)-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-ylidene)hydrazinecarboxylate (24 mg, 0.04 mmol) in dry DMF (1 mL) was heated at 200 °C for 10 min under microwave irradiation. The reaction mixture was diluted with dichloromethane, filtered on celite and concentrated to dryness under reduced pressure. The resulting residue was purified by preparative HPLC
(method G, gradient 5 to 95% 0.1%FA/ACN in 0.1%FA H2O) to give the title compound (7 mg, 32%). 1H N R (500 MHz,CDCI3): 9.18 (1 H, br s), 8.54 (1 H, s), 7.32 (2H, d), 7.28- 7.26 (5H,m), 7.22-7.20 (2H,m), 5.32 (2H, s), 5.07 (1 H, br s), 2.55-2.25 (4H, m), 2.10 - 2.03 (1H, m), 1.86-1.78 (1H, m), 1.44-1.15 (9H, br).
Step 4: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-2.4-dihydro-1 H-pyridor2,3- blf 1.2,41triazolof4,3-din .41oxazin-1 -one
Following the procedure for 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8- dihydroquinolin-5(6H)-one, tert-butyl (1-(4-(1-oxo-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (7 mg, 0.01 mmol) was reacted to afford the title compound (7 mg, quantitative). LCMS (Method A): RT = 3.54 min, M+2H+ =413. H NMR (500 MHz, MeOD): 8.56 (1 H, s), 7.44 (2H, d), 7.38 (2H, d), 7.30-7.29 (3H, m), 7.23-7.21 (m, 2H), 5.41 (2H, s), 2.78-2.72 (m, 2H), 2.59-2.53 (m, 2H), 2.25-2.18 (m, 1 H), 1.99-1.91 (m, 1 H).
Example 39: 1-(4-(8-phenyl-4H-pyridor2.3-bin,2.4Uriazolor4,3-diri,4loxazin-7- yl)phenyl)cyclobutanamine
Figure imgf000075_0001
Step 1 : tert-butyl (1-(4-(2-hvdrazono-7-phenyl-2.3-dihvdro-1 H-pyrido[2,3-b1f1.4loxazin-6- yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(7-phenyl-2-thioxo-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (20 mg, 0.04 mmol) in THF (2mL) was added hydrazine.H20 (10 μΙ_, 0.20 mmol) under nitrogen. The resulting mixture was stirred at room temperature for 1.5 h. After cooled down to room temperature, the mixture was concentrated to dryness under reduced pressure to give the title compound which was used directly in the next step without any purification. Step 2: tert-butyl (1-(4-(8-phenyl-4H-pyridor2.3-biri.2.41triazolor4.3-dlf1.41oxazin-7- vDphenyOcyclobutyQcarbamate
A solution of tert-butyl (1-(4-(2-hydrazono-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (20 mg, 0.04 mmol) in triethyl orthoformate (1 mL) was heated at 150 °C for 10 min under microwave irradiation. The reaction mixture was concentrated to dryness under reduced pressure. The resulting residue was purified by silica gel chromatography (gradient 0 to 2% MeOH in
dichloromethane) to give the title compound (3 mg, 15%). Ή NMR (500 MHz,CDCI3): 8.68 (1 H, s), 7.75 (1 H, s), 7.34-7.26 (7H, m), 7.22-7.20 (2H,m), 5.70 (2H, s), 5.02 (1 H, br s), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86-1.78 (1 H, m), 1.44-1.15 (9H, br).
Step 3: 1-(4-(8-phenyl-4H-pyridof2.3-biri.2.41triazolo[4.3-dir .41oxazin-7- vDphenvDcvclobutanamine
Following the procedure 2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin- 5(6H)-one tert-butyl (1-(4-(8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (2 mg, 0.004 mmol) was reacted to afford the title compound (2 mg, quantitative). LCMS (Method A): RT = 3.60 min, M+2H+ =397. 1H NMR (500 MHz, MeOD): 9.31 (1 H, s), 8.30 (1 H, s), 7.44 (2H, d), 7.38 (2H, d), 7.32-7.31 (3H, m), 7.27-7.25 (m, 2H), 5.77 (2H, s), 2.78-2.72 (m, 2H), 2.59-2.53 (m, 2H), 2.25-2.18 (m, 1 H), 1.99-1.91 (m, 1 H).
Example 40: 1-(4-(1-methyl-7-phenyl-2,3-dihvdro-1H-pyridor2.3-b1M.41oxazin-6- vnphenvDcyclobutanamine
Figure imgf000076_0001
Step 1 : tert-butyl (1-(4-(1-methyl-7-phenyl-2.3-dihvdro-1 H-pyridor2.3-biri.4loxazin-6- vDphenvDcvclobutvDcarbamate
To a solution of tert-butyl (1 -(4-(7-phenyl-2,3-dihydro-1 H-pyhdo[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (20mg, 0.04 mmol) in dry DMF (1 ml) at 0°C was added sodium hydride (2 mg, 0.05 mmol) and methyl iodide (5 μΙ, 0.05 mmol) under nitrogen. The resulting mixture was stirred for 30 minutes at 0°C. A saturated solution of NH4CI was added and the mixture was extracted with DCM (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 20% EtOAc in cyclohexane) to give the title compound (8 mg, 44%). 1H NMR (500 MHz.CDCIs): 7.39 -7.19 (9H, m), 6.85 (1 H, s), 4.51-4.50 (2H, d), 3.45-3.43 (2H, d), 2.80- 2.70 (2H, m), 2.61-2.51 (4H, m), 2.01 -1.88 (1 H, m), 2.00 (3H, s), 1.40-1.10 (9H, br).
Step 2: 1-(4-(1-methyl-7-phenyl-2.3-dihvdro-1 H-pyrido[2.3-b1f1.4loxazin-6- yQphenyQcvclobutanamine
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin-2(3H)-one, 2-((1 r,3r)-1-(4- (1-ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)-3-hydroxy- 3-methylcyclobutyl)isoindoline-1 ,3-dione (21 mg, 0.04 mmol) was reacted to afford the title compound (3 mg, quantitative). LCMS (Method A): RT = 4.21 min, M+1 = 355. 1H NMR (500 MHz, MeOD): 7.36- 7.32 (4H, m), 7.29-7.20 (3H, m), 7.19 -7.15 (2H, m), 7.00 (1 H, s), 4.45 (2H, d), 3.45 (2H, d), 2.88 (3H, s), 2.75-2.70 (2H, m), 2.61-2.51 (2H, m), 2.28-2.17 (1 H, m), 2.01-1.88 (1H, m).
Example 41: 1-(6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-2.3-dihvdro-1H- pyridor2,3-blH ,4loxazin-1 -vDethanone
Figure imgf000077_0001
Step 1 : tert-butyl (1-(4-(1-acetyl-7-phenyl-2.3-dihvdro-1 H-pyridor2.3-b1f1.41oxazin-6- vQphenvDcvclobutyDcarbamate
To a solution of tert-butyl (1-(4-(7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (22 mg, 0.05 mmol) in dry DCM (1 ml) was added triethyl amine (10 μΙ, 0.07 mmol) and acetyl chloride (5 μΙ, 0.07 mmol) under nitrogen. The resulting mixture was stirred for 1h at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 80% EtOAc in cyclohexane) to give the title compound (20 mg, 83%). Ή NM (500 MHz,CDCI3): 8.50 (1 H, br s), 7.25 (2H, d), 7.19-7.17 (5H, m), 7.11-7.10 (2H,m), 4.95 (1 H, s), 4.43 (2H, t), 3.94 (2H, br s), 2.55-2.25 (7H, m), 2.06-1.98 (1 H, m), 1.82-1.73 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 1-(6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- b][1 ,4loxazin-1-yl)ethanone
Tert-butyl (1-(4-(1-acetyl-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (20 mg, 0.04 mmol) was dissolved in 2M HCI in Et20 (1 ml). The resulting mixture was stirred overnight at room temperature and evaporated under reduced pressure. The deprotected compound was taken back twice into diethyl ether. The remaining solvent was removed under reduced pressure and dried to afford the title compound (21 mg, quantitative). LCMS (Method A): RT = 3.87 min, M-16 =383. Ή NMR (500 MHz, MeOD): 8.70 (1 H, br s), 7.44-7.43 (4H,m), 7.30-7.28 (3H, m), 7.21 - 7.19 (2H, m), 4.63 (2H, t), 4.11 (2H, t), 2.75 -2.70 (2H, m), 2.56-2.50 (2H, m), 2.42 (3H, s), 2.24-2.15 (1 H, m), 1.97-1.88 (1 H, m).
Example 42: 1-((1H-imidazol-2-yl)rnethyl)-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl- 1 H-pyridor2,3-biri .41oxazin -one
Figure imgf000078_0001
Step 1 : tert-butyl (1 -(4-(1-((1 H-imidazol-2-yl)methyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- Pyridor2,3-b1f1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (26 mg, 0.05 mmol) in dry DMF (1 ml) was added sodium hydride (5 mg, 0.13 mmol) at 0 °C under nitrogen. After 1 hour at 0 °C, 2- (chloromethyl)-I H-imidazole.HCI (10 mg, 0.07 mmol) was added and the resulting mixture was stirred for an additional 1 h at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by preparative HPLC (method G, gradient 5 to 95% 0.1 %FA/ACN in 0.1 %FA/H2O) to give the title compound (5 mg, 17%). 1H N R (500 MHz,CDCI3): 7.83 (1 H, s), 7.22-7.15 (7H,m), 7.14-7.13 (2H,m), 6.94 (2H,s), 6.22 (1 H, br s), 5.14 (2H, s), 4.98 (1 H, br s), 4.83 (2H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.75 -1.70 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 1-(( H-imidazol-2-yl)methyl)-6-(4-(1-aminocvclobutyl)Dhenyl)-7-phenyl-1 H- pyridor2.3-blf1.4loxazin-2(3H)-one
Tert-butyl (1-(4-(1-((1 H-imidazol-2-yl)methyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (5 mg, 0.001 mmol) was dissolved in TFA (1 ml). The resulting mixture was stirred for 30 seconds at room temperature and evaporated under reduced pressure. The deprotected compound was taken back twice into diethyl ether and the solid formed was washed twice with DCM. The remaining solvent was removed under reduced pressure and dried to afford the title compound (5 mg, quantitative). LC S (Method A): RT = 2.80 min, M-NH2 =435. Ή NMR (500 MHz, MeOD): 7.44 (1 H, s), 7.41 (2H, s), 7.31-7.26 (4H,m), 7.18-7.16 (3H,m), 7.06 -7.05 (2H, m), 5.44 (2H, s), 4.97 (2H, s), 2.66 -2.60 (2H, m), 2.49-2.43 (2H, m), 2.14-2.10 (1 H, m), 1.89-1.82 (1 H, m).
Example 43: 2-(6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-2.3-dihvdro-1H- pyrido[2,3-biri ,41oxazin-1 -vQacetonitrile
Example 44: 2-(6-(4-( 1 -aminocvclobutyl)phenyl)-7-phenyl-2.3-dihvdro-1 H- pyridor2,3-biri.41oxazin-1-yl)acetamide
Figure imgf000079_0001
Step 1 : tert-butyl (1 -(4-(1-(cvanomethyl)-7-phenyl-2.3-dihvdro-1 H-pyrido[2.3- bl[1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (24 mg, 0.05 mmol) in dry DMF (1 ml) was added potassium carbonate (93 mg, 1.57 mmol) and bromoacetonitrile (110 μΙ, 1.57 mmol) under nitrogen. The resulting mixture was stirred for 4 hours at 80C. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were
concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 60% EtOAc in cyclohexane) to give the title compound (5 mg, 19%). Ή NMR (500 MHz,CDCI3): 7.22-7.11 (9H, m), 6.93 (1 H, s), 4.92 (1 H, s), 4.51-4.49 (2H, m), 4.15 (2H, s), 3.39-3.37 (2H, m), 2.55-2.25 (4H, m), 1.99 - 1.94 (1 H, m), 1.75 -1.63 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 2-(6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-2.3-dihydro-1 H-pyridof2.3- blf 1 ,4loxazin-1 -vDacetonitrile
2-(6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-2,3-dihvdro-1 H-pyrido[2,3-bl[1 ,4loxazin-1 - vDacetamide
Following the procedure for 1 -(6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-1-yl)ethanone, 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1-(cyanomethyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (5 mg, 0.01 mmol) was reacted to afford the two title compounds (1 mg, 25% and 2 mg, 41 %). LCMS: RT = 3.99 min, M-16 =380. H NMR (500 MHz, MeOD): 7.20-7.13 (8H, m), 7.09-7.07 (2H, m), 4.46 (2H, t), 3.54 (2H, s), 3.22 (2H, t), 2.46-2.40 (2H, m), 2.17-2.11 (2H, m), 1.97-1.94 (1 H, m), 1.66-1.62 (1 H, m). LCMS (Method A): RT = 3.99 min, M-16 =398. 1H NMR (500 MHz, D20): 8.39 (2H, s), 7.27-7.22 (7H, m), 7.1 -7.09 (2H, m), 6.89 (1 H, s), 4.78 (2H, s), 4.48 (2H, t), 4.00 (2H, s), 3.50 (2H, t), 2.65 -2.60 (2H, m), 2.53 -2.47 (2H, m), 2.09-2.05 (1 H, m), 1.86-1.79 (1 H, m).
Example 45: 6-(4-(1 -aminocyclobutyl)phenv0-7-phenyl-1 -(pyridin-4-ylmethylH H- pyridor2,3-biri,41oxazin-2(3H)-one
Figure imgf000080_0001
Step 1 : tert-butyl n-(4-(2-oxo-7-phenyl-1-(pyridin-4-ylmethyl)-2,3-dihvdro-1H-pyridor2,3- b)n,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (35 mg, 0.07 mmol) in dry DMF (1 ml) was added sodium hydride (7 mg, 0.18 mmol) and 4-(bromomethyl)pyridine.HBr (23 mg, 0.09 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 80% EtOAc in cyclohexane) to give the title compound (21 mg, 50%). 1H NMR (500 MHz,CDCI3): 8.54 (2H,s), 7.19 -7.12 (9H,m), 6.97 (1 H,s), 6.93-6.91 (2H,m), 5.1 1 (2H, s), 4.98 (1 H, br s), 4.95 (2H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.75 -1.70 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-4-ylmethyl)-1 H-pyridor2,3- bin .41oxazin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (2-oxo-7-phenyl-1-(pyridin-4-ylmethyl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (21 mg, 0.04 mmol) was reacted to afford the title compound (26 mg, quantitative). LCMS (Method A): RT = 3.25 min, M+1 = 464. NMR (500 MHz, MeOD): 8.57 (2H, s), 7.67 (2H, s), 7.28-7.24 (4H, m), 7.23 (1 H, s), 7.14-7.10 (3H, m), 6.96 -6.94 (2H, m), 5.37 (2H, s), 5.03 (2H, s), 2.66 -2.60 (2H, m), 2.49-2.43 (2H, m), 2.14-2.10 (1 H, m), 1.89-1.82 (1 H, m).
Example 46: 6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 -(pyridin-3-Ylmethyl)-1 H- pyridor2.3-bU1.41oxazin-2(3H)-one
Figure imgf000081_0001
Step 1 : tert-butyl n-(4-(2-oxo-7-phenyl-1-(pyridin-3-ylmethyl)-2.3-dihvdro-1 H-Pyridof2,3- b1f1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (l^^-oxo-y-phenyl^.S-dihydro-I H-pyrido^.S-blll^loxazin-
6- yl)phenyl)cyclobutyl)carbamate (51 mg, 0.11 mmol) in dry DMF (1 ml) was added sodium hydride (11 mg, 0.26 mmol) and 3-(bromomethyl)pyridine.HBr (33 mg, 0.13 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 80% EtOAc in cyclohexane) to give the title compound (23 mg, 38%). Ή NMR (500 MHz,CDCI3): 8.60- 8.50 (2H, br s), 7.60 (2H, d), 7.28 (1 H, s), 7.19 -7.15 (6H, m), 7.12 (1 H, s), 6.97 (2H, d), 5.13 (2H, s), 4.93 (3H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.75 -1.70 (1 H, m), 1.44- 1.15 (9H, br).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-3-ylmethyl)-1 H-pyhdof2,3- bin.41oxazin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (2-oxo-7-phenyl-1-(pyridin-3-ylmethyl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (23 mg, 0.04 mmol) was reacted to afford the title compound (27 mg, quantitative). LCMS (Method A): RT = 3.48 min, M+1 = 464. Ή NMR (500 MHz, MeOD): 8.75 (1 H, br s), 8.60 (1 H, br s), 8.12 (1 H, d), 7.67 (1 H, s), 7.49 (1 H, s), 7.40 -7.36 (4H, m), 7.28-7.24 (3H, m), 7.09 (2H, d), 5.42 (2H, s), 5.12 (2H, s), 2.77 - 2.72 (2H, m), 2.59-2.53 (2H, m), 2.25 -2.21 (1 H, m), 1.98-1.94 (1 H, m).
Example 47: 1-(4-(1-ethyl-8-phenyl-4H-pyridor2.3-bin,2,4ltriazolor4,3-diri.41oxazin-
7- yl)phenyl)cyclobutanamine
Figure imgf000082_0001
Step 1 : tert-butyl (1-(4-(7-phenyl-2-thioxo-2.3-dihvdro-1 H-pyridof2,3-b]ri.4loxazin-6- yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6yl)phenyl)cyclobutyl)carbamate (5 g, 10.60 mmol) in toluene (380 ml) was added Lawesson's reagent (4.29 g, 10.60 mmol). The resulting mixture was heated under reflux for 5 hours. After it was cooled down to room temperature, the mixture was concentrated to dryness under reduced pressure. The resulting residue was dissolved in the minimum amount of dichloromethane (500 ml) and treated with diisopropyl ether (800ml). A yellow solid crushed out. It was filtered and dried until constant weight (4.7g, 91%). LCMS (Method D): RT = 1.526 min, M+1 = 488.
Step 2: (E)-tert-butyl (1-(4-(2-hvdrazono-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- bl[1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate(2 g, 2.05 mmol) in THF (350 mL) was added hydrazine monohydrate (645 μΙ, 20.51 mmol) to give a yellow solution. The resulting mixture was stirred at room temperature for 2 hours. The mixture was filtered to remove insoluble black particles and concentrated to dryness under reduced pressure. The resulting yellow solid was used directly in the following step as crude without any further purification. LCMS (Method D): RT = 1.04 min, M+1 = 486.
Step 3: tert-butyl (1-(4-(9-phenyl-5.6-dihvdro-ri.2,4ltriazolof3.4-firi ,6lnaphthyridin-8- vDphenvQcvclobutvDcarbamate
A solution of (E)-tert-butyl (1-(4-(2-hydrazono-7-phenyl-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.103 mmol) in triethyl orthopropionate (1 ml) was heated to 150 °C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n- hexanes) to give the title compound (30 mg, 55%). LCMS (Method D): RT = 1.43 min, M+1= 524. 1H NMR (500 MHz, CDCI3): 7.80 (1 H, s), 7.27-7.26 (5H, m), 7.22 (2H, d), 7.15 (2H, m), 5.56 (2H, s), 3.22 (2H, q), 2.50-2.33 (4H, m), 2.07-1.94 (1 H, m), 1.81-1.68 (1 H, m), 1.52 (3H, t), 1.38-1.11 (9H, br).
Step 4: 1-(4-(1-ethyl-8-phenyl-4H-pyridof2.3-b1f1.2,4ltriazolof4.3-dlf1.4loxazin-7- vDphenvQcyclobutanamine
Tert-butyl (1 -(4-(9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4-f][1 ,6]naphthyridin-8- yl)phenyl)cyclobutyl)carbamate (30 mg, 0.057 mmol) was dissolved in TFA (2 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (15 mg, 61 % yield). LCMS (Method A) RT = 3.78 min, M+1 =425. 1H-NMR (500 MHz, CD3OD) δ 8.10 (1 H, s), 7.49 (2H, d), 7.35 (3H, m), 7.29 (4H, m), 5.66 (2H, s), 3.26 (2H, t), 2.84-2.71 (2H, m), 2.64-2.54 (2H, m), 2.31-2.14 (1 H, m), 2.04-1.93 (1 H, m), 1.51 (3H, t).
Example 48: 6-(4-(1-aminocvclobutyl)phenyl)-1 -(cyclobutylmethyl)-7-phenyl-1 H- pyridor2,3-b1M,41oxazin-2(3 -one
Figure imgf000084_0001
Step 1 : tert-butyl n-(4-(1-(cvclobutylmethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-Pyridor2.3- blf1.4loxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (44 mg, 0.09 mmol) in dry DMF (1 ml) was added sodium hydride (9 mg, 0.22 mmol) and (bromomethyl)cyclobutane (13 μΙ, 0.1 1 mmol) under nitrogen. The resulting mixture was stirred for 1 h at 80C. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 40% EtOAc in cyclohexane) to give the title compound (8 mg, 16%). 1H NMR (500 MHz,CDCI3): 7.23 -7.17 (8H, m), 7.13-7.1 1 (2H, m), 4.95 (1 H, s), 4.80 (2H, s), 3.96 (2H, d), 2.68-2.64 (1 H, m), 2.52-2.20 (4H, m), 2.00-1.95 (3H, m), 1.86 -1.50 (5H, m), 1.44-1.15 (9H, br). Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-(cvclobutylmethyl)-7-phenyl-1 H-pyridor2,3- bin,4loxazin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (1-(cyclobutylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (8 mg, 0.02 mmol) was reacted to afford the title compound (8 mg, 97%). LCMS (Method A): RT = 4.65 min, M-16 = 423. Ή NMR (500 MHz, MeOD): 7.56 (1 H,s), 7.42-7.37 (4H,m), 7.32 -7.30 (3H, m), 7.23-7.21 (2H, m), 4.97 (1 H, s), 4.59 (2H, s), 4.14 (2H, d), 2.80 -2.72 (3H, m), 2.59-2.53 (2H, m), 2.24-2.20 (1 H, m), 2.10-2.06 (2H, m), 1.98-1.87 (5H, m).
Example 49: ethyl 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyridor2,3-b1[1 ,41oxazin-1 -vDacetate
Figure imgf000085_0001
Step 1 : ethyl 2-(6-(4-(1-(te/ -butoxycarbonylamino)cvclobutyl)phenyl)-2-oxo-7-phenyl-2l3- dihvdro-1 H-pyridof2,3-biri ,41oxazin-1 -vDacetate
In a 15 mL reaction tube was added fert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro- H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give a red solution, then cooled to 0°C under a nitrogen atmosphere. Sodium hydride, 60% in oil (6.4 mg, 0.159 mmol) was added and the mixture stirred at 0°C for 15 minutes. Ethyl 2-bromoacetate (0.035 mL, 0.318 mmol) was added and the mixture stirred at 0°C for 30 minutes, allowed to warm to room temperature then stirred for one hour. The reaction mixture was quenched by the addition of saturated ammonium chloride solution (5 mL) and extracted into ethyl acetate (3 x 3 mL). The organic phase was washed with 50:50 waterbrine (2 x 5 mL) then brine (5 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as an off-white solid (22 mg, 39% yield). 1H-NMR (500 MHz, CDCI3) δ 7.13-7.28 (8H, m), 7.37-7.43 (2H, m), 5.24 (1H, br s), 5.18 (2H, s), 4.89 (2H, s), 4.49 (2H, q), 2.40-2.90 (4H, m), 2.21-2.35 (1 H, m), 1.90-2.10 (1 H, m), 1.20-1.75 (9H, br m), 1.53 (3H, t). LCMS (Method A) RT = 7.20 min, M+H+ = 558.13.
Step 2: ethyl 2-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- Pyridof2,3-b1f ,4loxazin-1-yl)acetate
Ethyl 2-(6-(4-(1-(ierf-butoxycarbonylamino)cyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)acetate (22 mg, 0.039 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (-2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (22 mg, 98% yield). 1H-NMR (500 MHz, MeOD) δ 7.37-7.48 (5H, m), 7.25-7.37 (3H, m), 7.15-7.25 (2H, m), 5.01 (2H, s), 4.85 (2H, s), 4.26 (2H, q), 2.70-2.83 (2H, m), 2.48-2.62 (2H, m), 2.15-2.30 (1 H, m), 1.90-2.05 (1 H, m), 1.28 (3H, t). LCMS (Method A) RT = 4.08 min, M+2H+ = 459.20.
Example 50: 7-(4-(1 -aminocyclobutyl)phenyl)-8-phenyl-4H-pyridor213- biri.2.41triazolor4,3-diri.4loxazin-1-amine
Figure imgf000086_0001
Step 1: tert-butyl (1-(4-(1-amino-8-phenyl-4H-pyridof2.3-blf1.2.41triazolor3,4- d1f1,4]oxazin-7-yl)phenyl)cvclobutyl)carbamate
A solution of (E)-tert-butyl (1-(4-(2-hydrazono-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.103 mmol), cyanogen bromide (33mg, 0.309 mmol) and sodium carbonate (22mg, 0.206 mmol) in a mixture of ethanol (1 ml) and 1 ,4-dioxane (1 ml) was stirred at room temperature for one hour. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 100% to 94:6 dichloromethane in methanol) to give the title compound (15 mg, 30%). LCMS (Method A): RT = 5.73 min, M+1 = 511. Ή NMR (500 MHz, CDCI3): 7.80 (1 H, s), 7.36-7.28 (7H, m), 7.23- 7.19 (2H, m), 5.48 (2H, s), 2.56-2.41 (4H, m), 2.13-2.02 (1 H, m), 1.89-1.77 (1 H, m), 1.46-1.21 (9H, bs).
Step 2: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4H-pyhdor2,3-biri .2,41triazolor4,3- dlf1 ,4loxazin-1 -amine
Tert-butyl (1-(4-(1-amino-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[3,4-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (15 mg, 0.029 mmol) was dissolved in TFA (2 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (10 mg, 80% yield). LCMS (Method A) RT = 3.40 min, M+1 = 412. 1H-NMR (500 MHz, CD3OD) δ 8.21 (1 H, s), 7.41- 7.36 (2H, d), 7.35- 7.30 (2H, d), 7.27-7.19 (5H, m), 5.44 (2H, s), 2.72-2.62 (2H, m), 2.53-2.43 (2H, m), 2.20-2.09 (1 H, m), 1.92-1.82 (1 H, m).
Example 51 : 6-(4-(1 -aminocvclobutyl)phenvn-1 -(2-methoxyethyl)-7-phenyl-1 H- pyridor2,3-biri,41oxazin-2(3 -one
Figure imgf000087_0001
Step 1 : tert-butyl (1-(4-(1-(2-methoxyethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- b1f1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (48 mg, 0.10 mmol) in dry DMF (1 ml.) was added sodium hydride (16 mg, 0.41 mmol) and 1-bromo-2-methoxyethane (28 mg, 0.20 mmol) under nitrogen. The resulting mixture was stirred for 2 hours at 80°C. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were
concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 40% EtOAc in cyclohexane) to give the title compound (10 mg, 18%). Ή NMR (500 MHz,CDCI3): 7.49 (1 H, s), 7.23 -7.17 (7H, m), 7.12-7.1 1 (2H, m), 4.95 (1 H, s), 4.82 (2H, s), 4.05 (2H, t), 3.61 (2H, t), 3.29 (3H, s), 2.52-2.20 (4H, m), 2.00-1 .95 (1 H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-methoxyethyl)-7-phenyl-1 H-pyrido[2,3- bin .41oxazin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (1-(2-methoxyethyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (10 mg, 0.02 mmol) was reacted to afford the title compound (7 mg, 68%). LCMS (Method D): RT = 4.1 1 min, M-16 = 423. Ή NMR (500 MHz, MeOD): 7.77 (1 H,s), 7.43-7.38 (4H,m), 7.33-7.28 (3H, m), 7.23-7.21 (2H, m), 4.96 (2H, s), 4.24-4.22 (2H, t), 3.72-3.70 (2H, t), 3.36 (3H, s), 2.77 -2.72 (2H, m), 2.59-2.53 (2H, m), 2.25 -2.21 (1 H, m), 1.98-1.94 (1 H, m).
Example 52: 6-(4-(1-aminocvclobutyl)phenyl)-1-ethyl-7-phenyl-1H-pyridor2t3- b1f1.4loxazin-2(3H)-one
Figure imgf000088_0001
Step 1 : terf-butyl 1-(4-(1-ethyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-Pyridof2,3-blf1 ,4loxazin-6- vDphenvDcyclobutylcarbamate
In a 15 mL reaction tube was added ierf-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give a yellow solution, then cooled to 0°C under a nitrogen atmosphere. Sodium hydride, 60% in oil (10 mg, 0.254 mmol) was added and the mixture stirred at 0°C for 15 minutes, lodoethane (10 μΙ, 0.127 mmol) was added and the mixture stirred at 0°C for 30 minutes, allowed to warm to room temperature then stirred for one hour. Further sodium hydride, 60% in oil (10.18 mg, 0.254 mmol) was added and the reaction mixture was heated to 40°C under a nitrogen atmosphere for one hour. Further iodoethane (10 μΙ, 0.127 mmol) was added and the mixture stirred at room temperature for 30 minutes. The reaction mixture was quenched by the addition of saturated sodium bicarbonate solution (5 ml_) and extracted into ethyl acetate (3 x 5 ml_). The organic phase was washed with 50:50 water:brine (2 x 10 mL) then brine (10 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography
(cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a pale yellow solid (7 mg, 13% yield). 1H-NMR (500 MHz, CDCI3) δ 7.10-7.28 (10H, m), 4.93 (1 H, br s), 4.81 (2H, s), 3.94 (2H, q), 2.10-2.55 (4H, m), 1.90-2.08 (1 H, m), 1.65-1.80 (1 H, m), 1.00-1.40 (9H, br m), 1.14 (3H, t). LCMS (Method A) RT = 7.29 min, M+H+ = 500.12.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-ethyl-7-phenyl-1 H-pyrido[2,3-blf1 ,41oxazin- 2(3H)-one
ferf-Butyl 1-(4-(1-ethyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (7 mg, 0.014 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (6 mg, 83% yield). H-NMR (500 MHz, MeOD) δ 7.57 (1 H, s), 7.42 (2H, d), 7.38 (2H, d), 7.28-7.35 (3H, m), 7.20-7.28 (2H, m), 4.95 (2H, s), 4.10 (2H, q), 2.68-2.85 (2H, m), 2.50-2.65 (2H, m), 2.15-2.30 (1 H, m), 1.90- 2.05 (1 H, m), 1.30 (3H, t). LCMS (Method A) RT = 4.04 min, M+2H+ = 401.18.
Example 53: 1-((1 H-imidazol-5-yl)methvn-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl- 1 H-pyridf>r2.3-b]ri .41oxazin- -one
Figure imgf000090_0001
Step 1 : tert-butyl (1-(4-(1-((1 H-imidazol-5-yl)methyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof2,3-biri,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
In a round bottom flask was added tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (150 mg, 0.318 mmol) in DMF (Volume: 8 ml) to give a brown solution. sodium hydride (30.5 mg, 0.763 mmol) and 5- (chloromethyl)-I H-imidazole hydrochloride salt (58.4 mg, 0.501 mmol) was added. The resulting mixture was stirred at room temperature for 2 h. further sodium hydride (15.2 mg) and imidazole (29 mg) were added and the resulting mixture strried for another hour. The reaction mixture was concentrated to dryness. The residue was partitioned between ethyl acetate (35ml) and water (30ml). The organic phase was separated and washed with water (2X25ml), brine and concentrated. The residue was purified by column (biotage 25g) to give product (14mg). LCMS (Method A) RT = 4.58 min, M+H+ = 552.1.
Step 2: 1-((1 H-imidazol-5-yl)methyl)-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1 H- pyridor2.3-biri.41oxazin-2(3H)-one
tert-butyl (1-(4-(1-((1 H-imidazol-5-yl)methyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.022 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound (7.5 mg). 1H-NMR (500 MHz, MeOD) δ 8.81 ( H, s), 7.62 (2H, br), 7.39-7.41 (m, 4H), 7.28-7.30 (3H, m), 7.16-7.18 (2H, m), 5.39 (2H, s), 5.07 (2H, s), 2.7-2.8 (2H, m), 2.55-2.65 (2H, m), 2.2-2.3 (1 H, m), 1.9-2.05 (1 H, m), LCMS (Method A) RT = 2.83 min, M+H+ = 453.1. Example 54: 2-(e-(4-f1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H- pyridor2,3-biri,41oxazin-1-yl)propanenitrile
Figure imgf000091_0001
Step 1 : tert-butyl (1-(4-(1-(1-cyanoethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- blf1 ,4loxazin-6-vnphenyl)cyclobutyl)carbamate
In a 50ml round bottom flask was added tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50mg, 0.106 mmol) in DMF (Volume: 4 ml) to give a brown solution, sodium hydride (10.18 mg, 0.254 mmol) and 2- bromopropanenitrile (17.05 mg, 0.127 mmol) were added. The reaction mixture was stirred at 50 degree for 1 h then room temperature overnight. The reaction mixture was partitioned between DCM (20ml) and water (20ml). The organic phase was separated and concentrated. The residue was purified by column (biotage, 25g) eluted with ethyl acetate/cyclohexane (0-50%) to give product (19mg). LCMS (Method A): RT = 7.14 min, M+H+ = 525.0.
Step 2: 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- biri,4loxazin-1-yl)propanenitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(1-cyanoethyl)- 2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (7mg, 0.013 mmol) was reacted to afford the title compound (5.5mg). 1H NMR (500 MHz, CH3OD) 7.77 (1 H, s), 7.44 (2H, d), 7.40 (2H, d), 7.30-7.34 (3H, m), 7.25-7.28 (2H, m), 6.03 (1 H, q), 4.99 (2H, s), 2.7-2.85 (2H, m), 2.55-2.65 (2H, m), 2.2-2.3 (1 H, m), 1.9- 2.1 (1 H, m), 1.81 (3H, d). LCMS (Method A): RT = 4.1 1 min, M+H+ = 426.1. Example 55: 6-(4-(1 -aminocvclobutyl)phenyl)-1 -(cyclopropylmethyl)-7-phenyl-1 H- pyridor2.3-biri.41oxazin-2f3 -one
Figure imgf000092_0001
Step 1 : te/t-butyl 1-(4-(1-(cvclopropylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyridor2,3- biri,41oxazin-6-yl)phenyl)cyclobutylcarbamate
In a 15 mL reaction tube was added fert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyhdo[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol), potassium carbonate (44 mg, 0.318 mmol) and (bromomethyl)cyclopropane (0.031 mL, 0.318 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give an orange suspension. The reaction mixture was stirred at 80°C for one hour. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (40 mg, 72% yield). 1H-NMR (500 MHz, CDCI3) δ 7.32 ( H, s), 7.10-7.28 (9H, m), 4.98 (1 H, br s), 4.82 (2H, s), 3.79 (2H, d), 2.05-2.60 (4H, m), 1.90-2.05 (1 H, m), 1.65-1.79 (1 H, m), 1.00-1.45 (10H, br m), 0.45-0.57 (2H, m), 0.35-0.45 (2H, m). LCMS (Method A) RT = 7.79 min, M+H+ = 526.18.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-(cyclopropylmethyl)-7-phenyl-1 H-pyrido[2,3- blH ,41oxazin-2(3H)-one
tert-Butyl 1-(4-(1-(cyclopropylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (40 mg, 0.076 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (30 mg, 73% yield). 1H-N R (500 MHz, MeOD) δ 7.67 (1 H, s), 7.43 (2H, d), 7.40 (2H, d), 7.28-7.36 (3H, m), 7.20-7.25 (2H, m), 4.96 (2H, s), 3.98 (2H, d), 2.70-2.80 (2H, m), 2.52-2.63 (2H, m), 2.18- 2.30 (1 H, m), 1.90-2.03 (1 H, m), 1.17-1.30 (1 H, m), 0.55-0.65 (2H, m), 0.42-0.50 (2H, m). LCMS (Method A) RT = 4.44 min, M+2H+ = 427.20.
Example 56: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-2-ylmethyl)-1 H- Pyridor2.3-blM.41oxazin-2(3H)-one
Figure imgf000093_0001
Step 1 : tert-butyl (1 -(4-(2-oxo-7-phenyl-1-(pyridin-2-ylmethyl)-2,3-dihvdro-1H-pyridof2.3- blH ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (75 mg, 0.16 mmol) in dry DMF (1 mL) was added sodium hydride (15 mg, 0.38 mmol) and 2-(bromomethyl)pyridine.HBr (48 mg, 0.19 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 40% EtOAc in cyclohexane) to give the title compound (1 1 mg, 12%). H NMR (500 MHz,CDCI3): 8.49 (1 H, d), 7.63 (1 H, t), 7.42 (1 H, s), 7.27 (1 H, d), 7.19 -7.13 (8H, m), 7.01-6.98 (2H, d), 5.23 (2H, s), 4.92 (2H, s), 4.91 (1 H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.75 -1.70 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-2-ylmethyl)-1 H-pyridor2,3- blH .41oxazin-2(3H)-one
Following the procedure for 1-(( H-imidazol-2-yl)methyl)-6-(4-(1 - aminocyclobutyl)phenyl)-7-phenyl- H-pyrido[2,3-b][ ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (2-oxo-7-phenyl-1-(pyridin-2-ylmethyl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (16 mg, 0.03 mmol) was reacted to afford the title compound (17 mg, 87%). LCMS (Method A): RT = 4.11 min, M-16 = 446. Ή NMR (500 MHz, MeOD): 8.54 (1 H, d), 7.91-7.87 (1 H, t), 7.53 (1 H, d), 7.42-7.35 (6H, m), 7.26-7.21 (3H, m), 7.04 (2H, d), 5.39 (2H, s), 5.1 1 (2H, s), 2.77 -2.72 (2H, m), 2.59-2.53 (2H, m), 2.25 -2.21 (1 H, m), 1.98-1.94 (1 H, m).
Example 57; 6-(4-(1-aminocvclobutyl)phenvn-1-isopropyl-7-phenyl-1H-pyridor213- bin.41oxazin-2(3H)-one
Figure imgf000094_0001
Step 1 : fert-butyl 1 -(4-(1-isopropyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2.3- blfl ^loxazin-e-vDphenvDcyclobutylcarbamate
In a 15 mL reaction tube was added tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (25 mg, 0.053 mmol), potassium carbonate (22 mg, 0.159 mmol) and 2-iodopropane (16 μΙ, 0.159 mmol) in anhydrous N,N-dimethylformamide (500 μΙ) to give an orange suspension. The reaction mixture was stirred at 80°C for one hour. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 ml) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexaneiethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (13 mg, 48% yield). 1H-NMR (500 MHz, CDCI3) δ 7.37 (1 H, s), 7.10-7.28 (9H, m), 4.95 (1 H, br s), 4.65-4.77 (3H, m), 2.05-2.60 (4H, m), 1.92-2.06 (1 H, m), 1.65-1.80 (1 H, m), 1.50 (6H, d), 1.00-1.45 (9H, br m). LCMS (Method A) RT = 7.73 min, M+H+ = 514.15.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1 -isopropyl-7-phenyl-1 H-pyridof2.3- blH ,41oxazin-2(3H)-one
fert-Butyl 1 -(4-(1-isopropyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (7 mg, 0.014 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (7 mg, 97% yield). 1H-N R (500 MHz, MeOD) δ 7.64 (1 H, s), 7.41 (2H, d), 7.36 (2H, d), 7.26-7.32 (3H, m), 7.19-7.26 (2H, m), 4.80 (2H, s), 4.75 (1 H, sep), 2.68-2.80 (2H, m), 2.50-2.62 (2H, m), 2.13-2.29 (1 H, m), 1.89-2.00 (1 H, m), 1.48 (6H, d). LCMS (Method A) RT = 4.20 min, M+2H+ = 415.19.
Example 58: 6-(4-(1-aminocvclobutyl)phenyl)-1-cvclopentyl-7-phenyl-1H-pyridor2.3- blM ,41oxazin-2(3H)-one
Figure imgf000095_0001
Step 1 : tert-butyl 1-(4-(1-cvclopentyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- blf1 ,4loxazin-6-yl)phenyl)cvclobutylcarbamate
In a 15 mL reaction tube was added terf-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol), potassium carbonate (44 mg, 0.318 mmol) and bromocyclopentane (0.034 mL, 0.318 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give an orange suspension. The reaction mixture was stirred at 80°C for one hour. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 3 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (12 mg, 21% yield). 1H-NMR (500 MHz, CDCI3) δ 7.32 (1 H, s), 7.17-7.28 (7H, m), 7.10-7.17 (2H, m), 4.94 (1 H, br s), 4.83 (1 H, quin), 4.72 (2H, s), 2.05-2.60 (6H, m), 1.83-2.03 (5H, m), .54-1.82 (3H, m), 1.00-1.45 (9H, br m). LCMS (Method A) RT = 8.25 min, M+H+ = 540.19. Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-cvclopentyl-7-phenyl-1 H-pyridor2,3- b1f1,41oxazin-2(3H)-one
tert-Butyl 1 -(4-(1 -cyclopentyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (12 mg, 0.022 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (6 mg, 49% yield). 1H-NMR (500 MHz, MeOD) δ 7.60 (1 H, s), 7.40 (2H, d), 7.36 (2H, d), 7.25-7.36 (3H, m), 7.18-7.25 (2H, m), 4.91 (1 H, quin), 4.84 (2H, s), 2.66-2.31 (2H, m), 2.47-2.61 (2H, m), 2.12-2.31 (3H, m), 1.83-2.12 (5H, m), 1.63-1.81 (2H, m). LCMS (Method A) RT = 4.55 min, M+2H+ = 441.19.
Example 59: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(thiazol-4-ylmethyl)-1H- Pyridor2,3-b1H,41oxazin-2(3H)-one
Figure imgf000096_0001
Step 1 : ferf-butyl 1-(4-(2-oxo-7-phenyl-1-(thiazol-4-ylmethvn-2.3-dihvdro-1 H-pyridor2,3- biri,41oxazin-6-yl)phenyl)cvclobutylcarbamate
In a 15 mL reaction tube was added ferf-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol), potassium carbonate (59 mg, 0.424 mmol) and 4-(chloromethyl)thiazole HCI salt (54 mg, 0.318 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give an orange suspension. The reaction mixture was stirred at 80°C for one hour. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (54 mg, 90% yield). 1H-NMR (500 MHz, CDCI3) δ 8.70 (1 H, s), 7.61 (1 H, s), 7.28 (1 H, s), 7.12-7.27 (7H, m), 7.03-7.12 (2H, m), 5.23 (2H, s), 4.99 (1 H, br s), 4.86 (2H, s), 2.10- 2.60 (4H, m), 2.41-2.06 (1 H, m), 1.65-1.79 (1 H, m), 1.00-1.45 (9H, br m). LCMS (Method A) RT = 7.1 1 min, M+H+ = 569.14.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(thiazol-4-ylmethyl)-1 H-pyridof2,3- bin .41oxazin-2(3H)-one
tert-Butyl 1-(4-(2-oxo-7-phenyl-1-(thiazol-4-ylmethyl)-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (54 mg, 0.095 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (30 mg, 54% yield). 1H-NMR (500 MHz, MeOD) δ 8.97 (1 H, s), 7.63 (1 H, s), 7.60 ( H, s), 7.32-7.41 (4H, m), 7.20-7.29 (3H, m), 7.05-7.15 (2H, m), 5.39 (2H, s), 5.03 (2H, s), 2.66-2.88 (2H, m), 2.47- 2.38 (2H, m), 2.13-2.28 (1 H, m), 1.85-1.98 (1 H, m). LCMS (Method A) RT = 3.81 min, M+2H+ = 470.18.
Example 60: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(2,2,2-trifluoroethyl)-1H- pyridor2.3-b 1.41oxazin-2(3H)-one
Figure imgf000097_0001
Step 1 : tert-butyl 1-(4-(2-oxo-7-phenyl-1-(2,2.2-trifluoroethyl)-2.3-dihvdro-1 H-pyridor2.3- biri,41oxazin-6-yl)phenyl)cyclobutylcarbamate
In a 15 mL reaction tube was added fert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol), potassium carbonate (44 mg, 0.318 mmol) and 2-bromo-1 , 1 ,1-trifluoroethane (0.193 mL, 2.121 mmol) in anhydrous N.N-dimethylformamide (1 mL) to give an orange suspension. The reaction mixture was stirred at 80°C for one hour. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na2S04) filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (31 mg, 53% yield). H-NM (500 MHz, CDCI3) δ 7.38 (1 H, s), 7.25-7.37 (7H, m), 7.17-7.23 (2H, m), 5.05 (1 H, br s), 4.97 (2H, s), 4.63 (2H, q), 2.20-2.58 (4H, m), 2.00-2.15 (1 H, m), 1.75-1.88 (1 H, m), 1.10-1.50 (9H, br m). LCMS (Method A) RT = 7.65 min, M+H+ = 554.05.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1 -(2.2,2-trifluoroethyl)-1 H-pyridor2,3- biri.41oxazin-2(3H)-one
iert-Butyl 1-(4-(2-oxo-7-phenyl-1-(2,2,2-trifluoroethyl)-2,3-dihydro- H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (31 mg, 0.056 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (6 mg, 19% yield). 1H-NMR (500 MHz, MeOD) δ 7.74 (1 H, s), 7.41 (2H, d), 7.37 (2H, d), 7.25-7.33 (3H, m), 7.17-7.25 (2H, m), 5.01 (2H, s), 4.90 (2H, q), 2.65-2.82 (2H, m), 2.50-2.63 (2H, m), 2.13- 2.30 (1 H, m), 1.85-2.02 (1 H, m). LCMS (Method A) RT = 4.22 min, M+2H+ = 455.19.
Example 61: 6-(4-(1-aminocvclobutyl)phenyl)-1-cvclobutyl-7-phenyl-1H-pyridor2.3- blM .41oxazin-2(3H)-one
Figure imgf000098_0001
Step V. tert-butyl 1-(4-(1-cvclobutyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyrido[2,3- b1f1 ,4loxazin-6-yl)phenyl)cyclobutylcarbamate
In a 15 mL reaction tube was added tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.106 mmol), potassium carbonate (44 mg, 0.318 mmol) and bromocyclobutane (0.030 mL, 0.318 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give an orange suspension. The reaction mixture was stirred at 80°C for one hour, then 100°C for 2 hours. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage
chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as an off-white solid (15 mg, 27% yield). 1H-NMR (500 MHz, CDCI3) δ 7.25-7.37 (8H, m), 7.16-7.25 (2H, m), 5.03 (1 H, br s), 4.78 (2H, s), 4.53 (1 H, quin), 2.30-2.67 (8H, m), 2.02-2.14 (1 H, m), 1.76-1.97 (3H, m), 1.10-1.50 (9H, br m). LCMS (Method A) RT = 7.76 min, M+H+ = 526.16.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-cvclobutyl-7-phenyl-1 H-pyridoi2,3- bin ,41oxazin-2(3H)-one
iert-Butyl 1 -(4-(1 -cyclobutyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (31 mg, 0.059 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (1 1 mg, 35% yield). H-NMR (500 MHz, MeOD) δ 7.35-7.58 (5H, m), 7.25-7.35 (3H, m), 7.10-7.25 (2H, m), 4.80 (2H, s), 4.59 (1 H, quin), 2.65-2.83 (2H, m), 2.40-2.65 (6H, m), 2.13-2.29 (1 H, m), 1.79-2.02 (3H, m). LCMS (Method A) RT = 4.58 min, M+2H+ = 427.21. Example 62: 3-(6-(4-(1 -aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- pyrido[2,3-b1[1 ,41oxazin-1 -vDpropanenitrile
Figure imgf000100_0001
Step 1 : tert-butyl (1-(4-(1-(2-cvanoethvn-2-oxo-7-prienyl-2.3-dihvdro-1 H-pyridor2.3- b1H ,4]oxazin-6-yl)phenyl)cvclobutyl)carbamate
A mixture of tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (100mg, 0.212 mmol), 3-bromopropanenitrile (85 mg, 0.636 mmol) and potassium carbonate (88mg) in DMF (4ml) was heated at 80 degree for 16 h.
The reaction mixture was partitioned between dichloromethane (20ml) and water (20ml). The organic phase was separated using phase separator and concentrated. The crude product was purified by column (biotage, 25g) eluted with ethyl acetate/cyclohexane to afford product (45mg). Ή NMR (500 MHz, CDCI3): 7.39 (1 H, s), 7.16-7.39 (9H, m), 5.1 (1 H, br), 4.91 (,2H, s), 4.25 (2H, t), 2.83 (2H, t), 2.3-2.5 (4H, m), 2.04-2.09 (1 H, m), 1.79- 1.84 (1 H, m), 1.27-1.36 (9H, br).
Step 2: 3-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- blf 1 ,4]oxazin-1 -vDpropanenitrile
tert-butyl (1 -(4-(1 -(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (15 mg, 0.029 mmol) was dissolved in TFA (0.5 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (1 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (0.5 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid ( 2 mg). 1H NMR (500 MHz, CH3OD) 7.77 (1 H, s), 7.43 (2H, d), 7.39 ( 2H, d), 7.3 (3H, m), 7.27 (2H, m), 4.99 (2H, s), 4.37 (2H, t), 2.94 (2H, t), 2.73-2.79 (2H, m), 2.55-2.61 (2H, m), 2.2-2.27 (1 H, m), 1.93- 1.99 (1 H, m). LCMS (Method A): RT = 3.73 min, M+H+ = 425.
Example 63: 1-(4-(1-methyl-8-phenyl-4H-pyridor2.3-biri,2.4ltriazolor4,3- diri.41oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000101_0001
Step 1 : Tert-butyl (1-(4-(1 -methyl-8-phenyl-4H-pyridor2.3-biri,2.4ltriazolor4.3- d]f ,4loxazin-7-yl)phenyl)cvclobutyl)carbamate
A solution of (E)-tert-butyl (1 -(4-(2-hydrazono-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.103 mmol) in 1 ,1 ,1- trimethoxyethane (1 ml) was heated to 150 °C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n-hexanes) to give the title compound (24 mg, 47%). LCMS (Method A): RT = 6.71 min, M+1 = 510. Ή NMR (500 MHz, CDCI3): 7.84 (1 H, s), 7.37-7.33 (5H, m), 7.30- 7.27 (2H, m), 7.24- 7.19 (2H, m), 5.59 (2H, s), 2.87 (3H, s), 2.54-2.40 (4H, m), 2.13-2.02 (1 H, m), 1.87-1.78 (1 H, m), 1.45-1.27 (9H, br s).
Step 2: 1-(4-(1-methyl-8-phenyl-4H-pyridor2,3-biri,2.4ltriazolor4.3-diri,4loxazin-7- vDphenvQcvclobutanamine
Tert-butyl (1-(4-(1-methyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (1 1 mg, 0.022 mmol) was dissolved in TFA (1 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (5 mg, 55% yield). LCMS (Method A): RT = 3.71 min, M+1 =411. 1H-NMR (500 MHz, CD3OD) δ 8.11 (1 H, s), 7.49- 7.44 (2H, d), 7.42- 7.37 (2H, d), 7.35- 7.30 (3H, m), 7.29-7.25 (2H, m), 5.63 (2H, s), 2.84 (3H, s), 2.80-2.70 (2H, m), 2.61-2.51 (2H, m), 2.28-2.17 (1 H, m), 2.01-1.88 (1 H, m).
Example 64: 4-(6-(4-(1 -aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof2,3-b1M .41oxazin-1 -vDbutanenitrile
Figure imgf000102_0001
Step 1 : tert-butyl (1 -(4-(1-(3-cvanopropyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- blf1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1-(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-bj[1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (1 OOmg, 0.212 mmol) was reacted with 4-bromobutanenitrile (94mg, 0.636 mmol) to afford the title compound (68mg. Ή NMR (500 MHz, CDCI3): 7.20-7.32 (10H, m), 5.1 (1 H, br), 4.90 (2H, s), 4.11 (2H, m), 2.44-2.50 (6H, m), 2.05-2.13 (3H, m), 1.79-1.84 (1 H, m), 1.27-1.36 (9H, br).
Step 2: 4-(6-(4-(1 -aminocvclobutvnphenyl)-2-oxo-7-phenyl-2,3-dihvdro-1H-pyrido[2.3- blf 1 ,4loxazin-1 -vDbutanenitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(3- cyanopropyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (35mg, 0.065 mmol) was reacted to afford the title compound (28mg). 1H NMR (500 MHz, CH3OD) 7.62 (1 H, s), 7.41 (2H, d), 7.39 (2H, d), 7.3 (3H, m), 7.25 (2H, m), 4.98 (2H, s), 4.19 (2H, t), 2.73-2.79 (2H, m), 2.55-2.61 (4H, m), 2.22-2.25 (1 H, m), 2.10-2.25 (2H, m), 1.95- 1.99 (1 H, m). LCMS (Method A): RT = 4.01 min, M+H+ = 440.2. Example 65: e-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(tetrahvdro-2H-pyran-4- v))-1H-Pyridor2.3-b1i1.41oxazin-2(3H)-one
Figure imgf000103_0001
Step 1 : tert-butyl (1-(4-(6-hvdroxy-3-phenyl-5-((tetrahvdro-2H-pyran-4-yl)amino)pyridin-2- vDphenvDcyclobutvDcarbamate
To a solution of tert-butyl (1-(4-(5-amino-6-hydroxy-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (200 mg, 0.46 mmol) in dry DCE (2ml) was added dihydro-2H-pyran-4(3H)-one (88 mg, 0.88 mmol), AcOH (0.16 ml, 2.78 mmol), sodium triacetoxyborohydride (275 mg, 1.30 mmol) under nitrogen. The resulting mixture was stirred for 3h at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with AcOEt (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was used in the next step without any purification.
Step 2: tert-butyl (1-(4-(2-oxo-7-phenyl-1-(tetrahvdro-2H-pyran-4-yl)-2,3-dihvdro-1 H- Pyrido[2,3-bl[1 ,4loxazin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(6-hydroxy-3-phenyl-5-((tetrahydro-2H-pyran-4- yl)amino)pyridin-2-yl)phenyl)cyclobutyl)carbamate (240 mg, 0.46 mmol) in dry THF (11 mL) was added N,N-diisopropylethylamine (0.41 mL, 2.32 mmol). 2-chloroacetyl chloride (0.19 mL, 2.32 mmol) was added dropwise at 0C. The resulting mixture was stirred overnight at room temperature. A saturated solution of NaHC03 (8 mL) was added and the mixture was stirred under reflux for 2 hours. After allowing to cool to room
temperature, the organic phase was separated and the aqueous phase extracted with AcOEt (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 50% EtOAc in cyclohexane) to give the title compound (154 mg, 59%). Ή NMR (500 Hz,CDCI3): 7.45 (1 H, s), 7.24 -7.22 (5H, m), 7.19-7.17 (2H, m), 7.14-7.12 (2H, m), 4.94 (1 H, s), 4.71 (2H, s), 4.60-4.55 (1 H, m), 4.07-4.03 (2H, m), 3.45 (2H, t), 2.63-2.54 (2H, m), 2.52-2.20 (4H, m), 2.00-1.95 (1H, m), 1.79 -1.65 (3H, m), 1.40-1.10 (9H, br).
Step 3: 6-(4-(1-arninocvclobutyl)phenyl)-7-phenyl-1-(tetrahvdro-2H-pyran-4-yl)-1 H- pyridof2.3-bin ,41oxazin-2(3H)-one
Following the procedure for 1 -((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][ ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (2-oxo-7-phenyl-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (55 mg, 0.10 mmol) was reacted to afford the title compound (53 mg, 93%). LCMS (Method A): RT = 4.19 min, M+1 =457. 1H NMR (500 MHz, MeOD): 7.76 (1 H,s), 7.44-7.38 (4H,m), 7.33-7.31 (3H, m), 7.27-7.25 (2H, m), 4.90 (2H, s), 4.59-4.53 (1 H, m), 4.09-4.06 (2H, m), 3.60 (2H, t), 2.80-2.72 (4H, m), 2.59-2.53 (2H, m), 2.25 -2.21 (1 H, m), 1.98-1.94 (1 H, m), 1.81-1.79 (2H,m).
Example 66: 5-(6-(4-(1 -aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2.3-dihydro-1 H- pyridor2.3-biri ,41oxazin-1 -vQpentanenitrile
Figure imgf000104_0001
Step 1 : tert-butyl (1-(4-(1-(4-cvanobutyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2,3- blf1.41oxazin-6-yl)phenyl)cvclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1-(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (1 OOmg, 0.212 mmol) was reacted with 5-bromopentanenitrile (103mg, 0.636 mmol) to afford the title compound (82mg. Ή NMR (500 MHz, CDCI3): 7.20-7.33 (10H, m), 5.03 (1 H, br), 4.91 (2H, s), 4.05 (2H, t), 2.45-2.51 (6H, m), 2.05-2.10 (1 H, m), 1.88-1.94 (2H, m), 1.67- 1.85 (3H, m), 1.27-1.38 (9H, br). Step 2: 5-(6-(4-(1-aminocvclobutyl)phenyl)-2-oxo-7-phenyl-2.3-dihyclro-1 H-pyridof2.3- b1f1 ,41oxazin-1-yl)pentanenitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1 -yl)propanenitrile, tert-butyl (1 -(4-(1 -(4-cyanobutyl)- 2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (25mg, 0.045 mmol) was reacted to afford the title compound (14.5mg). 1H NMR (500 MHz, CH3OD) 7.60 (1 H, s), 7.42 (2H, d), 7.39 ( 2H, d), 7.31 (3H, m), 7.25 (2H, m), 4.97 (2H, s), 4.1 1 (2H, t), 2.73-2.79 (2H, m), 2.54-2.61 (4H, m), 2.22-2.25 (1 H, m), 1.95-1.97 (1 H, m), 1.84-1.89 (2H, m), 1.74-1.78 ( 2H, m). LCMS (Method A): RT = 4.19 min, M+H+ = 453.5.
Example 67: 1-allyl-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1H-pyridor2,3- bin.41oxazin-2(3H)-one
Figure imgf000105_0001
Step 1 : tert-butyl 1-(4-(1-allyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3-biri,4loxazin-6- vDphenyQcyclobutylcarbamate
In a sealed tube was added tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100mg, 0.212 mmol) in DMF to give a brown solution, bromocyclopropane (76.9652 mg, 0.636 mmol) and potassium carbonate were added. The reaction mixture was heated at 80 degree for 16 h. The reaction mixture was then heated under microwave 100 degree for 2h and 130 degree for 8h. The reaction mixture was partitioned between dichloromethane (20ml) and water (20ml). The organic phase was separated using phase separator and concentrated. The crude product was purified by column (biotage, 25g) eluted with ethyl acetate/cyclohexane to afford product (12mg). 1H NMR (500 MHz, CDCI3) 7.17-7.23 (m, 8H), 7.07-7.10 (m, 2H), 5.77-5.84 (m, 1 H), 5.18-5.24 (m, 2H), 4.51 (d, 2H), 2.2-2.5 (m, 4H), 1.9-2.0 (m, 1 H), 1.69- 1.79 (m, 1 H), 1.1-1.35 (br, 9H). LCMS (Method A): RT = 7.9 min, M+H+ = 512.17. Step 2: 1-allyl-6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1 H-pyridor2,3-bl[1 ,41oxazin- 2(3H)-one
to a round bottomed flask containing tert-butyl 1-(4-(1-allyl-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (6.5 mg, 0.013 mmol) was added trifluoroacetic acid (0.5ml). The resulting solution was stirred for 60 seconds then concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (1 ml) and concentrated to dryness three times to give the trifluoroacetic acid salt of the product (3.55mg) (3.55mg). 1H N R (500 MHz, CD3OD) 7.49 (s, 1 H), 7.38-7.43 (m, 4H), 7.29-7.30 (m, 3H), 7.18-7.20 (m, 2H), 5.9-6.0 (m, 1 H), 5.25-5.35 (m, 2H), 5.01 (s, 2H), 4.68 (d, 2H), 2.7-2.8 (m, 2H), 2.55-2.65 (m, 2H), 2.21 -2.27 (m, 1 H), 1.9-2.0 (m, 1 H), LCMS (Method A): RT = 4.38 min, M+H* = 413.18.
Example 68: 6-(4-(1-aminocvclobutvnphenyl)-1-(2-hvdroxyethyl -7-phenyl-1 H-
Figure imgf000106_0001
Step 1 tert-butyl (1-(4-(1-(2-hvdroxyethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyrido[2,3- bl[1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1 -(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100mg, 0.212 mmol) was reacted with 2-bromoethanol (80mg, 0.636 mmol) to afford the title compound (75mg. Ή NMR (500 MHz, CDCI3): 7.47 (s, 1 H), 7.23-7.29 (m, 7H), 7.16-7.19 (m, 2H), 5.04 (br, 1 H), 4.89 (s, 2H), 4.12 (m, 2H), 3.94 (m, 2H), 2.37-2.48 (m, 4H), 2.37 (t, 1 H), 2.05 (m, 1 H), 1.81 (m, 1 H), 1.2-1.43 (br, 9H).
Step 2: 6-(4-(1-aminocyclobutyl)phenyl)-1-(2-hvdroxyethyl)-7-phenyl-1 H-pyrido[2,3- blM ,41oxazin-2(3H)-one
to a round bottomed flask containing tert-butyl 1-(4-(1-(2-hydroxyethyl)-2-oxo-7-phenyl- 2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (25mg, 0.048 mmol) was added dichloromethane (2.5ml) followed with hydrochloric acid (2.5ml, 2M in diethyl ether). The resulting solution was stirred for 16 h. The precipitate was filtered and washed with ether (2X10ml) and dried to afford the product (15.5mg). 1H NMR (500 MHz, CH3OD) 7.78 (s, 1H), 7.42 (d, 2H), 7.38 (d, 2H), 7.30 (m, 3H), 7.22 (m, 2H), 4.96 (s, 2H), 4.16 (t, 2H), 3.86 (t, 2H), 2.73-2.79 (m, 2H), 2.54-2.60 (m, 2H), 2.21-2.26 (m, 1 H), 1.94- 1.99 (m, 1H). LCMS (Method D): RT = 1.99 min, M+H+ = 416.2.
Example 69: 1-(4-(8-phenyl-1-propyl-4H-pyridor2,3-b1[1,2.41triazolo[4,3- diri,41oxazin-7-yl)phenyl)cvclobutanamine
Figure imgf000107_0001
Step 1 : Tert-butyl (1-(4-(8-phenyl-1-propyl-4H-pyridor2.3-biri .2,41triazolor4.3- dlf1 ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A solution of (E)-tert-butyl (1-(4-(2-hydrazono-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (60 mg, 0.124 mmol) in 1 ,1 ,1 - trimethoxybutane (1 ml) was heated to 150 °C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n-hexanes) to give the title compound (30 mg, 45%). LCMS (Method A): RT = 7.34 min, M+1= 538. 1H NMR (500 MHz, CDCI3): 7.80 (1 H, s), 7.36-7.32 (5H, m), 7.31- 7.27 (2H, m), 7.23- 7.19 (2H, m), 5.57 (2H, s), 3.08 (2H, t), 2.55-2.40 (4H, m), 2.15-2.03 (1 H, m), 2.00 (2H, q), 1.89-1.77 (1 H, m), 1.41-1.29 (9H, br), 1.12 (3H, t).
Step 2: 1-(4-(8-phenyl-1-propy|-4H-pyridor2.3-b1[1.2,41triazolor4.3-dlf1.4loxazin-7- yl)phenyl)cyclobutanamine
Tert-butyl (1-(4-(8-phenyl-1-propyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (39 mg, 0.073 mmol) was dissolved in TFA (1 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (10 mg, 30% yield). LCMS (Method A): RT = 3.85 min, M+1 =439. H-NMR (500 MHz, CD3OD) δ 8.09 (1 H, s), 7.50 (2H, d), 7.43 (2H, d), 7.37- 7.34 (3H, m), 7.31-7.27 (2H, m), 5.66 (2H, s), 3.2 (2H, t), 2.83-2.71 (2H, m), 2.64-2.54 (2H, m), 2.32-2.19 (1 H, m), 2.02-1.90 (1 H, m + 2H, t), 1.16-1.09 (3H, t).
Example 70: 1-(4-(1-(methylsulfonyl)-7-phenyl-2,3-dihydro-1 H-pyridor2,3- biri.41oxazin-6-yl)phenyl)cvclobutanamine
Figure imgf000108_0001
Step 1 : tert-butyl f1-(4-(1-(methylsulfonyl)-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- b1f1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (50 mg, 0.1 1 mmol) in dry DCM (1 ml) was added triethyl amine (46 μΙ, 0.33 mmol) and methanesulfonyl chloride (26 μΙ, 0.33 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. A saturated solution of NaHC03 was added and the mixture was extracted with dichloromethane (3 x 10ml) using a phase separator (Isolute® SPE). The combined organic phases were concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 50% EtOAc in cyclohexane) to give the title compound (45 mg, 77%). H NMR (500 MHz,CDCI3): 8.00 (1 H, s), 7.25 (2H, d), 7.19- 7.17 (5H, m), 7.1 1-7.10 (2H,m), 4.95 (1 H, s), 4.43 (2H, t), 3.90 (2H,t), 2.55-2.25 (7H, m), 2.06-1.98 (1 H, m), 1.82-1.73 (1 H, m), 1.44-1.15 (9H, br). Step 2: 1-(4-(1-(rnethylsulfonyl)-7-phenyl-2.3-dihvdro-1 H-pyridor2,3-b1f1.41oxazin-6- vDphenvOcyclobutanamine
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (1-(methylsulfonyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (45 mg, 0.08 mmol) was reacted to afford the title compound (44 mg, 95%). LCMS (Method A): RT = 2.34 min, M+1 = 437. Ή NMR (500 MHz, MeOD): 8.15 (1 H, s), 7.43-7.38 (4H,m), 7.30-7.28 (3H, m), 7.21-7.19 (2H, m), 4.57 (2H, t), 4.01 (2H, t), 3.19 (3H, s), 2.79 -2.73 (2H, m), 2.60 -2.54 (2H, m), 2.24-2.21 (1 H, m), 1.99-1.94 (1 H, m).
Example 71 : 6-(4-(1-aminocyclobutyl)phenyl)-3-(2-hvdroxyethyl)-7-phenyl-1 H- pyridor2.3-biri.41oxazin- -one
Figure imgf000109_0001
Step 1 : tert-butyl (1 -(4-(3-(2-hvdroxyethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3- biri,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
In a 100ml_ round bottom flask was added tert-butyl 1-(4-(5-amino-6-oxo-3-phenyl-1 , 6- dihydropyridin-2-yl)phenyl)cyclobutylcarbamate (0.5 g, 1.159mmol) in THF (Volume: 25ml) to give a red suspension followed with the addition of DIPEA (1.0ml). 2, 4- dibromobutanoyl chloride (0.735g, 2.78mmol) in THF (5ml) was added in drop wise at 0 degree. The resulted mixture was stirred at room temperature for 20 hours. The reaction mixture was transferred to a 250ml round bottom flask and sodium bicarbonate saturated solution (40 ml) and sodium bicarbonate (1g) was added. The resulted mixture was heated to reflux for 24 h. The reaction mixture was cooled down to room temperature. The organic phase was separated. The aqueous phase was extracted with ethyl acetate (45ml). The organic phases were combined and washed with water (30mlX2), brine (25ml) then concentrated at reduced pressure. The residue was suspended in DCM (10ml). The precipitate was collected by filtration and washed with DCM (5 ml) to afford crude product which was purified by biotage column (25g silicon) to afford tert-butyl 1-(4- (3-(2-hydroxyethyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate(45mg) and tert-butyl 1-(4-(7-phenyl-3,3a-dihydro-2H- furo[2,3-e]pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (0.35g). Ή NMR (500 MHz, CD3OD): 7.50 (s, 1 H), 7.31 (m, 2H), 7.26 (m, 5H), 7.16 (m, 2H), 5.38 (dd, 1 H), 4.72 (t, 1H), 4.55 (m, 1H), 2.92 (m, 1H), 2.67 (m, 1 H), 2.43-2.47 (m, 4H), 2.07 (m, 1H), 1.85 (m, 1 H), 1.23- .39 (br, 9H). LCMS (Method A): RT = 6.89 min, M+H+ = 498.12. A solution of tert-butyl 1-(4-(7-phenyl-3,3a-dihydro-2H-furo[2,3-e]pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutylcarbamate (0.35g, 0.703 mmol) in DCM (15ml) and acetic acid (10ml) was stirred at room temp for 48h.The reaction mixture was concentrated and partioned between ethyl acetate (40ml) and water (30ml). The separated organic phase was washed with water (2X25ml), brine and concentrated to give product 0.36g (45mg). 1H NMR (500 MHz, CD3OD): 7.23-7.31 (m, 8H), 7.16 (m, 2H), 5.05 (dd, 1H), 3.91 (m, 1 H), 3.85 (m, 1 H), 2.45 (m, 4H), 2.26 (m, 1 H), 2.15 (m, 1 H), 2.07 (m, 1 H), 1.86 (m, 1 H), 1.23-1.39 (br, 9H). LCMS (Method A): RT = 5.92 min, M+H+ = 516.04.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-3-(2-hvdroxyethyl)-7-phenyl-1 H-pyridof2,3- blf .4loxazin-2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(4-cyanobutyl)- 2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (15mg, 0.029 mmol) was reacted to afford the title compound (8.5mg). Ή NMR (500 MHz, CD3OD): 7.28-7.42 (m, 8H), 7.18 (m, 2H), 5.08 (dd, 1H), 3.84-3.93 (m, 2H), 2.73-2.79 (m, 2H), 2.55-2.61 (m, 2H), 2.25-2.29 (m, 2H), 2.15-2.24 (m, 1H), 1.95-1.97 (m, 1H). LCMS (Method A): RT = 3.49 min, M+H+ = 416.18.
Example 72: 6-(4-(1 -aminocvclobutyl)phenyl)-4-methyl-7-phenyl-3,4- dihvdropyridor2.3-blpyrazin-2(1H)-one
Figure imgf000110_0001
Step 1 : methyl 2-((6-(4-(1-((fer -butoxycarbonyl)amino)cvclobutyl)phenyl)-3-nitro-5- phenylpyridin-2-yl)(methyl)amino)acetate
In a microwave vial were added fert-butyl (1-(4-(6-chloro-5-nitro-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (200 mg, 0.417 mmol), methyl 2-(methylamino)acetate hydrochloride (58 mg, 0.417 mmol) and triethylamine (58 μΙ, 0.4 7 mmol) in methanol (2 ml). The solution was heated to 80°C under microwave irradiation for 1 hour. The reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 100:0 to 0: 100) to afford the title compound (103 mg, 65%). 1H NMR (500 MHz, CDCI3): 8.29 (1 H, s), 7.25-7.38 (7H, m), 7.15-7.23 (2H, m), 5.05 (1 H, br s), 4.39 (2H, s), 3.83 (3H, s), 3.1 1 (3H, s), 2.25-2.65 (4H, m), 2.05-2.20 (1 H, m), 1.80-1.95 (1 H, m), 1.10-1.50 (9H, br).
Step 2: tert-butyl (1-(4-(4-methyl-2-oxo-7-phenyl-1 ,2,3,4-tetrahvdropyridor2,3-blpyrazin-6- yl)phenyl)cyclobutyl)carbamate
In a glass autoclave were added methyl 2-((6-(4-(1-((iert- butoxycarbonyl)amino)cyclobutyl)phenyl)-3-nitro-5-phenylpyridin-2- yl)(methyl)amino)acetate (800 mg, 1.46 mmol) and 10% palladium on carbon (300 mg, 0.28 mmol) in THF (100 ml). The solution was hydrogenated at room temperature under 1.5 atm hydrogen for 20 hours. The reaction mixture was filtered through celite, then concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 100:0 to 0: 100) to afford the title compound (270 mg, 38%). 'H NMR (500 MHz, CDCI3): 7.35 (2H, d), 7.20- 7.33 (5H, m), 7.15 (2H, d), 6.98 (1 H, s), 5.05 (1 H, br s), 4.11 (2H, s), 3.18 (3H, s), 2.20- 2.70 (4H, br m), 2.00-2.14 (1 H, m), 1.77-1.87 (1 H, m), 1.10-1.50 (9H, br).
Step 3: 6-(4-( 1 -aminocvclobutyl)phenyl)-4-methyl-7-phenyl-3,4-dihvdropyridor2,3- b]pyrazin-2(1 H)-one
Following the procedure for 1 -((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl 1 -(4- (4-methyl-2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[2,3-b]pyrazin-6- yl)phenyl)cyclobutylcarbamate (29 mg, 0.060 mmol) was reacted to afford the title compound (29 mg, 97%). LCMS (Method A): RT = 3.69 min, M+2H+ = 386. 1H NMR (500 MHz, MeOD): 7.44 (2H, d), 7.32 (2H, d), 7.20-7.28 (3H, m), 7.12 (2H, d), 7.02 (1 H, s), 4.08 (2H, s), 3.11 (3H, s), 2.70-2.80 (2H, m), 2.50-2.63 (2H, m), 2.13-2.27 (1 H, m), 1.88- 2.02 (1 H, m). - Ill -
Example 73: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-phenyl-1H-pyridof3.2- e]f1 3,41oxathiazine 2,2-dioxide
Figure imgf000112_0001
Step 1 : fe/f-butyl (1 -(4-(2,2-dioxido-7-phenyl-1 H-pyridof3,2-e1f1.3,4loxathiazin-6- vQphenvQcvclobutvOcarbamate
In a 40 mL reaction tube was added fert-butyl 1-(4-(5-amino-6-oxo-3-phenyl-1 ,6- dihydropyridin-2-yl)phenyl)cyclobutylcarbamate (500 mg, 1.159 mmol) in anhydrous pyridine (5 mL) to give a yellow solution. After stirring at room temperature for 15 minutes, chloromethanesulfonyl chloride (126 μΙ, 1.390 mmol) was added and the resulting solution stirred at room temperature for 3 hours. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in anhydrous methanol (5 mL), followed by the addition of potassium carbonate (641 mg, 4.63 mmol) and heating to 60°C overnight. The reaction mixture was allowed to cool to room temperature and concentrated to dryness under reduced pressure. The residue was redissolved in ethyl acetate (5 mL) and washed with 50:50 waterbrine (2 x 5 mL), dried over Na2S0 , filtered and concentrated to dryness under reduced pressure to give a brown oil. This was purified by Biotage chromatography (cyclohexane:ethyl
acetate:dichloromethane:methanol, gradient elution from 80:10:10:0 to 0:90:10:0 then to 0:90: 10) to give the desired product as an off-white solid (330 mg, 56% yield). H-NMR (500 MHz, de-Acetone) δ 7.35 (1 H, s), 7.23-7.32 (7H, m), 7.16-7.32 (2H, m), 5.39 (2H, s), 2.30-2.60 (4H, m), 2.00-2.12 (1 H, m), 1.75-1.88 (1 H, m), 1.05-1.40 (9H, br m). LCMS (Method A) RT = 6.68 min, M+H+ = 508.01.
Step 2: fe/t-butyl (1-(4-(1-methyl-2,2-dioxido-7-phenyl-1 H-pyridof3.2-ein .3.41oxathiazin- 6-yl)phenyl)cvclobutyl)carbamate
In a 15 mL reaction tube was added tert-butyl (1 -(4-(2,2-dioxido-7-phenyl-1 H-pyrido[3,2- e][1 ,3,4]oxathiazin-6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.099 mmol), potassium carbonate (41 mg, 0.296 mmol) and iodomethane (7.5 μί, 0.1 18 mmol) in anhydrous Ν,Ν-dimethylformamide (1 mL) to give an orange suspension. The reaction mixture was stirred at room temperature for 2 hours, then diluted with saturated sodium bicarbonate solution (5 ml.) and extracted into dichloromethane (3 x 3 ml_). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography
(cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as a colourless oil (50 mg, 97% yield). 1H-NMR (500 MHz, CDCI3) δ 7.24-7.47 (8H, m), 7.16-7.23 (2H, m), 5.23 (2H, s), 5.05 (1H, br s), 3.36 (3H, s), 2.20-2.65 (4H, m), 2.01-2.15 (1 H, m), 1.77-1.90 (1 H, m), 1.10-1.50 (9H, br m). LCMS (Method A) RT = 7.16 min, M+H+ = 522.01.
Step 3: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-phenyl-1 H-pyridof3,2- e]f1 ,3,41oxathiazine 2,2-dioxide
tert-Butyl (1-(4-(1-methyl-2,2-dioxido-7-phenyl-1 H-pyrido[3,2-e][1,3,4]oxathiazin-6- yl)phenyl)cyclobutyl)carbamate (50 mg, 0.096 mmol) was dissolved in TFA (3 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (32 mg, 62% yield). 1H-NMR (500 MHz, MeOD) δ 7.58 (1 H, s), 7.41 (2H, d), 7.37 (2H, d), 7.25-7.33 (3H, m), 7.17-7.25 (2H, m), 5.45 (2H, s), 3.35 (3H, s), 2.66-2.78 (2H, m), 2.46-2.57 (2H, m), 2.12-2.25 (1 H, m), 1.85- 1.97 (1 H, m). LCMS (Method A) RT = 3.98 min, M+2H+ = 423.13.
Example 74; 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-3-yl)-1H- pyridor3,2-e1H ,3.41oxathiazine 2,2-dioxide
Figure imgf000113_0001
Step 1 : te/t-butyl (1-(4-(2.2-dioxido-7-phenyl-1-(pyridin-3-yl)-1 H-pyridof3.2- e1f1 ,3,4loxathiazin-6-yl)phenyl)cvclobutyl)carbamate
In a 15 mL reaction tube was added fert-butyl (1-(4-(2,2-dioxido-7-phenyl-1H-pyrido[3,2- e][1 ,3,4]oxathiazin-6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.099 mmol), pyridin-3- ylboronic acid (24 mg, 0.197 mmol), copper(ll) acetate (27 mg, 0.148 mmol) and 5A molecular sieves (30 mg) in anhydrous dichloromethane (1 ml). To this was added anhydrous pyridine (16 μΙ, 0.197 mmol) and the resulting black suspension stirred at room temperature under air for 24 hours. Further pyridine (1 mL) was added and the resulting black solution stirred at room temperature for 24 hours. Further pyridin-3- ylboronic acid (24 mg, 0.197 mmol) and copper (II) acetate (27 mg, 0.148 mmol) were added and the mixture stirred at room temperature for 72 hours. The reaction mixture was concentrated to dryness under reduced pressure and the residue purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as an off-white solid (40 mg 70% yield). 1H-NMR (500 MHz, CDCI3) δ 8.67-8.76 (2H, m), 7.89 (1 H, d), 7.54 (1 H, dd), 7.20-7.35 (7H, m), 7.06 (2H, d), 7.03 (1 H, s), 5.38 (2H, s), 5.07 (1 H, br s), 2.20-2.60 (4H, m), 2.00-2.15 (1 H, m), 1.78-1.90 (1 H, m), 1.10-1.50 (9H, br s). LCMS (Method A) RT = 6.98 min, M+H+ = 584.99.
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-3-yl)-1 H-pyridor3,2- e1f1 ,3,41oxathiazine 2,2-dioxide
fert-Butyl (1-(4-(2,2-dioxido-7-phenyl-1-(pyridin-3-yl)-1 H-pyrido[3,2-e][1 ,3,4]oxathiazin-6- yl)phenyl)cyclobutyl)carbamate (40 mg, 0.068 mmol) was dissolved in TFA (3 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (23 mg, 56% yield). 1H-NMR (500 MHz, MeOD) δ 8.70 (1 H, d), 8.65 (1 H, dd), 8.00 (1H, dd), 7.61 (1 H, dd), 7.44 (2H, d), 7.38 (2H, d), 7.17-7.25 (3H, m), 7.09 (1 H, s), 7.06 (2H, d), 5.67 (2H, s), 2.68-2.77 (2H, m), 2.50- 2.59 (2H, m), 2.14-2.25 (1 H, m), 1.87-2.00 (1 H, m). LCMS (Method A) RT = 4.00 min, M+2H+ = 486.07. Example 75: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-(pyridin-3-yl)-1 H- pyridor2.3-biri.41oxazin-2(3 -one
Figure imgf000115_0001
Step 1 : teft-butyl 1-(4-(2-oxo-7-phenyl-1-(pyridin-3-yl)-2.3-dihvdro-1 H-Pyridor2,3- b1f1 ,4loxazin-6-yl)phenyl)cvclobutylcarbamate
In a 15 mL reaction tube was added tert-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100 mg, 0.212 mmol), pyridin- 3-ylboronic acid (52 mg, 0.424 mmol), copper(ll) acetate (58 mg, 0.318 mmol) and 5A molecular sieves (60 mg) in anhydrous dichloromethane (2 ml). To this was added anhydrous pyridine (34 μΙ, 0.424 mmol) and the resulting blue suspension stirred at room temperature under air for 24 hours. Further pyridine (1 mL) was added and the resulting deep blue solution stirred at room temperature for 24 hours. Further pyridin-3-ylboronic acid (52 mg, 0.424 mmol) and copper(ll) acetate (58 mg, 0.318 mmol) were added and the mixture stirred at room temperature for 72 hours. The reaction mixture was concentrated to dryness under reduced pressure and the residue purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 88: 12 to 0:100) to give the desired product as an off-white solid (32 mg 28% yield). 1H-NMR (500 MHz, CDCI3) δ 8.50-8.80 (2H, br m), 7.71 (1 H, d), 7.50 (1 H, br s), 7.09-7.26 (7H, m), 6.92-7.02 (2H, d), 6.71 (1 H, s), 5.00 (3H, s), 2.10-2.55 (4H, m), 1.92-2.05 (1 H, m), 1.66-1.80 (1 H, m), 1.00- 1.45 (9H, br m). LCMS (Method A) RT = 6.61 min, M+H+ = 549.09.
Step 2: 6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(pyridin-3-yl)-1 H-pyridof2,3- blf 1 ,4loxazin-2(3H)-one
tert-Butyl 1 -(4-(2-oxo-7-phenyl-1 -(pyridin-3-yl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (30 mg, 0.055 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (15 mg, 49% yield). 1H-NMR (500 MHz, MeOD) δ 8.62-8.77 (2H, m), 7.98 (1 H, d), 7.68 (1H, dd), 7.36 (2H, d), 7.39 (2H, d), 7.15- 7.24 (3H, m), 6.97-7.05 (2H, m), 6.77 (1 H, s), 5.14 (2H, s), 2.66-2.77 (2H, m), 2.46-2.58 (2H, m), 2.13-2.25 (1 H, m), 1.86-1.97 (1 H, m). LCMS (Method A) RT = 3.71 min, M+2H+ = 450.20.
Example 76: 2-(6-(4-(1-aminocvclobutyl)phenyl)-2,2-dioxido-7-phenyl-1H- pyridor3,2-eiri.3,41oxathiazi -1-yl)acetonitrile
Step 1 : terf-butyl (1-(4-(1-(cvanomethyl)-2,2-dioxido-7-phenyl-1 H-pyridor3,2- e1f1 ,3.41oxathiazin-6-yl)phenyl)cvclobutyl)carbamate
In a 15 mL reaction tube was added fert-butyl (1-(4-(2,2-dioxido-7-phenyl-1 H-pyrido[3,2- e][1 ,3,4]oxathiazin-6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.099 mmol), potassium carbonate (41 mg, 0.296 mmol) and 2-bromoacetonitrile (0.021 mL, 0.296 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give a yellow suspension. The reaction mixture was stirred at room temperature for one hour. The reaction mixture was allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (5 mL) and extracted into dichloromethane (3 x 3 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as a colourless oil (32 mg, 59% yield). 1H-NMR (500 MHz, CDCI3) δ 8.04 (1 H, s), 7.25-7.38 (7H, m), 7.17-7.25 (2H, m), 5.32 (2H, s), 5.05 (1 H, br s), 4.73 (2H, s), 2.20-2.65 (4H, m), 2.00-2.18 (1 H, m), 1.77-1.90 (1 H, m), 1.05-1.55 (9H, br m). LCMS (Method D) RT = 1.60 min, M+H+ = 547.10. Step 2: 2-(6-(4-( 1 -aminocvclobutyl)phenyr)-2.2-dioxido-7-phenyl-1 H-pyridor3,2- elf1.3,4loxathiazin-1-yl)acetonitrile
terf-Butyl (1-(4-(1-(cyanomethyl)-2,2-dioxido-7-phenyl-1 H-pyrido[3,2-e][1 ,3,4]oxathiazin- 6-yl)phenyl)cyclobutyl)carbamate (32 mg, 0.059 mmol) was dissolved in TFA (3 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (16 mg, 49% yield). 1H-NMR (500 MHz, eOD) δ 7.83 (1 H, s), 7.44 (2H, d), 7.39 (2H, d), 7.26-7.35 (3H, m), 7.20-7.26 (2H, m), 5.53 (2H, s), 5.01 (2H, s), 2.68-2.78 (2H, m), 2.50-2.60 (2H, m), 2.14-2.27 (1 H, m), 1.87- 1.99 (1 H, m). LCMS (Method A) RT = 4.00 min, M+2H+ = 448.12.
Example 77: 6-(4-(1-aminocvclobutyl)phenyl)-3-(2-hvdroxyethyl)-1-methyl-7-phenyl- 1 H-pyridoi2.3-biri ,41oxa -one
Figure imgf000117_0001
Step 1 : tert-butyl (1-(4-(3-(2-hvdroxyethyl)-1-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H- pyrido[2,3-b1f1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
A solution of tert-butyl 1 -(4-(3-(2-hydroxyethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (40mg, 0.078 mmol) in DMF(1 ml) was added potassium carbonate (32mg) and methyl iodide (13.2mg, 0.093mml) were added. The reaction mixture was stirred at room temperature for 4 h.
The reaction mixture was partitioned between dichloromethane (20ml) and water (20ml). The organic phase was separated using phase separator and concentrated. The crude product was purified by column (biotage, column size: 25g) eluted with ethyl
acetate/cyclohexane to afford product (18mg) Ή NMR (500 MHz, CDCI3): 7.25-7.30 (m, 8H), 7.19 (m, 2H), 5.04 (dd, 1 H), 5.01 (br, 1 H), 3.95 (m, 2H), 3.40 (s, 3H), 2.45 (m, 4H), 2.45 (m, 2H), 2.02 (m, 1 H), 1.79 (m, 1 H), 1.15-1.35 (m, 9H). Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-3-(2-hvdroxyethyl)-1-methyl-7-phenyl-1 H- pyridof 2.3-bU ,41oxazin-2(3H Vone
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1 -yl)propanenitrile, tert-butyl (1 -(4-(3-(2- hydroxyethyl)-1 -methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (12.5mg, 0.024 mmol) was reacted to afford the title compound (5.5mg). 1H N R (500 MHz, CH3OD) 7.64 (s, 1 H), 7.41 (m, 4H), 7.28 (m, 3H), 7.18 (m, 2H), 5.09 (dd, 1H), 3.80-3.92 (m, 2H), 3.41 (s, 1 H), 2.72 (m, 2H), 2.54 (m, 2H), 2.24 (m, 2H), 2.13 (m, 1 H), 1.94 (m, 1 H). LCMS (Method A): RT = 3.76 min, M+H+ = 413.16.
Example 78; 2-(6-(4-(1 -aminocvclobutyl)phenyl)-3-(2-hvdroxyethvn-2-oxo-7-phenyl-
2,3-dihvdro-1H-pyridor2, -b1[1,41oxazin-1-yl)acetonitrile
Figure imgf000118_0001
Step 1: tert-butyl (1-(4-(1-(cvanomethyl)-3-(2-hvdroxyethyl)-2-oxo-7-phenyl-2.3-dihvdro- 1 H-pyrido[2,3-biri ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
A solution of tert-butyl 1-(4-(3-(2-hydroxyethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (51 mg, 0.099 mmol) in DMF(2ml) was added potassium carbonate (41mg) and 2-bromoacetonitrile (35.5mg, 0.297mmol). The reaction mixture was stirred at 50 degree for 3 h.
The reaction mixture was partitioned between dichloromethane (20ml) and water (20ml). The organic phase was separated using phase separator and concentrated. The crude product was purified by column (biotage, column size:25g) eluted with ethyl
acetate/cyclohexane to afford product (18mg). LCMS (Method A): RT = 6.45 min, M+H: 555.0
Step 2: 2-(6-(4-(1-aminocvclobutyl)phenyl)-3-(2-hvdroxyethyl)-2-oxo-7-phenyl-2.3- dihvdro-1 H-pyridof2,3-blf 1 ,4loxazin-1 -vDacetonitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1 -yl)propanenitrile, tert-butyl (1 -(4-(1 -(cyanomethyl)- 3-(2-hydroxyethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (15mg, 0.027 mmol) was reacted to afford the title compound (7.5mg). 1H NMR (500 MHz, CH3OD) 7.67 (s, 1 H), 7.41 (d, 2H), 7.38 (d, 2H), 7.30 (m, 3H), 7.24 (m, 2H), 5.18 (dd, 1H), 5.11 (s, 2H), 3.81-3.89 (m, 2H), 2.71-2.77 (m, 2H), 2.52-2.58 (m, 2H), 2.27-2.30 (m, 1H), 2.16-2.23 (m, 2H), 1.93-2.10 (m, 1 H). LCMS (Method A): RT = 3.78 min, M+H+ = 456.2.
Example 79: 7-(4-((1 r,3r)-1 -amino-3-hydroxy-3-methylcvclobutyl)phenyl)-8-phenyl- 2.4-dihvdro-1 H-pyridor2,3-bin ,2.41triazolor4,3-din ,4loxazin-1 -one
Figure imgf000119_0001
Step 1 : 2-((1r,3r)-3-hvdroxy-3-methyl-1-(4-(1-oxo-8-phenyl-2.4-dihvdro-1 H-pyrido[2.3- b1f1.2,41triazolof4.3-din ,41oxazin-7-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione
In a 15 mL round-bottomed flask was 7-bromo-8-phenyl-2,4-dihydro-1H-pyrido[2,3- b][1,2,4]triazolo[4,3-d][1 ,4]oxazin-1-one (30mg, 0.087 mmol), 2-((1r,3r)-1-(4-(5,5- dimethyl-1 ,3,2-dioxaborinan-2-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline-1,3- dione (30.4 mg, 0.072 mmol), and cesium carbonate (118 mg, 0.362 mmol) in 1 ,4- dioxane (1.55 ml) and water (0.5ml) to give a yellow solution. The reaction mixture was degassed for 15 minutes, following by the addition of PdCl2(dppf)-CH2Cl2 adduct (20mg) and degassing for further 15 minutes. The reaction mixture was heated to 50OC overnight under nitrogen atmosphere.
The reaction mixture was allowed to cool to room temperature and water was added (7ml). The reaction mixture was extracted using ethyl acetate (10ml x 3). The combined organic layers were concentrated under reduced pressure.
The crude residue was purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n-hexanes) to give the title compound (5mg, 12%). LCMS (Method D): RT 1.167 min, M+1= 572. Ή NMR (500 MHz, CD3OD): 8.53 (1 H, s), 7.84- 7.78 (4H, m), 7.71 -7.64 (2H, m), 7.51- 7.45 (3H, m), 7.30- 7.22 (4H, m), 5.38 (2H, s), 3.50-3.45 (2H, m), 3.36 (3H, s), 3.02-2.96 (2H, m).
Step 2: 7-(4-((1r,3r)-1-amino-3-hvdroxy-3-methylcyclobutyl)phenyl)-8-phenyl-2,4-dihvdro- 1 H-pyridor2,3-biri ,2.41triazolor4,3-d1M ,41oxazin-1 -one
In a 10 ml microwave vial was 2-((1r,3r)-3-hydroxy-3-methyl-1-(4-(1-oxo-8-phenyl-2,4- dihydro-1 H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (5mg, 8.75 μηιοΙ), hydrazine monohydrate (2.75 μΙ, 0.087 mmol) in 1 ,4-dioxane (0.3 ml) and ethanol (0.1 ml) to give a colourless solution. The reaction mixture was heated to 120°C for one hour.
The reaction mixture was cooled to room temperature and filtered to remove the solid. This was washed thoroughly with ethanol. The filtrates were combined and concentrated under reduced pressure.
The crude residue (5mg) was purified by preparatory HPLC. The combined product fractions were concentrated to dryness under reduced pressure to give an off-white solid as the title compound (3mg, 78%). LCMS (Method D) RT = 0.578 min, M+1= 442.
1H-NMR (500 MHz, CD3OD) δ 8.43 (1 H, s), 7.27-7.17 (7H, m), 7.15- 7.10 (2H, m), 5.29 (2H, s), 2.59-2.52 (2H + 3H, m), 2.32-2.24 (2H, m).
Example 80: 6-(4-(1 -aminocvclobutyl)phenyl)-3-methyl-7-phenyl-1 H-pyridof2.3- bir ,41oxazin-2(3H)-one
Figure imgf000120_0001
Step 1 : tert-butyl (1-(4-(3-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3-biri,4loxazin- 6-yl)phenyl)cyclobutyl)carbamate
A solution of tert-butyl 1-(4-(5-amino-6-oxo-3-phenyl-1 ,6-dihydropyridin-2- yl)phenyl)cyclobutylcarbamate (0.5g, 1.159 mmol) in THF (Volume: 20ml) was added DIPEA (1.01 ml) followed by the addition of 2-chloropropanoyl chloride (0.235 g, 1.854 mmol) in THF (5ml) in dropwise at 0 degree over 1 h. The resulted mixture was stirred at r.t. over night. Sodium bicarbonate saturated solution (30 ml) was added. The resulted mixture was heated to reflux for 16 h. The reaction mixture was cooled down to room temperature. The organic phase was separated. The aqueous phase was extracted with ethyl acetate (40ml). The organic phase was combined and washed with water (30mlX2), brine (25ml) then concentrated at reduced pressure. The residue was suspended in DCM (10ml). The precipitate was collected by filtration and washed with DCM (5 ml) to afford product (439mg). H NMR (500 MHz, CD3OD): 7.20-7.29 (m, 8H), 7.13-7.15 (m, 2H),4.98 (q, 1 H), 2.41-2.47 (m, 4H), 2.05 (m, 1 H), 1.86 (m, 1 H), 1.64 (d, 3H), 1.21-1.36 (br, 9H).
Step 2: 6-(4-(1 -aminocyclobutyl)phenyl)-3-methyl-7-phenyl-1 H-pyridof2,3-b][1 ,41oxazin- 2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][ ,4]oxazin-1 -yl)propanenitrile, tert-butyl (1 -(4-(3-methyl-2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (25mg, 0.051 mmol) was reacted to afford the title compound (18.5mg). Ή NMR (500 MHz, CD3OD): 7.37-7.42 (m, 4H), 7.34 (s, 1 H), 7.28-7.30 (m, 3H), 7.18-7.20 (m, 2H), 5.03 (q, 1 H), 2.73-2.79 (m, 2H), 2.56-2.60 (m, 2H), 2.20-2.26 (m, 1 H), 1.96-1.99 (m, 1 H), 1.65 (d, 3H). LCMS (Method D): RT = 0.73 min, M+H+ = 386.2
Example 81 : 2-(6-(4-(1-aminocvclobutyl)phenyl)-3-methyl-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2.3-blli ,41oxazin-1 -vDacetonitrile
Figure imgf000121_0001
Step 1 : tert-butyl (1-(4-(1-(cvanomethyl)-3-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H- pyridor2,3-b1[1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
A solution of tert-butyl 1-(4-(3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50mg, 0.103 mmol) in DMF (2 ml) was added potassium carbonate (42.7 mg) and 2-bromoacetonitrile (22 μΙ, 0.309 mmol). The reaction mixture was stirred at room temperature for 48 h then was diluted with ethyl acetate (20 ml). Washed with water, brine, and dried with Na2S04, filtered and concentrated to give crude product which was purified by column (biotage, 25g) eluted with ethyl acetate and cyclohexane to afford product (45mg). 1H N R (500 MHz, CDCI3): 7.35 (s, 1H), 7.20-7.32 (m, 9H), 4.98 (q, 1 H), 4.87 (dd, 2H), 2.44-2.49 (m, 4H), 2.06 (m, 1 H), 1.80 (m, 1 H), 1.75 (d, 3H), 1.24-1.36 (br, 9H).
Step 2: 2-(6-(4-i1-aminocvclobutyl)phenyl)-3-methyl-2-oxo-7-phenyl-2.3-dihydro-1 H- pyrido[2,3-blf1 ,4loxazin-1-yl)acetonitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(cyanomethyl)- 3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (25mg, 0.048 mmol) was reacted to afford the title compound (20mg). H NMR (500 MHz, CH3OD) 7.71 (s, 1 H), 7.45 (d, 2H), 7.39 (d,2H), 7.32 (m, 3H), 7.26 (m, 2H), 5.14 (s, 2H), 5.10 (q, 1 H), 2.74-2.80 (m, 2H), 2.55-2.61 (m, 2H), 2.22-2.26 (m, 1 H), 1.96-1.99 (m, 1 H), 1.71 (d, 3H). LCMS (Method D): RT = 0.81 min, M+H+ = 425.2.
Example 82: 6-(4-(1 -aminocvclobutyl)phenyl)-1 -(methylsulfonylmethyl)-7-phenyl-
1 H-pyridor2,3-biri .41oxazin- -one
Figure imgf000122_0001
Step 1 : tert-butyl 1-(4-(1-(methylthiomethyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2,3- b1f1 ,41oxazin-6-yl)phenyl)cvclobutylcarbamate
In a 15 mL reaction tube was added terf-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (150 mg, 0.318 mmol), potassium carbonate (132 mg, 0.954 mmol) and chloromethyl methyl sulfide (0.079 mL, 0.954 mmol) in anhydrous N,N-dimethylformamide (2 mL) to give a yellow suspension. The reaction mixture was heated to 80°C under a nitrogen atmosphere for 2 hours, then allowed to cool to room temperature, diluted with saturated sodium bicarbonate solution (10 mL) and extracted into dichloromethane (3 x 5 mL). The combined organic phases were dried over Na2S0 , filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 80:20) to give the desired product as a white solid (75 mg, 44% yield). 1H-NMR (500 MHz, CDCI3) δ 7.43 (1H, s), 7.23-7.35 (7H, m), 7.19- 7.23 (2H, m), 5.09 (3H, br s), 4.94 (2H, s), 2.30-2.55 (4H, m), 2.26 (3H. s), 2.00-2.15 (1 H, m), 1.75-1.90 (1 H, m), 1.10-1.50 (9H, br s). LCMS (Method A) RT = 7.42 min, M+H+ = 532.14.
Step 2: te/t-butyl 1-(4-(1-(methylsulfonylmethyl)-2-oxo-7-phenyl-2.3-dihydro-1 H- PVridof2,3-biri,41oxazin-6-yl)phenyl)cvclobutylcarbamate
In a 15 mL reaction tube was added tert-butyl 1-(4-(1 -(methylthiomethyl)-2-oxo-7-phenyl- 2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50 mg, 0.094 mmol) in a mixture of methanol (1500 μΙ) and tetrahydrofuran (1500 μΙ) to give a yellow solution. To this was added dropwise a solution of oxone monopersulfate compound (463 mg, 0.752 mmol) in water (1500 pi) and the resulting suspension stirred at room temperature overnight. The reaction mixture was quenched by the careful addition of saturated sodium bicarbonate solution (3 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were washed with saturated sodium bicarbonate solution (5 mL), brine (5 mL), dried over Na2S0 , filtered and concentrated to dryness under reduced pressure to give an off-white solid. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a white solid (30 mg, 57%). 1H-NMR (500 MHz, CDCI3) δ 7.67 (1 H, s), 7.25- 7.34 (7H, m). 7.17-7.23 (2H, m), 5.21 (2H, br s), 5.08 (1 H, br s), 4.99 (2H, s). 3.08 (3H, s), 2.20-2.60 (4H, m), 2.00-2.15 (1 H, m), 1.75-1.90 (1 H, m), 1.10-1.50 (9H, br m). LCMS (Method D) RT = 1.44 min, M+H+ = 564.00.
Step 3: 6-(4-(1-aminocvclobutyl)phenyl)-1 -(methylsulfonylmethyl)-7-phenyl-1 H- pyridof2,3-biri,41oxazin-2(3H)-one
ferf-Butyl 1-(4-(1-(methylsulfonylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (28 mg, 0.050 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and freeze-dried over the weekend to give the desired product as an off-white solid (28 mg, 98% yield). 1H-NMR (500 MHz, MeOD) δ 7.78 (1 H, s), 7.32 (2H, d), 7.28 (2H, d), 7.13-7.23 (3H, m), 7.08-7.13 (2H, m), 5.43 (2H, s), 5.01 (2H, s), 2.86 (3H, s), 2.58-2.70 (2H, m), 2.40-2.52 (2H, m), 2.03-2.17 (1 H, m), 1.76-1.90 (1 H, m). LCMS (Method D) RT = 0.87 min, M+H+ = 464.00.
Example 83: 6-(4-(1 -aminocyclobutyl)phenyl)-1 -(1 -methyl-1 H-pyrazol-4-yl)-7- phenyl-1 H-pyridor2.3-b1H ,41oxazin-2(3H)-one
Figure imgf000124_0001
Step 1 : tert-butyl 1-(4-(1-(1-methyl-1 H-pyrazol-4-yl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyrido[2,3-blf1 ,4loxazin-6-yl)phenyl)cyclobutylcarbamate
In a 15 mL reaction tube was added terf-butyl 1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100 mg, 0.212 mmol), 1- methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1 H-pyrazole (44 mg, 0.212 mmol), copper(ll) acetate (58 mg, 0.318 mmol) and 5A molecular sieves (60 mg) in anhydrous dichloromethane (2 mL). To this was added anhydrous pyridine (1 mL, 12.36 mmol) and the resulting deep purple solution stirred at room temperature under air for 48 hours. Further 1-methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1 H-pyrazole (44 mg, 0.212 mmol) and copper(ll) acetate (58 mg, 0.318 mmol) were added and the reaction mixture stirred at room temperature for 72 hours, then heated to 40°C overnight. To the reaction mixture was added further 1-methyl-4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)-1 H-pyrazole (44 mg, 0.212 mmol) and copper(ll) acetate (58 mg, 0.318 mmol) and heated at 40°C for 6 days. The reaction mixture was allowed to cool to room temperature and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane.ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as an off-white solid (30 mg, 51% yield). 1 H-NMR (500 MHz, CDCI3) δ 7.53 (1 H, s), 7.49 (1 H, s), 7.12-7.27 (7H, m), 6.98-7.08 (3H, m), 4.93 (2H, s), 3.90 (3H, s), 2.10-2.50 (4H, m), 1.92-2.04 (1 H, m), 1.65-1.79 (1 H, m), 1.00-1.45 (9H, br m). LCMS (Method D) RT = 1.45 min, M+H+ = 552.20. Step 2: 6-(4-(1-aminocyclobutyl)phenyl)-1-(1-methyl-1 H-pyrazol-4-yl)-7-phenyl-1 H- Pyridof2.3-bU1.41oxazin-2(3H -one
terf-Butyl 1-(4-(1-(1-methyl-1 H-pyrazol-4-yl)-2-oxo-7-phenyl-2)3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (30 mg, 0.054 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (29 mg, 94% yield). 1H-NMR (500 MHz, MeOD) δ 7.89 (1 H, s), 7.62 (1 H, s), 7.42-7.51 (4H, m), 7.40 (2H, d), 7.37 (2H, d), 7.18-7.27 (3H, m), 7.03-7.12 (3H, m), 5.06 (2H, s), 3.96 (3H, s), 2.66-2.78 (2H, m), 2.48-2.58 (2H, m), 2.13-2.26 (1 H, m), 1.85-1.98 (1 H, m). LCMS (Method D) RT = 0.86 min, M+H+ = 452.20.
Example 84: 6-(4-(1 -aminocvclobutvQphenvO-1 ,3-dimethyl-7-phenyl-1 H-pyridor2,3- bin,41oxazin-2(3H)-one
Figure imgf000125_0001
Step 1 : tert-butyl n-(4-(1 ,3-dimethyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2.3- b1f1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
A solution of tert-butyl 1 -(4-(3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (50mg, 0.103 mmol) in DMF (2 ml) was added potassium carbonate (42.7 mg) and iodomethane (7.69 μΙ, 0.124 mmol). The reaction mixture was stirred at room temperature for 48 h then was diluted with ethyl acetate (20 ml). Washed with water, brine, and dried with Na2S04, filtered and concentrated to give product 43mg. 1H NMR (500 MHz, CDCI3): 7.19-7.32 (m, 10H), 4.96 (q, 1 H), 3.39 (s, 3H), 2.43-2.48 (m, 4H), 2.04 (m, 1 H), 1.79 (m, 1 H), 1.70 (d, 3H), 1.24- 1.36 (m, 9H). Step 2: 6-(4-(1 -aminocvclobutyl)phenyl)-1 ,3-dimethyl-7-phenyl-1 H-pyrido[2,3- bin.41oxazin-2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1 ,3-dimethyl-2- oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (40mg, 0.08 mmol) was reacted to afford the title compound (34.5mg). 1H NMR (500 MHz, CH3OD) 7.56 (s, 1 H), 7.42 (d, 2H), 7.38 (d,2H), 7.31 (m, 3H), 7.24 (m, 2H), 5.06 (q, 1 H), 3.44 (s, 3H), 2.73-2.77 (m, 2H), 2.53-2.59 (m, 2H), 2.20-2.26 (m, 1 H), 1.95- 1.97 (m, 1 H), 1.67 (d, 3H). LCMS (Method D): RT = 0.85 min, M+H+ = 400.2.
Example 85: 6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2,2-difluoroethyl)-7-phenyl-1 H- pyridor2.3-bin.41oxazin-2(3 -one
Figure imgf000126_0001
Step 1 tert-butyl (1-(4-(1-(2.2-difluoroethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- blf1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-py do[2,3-b][1,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1-(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl) cyclobutylcarbamate (100mg, 0.212 mmol) was reacted with 1 ,1-difluoro-2-iodoethane (122mg, 0.636 mmol) to afford the title compound (85mg. 1H NMR (500 MHz, CDCI3): 7.42 (1H, s), 7.24-7.30 (7H, m), 7.17-7.19 (2H, m), 6.11 (1H, tt), 5.05 (1H, br), 4.92 (s, 2H), 4.28 (dt, 2H), 2.44- 2.49 (m, 4H), 2.05 (m, 1H), 1.81 (m, 1 H), 1.2-1.43 (br, 9H).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-(2,2-difluoroethyl)-7-phenyl-1H-pyrido|2,3- bin .41oxazin-2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(2,2- difluoroethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (20mg, 0.037 mmol) was reacted to afford the title compound (16mg). 1H NMR (500 MHz, CH3OD) 7.71 (s, 1 H), 7.42 (d, 2H), 7.38 (d, 2H), 7.31 (m, 3H), 7.22 (m, 2H), 6.21 (tt, 1 H), 5.01 (s, 2H), 4.50 (dt, 2H), 2.73-2.78 (m, 2H), 2.54-2.59 (m, 2H), 2.21-2.25 (m, 1 H), 1.93-1.99 (m, 1 H). LCMS (Method D): RT = 0.84 min, M+H+ = 436.
Example 86: 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-fluoroethyl)-7-phenyl-1H- pyridor2,3-b1f1.4loxazin-2(3 -one
Figure imgf000127_0001
Step 1 tert-butyl (1 -(4-(1-(2-fluoroethvn-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2,3- b1f1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1-(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (1 OOmg, 0.212 mmol) was reacted with 1-fluoro-2-iodoethane (111 mg, 0.636 mmol) to afford the title compound (85mg. Ή NMR (500 MHz, CDCI3): 7.45 (1H, s), 7.24-7.31 (7H, m), 7.17- 7.20 (2H, m,), 5.01 (1 H, s), 4.94 (2H, s), 4.80 (1 H, t), 4.70 (1 H, t), 4.27 (1H, t), 4.22 (1 H, t), 2.44-2.49 (4H, m), 2.06 (1 H, m), 1.86 (1 H, m), 1.2-1.43 (9H, br).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-(2-fluoroethyl)-7-phenyl- H-pyridor2,3- blf 1.41oxazin-2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(2-fluoroethyl)- 2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (60mg, 0.1 16 mmol) was reacted to afford the title compound (50.5mg). 1H NMR (500 MHz, CH3OD) 7.69 (1 H, s), 7.42 (2H, d), 7.38 (2H, d), 7.30 (3H, m), 7.22 (2H, m), 4.99 (2H, s), 4.80 (1 H, t), 4.70 (1 H, t), 4.40 (1 H, t), 4.35 (1 H, t), 2.73-2.79 (2H, m), 2.54-2.60 (2H, m), 2.20-2.25 (1 H, m), 1.95-1.97 (1 H, m). LCMS (Method D): RT = 0.83 min, M+H+ = 418. Example 87: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-3.4-dihydropyridor3,2- dlpyrimidin-2(1 H)-one
Figure imgf000128_0001
Step 1 : tert-butyl (1-(4-(5-nitro-6-(nitromethyl)-3-phenylpyridin-2- vQphenvOcyclobutvDcarbamate
To a solution of potassium tert-butoxide (468 mg, 4.17 mmol) in dry DMSO (20 ml) was slowly added nitromethane (225 ul, 4.17 mmol) under nitrogen. The resulting mixture was stirred for 5 min at room temperature before tert-butyl (1-(4-(6-chloro-5-nitro-3- phenylpyridin-2-yl)phenyl)cyclobutyl)carbamate (1 g, 2.08 mmol) was added. The resulting mixture was stirred for 1 hour at room temperature. A saturated solution of NH4CI was added and the mixture was extracted with AcOEt (3 x 50ml). The combined organic phases were washed with water (3 x 20ml), dried over Na2S0 and concentrated to dryness under reduced pressure. The resulting residue was used directly without purification. 1H NMR (500 MHz,CDCI3): 8.60 (1 H, s), 7.44 -7.35 (7H, m), 7.30-7.28 (2H, m), 6.21 (2H, s), 5.09 (1H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br).
Step 2: tert-butyl (1-(4-(5-amino-6-(aminomethyl)-3-phenylpyridin-2- yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(5-nitro-6-(nitromethyl)-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (74 mg, 0.15 mmol) in EtOH (50 mL) was added Raney Nickel (17 mg, 0.15 mmol). The resulting mixture was purged with nitrogen and with H2 and then stirred under H2 (1.5 bars) overnight at room temperature. The black mixture was filtered through celite, rinsed few times with EtOH (making sure that the cake remained wet at all times), dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was used directly without purification.
Step 3: tert-butyl (1-(4-(2-oxo-7-phenyl-1 ,2,3,4-tetrahvdropyridor3.2-d1pyrimidin-6- yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (l-(4-(5-amino-6-(aminomethyl)-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (113 mg, 0.25 mmol) in dry THF (2 ml) was added CDI (41 mg, 0.25 mmol) under nitrogen. The resulting mixture was stirred for 1h at 60C and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 5% MeOH inDCM) to give the title compound (71 mg, 59%). Ή NMR (500 Hz,CDCI3): 8.91 (1H, br s), 7.19-7.11 (7H, m), 7.00-6.97 (3H, m), 5.73 ( H, s), 4.93 ( H, s), 4.65 (2H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br)
Step 4: 6-(4-(1 -aminocvclobutyl)phenyl)-7-phenyl-3,4-dihydropyridor3,2-dlpyrimidin- 2(1 H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[3,2-d]pyrimidin-6- yl)phenyl)cyclobutyl)carbamate (12 mg, 0.03 mmol) was reacted to afford the title compound (12 mg, 97%). LCMS (Method D): RT = 0.684 min, M+1 =371. 1H NMR (500 MHz, MeOD): 7.42-7.38 (4H, m), 7.31-7.28 (3H, m), 7.23 (1 H, s), 7.21-7.18 (2H, m), 4.66 (2H, s), 2.81 -2.75 (2H, m), 2.63-2.57 (2H, m), 2.26 -2.24 (1 H, m), 1.99-1.97 (1 H, m).
Example 88: 6-(4-(1-aminocvclobutyl)phenyl)-N-ethyl-7-phenyl-2,3-dihvdro-1H- pyridor2,3-bl[1,41oxazine-1-carboxamide
Figure imgf000129_0001
Step 1 : Tert-butyl (1-(4-(ethylcarbamovn-7-phenyl-2.3-dihvdro-1 H-pyrido[2.3- b1f1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
Tert-butyl 1-(4-(7-phenyl-2,3-dihydro-1 H-pyrido-[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (50mg, 0.109 mmol) and isocyanatoethane (40mg ml, 0.546 mmol) in THF (1.5 mL) were heated to 50°C for 24 hrs. The resulting reaction mixture was concentrated under reduced pressure. The obtained crude was purified by Biotage silica gel chromatography (gradient 100% to 80% n-hexane in ethyl acetate) to give the title compound (40 mg, 70%). Ή NMR (500 MHz, CDCI3): 7.83 (1H, s), 7.33- 7.29 (2H, d), 7.7-7.21 (4H, m), 7.18- 7.13 (4H, m), 4.47 (2H, t), 3.93 (3H, t), 3.38 (2H, q), 2.53-2.38 (4H, m), 2.12-1.99 (1 H, m), 1.96- 1.86 (1 H, m), 1.48-1.28 (9H, br), 1.29 (3H, t). LCMS (Method D): RT = 1.508 min, +1 = 529.
Step 2: 6 -(4-(1-aminocvclobutyl)phenyl)-N-ethyl-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- blf 1 ,41oxazine-1 -carboxamide
Tert-butyl (1-(4-(ethylcarbamoyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (45 mg, 0.093 mmol) was dissolved in TFA (1 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (13 mg, 53% yield). 1H-NMR (500 MHz, CD3OD) δ 8.24 (1 H, s), 7.41 (2H, d), 7.38 (2H, d), 7.29- 7.25 (3H, m), 7.23-7.18 (2H, m), 4.50 (2H, t), 3.89 (2H, t), 3.30 (2H, q), 2.81-2.71 (2H, m), 2.63-2.52 (2H, m), 2.29-2.19 (1 H, m), 2.02-1.90 (1 H, m), 1.21 (3H, t). LCMS (Method D) RT = 0.779 min, M+1 =429.
Example 89: 6-(4-(1-aminocvclobutyl)phenyl)-1 ,3-dimethyl-7-phenyl-3,4- dihydropyridor3.2-d1pyrimidin-2(1H)-one
Figure imgf000130_0001
Step 1 : tert-butyl (1-(4-(1.3-dimethyl-2-oxo-7-phenyl-1 ,2,3,4-tetrahvdropyridor3.2- dlpyrimidin-6-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[3,2-d]pyrimidin- 6-yl)phenyl)cyclobutyl)carbamate (46 mg, 0.10 mmol) in dry DMF (1 ml) at -78C was added sodium hydride (3.9 mg, 0.10 mmol) and methyliodide (6.1 μΙ, 0.10 mmol) under nitrogen. The bath was removed and the resulting mixture was stirred for 1 h at room temperature. The mixture was cooled down to -78C. Sodium hydride (3.9 mg, 0.10 mmol) and methyliodide (6.1 μΙ, 0.10 mmol) were added under nitrogen. The bath was removed and the resulting mixture was stirred for 1 h at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 50% EtOAc in cyclohexane) to give the title compound (30 mg, 61%). Ή NMR (500 MHz,CDCI3): 7.20- 7.19 (7H, m), 7.13-7.1 1 (2H, m), 7.02 (1H, s), 5.00 (1 H, br s), 4.56 (2H, s), 3.27 (3H, s), 3.03 (3H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br)
Step 2: 6-(4-(1 -aminocvclobutyl)phenyl)-1 ,3-dimethyl-7-phenyl-3,4-dihvdropyridor3,2- dlpyrimidin-2(1 H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1 - aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1 ,3-dimethyl-2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[3,2-d]pyrimidin-6- yl)phenyl)cyclobutyl)carbamate (30 mg, 0.06 mmol) was reacted to afford the title compound (28 mg, 91 %). LCMS (Method D): RT = 0.798 min, M-NH2 = 382. Ή NMR (500 MHz, MeOD): 7.45-7.39 (4H, m), 7.34 ( H, s), 7.32-7.30 (3H, m), 7.26-7.24 (2H, m), 4.67 (2H, s), 3.36 (3H, s), 3.1 1 (3H, s), 2.81 -2.75 (2H, m), 2.63-2.57 (2H, m), 2.26 -2.24 (1 H, m), 1.99-1.97 (1 H, m).
Example 90: 6-(4-(1 -Aminocyclobutyl)phenyl)-1 -methyl-7-phenyl-1 H-pyridor2,3- b1f1.41oxazin-2(3H)-one O-methyl oxime
Figure imgf000131_0001
Step 1 : tert-Butyl (1-(4-(2-(methoxyimino)-1-methyl-7-phenyl-2.3-dihvdro-1 H-pyridof2,3- blf1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
In a round-bottomed flask was added tert-butyl 1-(4-(1-methyl-7-phenyl-2-thioxo-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (0.1 g, 0.199 mmol) and o-methylhydroxylamine hydrochloride (0.067 g, 0.797 mmol) in Pyridine (Volume: 2 ml). The resulting solution was stirring and heating at 70 degree for 18h. The reaction mixture was concentrated to dryness and partitioned between ethyl acetate (20ml) and water (20ml). The organic phase was separated and washed with water (2X15ml), brine (15ml) concentrated to give the crude product which was purified by column (biotage, 25g) eluted with ethyl acetate/cyclohexane (0-50%) to give product (21 mg) 1H NMR (500 MHz, CDCI3) 7.20-7.29 (m, 9H), 7.10 (s, 1 H), 5.19 (s, 2H), 4.95 (s, 1 H), 3.81 (s, 3H), 3.31 (s, 3H), 2.2-2.6 (m, 4H), 1.95-2.15 (m, 1 H), 1.75-1.85 (m, 1 H), 1.2-1.4 (br, 9H).
Step 2: 6-(4-(1-Aminocvclobutyl)phenyl)-1-methyl-7-phenyl-1 H-pyridor2,3-blf1 ,4loxazin- 2(3H)-one O-methyl oxime
to a round bottomed flask containing tert-butyl 1-(4-(2-(methoxyimino)-1 -methyl-7-phenyl- 2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (28mg, 0.054 mmol) was added trifluoroacetic acid (1 ml). The resulting solution was stirred for 60 seconds then concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (1 ml) and concentrated to dryness three times to give the trifluoroacetic acid salt of the product (14.5mg). 1H NMR (500 MHz, CD3OD) 7.49 (s, 1 H), 7.35-7.39 (m, 5H), 7.28-7.30 (m, 3H), 7.21-7.23 (m, 2H), 5.24 (s, 2H), 3.83 (s, 3H), 3.35 (s, 3H), 2.72-2.78 (m, 2H), 2.54-2.60 (m, 2H), 2.2-2.3 (m, 1 H), 1.9-2.0 (m, 1 H), LCMS (Method D): RT = 0.96 min, M+H+ = 415.2.
Example 91: 1-(4-f7-phenyl-1-(pyridin-3-yl)-2.3-dihvdro-1H-pyridor2.3-bin.41oxazin-
6-yl)phenyl)cvclobutanamine
Figure imgf000132_0001
Step 1 : terf-butyl 1-(4-(7-phenyl-1-(pyridin-3-yl)-2.3-dihvdro-1 H-pyridor2.3-b1f1.41oxazin- 6-yl)phenyl)cyclobutylcarbamate
In a 15 mL reaction tube was added tert-butyl 1-(4-(2-oxo-7-phenyl-1 -(pyridin-3-yl)-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (90 mg, 0.164 mmol) in anhydrous tetrahydrofuran (3 ml) to give a colourless solution. This was cooled to 0°C followed by the addition of sodium borohydride (15 mg, 0.394 mmol) and boron trifluoride diethyl etherate (50 μΙ, 0.394 mmol). The reaction mixture was stirred at 0°C under a nitrogen atmosphere for 3 hours then heated to 40°C overnight. The reaction mixture was diluted with ethyl acetate (3 mL) and quenched by washing with saturated ammonium chloride solution (2 x 3 mL). The organic phase was dried over Na2S0 , filtered, concentrated to dryness under reduced pressure and purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) then Biotage chromatography (dichloromethane:methanol, gradient elution from 99:1 to 90:10) then Biotage chromatography (dichloromethane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a yellow oil (30 mg, 34% yield). 1H-NMR (500 MHz, CDCI3) δ 8.60 (1 H, br s), 8.24 (1 H, br s), 8.12 (1 H, d), 7.65 (1 H, br s), 7.36 (1 H, s), 7.13-7.28 (7H, m), 7.02-7.08 (2H, m), 4.94 (1H, br s), 4.51 (2H, t), 3.86 (2H, t), 2.20-2.50 (4H, m), 1.95-2.05 (1 H, m), 1.70-1.80 (1 H, m), 1.00-1.38 (9H, br m). LCMS (Method D) RT = 1.45 min, M+H+ = 535.20.
Step 2: 1-(4-(7-phenyl-1-(pyridin-3-yl)-2.3-dihvdro-1H-pyridor2,3-biri,41oxazin-6- yDphenvDcvclobutanamine
ferf-Butyl 1 -(4-(7-phenyl-1 -(pyridin-3-yl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutylcarbamate (10 mg, 0.019 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (-2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (4 mg, 32% yield). 1H-NMR (500 MHz, MeOD) δ 8.57 (1 H, br s), 8.28 (1 H, br s), 7.80 (1 H, d), 7.43 (1 H, br s), 7.26 (2H, d), 7.24 (2H, d), 7.04-7.16 (4H, m), 6.90-7.00 (2H, m), 4.51 (2H, t), 3.80 (2H, t), 2.56-2.70 (2H, m), 2.37-2.52 (2H, m), 2.02-2.16 (1 H, m), 1.74-1.90 (1 H, m). LCMS (Method D) RT = 0.77 min, M+H+ = 435.20.
Example 92: 1-f4-(1-bromo-8-phenyl-4H-pyridor2,3-biri.2.41triazolor4.3- d1M.41oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000133_0001
Step 1 : tert-butyl (1 -(4-(1-bromo-8-phenyl-4H-pyridof2.3-bin .2.41triazolor4,3- d1[1 ,4loxazin-7-yl)phenyl)cyclobutyl)carbamate
A mixture of tert-butyl 1-(4-(8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutylcarbamate (200mg, 0.404 mmol) and 1-bromopyrrolidine-2,5-dione (114.9275 mg, 0.646 mmol) in DCM (12 ml) and CCI4 (18 ml) was heating at 50 degree for 24h.The mixture was concentrated and the residue was partitioned between ethyl acetate (40ml) and water (40ml). The organic phase was separated and the aqueous phase was extracted with ethyl acetate (30ml). The combined organic phase was washed with water (50ml) brine (30ml) and concentrated to give product 159mg. 1H NMR (500 MHz, CDCI3): 8.55 (s, 1 H), 7.22-7.35 (m, 9H), 5.58 (s, 2H), 5.1 (br, 1 H), 2.4- 2.5 (m, 4H), 2.01-2.1 1 (m, 1 H), 1.79-1.89 (m, 1 H), 1.24-1.43 (br, 9H).
Step 2: 1-(4-(1-bromo-8-phenyl-4H-pyridor2.3-blf1 ,2.41triazolor4,3-d1f1.4loxazin-7- vDphenvQcyclobutanamine
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(1-(cyanomethyl)- 3-(methoxymethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (18mg, 0.031 mmol) was reacted to afford the title compound (4.5mg). 1H NMR (500 MHz, CH3OD) 8.71 (s, 1 H), 7.50 (d, 2H), 7.43 (d, 2H), 7.34-7.36 (m, 3H), 7.28-7.31 (m, 2H), 5.70 (s, 2H), 2.76-2.80 (m, 2H), 2.57-2.61 (m, 2H), 2.2-2.27 (m, 1 H), 1.9- 2.0 (m, 1 H), LCMS (Method D): RT = 0.774 min, M+ =474.2, M+2 = 476.2
Example 93: (R)-2-(6-(4-(1 -aminocvclobutyl)phenyl)-3-methyl-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b1M ,4loxazin-1 -vQacetonitrile
Figure imgf000134_0001
Step 1 : (R) tert-butyl (1 -(4-(1-(cvanomethyl)-3-methyl-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof 2 , 3-bU 1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
A solution of (R)-tert-butyl 1 -(4-(3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100mg, 0.206 mmol) in DMF (2 ml) was added potassium carbonate (85 mg) and 2-bromoacetonitrile (44 μΙ, 0.618 mmol). The reaction mixture was stirred at room temperature for 48 h then was diluted with ethyl acetate (20 ml). Washed with water, brine, and dried with Na2S04, filtered and concentrated to give crude product which was purified by column (biotage, 25g) eluted with ethyl acetate and cyclohexane to afford product (55mg). H NMR (500 MHz, CDCI3): 7.35 (s, 1 H), 7.20-7.32 (m, 9H), 4.98 (q, 1 H), 4.87 (dd, 2H), 2.44-2.49 (m, 4H), 2.06 (m, 1 H), 1.80 (m, 1 H), 1.75 (d, 3H), 1.24-1.36 (br, 9H).
Step 2: (R)-2-(6-(4-(1-aminocvclobutyl)phenyl)-3-methyl-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridoi2,3-b1i1 ,4]oxazin-1 -vOacetonitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, (R)-tert-butyl (1 -(4-(1- (cyanomethyl)-3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (41 mg, 0.078 mmol) was reacted to afford the title compound (22mg). 1H NMR (500 MHz, CH3OD) 7.71 (s, 1 H), 7.45 (d, 2H), 7.39 (d,2H), 7.32 (m, 3H), 7.26 (m, 2H), 5.14 (s, 2H), 5.10 (q, 1 H), 2.74-2.80 (m, 2H), 2.55-2.61 (m, 2H), 2.22-2.26 (m, 1 H), 1.96-1.99 (m, 1 H), 1.71 (d, 3H). LCMS (Method D): RT = 0.81 min, M+H+ = 425.2.
Example 94: (R)-6- 4-(1-aminocvclobutyl)phenyl)-1,3-dimethyl-7-phenyl-1H- pyridor2.3-b1M,41oxazin-2(3 -one
Figure imgf000135_0001
Step 1 : (R)-tert-butyl (1-(4-(3-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2.3- b 1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
(S)-2-Chloropropanoic acid (0.20 g, 1.854 mmol) in a round-bottomed flask was mixed with thionyl chloride (0.149 ml, 2.039 mmol). A drop of DMF was added and the mixture was stirred for 0.5h at room temperature then 80 degree for 2h. The resulted light yellow liquid was diluted with THF (5ml).
At 0 degree, in a 100 mL round-bottomed flask was tert-butyl 1-(4-(5-amino-6-oxo-3- phenyl-1 ,6-dihydropyridin-2-yl)phenyl)cyclobutylcarbamate (0.5g, 1.159 mmol) and N- ethyl-N-isopropylpropan-2-amine (1.015 ml, 5.79 mmol) in THF (25ml) to give a yellow suspension. The above acid chloride in THF solution was added in dropwise to the suspension in 30 min. The reaction mixture was stirred at room temperature for 16h. The reaction mixture was transferred to a 250ml round bottom flask. Sodium bicarbonate saturated solution (30 ml) and sodium bicarbonate (1g) were added. The resulted mixture was heated to reflux for 24 h. The reaction mixture was cooled down to room
temperature. The organic phase was separated. The aqueous phase was extracted with ethyl acetate (40ml). The organic phase was combined and washed with water (30mlX2), brine (25ml) then concentrated at reduced pressure. The residue was suspended in DCM (10ml). The precipitate was collected by filtration and washed with DCM (5 ml) to afford product 0.23g. 1H NMR (500 MHz, CD3OD): 7.20-7.32 (m, 8H), 7.13-7.15 (m, 2H), 4.98 (q, 1 H), 2.41-2.47 (m, 4H), 2.05 (m, 1 H), 1.86 (m, 1 H), 1.64 (d, 3H), 1.21 -1.36 (br, 9H).
Step 2: (RHert-butyl (1-(4-(1 ,3-dimethyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2.3- bli1.41oxazin-6-yl)phenyl)cvclobutyl)carbamate
A solution of (R)-tert-butyl 1-(4-(3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100mg, 0.206 mmol) in DMF (2 ml) was added potassium carbonate (85 mg) and iodomethane (15μΙ, 0.247 mmol). The reaction mixture was stirred at room temperature for 16 h then was diluted with ethyl acetate (20 ml). Washed with water, brine, and dried with Na2S04, filtered and concentrated to give crude product which was purified by column (Biotage, size: 25g) eluted with ethyl acetate/cyclohexane (0-50%) to afford product (33mg). Ή NMR (500 MHz, CDCI3): 7.19- 7.32 (m, 10H), 4.96 (q, 1 H), 3.39 (s, 3H), 2.43-2.48 (m, 4H), 2.04 (m, 1 H), 1.80 (m, 1 H), 1.70 (d, 3H), 1.24-1.36 (m, 9H). Step 3: 6-(4-(1-aminocvclobutyl)phenyl)-1 ,3-dimethyl-7-phenyl-1 H-pyridof2,3- bin .41oxazin-2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, (R)-tert-butyl (1-(4-(1 ,3-dimethyl- 2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b]t1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (16mg, 0.032 mmol) was reacted to afford the title compound (6.5mg). 1H NMR (500 MHz, CH3OD) 7.56 (s, 1 H), 7.42 (d, 2H), 7.38 (d,2H), 7.31 (m, 3H), 7.24 (m, 2H), 5.06 (q, 1 H), 3.44 (s, 3H), 2.73-2.77 (m, 2H), 2.53-2.59 (m, 2H), 2.20-2.26 (m, 1 H), 1.95- 1.97 (m, 1 H), 1.67 (d, 3H). LCMS (Method D): RT = 0.85 min, M+H+ = 400.2.
Example 95: 2-(6-(4-(1-aminocvclobutyl)phenyl)-3-(methoxymethyl)-2-oxo-7-phenyl-
2,3-dihydro-1H-pyridor2,3-biri,41oxazin-1-yl)acetonitrile
Figure imgf000137_0001
Step 1 : tert-butyl (1 -(4-(1-(cvanomethyl)-3-(methoxymethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyridor2,3-bin ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
A solution of tert-butyl (1-(4-(3-(methoxymethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (60mg, 0.1 16 mmol) in DMF (2 ml) was added potassium carbonate (48 mg) and 2-bromoacetonitrile (41.9mg, 0.349 mmol). The reaction mixture was stirred at room temperature for 16 h then was diluted with ethyl acetate (20 ml). Washed with water, brine, and dried with Na2S04, filtered and concentrated to give crude product which was purified by column (biotage, 25g) eluted with ethyl acetate and cyclohexane to afford product (35mg). Ή NMR (500 MHz, CDCI3): 7.20-7.33 (m, 10H), 5.10 (t, 1 H), 4.97 (s, 1 H), 4.82 (dd, 2H), 4.03 (dd, 2H), 3.36 (s, 3H), 2.44-2.50 (m, 4H), 2.06 (m, 1 H), 1.80 (m, 1 H), 1.24-1.36 (br, 9H).
Step 2: 2-(6-(4-(1 -aminocvclobutyl)phenyl)-3-(methoxymethyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyridor2,3-b][1 ,41oxazin-1-yl)acetonitrile
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-I H-pyrido^.S-bJII ^loxazin-l-y propanenitrile, tert-butyl (1-(4-(1-(cyanomethyl)- 3-(methoxymethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (35mg, 0.063 mmol) was reacted to afford the title compound (18.5mg). 1H NMR (500 MHz, CH3OD) 7.63 (s, 1 H), 7.45 (d, 2H), 7.39 (d,2H), 7.32 (m, 3H), 7.26 (m, 2H), 5.28 (t, 1 H), 5.16 (dd, 2H), 4.01 (dd, 2H), 2.74-2.79 (m, 2H), 2.55-2.61 (m, 2H), 2.22-2.26 (m, 1 H), 1.96-1.99 (m, 1 H). LCMS (Method D): RT = 0.78 min, M+H+ = 456.2.
Example 96: 6-(4-(1 -aminocvclobutyl)phenyl)-3-(methoxymethyl)-1 -methyl-7- phenyl-1 H-pyridor2,3-b][1 ,41oxazin-2(3H)-one
Figure imgf000138_0001
Step 1 : tert-butyl (1-(4-(3-(methoxymethyl)-1-methyl-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof2,3-bU1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
2-chloro-3-methoxypropanoic acid (0.257 g, 1.854 mmol) In a round-bottomed flask was mixed with thionyl chloride (0.149 ml, 2.039 mmol). A drop of DMF was added and the mixture was stirred for 0.5h at room temperature then 80 degree for 2h. The resulted light yellow liquid was diluted with THF (5ml).
At 0 degree, in a 100 mL round-bottomed flask was tert-butyl 1-(4-(5-amino-6-oxo-3- phenyl-1 ,6-dihydropyridin-2-yl)phenyl)cyclobutylcarbamate (0.5g, 1.159 mmol) and N- ethyl-N-isopropylpropan-2-amine (1.015 ml, 5.79 mmol) in THF (25ml) to give a yellow suspension. The above acid chloride in THF solution was added in dropwise to the suspension in 30 min. The reaction mixture was stirred at room temperature for 16h. The reaction mixture was transferred to a 250ml round bottom flask. Sodium bicarbonate saturated solution (30 ml) and sodium bicarbonate (1g) were added. The resulted mixture was heated to reflux for 24 hours. The reaction mixture was cooled down to room temperature. The organic phase was separated. The aqueous phase was extracted with ethyl acetate (40ml). The organic phase was combined and washed with water (30mlX2), brine (25ml) then concentrated at reduced pressure. The residue was suspended in DCM (10ml). The precipitate was collected by filtration and washed with DCM (5 ml) to afford product 0.26g. ).Ή NMR (500 MHz, CDCI3): 8.11 (s, 1 H), 7.23-7.31 (m, 6H), 7.15 (m, 2H), 7.07 (s, 1H), 5.06 (t, 1H), 5.01 (br, 1H), 4.08 (dd, 1 H), 3.95 (dd, 1H), 3.39 (s, 3H), 2.41-2.47 (m, 4H), 2.05 (m, 1 H), 1.80 (m, 1 H), 1.21-1.36 (br, 9H).
Step 2: tert-butyl (1-(4-(3-(methoxymethyl)-1-rnethyl-2-oxo-7-phenyl-2.3-dihvdro-1H- PVridof2,3-b1[1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
A solution of tert-butyl (1-(4-(3-(methoxymethyl)-1-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (60mg, 0.116 mmol) in DMF (1 ml) was added potassium carbonate (48 mg) and iodomethane (8.7μΙ, 0.14 mmol). The reaction mixture was stirred at room temperature for 16 h then was diluted with ethyl acetate (20 ml). Washed with water, brine, and dried with Na2S04, filtered and
concentrated to give crude product which was purified by column (Biotage, size: 25g) eluted with ethyl acetate/cyclohexane (0-50%) to afford product (27mg). Ή NMR (500 MHz, CDCI3): 7.19-7.32 (m, 10H), 5.05(t, 1 H), 4.97 (br, 1 H), 4.06 (dd, 1 H), 3.94 (dd, 1 H), 3.40 (s, 3H), 3.37 (s, 3H), 2.43-2.48 (m, 4H), 2.04 (m, 1 H), 1.79 (m, 1 H), 1.24-1.36 (m, 9H).
Step 3: 6-(4-(1-aminocvclobutyl)phenyl)-3-(methoxymethyl)-1-methyl-7-phenyl-1 H- pyridof2,3-biri.41oxazin-2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(3- (methoxymethyl)-1-methyl-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (35mg, 0.066 mmol) was reacted to afford the title compound (28mg). 1H NMR (500 MHz, CH3OD) 7.49 (s, 1H), 7.42 (d, 2H), 7.38 (d,2H), 7.31 (m, 3H), 7.25 (m, 2H), 5.19 (t, 1 H), 4.02 (dd, 1 H), 3.92 (dd, 1 H), 3.44 (s, 3H), 2.74- 2.79 (m, 2H), 2.56-2.60 (m, 2H), 2.21-2.26 (m, 1 H), 1.94-1.99 (m, 1H). LCMS (Method D): RT = 0.79 min, M+H+ = 430.2. Example 97: 6-(4-((1s.3s)-1-amino-3-hvdroxycvclobutyl)phenyl)-1-methyl-7-phenyl- 1 H-pyridof2.3-bin .41oxazin-2(3H)-one
Figure imgf000140_0001
Step 1 : tert-butyl (1s,3s)-3-hvdroxy-1-(4-(4,4,5,5-tetramethyl-1 ,3.2-dioxaborolan-2- yQphenvDcyclobutylcarbamate
In a 40 mL reaction tube was added tert-butyl (1 s,3s)-1-(4-bromophenyl)-3- hydroxycyclobutylcarbamate (0.25 g, 0.731 mmol) in anhydrous tetrahydrofuran (14 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 20 minutes, followed by the addition of [1 ,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (60 mg, 0.073 mmol). After bubbling nitrogen for a further 15 minutes, potassium acetate (143 mg, 1.461 mmol) and bis(pinacolato)diboron (223 mg, 0.877 mmol) were added. The reaction mixture was heated to reflux overnight then concentrated to dryness under reduced pressure and purified by Biotage
chromatography (cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give the desired product as a colourless oil that solidified upon standing (240 mg, 84% yield). 1H-NMR (500 MHz, CDCI3) δ 7.71 (2H, d), 7.44 (2H, d), 4.15 (1 H, br s), 2.87-2.98 (2H, m), 2.27-2.44 (2H, m), 1.22-1.49 (21 H, br m).
Step 2: tert-butyl (1s.3s)-3-hvdroxy-1-(4-(1-methyl-2-oxo-7-phenyl-2,3-dihvdro- H- pyridor2,3-biri,41oxazin-6-yl)phenyl)cyclobutylcarbamate
In a 10 mL microwave tube was added 6-bromo-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (50 mg, 0.157 mmol), fert-butyl (1 s,3s)-3-hydroxy-1-(4-(4,4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutylcarbamate (73 mg, 0.188 mmol) and terf-butyl (1s,3s)-3-hydroxy-1-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl)cyclobutylcarbamate (73 mg, 0.188 mmol) in a mixture of 1 ,4-dioxane (3.6 mL) and water (1.2 mL) to give a yellow solution. This was degassed by bubbling nitrogen for 15 minutes, then heated to 100°C under microwave irradition for 15 minutes. The reaction mixture was diluted with water (10 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 100:0 to 0: 100) then Biotage chromatography (dichloromethane:methanol, gradient elution from 100:0 to 90:10) to give the desired product as a white solid (8 mg, 10% yield). 1H-N R (500 MHz, CDCI3) δ 7.18-7.36 (10H, m), 4.91 (3H, s), 4.06 (1 H, br s), 3.40 (3H, s), 3.00 (2H, br s), 2.77 (2H, br s), 1.12-1.52 (9H, br m). LCMS (Method D) RT = 1.19 min, M+H+ = 502.20.
Step 3: 6-(4-((1s,3s)-1-amino-3-hvdroxycvclobutyl)phenyl)-1 -methyl-7-phenyl-1 H- Pyridor2.3-biri.41oxazin-2(3H)-one
In a 10 mL round-bottomed flask was added terf-butyl (1 s,3s)-3-hydroxy-1-(4-(1-methyl- 2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (5 mg, 9.97 μπιοΙ) in 1 ,4-dioxane (1 mL) to give a colourless solution. To this was added dropwise 4M HCI in 1 ,4-dioxane (1 mL, 4.00 mmol) and the resulting solution stirred at room temperature for 6 hours. The reaction mixture was diluted by the dropwise addition of diethyl ether (2 mL) and stirred for 10 minutes to give a white precipitate. The layers were allowed to settle and the supernatant solution removed. The solid was washed twice more with diethyl ether (2 mL), allowing to settle and removing the supernatant solvent each time. The remaining solvent was removed by concentration to dryness under reduced pressure, then freeze-drying over the weekend to give the desired product as a white solid (4 mg, 92% yield). 1H-NMR (500 MHz, MeOD) δ 7.53 (1 H, s), 7.38-7.45 (4H, m), 7.24-7.32 (3H, m), 7.18-7.24 (2H, m), 4.93 (2H, s), 4.05 (1 H, quin), 3.42 (3H, s), 3.00-3.12 (2H, m), 2.37-2.48 (2H, m). LCMS (Method D) RT = 0.71 min, M+H+ = 402.20.
Example 98: 1-(4-(1-(difluoromethvn-8-phenyl-4H-pyridor2.3-bin.2.4Uriazolor4.3- dlH ,41oxazin-7-yl)phenyl)cylobutanamine
Figure imgf000141_0001
Step 1 : (E)-Tert-butyl (1-(4-(2-(2-(2.2-difluoroacetyl)hvdrazono)-7-phenyl-2.3-dihvdro-1 H- Pyridof2,3-b1f1 ,4loxazin-6-yl)phenyl)cvclobutyl)carbamate
Ethyl 2,2-difluoroacetate (0.43ml, 4.10 mmol) and hydrazine monohydrate were heated to 60°C in N,N-dimethylformamide (1 ml) for 30 minutes to give a colourless solution. Tert-butyl (IWy-phenyl- -thioxo-Z.S-dihydro-IH-pyrido^.S-bltl ^loxazin-e- yl)phenyl)cyclobutyl)carbamate (100mg, 0.205mmol) was added to the previous solution at room temperature. The resulting mixture was heated to 70°C for 20 hours. The reaction mixture was concentrated under reduced pressure and the crude material (100mg, quantitative yield) was used for the next step without further purification. LCMS (Method D): RT = 1.37 min, M+1= 564.
Step 2: Tert-butyl (1-(4-(1-(difluoromethyl)-8-phenyl-4H-pyridor2.3-b1f1.2,4ltriazolof4.3- d1f1.4loxazin-7-yl)phenyl)cyclobutyl)carbamate
A solution of (E)-tert-butyl (1-(4-(2-(2-(2,2-difluoroacetyl)hydrazono)-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (80 mg, 0.142 mmol) in p-xylene (1 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 2% methanol in dichloromethane) to give the title compound (15 mg, 20%). LCMS (Method D): RT = 1.529 min, M+1= 546.
Step 3: 1-(4-(1-(difluoromethvn-8-phenyl-4H-pyrido[2.3-bl[1.2,41triazolof4.3-dlf1.41oxazin- 7-yl)phenyl)cylobutanamine
Tert-butyl (1-(4-(1-(difluoromethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (15 mg, 0.073 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The residue was then slurred in n- hexane (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (8 mg, 65% yield). LCMS (Method D): RT = 0.803 min, M+1 =446. 1H-NMR (500 MHz, CD3OD) δ 8.20 (1 H, s), 7.53-7.47 (3H, m), 7.46-7.41 (2H, m), 7.39- 7.33 (3H, m), 7.30-7.23 (2H, m), 5.77 (2H, s), 2.83-2.70 (2H, m), 2.63-2.53 (2H, m), 2.30-2.19 (1 H, m), 2.06-1.90 (1 H, m). Example 99: 7-(4-(1-aminocvclobutyl)phenyl)-2-methyl-8-phenyl-2.4-dihvdro-1 H- pyridof2,3- iri ,2,41triazolor -diri ,41oxazin-1 -one
Figure imgf000143_0001
Step 1 : tert-butyl (1-(4-(2-methyl-1-oxo-8-phenyl-2,4-dihvdro-1 H-pyridor2.3- biri.2,4ltriazolo[4,3-dl[1 ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(1-oxo-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (26 mg, 0.05 mmol) in dry DMF (1 ml) was added potassium carbonate (21 mg, 0.15 mmol) and methyliodide (10 μΙ, 0.15 mmol) under nitrogen. The resulting mixture was stirred overnight at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 5ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel
chromatography (gradient 0 to 40% EtOAc in cyclohexane) to give the title compound (22 mg, 82%). 1H N R (500 MHz,CDCI3): 8.50 (1 H, s), 7.25 (2H, d), 7.20-7.19 (5H,m), 7.15- 7.12 (2H,m), 5.21 (2H, s), 5.00 (1 H, br s), 3.46 (3H, s), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86-1.78 ( H, m), 1.44-1.15 (9H, br).
Step 2: 7-(4-(1-aminocyclobutyl)phenyl)-2-methyl-8-phenyl-2,4-dihydro- H-pyridof2,3- biri,2.4ltriazolof4,3-din,4loxazin-1-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (2-methyl-1-oxo-8-phenyl-2,4-dihydro-1 H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (26 mg, 0.05 mmol) was reacted to afford the title compound (18 mg, 56%). LCMS (method D): RT = 0.764 min, M-NH2 = 409. 1H NMR (500 MHz, MeOD): 8.56 (1 H, s), 7.45 (2H, d), 7.41 (2H, d), 7.33-7.31 (3H, m), 7.24-7.23 (2H, m), 5.43 (2H, s), 3.53 (3H, s), 2.78-2.72 (2H, m), 2.59-2.53 (2H, m), 2.25-2.18 (1 H, m), 1.99-1.91 (1 H, m). Example 100: 1-(4-(8-phenyl-1-(trifluoromethyl)-4H-pyridor2,3-b 1,2,41triazolof4,3- diri,41oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000144_0001
Step 1 : Tert-butyl (1-(4-(8-phenyl-1-(trifluoromethyl)-4H-pyridor2.3-blf1,2.41triazolor4.3- d1f1 ,4loxazin-7-yl)phenyl)cyclobutyl)carbannate
A solution of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (70 mg, 0.144 mmol) and 2,2,2-trifluoroacetohydrazide (7.3mg, 0.057 mmol) in p-xylene (1 ml) was heated to 150°C for 30 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (7 mg, 20%). LC S (Method D): RT = 1.633 min, M+1 = 564. Ή NMR (500 MHz, CDCI3): 7.96 (1 H, s), 7.36-7.27 (7H, m), 7.21- 7.17 (2H, m), 5.63 (2H, s), 2.55-2.40 (4H, m), 2.14-1.98 (1 H, m), 1.88-1.78 (1 H, m), 1.37-1.20 (9H, bs).
Step 2: 1-(4-(8-phenyl-1-(trifluoromethyl)-4H-pyridor2.3-biri,2.4ltriazolor4,3-dlf1.41oxazin- 7-yl)phenyl)cyclobutanamine
Tert-butyl (1-(4-(8-phenyl-1-(trifluoromethyl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (7 mg, 0.012 mmol) was dissolved in TFA (1 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (1.5 mg, 26% yield). LCMS (Method D): RT = 0.877 min, M+1 =465. H-NMR (500 MHz, CD3OD) δ
8.02 (1 H, s), 7.50 (2H, d), 7.43 (2H, d), 7.38-7.36 (3H, m), 7.25 (2H, m), 5.77 (2H, s), 2.75-2.69 (2H, m), 2.55-2.42 (2H, m), 2.29-2.17 (1 H, m), 2.05-1.90 (1 H, m). Example 101: 2-(7-(4-(1 -aminocvclobutyl)phenyl)-1 -oxo-8-phenyl-1 H-pyridor2,3- biri,2,41triazolor4,3-diri,41oxazin-2(4H)-yl)acetoriitrile
Figure imgf000145_0001
Step 1 : tert-butyl (1-(4-(2-(cvanomethyl)-1-oxo-8-phenyl-2,4-dihvdro-1 H-pyridor2.3- b1[1,2,4ltriazolo[4,3-dlf1 ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1-(4-(1-oxo-8-phenyl-2,4-dihydro-1H-pyrido[2,3- b][1,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (70 mg, 0.14 mmol) in dry DMF (1 ml) was added potassium carbonate (57 mg, 0.41 mmol) and
bromoacetonitrile (29 μΙ, 0.41 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 35% EtOAc in cyclohexane) to give the title compound (45 mg, 60%). Ή NMR (500 Hz,CDCI3): 8.43 (1 H, s), 7.26-7.19 (7H,m), 7.14-7.12 (2H, m), 5.25 (2H, s), 4.96 (1H, br s), 4.72 (2H, s), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86- 1.78 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 2-(7-(4-( 1 -aminocyclobutyl)phenyl)-1 -oxo-8-phenyl-1 H-pyridof2,3- bin .2.4ltriazolof4,3-dl[1 ,41oxazin-2(4H)-yl)acetonitrile
Following the procedure for 1-((1H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (2-(cyanomethyl)-1-oxo-8-phenyl-2,4-dihydro-1H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (45 mg, 0.08 mmol) was reacted to afford the title compound (34 mg, 61%). LCMS (Method D): RT = 0.777 min, M-NH2 =434. 1H NMR (500 MHz, MeOD): 8.54 (1H, s), 7.45 (2H, d), 7.41 (2H, d), 7.33-7.31 (3H, m), 7.24- 7.23 (m, 2H), 5.47 (2H, s), 5.01 (2H, s), 2.78-2.72 (m, 2H), 2.59-2.53 (m, 2H), 2.25-2.18 (m, 1 H), 1.99-1.91 (m, 1 H). Example 102: 6-(4-(1 -aminocvclobutvDphenylH -isobutyl-7-phenyl-1 H-pyridor2,3- bin,41oxazin-2(3H)-one
Figure imgf000146_0001
Step 1 tert-butyl (1-(4-n-isobutyl-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridor2,3-biri.41oxazin- 6-yl)phenyl)cvclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1 -(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (100mg, 0.212 mmol) was reacted with 1-bromo-2-methylpropane (87mg, 0.636 mmol) to afford the title compound (78mg). LCMS (method D) RT= 1.73 min, M+H= 528.2
Step 2: 6-(4-(1 -aminocvclobutyl)phenyl)-1-isobutyl-7-phenyl-1 H-pyridof2,3-blf1 ,41oxazin- 2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-I H-pyrido^.S-bltl^loxazin-l-yOpropanenitrile, tert-butyl (1-(4-(2-oxo-7-phenyl-1- propyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (50mg, 0.078 mmol) was reacted to afford the title compound (34.5mg). 1H NMR (500 MHz, CH3OD) 7.57 (1 H, s), 7.38-7.42 (4H, m), 7.30-32 (3H, m), 7.21-7.2 (2H, m), 4.97 (2H, s), 3.92 (2H, d), 2.73-2.79 (2H, m), 2.54-2.60 (2H, m), 2.22-2.25 (1 H, m), 2.13-2.16 (1 H, m), 1.95-1.99 (1 H, m), 1.0 (6H, d). LCMS (method D) RT: 0.93, M+H+ = 428.2.
Example 103: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-2-(2,2,2-trifluoroethyl)-2,4- dihvdro-1 H-pyridof2.3-bin .2,41triazolor4.3-dTM .41oxazin-1 -one
Figure imgf000146_0002
Step 1 : tert-butyl (1-(4-(1-oxo-8-phenyl-2-(2.2.2-trifluoroethyl)-2,4-dihvdro-1 H-pyridor2,3- blf1 ,2,41triazolor4,3-dl[1 ,4]oxa2in-7-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(1 -oxo-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1,2,4]triazolo[4,3-d][1 ,4]oxa2in-7-yl)phenyl)cyclobutyl)carbamate (98 mg, 0.19 mmol) in dry DMF (2 ml) was added potassium carbonate (79 mg, 0.57 mmol) and 2-bromo- 1 ,1 ,1-trifluoroethane (94 mg, 0.57 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at 80°C. Water was added and the mixture was extracted with EtOAc (3x). The combined organic phases were dried over Na2S0 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 27% EtOAc in cyciohexane) to give the title compound (45 mg, 40%). 1H NMR (500 MHz,CDCI3): 8.47 (1 H, s), 7.26-7.20 (7H,m), 7.14-7.12 (2H, m), 5.25 (2H, s), 4.96 (1 H, br s), 4.38 (2H, q), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86- 1.78 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-2-(2.2.2-trifluoroethyl)-2,4-dihvdro-1 H- pyrido[2,3-biri.2,4ltriazolor4.3-d1f1 ,4loxazin-1-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1 -(4- (1-oxo-8-phenyl-2-(2,2,2-trifluoroethyl)-2,4-dihydro-1 H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (45 mg, 0.08 mmol) was reacted to afford the title compound (34 mg, 61 %). LCMS (Method D): RT = 0.923 min, M-NH2 = 477. 1H NMR (500 MHz, MeOD): 8.43 (1 H, s), 7.34-7.28 (4H, m), 7.21-7.20 (3H, m), 7.13 -7.12 (2H, m), 5.34 (2H, s), 4.49 (2H, q), 2.70 -2.62 (2H, m), 2.4 9-2.43 (2H, m), 2.1 5-2.08 (1 H, m), 1.91-1.77 (1 H, m).
Example 104: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4,5-dihvdro-1 H-pyridor2,3- blH ,41diazepin-2(3H)-one
Figure imgf000147_0001
Step 1 : Ethyl 3-((6-(4-(1-((tert-butoxycarbonyl)amino)cvclobutyl)phenyl)-3-nitro-5- phenylpyridin-2-yl)amino)propanoate
To a solution of tert-butyl (1-(4-(6-chloro-5-nitro-3-phenylpyridin-2- yl)phenyl)cyclobutyl)carbamate (1.03 g, 2.14 mmol) in EtOH (8 ml) was added triethylamine (1.49 ml, 10.72 mmol) and ethyl 3-aminopropanoate.HCI (1.65 g, 10.72 mmol) under nitrogen. The resulting mixture was stirred for 1.5 hours at 80°C under microwave irradiation. Water was added and the mixture was extracted with AcOEt (3 x 15ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 30% EtOAc in cyclohexane) to give the title compound (1.2 g, quantitative). 1H NMR (500 MHz,CDCI3): 8.50 (1 H, s), 8.38 (1H, s), 7.32 (2H, d), 7.23 -7.18 (5H, m), 7.09 -7.07 (2H, m), 5.00 (1H, br s), 4.12 (2H, q), 3.98 (2H, q), 2.71- 2.68 (2H, t), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br), 1.20 (3H, t).
Step 2: 3-((6-(4-(1-((tert-butoxycarbonyl)amino)cvclobutyl)phenyl)-3-nitro-5- phenylpyridin-2-yl)amino)propanoic acid
To a solution of ethyl 3-((6-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-3-nitro-5- phenylpyridin-2-yl)amino)propanoate (558 mg, 0.99 mmol) in THF (4 ml) was added 1 NaOH (4 ml, 4 mmol). The resulting mixture was stirred overnight at 50°C. The organic was separated and concentrated to dryness under reduced pressure. The resulting residue was used directly without purification.
Step 3: 3-((3-amino-6-(4-(1 -((tert-butoxycarbonyl)amino)cvclobutyl)phenyl)-5- phenylpyridin-2-yl)amino)propanoic acid
To a solution of 3-((6-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-3-nitro-5- phenylpyridin-2-yl)amino)propanoic acid (200 mg, 0.38 mmol) in THF (100 mL) was added Pd/C (4 mg, 0.04 mmol). The resulting mixture was purged with nitrogen and with H2 and then stirred under H2 (2 bars) overnight at room temperature. The black mixture was filtered over celite, rinsed few times with THF (making sure the cake was wet at all times), dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was used directly without purification.
Step 4: tert-butyl (1-(4-(2-oxo-8-phenyl-2.3.4.5-tetrahvdro-1 H-pyridor2.3-bin .41diazepin- 7-yl)phenyl)cvclobutyl)carbamate
To a solution of 3-((3-amino-6-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-5- phenylpyridin-2-yl)amino)propanoic acid (100 mg, 0.20 mmol) in dry DMF (1 ml) was added EDC (153 mg, 0.80 mmol) and HOBT (122 mg, 0.796 mmol) under nitrogen. The resulting mixture was stirred overnight at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 55% EtOAc in cyclohexane) to give the title compound (68 mg, 71 %). 1H NMR (500 MHz,CDCI3): 8.38 (1 H, br s), 7.94 (1 H, br s), 7.72-7.12 (8H, m), 7.01-6.99 (2H, m), 5.00 (1 H, s), 3.68-3.66 (2H, m), 2.82-2.80 (2H, m), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br)
Step 5: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4,5-dihydro-1 H-pyridof2.3- blf 1 ,41diazepin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (2-oxo-8-phenyl-2,3,4)5-tetrahydro-1 H-pyrido[2,3-b][1 ,4]diazepin-7- yl)phenyl)cyclobutyl)carbamate (27 mg, 0.06 mmol) was reacted to afford the title compound (25 mg, 73%). LCMS (Method D): RT = 0.644 min, M+1 = 385. 1H NMR (500 MHz, MeOD): 7.47-7.43 (5H, m), 7.27-7.25 (3H, m), 7.16-7.14 (2H, m), 3.78-3.76 (2H, m), 2.90-2.88 (2H, m), 2.79-2.73 (2H, m), 2.62-2.56 (2H, m), 2.26 -2.22 (1 H, m), 1.99- 1.95 (1 H, m).
Example 105 : 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-propyl-1 H-pyridor2,3- bin.41oxazin-2(3H)-one
Figure imgf000149_0001
Step 1 tert-butyl (1-(4-(2-oxo-7-phenyl-1-propyl-2,3-dihvdro-1 H-pyrido[2.3-bin ,41oxazin- 6-yl)phenyl)cvclobutyl)carbamate
Following the procedure for tert-butyl (1-(4-(1-(2-cyanoethyl)-2-oxo-7-phenyl-2,3-dihydro- 1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate, tert-butyl 1 -(4-(2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (60mg, 0.127 mmol) was reacted with 1-iodopropane (32.4mg, 0.191 mmol) to afford the title compound (43mg. 1H N R (500 MHz, CD3OD): 7.53 (1 H, s), 7.21-7.32 (9H, m), 4.94 (2H, s), 4.02 (2H, t), 2.44-2.47 (4H, m), 2.05-2.10 (1H, m), 1.85-1.87 (1 H, m), 1.73-1.77 (2H, m), 1.02-1.48 (9H, br), 1.01 (3H, t).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-7-phenyl-1-propyl-1 H-pyridor2,3-b1i1.41oxazin- 2(3H)-one
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)propanenitrile, tert-butyl (1-(4-(2-oxo-7-phenyl-1- propyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (40mg, 0.078 mmol) was reacted to afford the title compound (16.5mg). 1H NMR (500 MHz, CH3OD) 7.55 (1 H, s), 7.41 (2H, d), 7.38 ( 2H, d), 7.32 (3H, m), 7.24 (2H, m), 4.96 (2H, s), 4.02 (2H, t), 2.73-2.78 (2H, m), 2.54-2.60 (2H, m), 2.20-2.28 (1H, m), 1.91-1.99 (1 H, m), 1.71-1.79 (2H, m), 1.01 (3H, t). LCMS (Method D): RT = 0.79 min, M+H+ = 414.2.
Example 106 : 6-(4-(1-am'inocvclobutyl)phenyl)-1-(3-hvdroxy-2-(hvdroxymethyl)-2- methylpropyl)-7-phenyl-1 -pyridor2.3-b1M ,41oxazin-2(3H)-one
Figure imgf000150_0001
Step 1 : tert-butyl (1-(4-(1-((3-methyloxetan-3-yl)methyl)-2-oxo-7-phenyl-2.3-dihvdro-1 H- Pyridof2,3-b1f1,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (50 mg, 0.11 mmol) in dry DMF (1 ml.) was added potassium carbonate (44 mg, 0.32 mmol) and 3-(chloromethyl)-3-methyloxetane (38 mg, 0.32 mmol) under nitrogen. The resulting mixture was stirred for 2.5 hours at 80°C. A saturated solution of NaHC03 was added and the mixture was extracted with AcOEt (3x). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 40% EtOAc in cyclohexane) to give the title compound (21 mg, 36%). 1H NMR (500 MHz,CDCI3): 7.23 -7.17 (8H, m), 7.09-7.07 (2H, m), 4.94 (2H, s), 4.83 (2H, s), 4.64 (2H, d), 4.23 (2H, d), 4.05 (2H, s), 2.52-2.20 (4H, m), 2.00-1.95 (1 H, m), 1.79 -1.65 (1 H, m), 1.36 (3H, s), 1.40-1.10 (9H, br).
Step 2: 6-(4-( 1 -aminocyclobutyl)phenyl)-1 -(3-hydroxy-2-(hvdroxymethyl)-2- methylpropyl)-7-phenyl-1 H-pyridor2,3-biri .41oxazin-2(3H)-one
Following the procedure for 1 -((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1 -((3-methyloxetan-3-yl)methyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (16 mg, 0.03 mmol) was reacted to afford the title compound (16 mg, 79%). LCMS (Method D): RT = 0.721 min, M+1 =474. 1H NMR (500 MHz, MeOD): 8.03 (1 H,s), 7.44-7.38 (4H,m), 7.30-7.24 (5H, m), 4.96 (2H, s), 4.11 (2H, s), 3.47-3.41 (4H, m), 2.77 -2.72 (2H, m), 2.59-2.53 (2H, m), 2.25 -2.21 (1 H, m), 1.98- 1.94 (1 H, m), 0.92 (3H, s).
Example 107: 7-(4-(1 -Aminocyclobutyl)phenyl)-8-phenyl-4H-pyridor213- biri.2,41triazolor4,3-diri,4loxazin-1-yl)phenyl)methanol
Figure imgf000151_0001
Step 1 : 2-Hvdroxyacetohydrazide
Ethyl glycolate (5g, 0.047 mol) and hydrazine monohydrate (3.6g, 0.071 mol) were heated to reflux for 15 hours. The reaction mixture was concentrated under reduced pressure and the residue was treated with toluene (3 x 50ml) to azeotrope hydrazine in excess.
This was repeated three times. The crude residue was used for the next step without further purification. Step 2: Tert-butyl (1-(4-(1-(hvdroxymethyl)-8-phenyl-4H-pyridor2.3-bl[1.2.4ltriazolor4.3- d1f1 ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (250 mg, 0.513 mmol) and 2- hydroxyacetohydrazide (230mg, 2.56 mmol) in p-xylene (1.5 ml) was heated to 150°C for 30 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (40 mg, 15%). LCMS (Method D): RT = 1.297 min, M+1 = 526. Ή NMR (500 MHz, CDCI3) δ 8.33 (1 H, s), 7.37-7.32 (2H, m), 7.31-7.27 (5H, m), 7.25-7.20 (2H, m), 5.59 (2H, s), 5.06 (2H, bs), 5.02 (1 H, bs), 2.53-2.43 (4H, m), 2.14-2.02 (1 H, m), 1.88-1.76 (1 H, m), 1.45-1.27 (9H, bs).
Step 3: 7-(4-(1-Aminocvclobutyl)phenyl)-8-phenyl-4H-pyridor2.3-biri ,2.41triazolof4.3- dlf 1 ,41oxazin-1 -vDphenvDmethanol
Tert-butyl (1 -(4-(1 -(hydroxymethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (5 mg, 9.51 μηηοΙ) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (2.5 mg, 60% yield). LCMS (Method D) RT = 0.747 min, M+1 =436. 1H-NMR (500 MHz, CD3OD) δ 8.49 (1 H, s), 7.42- 7.36 (2H, d), 7.34-7.29 (2H, d), 7.26-7.20 (3H, m), 7.19-7.14 (2H, m), 5.60 (2H, s), 4.90 (2H, s), 2.72-2.61 (2H, m), 2.53-2.43 (2H, m), 2.20-2.08 (1 H, m), 1.93- 1.82 (1 H, m), 2.01-1.92 (1 H, m).
E xample 108: 6-(4-(1 -aminocvclobutyl)phenyl)-1 -methyl-7-phenyl-3.4- dihvdropyridor3,2-d1pyrimi -one
Figure imgf000153_0001
Step 1 : tert-butyl 1-(4-(3-methyl-2-oxo-7-phenyl-1.2,3,4-tetrahvdropyhdor2.3-blpyrazin-6- vDphenvDcvclobutylcarbamate
To a solution of terf-butyl (1-(4-(2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[3,2-d]pyrimidin- 6-yl)phenyl)cyclobutyl)carbamate (71 mg, 0.15 mmol) in dry DMF (1 ml) was added potassium carbonate (21 mg, 0.15 mmol) and methyl iodide (9.4 μΙ, 0.15 mmol) under nitrogen. The resulting mixture was stirred overnight at room temperature. Potassium carbonate (42 mg, 0.30 mmol) and methyl iodide (18.8 μΙ, 0.30 mmol) were added under nitrogen. The resulting mixture was stirred for 1 hr at room temperature. Potassium carbonate (146 mg, 1.05 mmol) and methyl iodide (66 μΙ, 1.05 mmol) were added under nitrogen. The resulting mixture was stirred for 4 hr at 60°C. Saturated sodium
bicarbonate solution was added and the mixture was extracted with ethyl acetate (3 x 10ml). The combined organic phases were dried over Na2S0 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 100% EtOAc in cyclohexane) to give the title compound (31 mg, 42%). LCMS (Method D): RT = 1.437 min, M+H+ = 485. 1H NMR (500 MHz, CDCI3): 7.25-7.38 (7H, m), 7.20-7.25 (2H, m), 7.16 (1 H, s), 5.47 (1 H, br s), 5.10 (1 H, br s), 4.73 (2H, s), 3.35 (3H, s), 2.25-2.70 (4H, br m), 2.00-2.13 (1 H, m), 1.75-1.85 (1 H, m), 1.10-1.40 (9H, br).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-phenyl-3,4-dihvdropyrido[3,2- dlpyrimidin-2(1 H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl 1 -(4- (1-methyl-2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[3,2-d]pyrimidin-6- yl)phenyl)cyclobutylcarbamate (31 mg, 0.064 mmol) was reacted to afford the title compound (5 mg, 20%). LCMS (Method D): RT = 0.763 min, M-NH2 = 368. Ή NMR (500 MHz, MeOD): 7.20-7.40 (10H, m), 4.62 (2H, s), 3.30 (3H, s), 2.48-2.13 (2H, m), 2.20-2.35 (2H, m), 2.00-2.13 (1 H, m), 1.70-1.80 (1 H, m).
Example 109: 6-(4-(1-aminocvclobutyl)phenyl)-1,3-bis(2-fluoroethyl)-7-phenyl-3,4- dihydropyrido[3,2-dlpyrimidin-2(1H)-one
Figure imgf000154_0001
Step V. tert-butyl (1 -(4-(1.3-bis(2-fluoroethyl)-2-oxo-7-phenyl-1 ,2.3.4- tetrahydroPYhdof3,2-dipyrirnidin-6-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-7-phenyl-1 ,2,3,4-tetrahydropyrido[3,2-d]pyrimidin- 6-yl)phenyl)cyclobutyl)carbamate (41 mg, 0.09 mmol) in dry DMF (1 ml) at room temperature was added sodium hydride (3.5 mg, 0.09 mmol) and 1-fluoro-2-iodoethane (7.6 μΙ, 0.09 mmol) under nitrogen. The resulting mixture was stirred for 1h at room temperature. Sodium hydride (3.5 mg, 0.09 mmol) and 1 -fluoro-2-iodoethane (7.6 μΙ, 0.09 mmol) were added under nitrogen. The resulting mixture was stirred for 1 h at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 5ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel
chromatography (gradient 0 to 35% EtOAc in cyclohexane) to give the title compound (30 mg, 61%). 1H NMR (500 MHz,CDCI3): 7.24 (1H, s), 7.20-7.17 (7H, m), 7.12-7.10 (2H, m), 4.96 (1 H, br s), 4.74 (2H, s), 4.72 (2H, dt), 4.62 (2H, dt), 4.12 (2H, dt), 3.71 (2H, dt), 2.52- 2.20 (4H, m), 2.00-1.95 (1H, m), 1.79 -1.65 (1 H, m), 1.40-1.10 (9H, br)
Step 2: 6-(4-(1-aminocyclobutyl)phenyl)-1 ,3-bis(2-fluoroethyl)-7-phenyl-3.4- dihvdropyhdof3,2-dlpyrimidin-2(1 H)-one
Following the procedure for 1 -((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1 ,3-bis(2-fluoroethyl)-2-oxo-7-phenyl-1
yl)phenyl)cyclobutyl)carbamate (15 mg, 0.03 mmol) was reacted to afford the title compound (5 mg, 27%). LCMS (Method D): RT = 0.872 min, M-NH2 = 446. Ή NMR (500 MHz, MeOD): 7.52 (1H, s), 7.45-7.39 (4H, m), 7.32-7.30 (3H, m), 7.24-7.22 (2H, m), 4.81-4.77 (4H, m), 4.68 (2H, dt), 4.28 (2H, dt), 3.83 (2H, dt), 2.81-2.75 (2H, m), 2.63-2.57 (2H, m), 2.26 -2.24 (1 H, m), 1.99-1.97 (1 H, m).
Example 110: 1-(4-(1-cvclopropyl-8-phenyl-4H-pyridor2,3-bU1,2141triazolor4,3- d1[1,4]oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000155_0001
Step 1 : Tert-butyl (1-(4-(1-cvclopropyl-8-phenyl-4H-pyridor2,3-b1H .2,4ltriazolo[4,3- d1f1 ,4loxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (150 mg, 0.308 mmol) and
cyclopropanecarbohydrazide (41 mg, 0.410 mmol) in p-xylene (1.5 ml) was heated to 150 °C for 20 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (36 mg, 32%). LCMS (Method D): RT = 1.457 min, M+1 = 536.
1H NMR (500 MHz, CDCI3): 8.24 (1 H, s), 7.37-7.27 (7H, m), 7.24-7.19 (2H, m), 5.57 (2H, s), 2.55-2.40 (4H, m), 2.12-2.05 (1 H, m), 1.87-1.78 (1 H, m), 1.43-1.31 (9H, bs), 1.34-1.29 (3H, m), 1.23-1.17 (2H, m).
Step 2: 1-(4-(1-cvclopropyl-8-phenyl-4H-pyridor2,3-b1f1.2.4ltriazolor4.3-d1f1 ,41oxazin-7- vDphenvDcvclobutanamine
Tert-butyl (1-(4-(1 -cyclopropyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin- 7-yl)phenyl)cyclobutyl)carbamate (36 mg, 0.012 mmol) was dissolved in TFA (1 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and the residue was dried to give the desired product as an off-white solid (24 mg, 82% yield). LCMS (Method D) RT = 0.747 min, M+1 =436. H-NMR (500 MHz, CD3OD) δ 8.48 (1 H, s), 7.53-7.47 (2H, m), 7.45-7.41 (2H, d), 7.38-7.33 (3H, m), 7.32-7.26 (2H, m), 5.66 (2H, s), 2.82-2.72 (2H, m), 2.63-2.55 (2H, m), 2.44-2.36 (1 H, m), 2.30- 2.19 (1 H, m), 2.01-1 .92 (1 H, m), 1.28-1.17 (4H, overlapped ddd).
Example 111 : 7-(4-(1-aminocvclobutyl)phenyl)-1,5-dimethyl-8-phenyl-4,5-dihvdro-
1 H-pyridor2,3-biri ,41diazepi -2(3H)-one
Figure imgf000156_0001
Step 1 : tert-butyl n-(4-(1 ,5-dimethyl-2-oxo-8-phenyl-2.3,4.5-tetrahvdro-1 H-pyridor2,3- b1[1 ,41diazepin-7-yl)phenyl)cvclobutyl)carbamate
To a solution of tert-butyl (1 -(4-(2-oxo-8-phenyl-2,3,4,5-tetrahydro-1 H-pyrido[2,3- b][1 ,4]diazepin-7-yl)phenyl)cyclobutyl)carbamate (41 mg, 0.09 mmol) in dry DMF (1 mL) was added potassium carbonate (1 17 mg, 0.85 mmol) and methyliodide (53 μΙ, 0.85 mmol) under nitrogen. The resulting mixture was stirred 1 hour at 80°C. Water was added and the mixture was extracted with EtOAc (3 x 15ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 60% EtOAc in cyclohexane) to give the title compound (15 mg, 35%). Ή NMR (500 MHz,CDCI3): 7.32-7.28 (3H, m), 7.21-7.18 (5H,m), 7.12-7.10 (2H,m), 4.94 (1 H, br s), 3.65 (2H, t), 3.28 (3H, s), 2.96 (3H, s), 2.61-2.59 (2H, t), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86-1.78 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 7-(4-(1-aminocyclobutyl)phenyl)-1 ,5-dimethyl-8-phenyl-4,5-dihvdro-1 H- pyridof2,3-b1[1 ,4ldiazepin-2(3H)-one
Following the procedure for 1 -((1 H-imidazol-2-yl)methyl)-6-(4-(1 - aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1 ,5-dimethyl-2-oxo-8-phenyl-2,3,4,5-tetrahydro-1 H-pyrido[2,3-b][1 ,4]diazepin-7- yl)phenyl)cyclobutyl)carbamate (15 mg, 0.03 mmol) was reacted to afford the title compound (12 mg, 64%). LCMS (Method D): RT = 0.880 min, M+1 = 413. Ή NMR (500 MHz, MeOD): 7.65 (1 H, s), 7.53 (2H, d), 7.39 (2H, d), 7.31-7.28 (3H, m), 7.24-7.22 (2H, m), 3.73 (2H, t), 3.37 (3H, s), 3.02 (3H, s), 2.81 -2.75 (2H, m), 2.68 (2H, t), 2.61-2.55 (2H, m), 2.25-2.18 (1 H, m), 1.99-1.91 (1 H, m).
Example 112: 1 -(4-(1 -isopropyl-8-phenyl-4H-pyridor2,3-b1M .2,41triazolor4.3- cflpl ,41oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000157_0001
Step 1 : Isobutyrohvdrazide
Ethylisobutyrate (5g, 0.043 mol) and hydrazine monohydrate (3.2g, 0.065 mol) were heated to reflux for 15 hours. The reaction mixture was concentrated under reduced pressure and the residue was treated with toluene(3 x 100ml) to azeotrope hydrazine in excess. The crude residue was used for the next step without further purification.
Step 2: Tert-butyl (1-(4-(1 -isopropyl-8-phenyl-4H-pyridor2.3-blf1 ,2.41triazolof4,3- d1[1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (150 mg, 0.308 mmol) and
isobutyrohydrazide (28mg, 0.308 mmol) in p-xylene (1.5 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure. The crude residue was purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (28 mg, 17%). LCMS (Method D): RT = 1.497 min, M+1 = 538. Ή NMR (500 MHz, CDCI3) δ 7.85 (1 H, s), 7.36-7.31 (5H, m), 7.28 (2H, m), 7.24- 7.19 (2H, m), 5.56 (2H, s), 3.46- 3.36 (1 H, m), 2.57-2.39 (4H, m), 2.13-2.02 (1 H, m), 1.88-1.76 (1 H, m), 1.57- 1.53 (6H, d), 1.44-1.29 (9H, bs). Step 3: 1-(4-(1-isopropyl-8-phenyl-4H-pyridor2,3-blM .2,41triazolof4.3-dl[1 ,41oxazin-7- vQphenvOcyclobutanamine
Tert-butyl (1-(4-(1-isopropyl-8-phenyl-4H-pyrido[2>3-b][1 ,2I4]triazolo[4>3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (24 mg, 0.045 mmol) was dissolved in TFA (1 ml) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (18 mg, 92% yield). LCMS (Method D) RT = 0.815 min, M+1 =438. 1H-NMR (500 MHz, CD3OD) δ 8.18 (1 H, s), 7.52-7.47 (2H, m), 7.45-7.41 (2H, d), 7.38-7.34 (3H, m), 7.31-7.26 (2H, m), 5.64 (2H, s), 2.82-2.72 (2H, m), 2.63-2.54 (2H, m), 2.30- 2.19 (1 H, m), 2.02-1.92 (1 H, m), 1.51 (6H, d).
Example 113: 6-(4-((1 r,3r)-1 -amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1 -methyl- 7-phenyl-1 H-pyridor2.3-bin.41oxazin-2(3H)-one
Figure imgf000158_0001
Step 1 : Ethyl 2-((5-bromo-3-nitropyridin-2-yl)oxy)acetate
To a suspension of sodium hydride (5.31 g, 133 mmol) in 1 ,4-dioxane (250 ml), ethyl glycolate (12.56 ml, 133 mmol) was added drop wise over a period of 30 minutes ensuring that the temperature was maintained below 30°C. The resulting thick suspension was stirred at room temperature for 15 minutes. In a separate 11 round- bottomed flask was added 5-bromo-2-chloro-3-nitropyridine (21 g, 88 mmol) in 1 ,4- dioxane (150 ml) to give a brown solution. The suspension of sodium hydride and ethyl glycolate was added drop wise over a period of 30 minutes at 0°C. The resulting reaction mixture was heated to 80°C overnight. The reaction mixture was concentrated under reduced pressure and the crude residue was purified by Biotage silica chromatography (gradient 0% to 10% ethyl acetate in n- hexanes) to give the title compound (1 .8g, 44%). Ή N R (500 MHz, CDCI3): 8.48 (1 H, s), 8.42 (1 H, s), 5.07 (2H, s), 4.28-4.24 (2H, q), 1.31-1.28 (3H, t).
Step 2: Ethyl 2-((3-nitro-5-phenylpyridin-2-yl)oxy)acetate
In a 11 round-bottomed flask was added ethyl 2-(5-bromo-3-nitropyridin-2-yloxy)acetate (18.33 g, 60.1 mmol), phenylboronic acid (10.99 g, 90 mmol), triphenylphosphine (4.73 g, 18.02 mmol) and cesium fluoride (45.6 g, 300 mmol) in 1 ,2-dimethoxyethane (300 ml) to give a yellow solution. The reaction mixture was degassed by bubbling nitrogen for 30 minutes. Palladium(ll) acetate (2.023 g, 9.01 mmol) was added and the mixture was heated to 75°C under a nitrogen atmosphere overnight. The reaction mixture was allowed to cool to room temperature and concentrated to dryness under reduced pressure to give a brown solid. This was re-dissolved in dichloromethane, filtered and concentrated to dryness under reduced pressure to give a brown solid The crude residue was purified via Biotage chromatography (gradient 5% to 60% ethyl acetate in n- hexanes) to give the title compound (6.9g, 38%). H NMR (500 MHz, CDCI3) δ 8.58 (1 H, s), 8.56 (1 H, s), 7.59-7.52 (2H, m), 7.48-7.46 (2H, m), 7.45-7.43 (1 H, m), 5.13 (2H, s), 4.30-4.26 (2H, q), 1.33-1.30 (3H, t).
Step 3: 7-Phenyl-1 H-pyridof2,3-biri.41oxazin-2(3H)-one
In a 500 ml round-bottomed flask was added ethyl 2-(3-nitro-5-phenylpyridin-2- yloxy)acetate (4.6 g, 15.22 mmol) in hydrochloric acid, 37% (40 ml) to give a yellow suspension. The mixture was cooled to 0-5°C followed by the portion wise addition of tin powder (9.94 g, 84 mmol). The addition proved to be exothermic. Caution should be taken while adding. The mixture was then stirred at room temperature for further 30 minutes until all foaming ceased. The reaction mixture was heated to 80°C under a nitrogen atmosphere for 3 hours.
The reaction mixture cooled to room temperature and diluted with water (800ml). The white precipitate was isolated by filtration, washed with water (100 ml) and sucked dry to give a white solid. The solid was azeotroped with toluene (3 x 30 ml) to give a white solid as the title compound (2.6g, 77%). 1H NMR (500 MHz, (CD3)2SO) δ 10.41 (1H, s), 8.10 (1 H, s), 7.59 (2H, d), 7.49-7.42 (2H, t), 7.39-7-38 (1H, d), 4.83 (2H, s). Step 4: 6-bromo-7-phenyl-1 H-pyridor2.3-bli1.4loxazin-2(3H)-one
In a 10ml microwave vial was 7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (50 mg, 0.221 mmol) and N-bromosuccinimide (78.6 mg, 0.441 mmol) in dimethylformamide (1 ml). The reaction mixture was heated to 80°C under microwave irradiation for 30 minutes. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (10ml). The organic solution was washed with water (2x10ml) and brine (2x10ml). The organic phase was dried over anhydrous Na2S04, filtered and concentrated under reduced pressure. The crude residue was purified via Biotage chromatography (gradient 0% to 5% methanol in dichloromethane) to give the title compound as a yellow solid (61 mg, 90%). Ή NMR (500 MHz, CD3OD) δ 7.48-7.32 (5H, m), 7.12 (1 H, s), 4.82 (2H, s).
Step 5: 2-((1 r,3r)-3-hvdroxy-3-methyl-1-(4-(1-methyl-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridor2,3-blf1 ,41oxazin-6-yl)phenyl)cvclobutyl)isoindoline-1 ,3-dione
In a 10 mL microwave tube was added 6-bromo-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (32 mg, 0.100 mmol), 2-(( r,3r)-3-hydroxy-3-methyl-1-(4- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (46 mg, 0.105 mmol) and sodium carbonate (21 mg, 0.201 mmol) in a mixture of 1 ,4-dioxane (3.50 mL) and water (1.20 mL) to give a colourless solution. This was degassed by bubbling nitrogen for 10 minutes followed by the addition of
tetrakis(triphenylphosphine)palladium(0) (12 mg, 10.03 Mmol). The reaction mixture was heated to 100°C under microwave irradiation for 15 minutes. Additional 2-((1r,3r)-3- hydroxy-S-methyl-I^^^.S.S-tetramethyl-I .S^-dioxaborolan^- yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (24 mg, 0.055 mmol) was added and the mixture heated to 100°C under microwave irradiation for a further 15 minutes. The reaction mixture was diluted with ethyl acetate (12 mL), washed with water (10 mL), brine (10 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow oil. This was purified by Biotage chromatography
(cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (18 mg, 33% yield). 1H-NMR (500 MHz, CDCI3) δ 7.74 (2H, dd), 7.66 (2H, dd), 7.51 (2H,d), 7.31 (2H, d), 7.22-7.29 (4H, m), 7.15-7.20 (2H, m), 4.87 (2H, s), 3.37 (3H, s), 3.32 (2H, d), 3.09 (2H, d), 1.40 (3H, s). LCMS (Method D) RT = 1.30 min, M+H+ = 546.00. Step 6: 6-(4-((1r,3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1-methyl-7-phenyl- 1 H-pyridor2,3-biri .41oxazin-2(3H)-one
In a 15 mL reaction tube was added 2-((1r,3r)-3-hydroxy-3-methyl-1-(4-(1-methyl-2-oxo- 7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)isoindoline-1 ,3- dione (18 mg, 0.033 mmol) and hydrazine monohydrate (8 μΙ, 0.165 mmol) in ethanol (1650 μΙ) to give a yellow solution. This was heated to 90°C under a nitrogen atmosphere for 5 hours then allowed to cool to room temperature. Further hydrazine monohydrate (16 μΙ, 0.330 mmol) was added and the reaction mixture heated to 100°C under microwave irradiation for 10 minutes, then to 120°C under microwave irradiation for 20 minutes. Further hydrazine monohydrate (16 μΙ, 0.330 mmol) was added and the mixture heated to 120°C under microwave irradiation for 60 minutes. The resulting precipitate was removed by filtration and washed well with ethanol. The combined filtrates were concentrated to dryness under reduced pressure to give a white solid. This was purified on an SCX catch-and-release column and freeze-dried to give the desired product as a white solid (10 mg, 73% yield). 1H-NMR (500 MHz, MeOD) δ 7.52 (1 H, s), 7.20-7.35 (9H, m), 4.94 (2H, s), 3.42 (3H, s), 2.66 (2H, d), 2.39 (2H, d), 1.55 (3H, s). LCMS (Method D) RT = 0.67 min, M+H+ = 416.20.
Example 114: 7-(4-(1-aminocvclobutyl)phenyl)-1-methyl-8-phenyl-4,5-dihvdro-1H- pyridor2,3-bin.41diazepin-2(3H)-one
Figure imgf000161_0001
Step 1 : tert-butyl (1-(4-(1-methyl-2-oxo-8-phenyl-2,3,4.5-tetrahydro-1H-pyridof2.3- biri.41diazepin-7-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-8-phenyl-2,3,4,5-tetrahydro-1H-pyrido[2,3- b][1 ,4]diazepin-7-yl)phenyl)cyclobutyl)carbamate (53 mg, 0.11 mmol) in dry DMF (1 ml) was added sodium hydride (4.4 mg, 0.11 mmol) and methyliodide (6.9 μΙ, 0.11 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 15ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 57% EtOAc in cyclohexane) to give the title compound (21 mg, 38%). Ή NMR (500 MHz,CDCI3): 7.39 (1 H, s), 7.27-7.23 (7H, m), 7.16-7.14 (2H,m), 5.04 (1H, s), 4.91 (1 H, br s), 3.88 (2H, br s), 3.39 (3H, s), 2.74 (2H, t), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86- 1.78 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 7-(4-(1-aminocvclobutyl)phenyl)-1-methyl-8-phenyl-4,5-d^
bin ,41diazepin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1 -methyl-2-oxo-8-phenyl-2,3,4,5-tetrahydro-1 H-pyrido[2,3-b][1 ,4]diazepin-7- yl)phenyl)cyclobutyl)carbamate (21 mg, 0.04 mmol) was reacted to afford the title compound (19 mg, 72%). LC S (Method D): RT = 0.806 min, M+1 = 399. 1H NMR (500 MHz, MeOD): 7.65 (1 H, s), 7.47 (2H, d), 7.41 (2H, d), 7.29-7.25 (3H, m), 7.20-7.19 (2H, m), 3.86-3.84 (4H, m), 3.41 (3H, s), 2.79 -2.73 (4H, m), 2.61-2.55 (2H, m), 2.25-2.18 (1 H, m), 1.99-1.91 (1 H, m).
Example 115: 1-(4-(1-isobutyl-8-phenyl-4H-pyridor2,3-birt2,4ltriazolor4.3- diri,41oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000162_0001
Step 1: 3-Methylbutanehydrazide
Ethylvalerate (5g, 0.038 mol) and hydrazine monohydrate (2.8g, 0.057 mol) were heated to reflux for 15 hours. The reaction mixture was concentrated under reduced pressure and the residue was treated with toluene (100ml x 3) to azeotrope hydrazine in excess. This was repeated three times. The crude residue was used for the next step without further purification.
Step 2: Tert-butyl (1-(4-(1-isobutyl-8-phenyl-4H-pyridor2.3-b1f1.2.4ltriazolor4.3- d1f1,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (200 mg, 0.410 mmol) and 3- methylbutanehydrazide (47mg, 0.410 mmol) in p-xylene (1.5 ml) was heated to 150 °C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (15 mg, 5%). LCMS (Method D): RT = 1.5687 min, M+1 = 552. Ή NMR (500 MHz, CDCI3): 7.80 (1 H, s), 7.36-7.31 (5H, m), 7.30- 7.27 (2H, m), 7.23- 7.19 (2H, m), 5.57 (2H, s), 2.98 (2H, d), 2.54-2.41 (4H, m), 2.33- 2.24 (1H, m), 2.13-2.01 (1H, m), 1.86- 1.76 (1 H, m), 1.44-1.26 (9H, bs), 1.11 (6H, d).
Step 3: 1-(4-(1-isobutyl-8-phenyl-4H-pyridor2.3-biri .2.41triazolor4,3-dlM ,41oxazin-7- vQphenvDcvclobutanamine
Tert-butyl (1-(4-(1-isobutyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (7 mg, 0.013 mmol) was dissolved in dichloromethane (1 mL); TFA (1 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the
supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (4 mg, 68% yield). LCMS (Method D): RT = 0.844 min, M+1= 452. 1H-NMR (500 MHz, CD3OD) δ 8.07 (1H, s), 7.52-7.45 (2H, d), 7.44-7.40 (2H, d), 7.38-7.33 (3H, m), 7.31-7.26 (2H, m), 5.66 (2H, s), 3.11 (2H, d), 2.81-2.68 (2H, m), 2.61-2.45 (2H, m), 2.32- 2.17 (1 H+ 1H, 2 m), 2.02-1.87 (1 H, m), 1.10 (6H, d).
Example 116: 7-(4-(1-aminocvclobutyl)phenyl)-N,N-dimethyl-8-phenyl-4H- pyrido[2,3-biri.2.41triazolor -diri,4loxazin-1 -amine
Figure imgf000163_0001
Step 1 : tert-butyl (1-(4-n-(dimethylamino)-8-phenyl-4H-pyridof2.3-b1f1.2.41triazolor4.3- diri,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A mixture of tert-butyl 1-(4-(1-bromo-8-phenyl-4H-pyrido[2>3-b][1 ,2,4]triazolo[4l3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutylcarbamate (41 mg, 0.071 mmol), dimethylamine hydrochloride (58.2 mg, 0.714 mmol), and triethylamine (0.099 ml, 0.714 mmol) in NMP (1 ml) was heating at 130 degree for 50 min under microwave condition. The mixture was diluted with DCM (8ml) and water (8ml). The organic phase was washed with water (8mlX3) and brine (5ml). Concentrated and purified by column eluted with MeOH/DCM (0-4%) to give product (8mg). 1H N R (500 MHz, CDCI3) 8.15 (s, 1 H), 7.15-7.34 (m, 9H), 5.51 (s, 2H), 5.01 (s, 1 H), 2.95 (s, 6H), 2.4-2.5 (m, 4H), 2.0-2.15 (m, 1 H), 1.75- 1.85 (m, 1 H), 1.2-1.35 (m, 9H).
Step 2: 7-(4-(1-aminocvclobutyl)phenyl)-N,N-dimethyl-8-phenyl-4H-pyrido[2,3- b]f 1 ,2,4|triazolof413-dlf 1 ,41oxazin-1 -amine
Following the procedure for 3-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1 -yl)propanenitrile, tert-butyl (1 -(4-(1 -(cyanomethyl)- 3-(methoxymethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (8mg, 0.015 mmol) was reacted to afford the title compound (5.5mg). 1H NMR (500 MHz, CH3OD) 8.26 (s, 1H), 7.48 (d, 2H), 7.42 (d, 2H), 7.35 (m, 3H), 7.26 (m, 2H), 5.6 (s, 2H), 2.96 (s, 6H), 2.7-2.8 (m, 2H), 2.55-2.65 (m, 2H), 2.2-2.3 (m, 1 H), 1.9- 2.0 (m, 1 H), LCMS (Method D): RT = 0.77 min, M+H+ = 439.2
Example 117: 7-(4-(1-aminocvclobutyl)phenyl)-1-(2-fluoroethyl)-8-phenyl-4.5- dihvdro-1 H-pyridor2.3-b1H .41diazepin-2(3H)-one
Figure imgf000164_0001
Step 1 : tert-butyl (1-(4-(1-(2-fluoroethvn-2-oxo-8-phenyl-2.3.4.5-tetrahvdro-1 H-pyridof2,3- b1i1 ,4ldiazepin-7-yl)phenyl)cyclobutyl)carbamate
To a solution of tert-butyl (1-(4-(2-oxo-8-phenyl-2,3,4,5-tetrahydro-1H-pyrido[2,3- b][1 ,4]diazepin-7-yl)phenyl)cyclobutyl)carbamate (101 mg, 0.21 mmol) in dry DMF (1 ml) was added sodium hydride (12 mg, 0.32 mmol) and 1-fluoro-2-iodoethane (27 μΙ, 0.32 mmol) under nitrogen. The resulting mixture was stirred for 1 hour at room temperature. Water was added and the mixture was extracted with EtOAc (3 x 10ml). The combined organic phases were dried over Na2S04 and concentrated to dryness under reduced pressure. The resulting residue was purified by Biotage silica gel chromatography (gradient 0 to 100% EtOAc in cyclohexane) to give the title compound (52 mg, 47%). 1H NMR (500 MHz,CDCI3): 7.62 (1 H, s), 7.31-7.22 (7H, m), 7.16-7.14 (2H,m), 5.01 (1 H, s), 4.75 (1 H, br s), 4.74 (2H, dt), 4.07 (2H, dt), 3.92-3.88 (2H, m), 2.73 (2H, t), 2.55-2.25 (4H, m), 2.10 -2.03 (1 H, m), 1.86-1.78 (1 H, m), 1.44-1.15 (9H, br).
Step 2: 7-(4-(1-aminocvclobutyl)phenyl)-1-(2-fluoroethyl)-8-phenyl-4,5-dihvdro-1 H- pyridor2.3-blf1 ,4ldiazepin-2(3H)-one
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin-2(3H)-one, tert-butyl (1-(4- (1-(2-fluoroethyl)-2-oxo-8-phenyl-2,3,4,5-tetrahydro-1 H-pyrido[2,3-b][1 ,4]diazepin-7- yl)phenyl)cyclobutyl)carbamate (15 mg, 0.03 mmol) was reacted to afford the title compound (12 mg, 64%). LCMS (Method D): RT = 0.808 min, M+1 = 431. 1H NMR (500 MHz, MeOD): 7.76 (1 H, s), 7.48 (2H, d), 7.41 (2H, d), 7.29-7.25 (3H, m), 7.20-7.18 (2H, m), 4.67 (2H, dt), 4.16 (2H, dt), 3.88-3.86 (2H, m), 2.79 -2.73 (4H, m , 2.61-2.55 (2H, m), 2.25-2.18 (1 H, m), 1.99-1.91 (1 H, m).
Example 118: 1 -(4-(8-phenyl-1 -(2.2.2-trifluoroethvn-4H-pyridor2.3- b1f1.2,41triazolor4.3-d]ri,41oxazin-7-yl)phenyl)cvclobutanamine
Figure imgf000165_0001
Step 1 : 3,3,3-Trifluoropropanehvdrazide
Ethyl-3,3,3-trifluoropropanoate (2.5g, 0.016 mol) and hydrazine monohydrate (1.2g, 0.024 mol) were heated to reflux for 1 hour. The reaction mixture was concentrated under reduced pressure and the residue was treated with toluene (100ml x 3) to azeotrope hydrazine in excess. The crude residue was used for the next step without further purification.
Step 2:Tert-butyl (1-(4-(8-phenyl-1-(2,2.2-trifluoroethvn-4H-pyrido[2.3- biri ,2,41triazolo[4,3-d1[1 ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (250 mg, 0.513 mmol) and 3,3,3- trifluoropropanehydrazide (145mg, 1.205 mmol) in p-xylene (2 ml) was heated to 150 °C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (38 mg, 13%). LCMS (Method D): RT = 1 499 min, M+1= 578.
1H NMR (500 MHz, CDCI3): 7.84 (1 H, s), 7.36-7.32 (5H, m), 7.31- 7.28 (2H, m), 7.23- 7.18 (2H, m), 5.60 (2H, s), 4.12 (2H, q), 2.52-2.42 (4H, m), 2.12-2.01 (1H, m), 1.87-1.78 (1 H, m), 1.44-1.26 (9H, bs).
Step 3: 1-(4-(8-phenyl-1-(2.2.2-trifluoroethyl)-4H-pyridof2,3-blf1.2.41triazolor4.3- d1[1 ,41oxazin-7-yl)phenyl)cvclobutanamine
Tert-butyl (1-(4-(8-phenyl-1-(2,2,2-trifluoroethyl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (25 mg, 0.043 mmol) was dissolved in dichloromethane (1 ml); TFA (1 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (16 mg, 77% yield). LCMS (Method D): RT = 0.791 min, M+1= 478. 1H-NMR (500 MHz, CD3OD) δ 8.19 (1 H, s), 7.52-7.48 (2H, d), 7.45-7.42 (2H, d), 7.38-7.34 (3H, m), 7.31-7.27 (2H, m), 5.69 (2H, s), 4.47 (2H, q), 2.82-2.73 (2H, m), 2.64-2.54 (2H, m), 2.31- 2.19 (1 H, m), 2.02-1.92 (1H, m). Example 119: 7-(4-(1-aminocvclobutyl)phenyl)-8-phenyl-4H-pyridof2,3- blH .2,41triazolor4.3-diri ,41oxazine-1-carboxamide
Figure imgf000167_0001
Step 1 : Ethylhydrazine carboxylate
Diethyl oxalate (5g, 0.042 mol) and hydrazine monohydrate (2.8g, 0.057 mol) in ethanol (3ml) were heated to reflux for 15 hour. The reaction mixture was concentrated under reduced pressure and the residue was treated with toluene (100ml x 3) to azeotrope hydrazine in excess. The crude residue was used for the next step without further purification.
Step 2: Ethyl 7-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-8-phenyl-4H- pyridof2,3-blf 1 ,2,4Uriazolor4,3-din ,41oxazine-1 -carboxylate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (250 mg, 0.513 mmol) and ethylhydrazine carboxylate (271 mg, 2.05 mmol) in p-xylene (2 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 30% ethyl acetate in n-hexanes) to give the title compound (38 mg, 13%). LCMS (Method D): RT = 1.531 min, M+1 = 568.
Step 3: Tert-butyl (1-(4-(1-carbamoyl-8-phenyl-4H-pyridof2.3-b]f1.2,41triazolof4.3- dlf1 ,41oxazin-7-yl)phenyl)cyclobutyl)carbamate
A suspension of ethyl 7-(4-(1-((tert-butoxycarbonyl)amino)cyclobutyl)phenyl)-8-phenyl- 4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 )4]oxazine-1 -carboxylate (70 mg, 0.123 mmol) and ammonium chloride (10mg, 0.19 mmol) in ammonium hydroxide (1 ml) was heated to 100 °C for 10 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 10% ethyl acetate in cyclohexane) to give the title compound (38 mg, 13%). LCMS (Method D): RT = 1.347 min, M+1 = 539. 1H-NMR (500 MHz, CD3OD) δ 9.10 (1H, s), 7.47-7.40 (1 H, bs), 7.40-7.37 (2H, d), 7.32-7.27 (6H, m), 5.59 (2H, s), 2.59-2.39 (4H, m), 2.14- 2.01 (1H, m), 1.88-1.76 (1H, m), 1.44-1.33 (9H, bs).
Step 4: 7-(4-(1-aminocvclobutyl)prienyl)-8-phenyl-4H-pyridof2,3-biri.2.4ltriazolor4.3- d1[1 ,4]oxazine-1 -carboxamide
Tert-butyl (1-(4-(1-carbamoyl-8-phenyl-4H-pyrido[2,3-b][1 I2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (10 mg, 0.019 mmol) was dissolved in dichloromethane (0.5 ml); this solution was added to a scx-2 catch and release cartridge (1g),
precalibrated using methanol (15ml) and dichloromethane (15ml). After allowing to binding for 15 hours, the column was washed with methanol (6ml) and then the product eluted with 7M ammonia in methanol (4ml x 2). The combined product fractions were concentrated to dryness under reduced pressure to give the desired product as a white solid (5 mg, 61% yield). LCMS (Method D) RT = 0.695 min, M+1= 440. 1H-NMR (500 MHz, CD3OD) δ 8.80 (1H, s), 7.27-7.24 (4H, bs), 7.22-7.17 (3H, m), 7.16-7.13 (2H, m), 5.57 (2H, s), 2.49-2.38 (2H, m), 2.19-2.10 (2H, m), 2.03- 1.92 (1 H, m), 1.71-1.60 (1 H, m).
Example 120: 1 -(4-(1-(fluoromethyl)-8-phenyl-4H-pyridor2.3-b1M ,2.41triazolor4.3- dU1 ,41oxazin-7-yl)phenyl)cvclobutanamine
Figure imgf000168_0001
Step 1 : Tert-butyl n-(4-(1-(fluoromethyl)-8-phenyl-4H-pyridor2.3-biri.2.4ltriazolor4.3- d1H ,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A solution of tert-butyl (1-(4-(1-(hydroxymethyl)-8-phenyl-4H-pyrido[2,3- b][1,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (40 mg, 0.019 mmol), triethylamine (2.65ml, 0.019 mmol) and DeoxoFluor® (50% in THF, 21.05mg, 0.048 mmol) in dichloromethane (1 ml) was heated to 30°C for 15 hours. The resulting reaction mixture was quenched with a saturated solution of sodium bicarbonate and extracted with dichloromethane (10ml x 3). The combined organic layers were concentrated to dryness under reduced pressure. The crude residue was purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in cyclohexane) to give the title compound (38 mg, 13%). LCMS (Method D): RT = 1.439 min, M+1= 528.
1H NMR (500 MHz, CDCI3): 8.01 (1 H, s), 7.37-7.27 (7H, m), 7.24- 7.20 (2H, dd), 5.83 (2H, s), 5.73 (1 H, s), 5.64 (1H, s), 2.54-2.41 (4H, m), 2.12-2.02 (1 H, m), 1.88-1.76 (1 H, m), 1.43-1.30 (9H, bs).
Step 2: 1-(4-(1-(fluoromethyl)-8-phenyl-4H-Dyridof2.3-biri ,2.41triazolof4.3-diri ,41oxazin-7- vOphenyOcvclobutanamine
Tert-butyl (1-(4-(1-(fluoromethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (3 mg, 5.69 μΐηοΙ) was dissolved in dichloromethane (1 ml); TFA (0.5 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (1.5 mg, 61% yield). LCMS (Method D) RT = 0.774 min, M+1= 428. 1H-NMR (500 MHz, CD3OD) δ 8.03 (1 H, s), 7.41- 7.35 (2H, d), 7.34-7.29 (2H, d), 7.27-7.23 (3H, m), 7.21-7.15 (2H, m), 5.86 (1H, s), 5.75 (1 H, s), 5.64 (2H, s), 2.71-2.58 (2H, m), 2.49-2.37 (2H, m), 2.18- 2.07 (1 H, m), 1.91-1.79 (1 H, m).
Example 121 : 1-(4-(1-(2-methoxyethyl)-8-phenyl-4H-pyridor2.3-bin.2.41triazolor4.3- d]M ,41oxazin-7-yl)phenyl)cvclobutanam8ne
Figure imgf000169_0001
Step 1 : Tert-butyl (1-(4-(1-(2-methoxyethyl)-8-Dhenyl-4H-pyridor2.3-b1f1 ,2,4ltriazolor4,3- dlf1 ,4loxazin-7-yl)phenyl)cyclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.205 mmol) and 3- methoxypropanehydrazide (48mg, 0.410 mmol) in p-xylene (2 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in cyclohexane) to give the title compound (33 mg, 30%). LCMS (Method D): RT = 1.414 min, M+1 = 554. 1H NMR (500 MHz, CDCI3) δ 8.16 (1 H, s), 7.27-7.36 (7H, m), 7.23- 7.18 (2H, m), 5.60 (2H, s), 3.99 (2H, t), 3.43 (3H, s), 3.38 (2H, t), 2.57-2.40 (4H, m), 2.12-2.01 (1 H, m), 1.87-1.77 (1 H, m), 1.49-1.27 (9H, bs).
Step 2: 1-(4-(1-(2-methoxyethyl)-8-phenyl-4H-pyridof2.3-biri .2.41triazolof4.3- d1f1.4loxazin-7-yl)phenyl)cyclobutanamine
Tert-butyl (1 -(4-(1 -(2-methoxyethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.022 mmol) was dissolved in dichloromethane (0.5 ml); TFA (0.5 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (5 mg, 50% yield). LCMS (Method D) RT = 0.741 min, M+1 = 454. 1H-NMR (500 MHz, CD3OD) δ 8.40 (1 H, s), 7.52- 7.47 (2H, d), 7.46-7.41 (2H, d), 7.38-7.34 (3H, m), 7.32-7.26 (2H, dd), 5.65 (2H, s), 3.94 (2H, t), 3.49 (2H, t), 3.35 (3H, s), 2.82-2.74 (2H, m), 2.63-2.54 (2H, m), 2.30- 2.19 (1H, m), 2.04-1.93 (1 H, m). Example 122: 1 -(4-(8-phenyl-1 -(pyridin-3-yl)-4H-pyridor2,3-biri .2.41triazolor4.3- d1H.41oxazin-7-yl)phenyl)cvclobutanamine
Figure imgf000171_0001
Step 1 : Tert-butyl (1-(4-(8-phenyl-1 -(pyridin-3-yl)-4H-pyridor2,3-b1f1.2.4ltriazolof4.3- dlf1.41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.205 mmol) and nicotinic acid hydrazide (84mg, 0.308 mmol) in p-xylene (2 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in cyclohexane) to give the title compound (33 mg, 25%). LCMS (Method D): RT = 1.364 min, M+1 = 573. Ή NMR (500 MHz, CDCI3): 9.00 (1 H, d), 8.87 (1 H, dd), 8.10 (1 H, dt), 7.56 (1H, dd), 7.36-7.27 (5H, m), 7.24- 7.17 (3H, m), 6.99- 6.93 (2H, m), 5.67 (2H, s), 2.56-2.39 (4H, m), 2.13-2.01 (1 H, m), 1.88-1.75 (1 H, m), 1.49-1.26 (9H, bs).
Step 2: -(4-(8-phenyl-1-(pyridin-3-yl)-4H-pyrido[2,3-biri .2,41triazolo[4,3-d1M .41oxazin-7- vDphenvDcyclobutanamine
Tert-butyl (1-(4-(8-phenyl-1-(pyridin-2-yl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 l4]oxazin- 7-yl)phenyl)cyclobutyl)carbamate (15 mg, 0.026 mmol) was dissolved in dichloromethane (1 ml); TFA (0.5 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (8 mg, 56% yield). LC S (Method A) RT = 3.73 min, M+1 = 474. Ή NMR (500 MHz, CDCI3): 8.99 (1 H, d), 8.86 (1 H, dd), 8.30 (1 H, dt), 7.75 (1 H, dd), 7.48 (2H, d), 7.42 (2H, d), 7.37 (1 H, s), 7.31-7.20 (3H, m), 7.01 (2H, d), 5.78 (2H, s), 2.83-2.72 (2H, m), 2.63- 2.54 (2H, m), 2.30-2.17 (1 H, m), 2.05-1.90 (1 H, m).
Example 123: 1-f4- 8-phenyl-1- pyridin-4-vn-4H-pyridor2,3-biri.2.4ltriazolor4.3- dlH ,41oxazin-7-yl)phenyl)cyclobutanamine
Figure imgf000172_0001
Step 1 : Tert-butyl (1 -(4-(8-phenyl-1-(pyridin-4-yl)-4H-pyridof2.3-biri.2,41triazolor4.3- din,41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1 -(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.205 mmol) and isonicotinic acid hydrazide (127mg, 0.923 mmol) in p-xylene (2 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (14 mg, 12%). LCMS (Method A): RT = 6.28 min, M+1 = 573. 1H NMR (500 MHz, CDCI3): 8.90 (2H, d), 7.70 (2H, d), 7.38 (1 H, s), 7.36-7.32 (2H, d), 7.31- 7.27 (2H, d), 7.25- 7.18 (3H, m), 6.98- 6.93 (2H, dd), 5.66 (2H, s), 2.55-2.41 (4H, m), 2.14-2.01 (1 H, m), 1.88-1.78 (1 H, m), 1.46-1.24 (9H, bs).
Step 2: 1-(4-(8-phenyl-1-(pyridin-4-yl)-4H-pyridof2.3-blf1,2.4ltriazolof4.3-d]f1.4loxazin-7- vDphenvDcvclobutanamine
Tert-butyl (1 -(4-(8-phenyl-1 -(pyridin-4-yl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin- 7-yl)phenyl)cyclobutyl)carbamate (14 mg, 0.024 mmol) was dissolved in dichloromethane (1 ml); TFA (0.5 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (10 mg, 87% yield). LCMS (Method A) RT = 3.66 min, M+H+= 474. Ή NMR (500 MHz, CDCI3): 8.77 (2H, d), 8.79 (2H, d), 7.41 (2H, d), 7.33 (1 H, s), 7.32 (2H, d), 7.18-7.13 (3H, m), 6.96 (2H, dd), 5.66 (2H, s), 2.70-2.61 (2H, m), 2.52- 2.43 (2H, m), 2.19-2.08 (1 H, m), 1.92-1.82 (1 H, m).
Example 124: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-(pyridin-3-yl)-1H- Pyridor2,3-biri.41oxazin-2(3 -one
Figure imgf000173_0001
Step 1 : 6,7-dibromo-1 H-pyridor2,3-biri,4loxazin-2(3H)-one
7-bromo-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (1.5g, 6.55mmol) and NBS (5.83, 32.7mmol) were heated to 80°C in DMF (30ml) for 2 hours. The resulting reaction mixture was diluted with ethyl acetate (30ml) and water (30ml). The aqueous phase was extracted with fresh ethyl acetate (50ml x 3) and the combined organic layers were washed with brine (50ml), dried on anhydrous Na2S0 , filtered and concentrated to dryness under reduced pressure to give the title compound (1.6g, 79%). It was used for the next step without further purification. LCMS (Method D): RT = 0.91 min, M+1 = 309.
Step 2: 6,7-dibromo-1-methyl-1 H-pyhdof2,3-blf 1 ,41oxazin-2(3H)-one
6,7-Dibromo-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (1.5g mg, 4.88mmol), potassium carbonate (2.02g, 14.61 mmol) and methyl iodide (2.07, 14.61 mmol) were stirred in DMF (35 ml) at room temperature for one hour. The resulting reaction mixture was quenched with water (100ml) and extracted with ethyl acetate (100ml x 3). The combined organic layers were washed with water (100ml), brine (100ml) and dried on anhydrous Na2S04, filtered and concentrated to dryness under reduced pressure to give the title compound (1.3g, 83%). It was used for the next step without further purification. LCMS (Method D) RT 1.053 min, M+1 =322. 1H-NMR (500 MHz, CD3OD) δ 7.41 (1 H, s), 4.85 (2H, s), 3.35 (3H, s).
Step 3: Tert-butyl (1-(4-(trimethylstannyl)phenyl)cvclobutyl)carbamate
Tert-butyl (1-(4-bromophenyl)cyclobutyl)carbamate (5.45g mg, 13.36mmol) was dissolved in anhydrous toluene (250ml). The solution was degassed by bubbling nitrogen for 15 minutes. Tetrakistriphenylphosphine palladium (0) (3.09g, 2.67mmol) was added and the resulting reaction mixture was degassed by bubbling nitrogen for 15 minutes. Hexamethylditin (6.57g, 4.19ml, 20.05 mmol) predissolved in anydrous toluene (30ml) was added to the previous mixture. The resulting reaction mixture was degassed by bubbling nitrogen for 10 minutes and heated to reflux for 2 hours.
The reaction mixture was cooled down and filtered through celite, using fresh ethyl acetate to wash the cake. The filtrate was concentrated to dryness under reduced pressure. The obtained crude was purified by Biotage silica gel chromatography
(gradient 0% to 90% ethyl acetate in n-hexanes; TLC stained with KMn04) to give the title compound (1.2g, 22%). Ή NMR (500 MHz, CDCI3): 7.53-7.36 (4H, m), 2.60-2.47 (4H, m), 2.13-2.03 (1 H, m), 1.90- 1.78 (1 H, m), 1.47-1.27 (9H, br).
Step 4: Tert-butyl (1-(4-(7-bromo-1-methyl-2-oxo-2.3-dihydro-1 H-pyridor2,3-bli1 ,4loxazin- 6-yl)phenyl)cvclobutyl)carbamate
6,7-Dibromo-1-methyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (0.628 g, 1.951 mmol) and tert-butyl 1-(4-(trimethylstannyl)phenyl)cyclobutylcarbamate (1.2 g, 2.93 mmol) was dissolved in DMF ( 0ml) to give a yellow solution. The reaction mixture was degassed by bubbling nitrogen for 30 minutes.
Bis(dimethylphosphino)palladium(IV) chloride (0.137g, 0.195 mmol) was added at room temperature. The resulting reaction was degassed and heated to 80oc.
The reaction mixture was quenched with water (80ml); the aqueous phase was extracted with ethyl acetate (80ml x 3); the combined organic layers were washed with water (80ml x 2) and brine (80ml x 2). The organic layer was dried on anhydrous Na2S0 , filtered and concentrated to dryness under reduced pressure. The crude was purified by Biotage silica gel chromatography (gradient 0% to 70% ethyl acetate in n-hexanes) to give the title compound (400mg, 42%).1 Ή NMR (500 MHz, CDCI3): 7.73-7.56 (2H, m), 7.56-7.46 (3H, m), 4.88 (2H, s), 3.33 (3H, s), 2.68-2.49 (4H, m), 2.20-2.04 (1 H, m), 1.94- 1.80 ( H, m), 1.47-1.27 (9H, br s). 1 This was a mixture of two regioisomers (7:3 in ratio).
Step 5: Tert-butyl (1-(4-(1-methyl-2-oxo-7-(pyridin-3-yl)-2.3-dihvdro-1 H-pyridof2.3- b]f1 ,41oxazin-6-yl)phenyl)cyclobutyl)carbamate
Tert-butyl (1 -(4-(7-bromo-1 -methyl-2-oxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (57mg, 0.117mmol), pyridin-3-yl boronic acid (21.52mg, 0.175 momol), triphenylphosphine (9.18mg, 0.035mmol), cesium fluoride (89mg,
0.584mmol) were dissolved in 1 ,2-dimethoxyethane (2ml) to give an orange solution. The reaction mixture was degassed by bubbling nitrogen for 10 minutes.
Palladium(ll) acetate (3.93 mg, 0.018 mmol) was added at room temperature. The resulting reaction was degassed and heated to 80°C for 18 hours.
The reaction mixture was concentrated to dryness under reduced pressure. The crude was purified by preparatory HPLC to give the title compound (12mg, 21%). LCMS
(Method D) RT 1.158 min, M+1 = 487. Ή N R (500 MHz, CDCI3): 7.64-7.52 (1H, m), 7.37-7.21 (8H, m), 4.94 (2H, s), 3.43 (3H, s), 2.59-2.36 (4H, m), 2.15-2.01 (1 H, m), 1.91- 1.77 (1H, m), 1.47- .20 (9H, br s).
Step 6: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-(pyridin-3-yl)-1 H-pyrido[2.3- bin .41oxazin-2(3H)-one
Tert-butyl (1-(4-(1-methyl-2-oxo-7-(pyridin-3-yl)-2,3-dihydro-1H-pyhdo[2,3-b][1,4]oxazin- 6-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.025 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (14 mg, 78% yield). LCMS (Method D) RT = 0.546 min, M+1 =387. 1H-NMR (500 MHz, CD3OD) δ 8.57-8.49 (1H, br s), 8.45-8.36 (1 H, br s), 7.90 (1 H, d), 7.67 (1 H, s), 7.61-7.48 (2H, d), 7.48-7.36 (4H, m), 5.00 (2H, s), 3.45 (3H, s), 2.84-2.72 (2H, m), 2.65-2.54 (2H, m), 2.32-2.16 (1 H, m), 2.06-1.90 (1 H, m). Example 125: 6- 4-n-aminocvclobutyl)phenyl)-1-methyl-7- thiophen-3-yl)-1H- pyridor2,3-biri.41oxazin-2(3 -one
Figure imgf000176_0001
Step 1 : Tert-butyl (1-(4-(1-methyl-2-oxo-7-(thiophen-3-yl)-2,3-dihvdro-1 H-pyridor2.3- biri,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
Tert-butyl (1-(4-(7-bromo-1-methyl-2-oxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (70mg, 0.143mmol), thiophen-3-yl boronic acid (30mg, 0.215 momol), triphenylphosphine (11mg, 0.043 mmol), cesium fluoride (109mg, 0.717mmol) were dissolved in 1 ,2-dimethoxyethane (3ml) to give an orange solution. The reaction mixture was degassed by bubbling nitrogen for 10 minutes.
Palladium(ll) acetate (5 mg, 0.021 mmol) was added at room temperature. The resulting reaction was degassed and heated to 80OC for 18 hours.
The reaction mixture was concentrated to dryness under reduced pressure. The crude was purified by Biotage silica gel chromatography (gradient 0% to 70% ethyl acetate in n- hexanes) to give the title compound as a mixture of two regioisomers (37mg, 52%, 7:3).3 H NMR (500 MHz, CDCI3): 7.39-7.30 (5H, m), 7.24-7.20 (1 H, dd), 7.15-7.12 (1 H, dd), 6.82-6.78 (1H, dd), 4.81(21-1, s), 3.39 (3H, s), 2.68-2.33 (4H, m), 2.15-2.05 (1H, m), 1.90- 1.78 (1 H, m), 1.47-1.20 (9H, br s). LCMS (Method D): RT 1.514 min, M+1= 492.
3 This was a mixture of two regioisomers (7:3 in ratio).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-(thiophen-3-yl)-1 H-pyridof2,3- b f 1 ,4loxazin-2(3H)-one
Tert-butyl (1-(4-(1-methyl-2-oxo-7-(thiophen-3-yl)-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (25 mg, 0.051 mmol) was dissolved in dichloromethane (0.5 ml); this solution was added to a scx-2 catch and release cartridge (5g), precalibrated using methanol (15ml) and dichloromethane (15ml). After aloowing binding for 12 hours, the column was washed with methanol (6ml), then the product eluted with 7M ammonia in methanol (4 x 2ml). The combined product fractions were concentrated to dryness under reduced pressure to give the desired product as a white solid (5.8 mg, 30% yield). LCMS (Method D): RT = 0.777 min, M+1 =393. 1H-NMR (500 MHz, CD3OD) δ 7.35-7.29 (3H, m), 7.28-7.25 (1 H, dd), 7.21-7.18 (1 H, s), 7.17-7.14 (1 H, dd), 7.07-7.03 (1 H, dd), 6.71 -6.67 (1 H, dd), 4.80 (2H, s), 3.32 (3H, s), 2.51-2.37 (4H, m), 2.25-2.13 (1 H, m).
Example 126: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-(thiophen-2-yl)-1H- pyridor2,3-b]H,41oxazin-2(3 -one
Figure imgf000177_0001
Step 1 : Tert-butyl (1-(4-(1-methyl-2-oxo-7-(thiophen-2-yl)-2.3-dihvdro-1 H-pyridof2,3- b1i1 ,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
Tert-butyl (1-(4-(7-bromo-1-methyl-2-oxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (70mg, 0.143mmol), thiophen-2-yl boronic acid (30mg, 0.215 momol), triphenylphosphine (11mg, 0.043 mmol), cesium fluoride (109mg, 0.717mmol) were dissolved in 1 ,2-dimethoxyethane (3ml) to give an orange solution. The reaction mixture was degassed by bubbling nitrogen for 10 minutes.
Palladium(ll) acetate (4 mg, 0.018 mmol) was added at RT. The resulting reaction was degassed, bubbling N2 and heated to 80OC for 18 hours.
The reaction mixture was concentrated to dryness under reduced pressure. The crude was purified by Biotage silica gel chromatography (gradient 0% to 70% ethyl acetate in n- hexanes) to give the title compound as a mixture of two regioisomers (15mg, 22%, 7:3).2 LCMS (Method D): RT 1.524 min, M+1 = 492. Ή NMR (500 MHz, CDCI3): 7.69-7.32 (7H, m), 6.81-6.76 (1 H, m), 4.93 (2H, s), 3.43 (3H, s), 2.58-2.48 (4H, m), 2.20-2.07 (1 H, m), 1.96- 1.81 (1 H, m), 1.46-1.20 (9H, br s).
2 This was a mixture of two regioisomers (7:3 in ratio).
Step 2: 6-(4-(1-aminocvclobutyl)phenyl)-1-methyl-7-(thiophen-2-yl)-1 H-pyrido[2.3- bin .41oxazin-2(3H)-one
Tert-butyl (1-(4-(1 -methyl-2-oxo-7-(thiophen-2-yl)-2,3-dihydro- H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (12 mg, 0.024 mmol) was dissolved in dichloromethane (1.5ml). TFA (1 ml) was added at room temperature and the reaction mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (14 mg, 78% yield). LCMS (Method D): RT = 0.781 min, M+1 =393. 1H-NMR (500 MHz, CD3OD) δ 7.66 (1 H, s), 7.53-7.49 (2H, dd), 7.48-7.44 (2H, dd), 7.42-7.39 (1 H, dd), 7.01-7.97 (1H, dd), 6.97-6.95 (1 H, dd), 4.97 (2H, s), 3.44 (3H, s), 2.84-2.74 (2H, m), 2.64-2.55 (2H, m), 2.31-2.21 (1 H, m), 2.07-1.93 (1 H, m).
Example 127: 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-6,8-dihvdro-5H-pyrano[3,4- blpyridin-5-one
Figure imgf000178_0001
Step 1 : fert-butyl (1-(4-(5-oxo-3-phenyl-6,8-dihvdro-5H-pyranor3,4-blPyridin-2- yl)phenyl)cyclobutyl)carbamate
In a 5 mL reaction tube was added acetic acid (5 ml) followed by (E)-ferf-butyl 1-(4-(3- (dimethylamino)-2-phenylacryloyl)phenyl)cyclobutylcarbamate (750 mg, 1.783 mmol), 2H-pyran-3,5(4H,6H)-dione (305 mg, 2.68 mmol), ammonium acetate (412 mg, 5.35 mmol) and molecular sieves (100 mg) to give a brown suspension. The reaction mixture was stirred at 100°C under a nitrogen atmosphere for 2 hours, then allowed to cool to room temperature and concentrated under reduced pressure. The residue was partitioned between water (10 mL) and dichloromethane (10 mL) and decanted from the molecular sieves. The layers were separated and the aqueous phase extracted into dichloromethane (2 x 10 mL). The combined organic phases were washed with saturated sodium bicarbonate solution (2 x 10 mL), brine (10 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow/brown solid. This was purified twice by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 93:7 to 60:40) and then preparative HPLC (Method F) to give the desired product as a white solid (12 mg, 1.4% yield). H-NMR (500 MHz, CDCI3) δ 8.29 (1 H, s), 7.16-7.41 (9H, m), 5.05 (2H, s), 5.02 (1 H, br s), 4.44 (2H, s), 2.21-2.66 (4H, br m), 2.01-2.16 (1 H, m), 1.74-1.88 (1 H, m), 1.10-1.50 (9H, br m).
Step 2: 2-(4-(1-aminocvclobutyl)phenyl)-3-phenyl-6,8-dihvdro-5H-pyranof3,4-b1pyridin-5- one
te/f-Butyl (1-(4-(5-oxo-3-phenyl-6,8-dihydro-5H-pyrano[3,4-b]pyridin-2- yl)phenyl)cyclobutyl)carbamate (12 mg, 0.026 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (10 mg, 77% yield). 1H-NMR (500 MHz, MeOD) δ 8.19 (1 H, s), 7.41 (2H, d), 7.32 (2H, d), 7.20-7.24 (3H, m), 7.11-7.15 (2H, m),
4.94 (2H, s), 4.37 (2H, s), 2.59-2.68 (2H, m), 2.39-2.49 (2H, m), 2.06-2.16 (1 H, m), 1.79-
1.95 (1 H, m). LCMS (Method D) RT = 0.86 min, M+H+ = 371.00.
Example 128: (1 r,3r)-3-amino-3-(4-(1 -(difluoromethvn-8-phenyl-4H-pyridor2.3- b")f1.2,41triazolor4,3-diri.41oxazin-7-yl)phenyl)-1-methylcvclobutanol
Figure imgf000179_0001
Step 1 : 6-bromo-7-phenyl-1 H-pyhdor2,3-b1M ,41oxazine-2(3H)-thione
A mixture of 6-bromo-7-phenyl-1H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (0.26 g, 0.852 mmol) and Lawesson's Reagent (0.34 g, 0.852 mmol) in toluene (10 ml) was heated at 120 degree for 16h. The reaction mixture was concentrated to dryness and was added DCM (5ml) and diisopropyl ether (20ml). The precipitate was filtered to give product (0.18g). Ή NMR (500 MHz, DMSO-d6): 12.9 (br, 1 H), 7.35-7.55 (m, 5H), 7.32 (s, 1 H), 5.16 (s, 2H).
Step 2: 7-bromo-1-(difluoromethyl)-8-phenyl-4H-pyridor2.3-biri ,2,41triazolor4,3- din,41oxazine
A mixture of 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazine-2(3H)-thione (0.21 g, 0.654 mmol) and 2,2-difluoroacetohydrazide (0.14 g, 1.3 mmol) in xylene (6 ml) was heated at 150 degree under microwave for 60 min. The reaction solution was concentrated and the residue was partitioned between ethyl acetate (10ml) and water (10ml). The organic phase was separated and the aqueous phase was extracted with ethyl acetate (10ml). The combined organic phase was concentrated and purified by column (biotage 25g) etuted with ethyl acetate/cyclohexane (0-60%) to afford product (55mg). 1H NMR (500 MHz, CDCI3) 8.09 (s, 1 H), 7.4-7.55 (m, 5H), 7.06 (t, 1 H), 5.66 (s, 2H).
Step 3: 2-((1 r,3r)-1-(4-(1 -(difluoromethyl)-8-phenyl-4H-pyridor2,3-biri .2,4ltriazolor4,3- d1f1 ,4loxazin-7-yl)phenyl)-3-hvdroxy-3-methylcvclobutyl)isoindoline-1 ,3-dione
A mixture of 7-bromo-1-(difluoromethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazine (30 mg, 0.079 mmol), 2-((1r,3r)-3-hydroxy-3-methyl-1 -(4-(4,4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (46mg, 0.106 mmol), and sodium carbonate (16.7721 mg, 0.158 mmol) in dioxane (Ratio: 4.00, Volume: 4 ml) and Water (Ratio: 1.000, Volume: 1 ml) to give a yellow suspension. The suspension was degassed for 5 min. Pd(PPh3) (6.4614 mg, 7.91 mol) was added and the mixture was heated under microwave condition at 100degree for 20 min. The reaction mixture was concentrated to dryness. The residue was partitioned between ethyl acetate (15ml) and water (15ml). The organic phase was separated. The aqueous phase was extracted with ethyl acetate (2X15ml). The combined organic phase was washed with water (2X30ml), brine (25ml) then concentrated. The crude product was purified by column (biotage 10g) eluted with ethyl acetate/cyclohexane (0-50%) to afford product (10mg). 1H NMR (500 MHz, CDCI3): 8.14 (s, 1 H), 7.74 (m, 2H), 7.69 (m, 2H), 7.54 (d, 2H), 7.33 (d, 2H), 7.27-7.31 (m, 3H), 7.05-7.19 (m, 3H), 5.61 (s, 2H), 3.35 (d, 2H), 3.09 (d, 2H), 1.4 (s, 3H). Step 4: (1r,3r)-3-amino-3-(4-n-(difluoromethyl)-8-phenyl-4H-pyridof2.3- bl[1 ,2,41triazolof4,3-din ,41oxazin-7-yl)phenyl)-1 -methylcvclobutanol
A mixture of 2-((1r,3r)-1-(4-(1-(difluoromethyl)-8-phenyl-4H-pyrido[2,3- b][1,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline- 1 ,3-dione (10mg, 0.017 mmol) and HYDRAZINE (0. ml, 3.19 mmol) hydrate in ethanol (3ml) was heated under microwave irradiation at 120 °C for 30 min. The mixture was concentrated to dryness. The residue was partitioned between ethyl acetate (10ml) and water (10ml). The organic phase was separated. The aqueous phase was extracted with ethyl acetate (2X10ml). The combined organic phase was washed with water (2X15ml), brine (15ml) and concentrated. The crude product was purified by SCX-2 column (2g) eluted with ammonia in methanol (2M) to give product (3mg). 1H NMR (500 MHz, CD3OD): 8.05 (s, 1 H), 7.4 (t, 1 H), 7.2-7.3 (m, 7H), 7.11-7.15 (m, 2H), 5.63 (s, 2H), 2.59 (d, 3H), 2.34 (d, 2H). 1.43 (s, 3H). LCMS (Method D): RT = 0.78 min, M+H+ = 476.0
Example 129: 1-(4-(1,8-diphenyl-4H-pyridor2.3-biri,2.4ltriazolor4,3-din.4loxazin-7- yl)phenyl)cvclobutanamine
Figure imgf000181_0001
Step 1 : Tert-butyl (1-(4-(1 ,8-diphenyl-4H-pyridor2,3-b1[1.2,4ltriazolof4.3-din.4loxazin-7- vQphenvDcvclobutvDcarbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.205 mmol) and benzoic acid hydrazide (84mg, 0.615 mmol) in p-xylene (2 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexane) to give the title compound (46 mg, 40%). LCMS (Method D): RT = 1.546 min, M+1= 572. 1H NMR (500 MHz, CDCI3): 7.77- 7.69 (2H, m), 7.64-7.57 (2H, m), 7.56-7.50 (1 H, t), 7.49- 7.41 (2H, t), 7.34 -7.30 (2H, m), 7.28 (1 H, s), 7.22- 7.14 (3H, m), 6.95- 6.90 (2H, dd), 5.65 (2H, s), 2.55-2.40 (3H, m), 2.15-2.01 (2H, m), 1.89-1.77 (1H, m), 1.46-1.24 (9H, bs).
Step 2: 1-(4-(1 ,8-diphenyl-4H-pyridor2.3-bin .2.41triazolor4.3-din .41oxazin-7- yQphenvQcvclobutanamine
Tert-butyl (1-(4-(1 ,8-diphenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutyl)carbamate (15 mg, 0.026 mmol) was dissolved in dichloromethane (1 ml); TFA (0.5 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexanes (2 ml) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure and dried to give the desired product as an off-white solid (5 mg, 40% yield). LCMS (Method D) RT = 0.846 min, M+1 = 472. H NMR (500 MHz, CDCI3): 7.91- 7.86 (2H, dd), 7.83- 7.76 (3H, m), 7.59- 7.54 (2H, d), 7.53- 7.48 (2H, d), 7.42 (1 H, s), 7.37-7.28 (3H, m), 7.09- 7.03 (2H, dd), 5.86 (2H, s), 2.91-2.81 (2H, m), 2.74- 2.63 (2H, m), 2.40-2.29 (1 H, m), 2.13- 2.02 (1H, m).
Example 130: 1-(4-(8-phenyl-1-(pyridin-2-yl)-4H-pyridor2.3-bin,2.4Uriazolor4,3- dlM ,41oxazin-7-yl)phenyl)cvclobutanamine
Figure imgf000182_0001
Step 1 : 2-Picolvnil hydrazide
Ethyl 2-picolinate (5g, 33.1 mmol) and hydrazine monohydrate (5.3g, 165 mmol) in ethanol (70ml) were heated to reflux for 15 hour. The reaction mixture was concentrated under reduced pressure and the residue was treated with toluene to azeotrope hydrazine in excess. This was repeated three times. The crude residue was used for the next step without further purification.
Step 2: Tert-butyl n-(4-(8-Dhenyl-1-(pyridin-4-yl)-4H-pyridor2.3-biri.2,41triazolof4.3- din.41oxazin-7-yl)phenyl)cvclobutyl)carbamate
A suspension of tert-butyl (1-(4-(7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (100 mg, 0.205 mmol) and 2- picolinylhydrazide (56.2mg, 0.410 mmol) in p-xylene (1 ml) was heated to 150°C for 15 minutes under microwave irradiation. The resulting reaction mixture was concentrated to dryness under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 4% methanol in dichloromethane) to give the title compound (15 mg, 13%). LCMS (Method A): RT = 6.95 min, M+1= 573. 1H NMR (500 MHz, CDCI3): 8.78- 8.74 (1 H, dd), 8.45 (1H, s), 8.27- 8.23 (1H, dd), 7.96-7.91 (1 H, dt), 7.53-7.48 (1H, dd), 7.38-7.33 (2H, dd), 7.31- 7.27 (3H, m), 7.25- 7.23 (2H, m), 7.19- 7.13 (2H,dd), 5.63 (2H, s), 2.56-2.41 (4H, m), 2.14-2.01 (1 H, m), 1.88-1.75 (1 H, m), 1.46-1.21 (9H, bs).
Step 3: 1-(4-(8-phenyl-1-(pyridin-2-yl)-4H-pyridor2.3-biri.2.4ltriazolof4.3-diri.4loxazin-7- vDphenvDcyclobutanamine
Tert-butyl (1-(4-(8-phenyl-1-(pyridin-2-yl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin- 7-yl)phenyl)cyclobutyl)carbamate (15 mg, 0.026 mmol) was dissolved in dichloromethane (1 ml); TFA (0.5 ml) was added at room temperature and the resulting mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The residue was then slurred in n-hexane (2 ml) and after settling the supernatant solvent was removed by pipette. This was repeated three times. The remaining solvent was removed under reduced pressure. The resulting residue was purified via preparative HPLC to give the desired product as a white solid (8 mg, 64% yield). LCMS (Method D) RT = 0.825min, M+H+= 474. Ή NMR (500 MHz, d6-DMSO): 8.79 (1 H, d), 8.44 (1 H, bs), 8.13 (1H, s), 8.11-8.05 (1 H, m), 7.62- 7.59 (1 H, m), 7.41 (2H, s), 7.42-7.38 (2H, d), 7.25- 7.27 (3H, m), 7.10 (2H, m), 5.71 (2H, s), 2.60-2.51 (4H, m), 2.27-2.19 (1 H, m), 1.88-1.76 (1H, m). Example 131 : 6-(4-(aminomethyl)phenyl)-1 -methyl-7-phenyl-1 H-pyridor2.3- blH .41oxazin-2(3H)-one
Figure imgf000184_0001
Step 1 : terf-butyl 4-(1-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyrido[2,3-bU1 ,4loxazin-6- yl) benzyl ca rba m ate
In a 15 mL reaction tube was added 6-bromo-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (40 mg, 0.125 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)benzylcarbamate (50 mg, 0.150 mmol) in 1 ,4-dioxane (2 ml), followed by a solution of sodium carbonate (40 mg, 0.3 mmol) in water (0.5 ml) to give a white suspension. This was degassed by bubbling nitrogen for 10 minutes, followed by the addition of [1 ,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (9 mg, 0.013 mmol). The reaction mixture was heated to 80°C overnight. The reaction mixture was allowed to cool to room temperature, diluted with brine (4 ml) and extracted into ethyl acetate (3 x 4 ml). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as a white solid (40 mg, 72% yield). 1H-NMR (500 MHz, CDCI3) δ 7.27-7.32 (5H, m), 7.25 (1 H, s), 7.16-7.20 (2H, m), 7.10 (2H, d), 4.89 (2H, s), 4.81 (1H, br s), 4.26 (2H, br s), 3.39 (3H, s), 1.23 (9H, br s). LCMS (Method D): RT = 1.40 min, M+hT = 446.20.
Step 2: 6-(4-(aminomethyl)phenyl)-1 -methyl-7-phenyl-1 H-pyrido[2,3-b1f1.4loxazin-2(3H)- one
tert-Butyl 4-(1-methyl-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6- yl)benzylcarbamate (40 mg, 0.090 mmol) was dissolved in TFA (2 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~2 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (2 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as a white solid (35 mg, 85% yield). 1H-NMR (500 MHz, eOD) δ 7.54 (1 H, s), 7.36 (2H, d), 7.31 (2H, d), 7.25-7.29 (3H, m), 7.18-7.23 (2H, m), 4.95 (2H, s), 4.07 (2H, s), 3.41 (3H, s). LCMS (Method D): RT = 0.69 min, M+hf = 346.20.
Example 132: 6-(4-(1-aminocvclobutyl)phenyl)-7-(4-fluorophenyl)-1-methyl-1H- pyridor2,3- lF1.41oxazin-2(3H)-one
Figure imgf000185_0001
Step 1 : tert-butyl 1-(4-(7-(4-fluorophenyl)-1-methyl-2,3-dihvdro-1 H-pyridor2,3- bin ,41oxazin-6-yl)phenyl)cyclobutylcarbamate
In a sealed tube was added tert-butyl 1-(4-(7-bromo-1-methyl-2-oxo-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutylcarbamate (82mg, 0.168mmol), 4- fluorophenylboronic acid (35.2mg, 0.252mmol), triphenylphosphine (13.21 mg, 0.05mmol) and cesium fluoride (128mg, 0.84mmol) in 1 ,2-dimethoxyethane (2ml). The reaction mixture was degassed by bubbling nitrogen for 10min and palladium acetate (5.65mg, 0.025mmol) was added and the mixture was heated at 80°C overnight. The reaction mixture was concentrated and the residue was diluted with ethyl acetate and the suspension was pass through sillica. The filtration was concentrated and to the residue was added methanol (2.5ml) to form a suspension. The solid was filtered and dried to give tert-butyl 1-(4-(7-(4-fluorophenyl)-1-methyl-2,3-dihydro-1H-pyrido[2,3-b][1,4]oxazin- 6-yl)phenyl)cyclobutylcarbamate (19mg). LCMS (Method D): RT = 1.54 min, M+H+ = 504.
Step 2: 6-(4-( 1 -aminocvclobutyl)phenyl)-7-(4-fluorophenyl)-1 -methyl-1 H-pyridor2.3- blf 1 ,41oxazin-2(3H)-one
TRIFLUOROACETIC ACID (0.5 ml, 6.49 mmol) was added to tert-butyl (l-(4-(7-(4- fluorophenyl)-1 -methyl-2-oxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (16mg, 0.032 mmol) in a RBF (5ml), The resulting solution was stirred for 60 seconds then concentrated to dryness under reduced pressure. The residue was suspended in diethyl ether (1 ml) and concentrated to dryness three times to give the trifluoroacetic acid salt of the product (7mg). LC S (Method D): RT = 0.78 min, M+H+ = 404. 1H NMR (500 MHz, CD3OD) 7.44 (s, 1 H), 7.29 (m, 4H), 7.12-7.15 (m, 2H), 6.91-6.95 (m, 2H), 4.85 (s, 2H), 3.31 (s, 3H), 2.62-2.69 (m, 2H), 2.43- 2.49 (m, 2H), 2.10-2.15 (m, 1H), 1.80-1.86 (m, 1 H).
Example 133: 6-(4-(1 -aminocyclobutyl)phenyl)-7-(cyclopropylmethYl)-1 -methyl-1 H- pyrido[2,3-b1H ,41oxazin-2(3 -one
Figure imgf000186_0001
Step 1 : Tert-butyl (1-(4-(7-allyl-1-methyl-2-oxo-2,3-dihydro-1 H-pyridof2,3-biri,4loxazin-6- yDphenvDcvclobutvDcarbamate
Tert-butyl (1-(4-(7-bromo-1-methyl-2-oxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (100mg, 0.205mmol) and allyl-tributyl stannane were dissolved in anhydrous dimethylformamide (1 ml) to give a white solution. The reaction mixture was degassed by bubbling nitrogen for 30 minutes.
Bis(triphenylphosphine)palladium(ll) dichloride (15mg, 0.020 mmol) was added at room temperature. The resulting reaction was degassed and heated to 80OC for 2 hours. The reaction mixture was diluted with ethyl acetate (3ml), washed with water (5ml) and brine (5ml); the organic phase was concentrated to dryness under reduced pressure. The crude was purified by Biotage silica gel chromatography (gradient 0% to 20% ethyl acetate in n-hexanes) to give the title compound (52mg, 56%). LCMS (Method A) RT 6.83 min, M+1 = 450. Ή NMR (500 MHz, CDCI3): 7.57-7.40 (4H, m), 7.15 (1 H, s), 6.04- 5.88 (1H, m), 5.20-5.13 (1H, m), 5.11-4.99 (2H, m), 4.85 (2H, s), 3.52- 3.41 (1 H, dd), 3.37 (3H, s), 2.67-2.41 (4H, m), 2.15-2.04 (1 H, m), 1.90- 1.80 (1 H, m), 1.47-1.31 (9H, br s).
Step 2: Tert-butyl (1-(4-(7-(cvclopropylmethyl)-1-methyl-2-oxo-2,3-dihvdro-1H-pyridor2l3- biri,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
In a 50 ml 3 necks round-bottomed flask was tert-butyl (1-(4-(7-allyl-1-methyl-2-oxo-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (52mg, 0.116 mmol), palladium acetate (32mg, 0.180 mmol) in diethyl ether (5ml) to give a yellow solution at room temperature under a nitrogen atmosphere. The reaction mixture was cooled to 0OC and trimethylsilyldiazomethane (2ml) was added via a plastic syringe. The reaction mixture was left stirring at 0°C for 10 minutes and warmed up to room temperature overnight.
The reaction was quenched with aqueous acetic acid (3ml).
The reaction was then diluted with ethyl acetate (5ml) and concentrated under reduced pressure.
The crude residue was dissolved in ethyl acetate (10ml) and filtered through celite. The filtrate was concentrated under reduced pressure. The crude residue was purified by preparatory HPLC (Method E).
Fractions containing product were combined and concentrated under reduced pressure to give the title compound (3mg, 5%). LCMS (Method D) RT 1.561 min, M+1 = 464. 1H NMR (500 MHz, CDCI3): 7.58-7.39 (4H, m), 7.36 (1 H, s), 4.85 (2H, s), 3.41 (3H, s), 2.64-2.60 (2H, m), 2.59-2.52 (4H, m), 2.15-2.04 (1 H, m), 1.91- 1.84 (2H, m), 1.46-1.35 (9H, br s), 1.33-1.27 (1 H, m), 0.95-0.84 (2H, m), 0.17-0.10 (1 H, m).
Step 3: 6-(4-(1 -aminocvclobutyl)phenyl)-7-(cyclopropylmethyl)-1-methyl-1 H-pyridor2.3- bin .41oxazin-2(3H)-one
Tert-butyl (1-(4-(7-(cyclopropylmethyl)-1-methyl-2-oxo-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (0.5 mg, 1.79 μιηοΙ) was dissolved in dichloromethane (1 ml). TFA (0.5 ml) was added at room temperature and the reaction mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (1 ml) and after settling the supernatant solvent was removed by pipette. This was repeated twice. The residue was then slurred in n- hexane (1 ml) and after settling the supernatant solvent was removed by pipette. This was repeated twice. The residue remaining solvent was dried to give the desired product as an off-white solid as the title compound (0.3 mg, 77% yield). LCMS (Method D) RT = 0.77 min, M+1 = 364. 1H-NMR (500 MHz, CD3OD) δ 7.66- 7.50 (4H, m), 7.47 (1 H, s), 4.48 (2H, s), 4.15-4.09 (2H,m), 3.06 (3H, s), 2.75-2.65 (2H, m), 2.51-2.42 (2H, m), 2.28- 2.20 (1 H, m), 1.97-1.90 (1 H, m), 1.76-1.71 (1 H, m), 1.66-1.55 (2H, m), 1.38-1.31 (2H, m).
Example 134: 6-(4-((1s,3s)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1- methyl-7-phenyl-1 H-pyridor -b1H ,41oxazin-2(3H)-one
Figure imgf000188_0001
Step 1 : 2-((1 s.3s)-3-Hvdroxy-3-methyl-1 -(4-(1 -methyl-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridof2,3-b1f1 ,4loxazin-6-yl)phenvQcyclobutyl)isoindoline-1.3-dione
In a 15 ml reaction tube was added 6-bromo-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (50mg, 0.157 mmol), 2-((1 s,3s)-3-hydroxy-3-methyl-1-(4- (4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (56.6 mg, 0.131 mmol) and cesium carbonate (213 mg, 0.652mmol) in a mixture of 1 ,4-dioxane (2ml) and water (667μΙ) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of PdCI2(dppf)-CH2CI2 adduct and degassing for a further 15 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for 1.5 hrs. The reaction mixture was cooled to room temperature and water was added (7ml). The resulting mixture was extracted with ethyl acetate (10ml x 3). The combined organic layers were dried over anhydrous Na2S0 , filtered and concentrated under reduced pressure. The crude was purified by Biotage silica gel chromatography (gradient 0% to 30% ethyl acetate in n-hexanes) to give the title compound (35mg, 50%). Ή NMR (500 MHz, CDCI3): 7.75-7.70 (2H, m), 7.66-J.63 (2H, m), 7.58-7.54 (2H, m), 7.32-7.29 (2H + 1 H, m), 7.25-7.21 (2H + 1 H, m), 7.18-7.14 (2H, m), 4.90 (2H, s), 3.37 (3H, s), 3.34-3.25 (2H, m), 3.10-3.01 (2H, m), 1.09 (3H, s). LCMS (Method D) RT 1.252 min, M+1= 546.
Step 2: 6-(4-((1s,3s)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1-methyl-7-phenyl- 1 H-pyridof2.3-bir .41oxazin-2(3H)-one
In a 10ml microwave vial was 2-((1s,3s)-3-hydroxy-3-methyl-1 -(4-(1-methyl-2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)isoindoline-1 ,3- dione (35mg, 0.064 mmol) and hydrazine monohydrate (0.040 ml, 1.2 mmol) in ethanol (1.5ml) and 1,4-dioxane (0.5ml) to give a yellow solution. The reaction mixture was heated to 120OC for 1 hr under microwave conditions.
The reaction mixture was cooled to room temperature and filtered to remove the solid. This was washed thoroughly with ethanol. The filtrates were combined and concentrated under reduced pressure.
The crude residue (20mg) was dissolved in 1ml of a solution of methanol:
dichloromethane (1 :1) and added to a scx-2 catch-and-release cartridge (5g). After allowing to binding for 15 minutes, the column was washed with methanol (4 x 2ml) then the product eluted with 2M ammonia in methanol (4x 2ml). The combined product fractions were concentrated to dryness under reduced pressure to give an off-white solid (15mg). This was re-dissolved in methanol/water (2ml, 1:3) and freeze dried overnight to give the title compound (13mg, 50%). 1H-NMR (500 MHz, CD3OD) δ 7.54(1 H, s), 7.39- 7.18 (4H, m), 4.95 (2H, s), 3.43 (3H, s), 2.76-2.68 (2H, m), 2.46-2.33 (2H, m), 1.14 (3H, s). LCMS (Method D) RT = 0.747 min, M+1= 416.
Example 135: 2-(6-(4-(aminomethyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- pyridor2,3-biri.41oxazin-1-yl)acetonitrile
Figure imgf000189_0001
Step 1 : tert-butyl 4-(2-oxo-7-phenyl-2.3-dihvdro-1H-pyridor2,3-blf1.41oxazin-6- vDbenzylcarbamate
In a reaction tube was added 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (0.19g, 0.623 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)benzylcarbamate (0.25 g, 0.750 mmol) in 1 ,4-Dioxane (10 ml), followed by a solution of sodium carbonate (0.198 g, 1.868 mmol) in water (2.5ml) to give a suspension. This was degassed followed by the addition of PdCI2(dppf)-CH2CI2 adduct (0.051 g, 0.062 mmol). The resulting mixture was heated at 80 °C for 20h.
The reaction mixture was cooled down to room temperature and diluted with saturated sodium bicarbonate solution (30ml) and extracted with ethyl acetate (20mlX3). The combined organic phase was washed with water, brine and concentrated to give product (0.21 g) used for next step without further
purification. LCMS (Method D): RT = 1.25 min, M+H+ = 433.2.
Step 2: tert-butyl 4-(1-(cvanomethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyridof2,3- b1f1,41oxazin-6-yl)benzylcarbamate
A mixture of tert-butyl 4-(2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1,4]oxazin-6- yl)benzylcarbamate (70mg, 0.162 mmol), 2-bromoacetonitrile (58.4 mg, 0.487 mmol) and potassium carbonate (67.3 mg, 0.487 mmol) in DMF (2ml) was stirred at 40 °C for 16h. The reaction mixture was cooled and diluted with water (10ml) and extracted with DCM (8mlX3). The combined organic phase was concentrated and the residue was purified by column (biotage, 10g) eluted with Ethyl acetate/cyclohexane 0-30% to give product 12mg. H NMR (500 MHz, CDCI3) 7.29 (s, 1H), 7.19-7.26 (m, 5H), 7.13 (m, 2H), 7.05 (d, 2H), 4.88 (s, 2H), 4.81 (s, 2H), 4.73 (br, 1 H), 4.21 (d, 2H), 1.38 (br, 9H). LCMS (Method D): RT = 1.35 min, M+H+ = 472.2.
Step 3: 2-(6-(4-(aminomethyl)phenyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2.3- b]f 1 ,41oxazin-1 -vDacetonitrile
to a round bottomed flask containing tert-butyl 4-(1-(cyanomethyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)benzylcarbamate (12mg, 0.026 mmol) was added TFA (0.5ml, 6.49 mmol). The resulting solution was stirred for 60 seconds then concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (1 ml) and concentrated to dryness three times to give the trifluoroacetic acid salt of the product (9.5mg). 1H NMR (500 MHz, CD3OD) 7.58 (s, 1 H), 7.29 (m, 2H), 7.19-7.23 (m, 5H), 7.13 (m, 2H), 5.03 (s, 2H), 4.93 (s, 2H), 3.98 (s, 2H). LCMS (Method D): RT = 0.65 min, M+H+ = 372.2. Example 136: (1s.3s¾-3-Amino-3-(4-(1-ethyl-8-phenyl-4H-pyridor2.3- blH ,2,41triazolor4,3-din ,41oxazin-7-yl)phenyl)cyclobutanol
Figure imgf000191_0001
Step 1 : 6-bromo-7-phenyl-1 H-pyrido[2,3-b1f1 ,41oxazine-2(3H)-thione
In a 500 ml round-bottomed flask were 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one (2 g, 6.55 mmol) and Lawesson's reagent (2.65 g, 6.55 mmol) in toluene (150 ml) to give an orange suspension.
The reaction mixture was heated to reflux for 2 hours.
The reaction mixture was cooled to room temperature and toluene was removed under reduced pressure.
The crude residue was then dissolved in minimum amount of dichloromethane. Di- isopropyl ether was added and a precipitate developed. It was filtered and dried until constant weigh to give the title compound (1.5g, 71 %). It was used for the next step without further purification.
LCMS (Method D): RT 1.274 min, +1 = 323.
Step 2: (E)-6-bromo-2-hydrazono-7-phenyl-2,3-dihvdro-1 H-pyridof2,3-blH ,41oxazine
In a 15 ml vial was 6-bromo-7-phenyl- H-pyrido[2,3-b][1 ,4]oxazine-2(3H)-thione (100mg, 0.311 mmol) and hydrazine monohydrate (0.049 ml, 1.557 mmol) in tetrahydrofuran (5ml) to give a brown solution. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and the crude residue was slurred in diethyl ether. The supernatant solvent was removed and the brown solid vas dried until constant weigh to afford the title compound (90mg, 90%). It was used for the next step without further purification. LCMS (Method D): RT 0.770 min, M+1 = 321. Step 3: 7-bromo-1-ethyl-8-phenyl-4H-pyridor2,3-b1f1.2,41triazolor4,3-dlf1.41oxazine
In a 10 ml microwave vial was (E)-6-bromo-2-hydrazono-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazine (90mg, 0.282 mmol) in 1,1 ,1-triethoxypropane to give a yellow suspension. The reaction mixture was heated to 120°C for 20 minutes. The crude residue was concentrated under reduced pressure and purified by Biotage silica gel chromatography (gradient 100% to 100% n-hexane in ethyl acetate) to give the title compound (41 mg, 40%). 1H NMR (500 MHz, CDCI3): 7.73 (1 H, s), 7.54-7.48 (3H, m), 7.45- 7.41 (2H, m), 5.56 (2H, s), 3.10-3.00 (2H, q), 1.53-1.48 (3H, t). LCMS (Method D) RT 1.117 min, M+1= 358.
Step 4: Tert-butyl ((1s.3s)-1-(4-(1-ethyl-8-phenyl-4H-pyridor2.3-blf1.2.41triazolor4.3- dlf1 ,4loxazin-7-yl)phenyl)-3-hvdroxycvclobutyl)carbamate
In a 15 ml reaction tube was 7-bromo-1-ethyl-8-phenyl-4H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazine (40mg, 0.112 mmol), tert-butyl ((1s,3s)-3-hydroxy-1- (4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (36.3 mg, 0.093 mmol), and cesium carbonate (152 mg, 0.467 mmol) in 1 ,4-dioxane (1.8ml) and water (0.6ml) to give an orange suspension. The reaction was degassed by bubbling nitrogen for 15 minutes. PdCI2(dppf)-CH2Cl2 adduct (15.24 mg, 0.019 mmol) was added and the reaction mixture degassed by bubbling nitrogen for further 15 minutes. The reaction mixture was heated to 50°C for 1 hr.
The reaction mixture was allowed to cool to RT and water was added (7ml). The reaction mixture was extracted using ethyl acetate (10ml x 3). The combined organic layers were concentrated under reduced pressure.
The crude was purified by Biotage silica gel chromatography (gradient 0% to 5% methanol in dichloromethane) to give the title compound (27mg, 53%). LCMS (Method D) RT 1.091 min, M+1= 530. 1H NMR (500 MHz, CDCI3): 7.82 (1 H, s), 7.39-.7.32 (5H, m), 7.25-7.19 (4H, m), 5.57 (2H, s), 3.53-3.46 (2H, d), 3.18-3.09 (2H, q), 3.08-2.96 (2H, bs), 2.19 (1H, s), 1.82-1.75 (3H, t), 1.59-1.53 (9H, bs Step 5: (1s,3s)-3-Amino-3-(4-(1-ethyl-8-phenyl-4H-pyridor2.3-biri ,2,4Uriazolo[4,3- d1f1 ,41oxazin-7-yDphenyl)cvclobutanol
Tert-butyl ((1s,3s)-1-(4-(1-ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin- 7-yl)phenyl)-3-hydroxycyclobutyl)carbamate (34mg, 0.063 mmol) was dissolved in dichloromethane (1ml). TFA (1 ml) was added at room temperature and the reaction mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (1 ml) and after settling the supernatant solvent was removed by pipette. This was repeated twice. The residue was then slurred in n- hexane (1 ml) and after settling the supernatant solvent was removed by pipette. This was repeated twice. The remaining solvent was removed under reduced pressure and the residue was dried until constant weigh. It was further purified by preparatory HPLC to give the desired product as an off-white solid (7 mg, 25% yield). LCMS (Method D) RT = 0.658 min, M+1= 440. Ή-NMR (500 MHz, CD3OD) 6 7.95 (1 H, s), 7.31-7.13 (8H, m), 5.52 (2H, s), 3.84-3.76 (1 H, m), 3.17-3.10 (2H, q), 2.75-2.65 (2H, m), 2.86-2.77 (2H, m), 2.13-2.02 (1H, m), 1.43-1.38 (3H, t).
Example 137: 6-(4-((1 r.3r)-1-amino-3-hvdroxycvclobutyl)phenyl)-1 -methyl-7-phenyl- 1 H-pyridor2.3-bin .41oxazin-2(3H)-one
Figure imgf000193_0001
Step 1 : 1-(4-bromophenyl)-3,3-dimethoxycyclobutanecarbonitrile
A suspension of sodium hydride (2 g, 50.0 mmol) in DMF (20 ml) was cooled to 0 degree before 2-(4-bromophenyl)acetonithle (4 g, 20.40 mmol) was added slowly. The suspension was stirred at 0 degree for another 10min before 1,3-dibromo-2,2- dimethoxypropane (2.62 g, 10.00 mmol) was added. The reaction mixture was stirred at
60 °C for 20h before cooled to room temperature, poured into water (75ml) and extracted with ethyl acetate (2X50ml). The combined organic phase was washed with water (75ml), brine (50ml) and concentrated. The crude product was purified by column (100g, biotage) eluted with ethyl acetate/cyclohexane (0-10%) to afford product 1.7g (59% yield). Ή NMR (500 MHz, DMSO-d6): 7.46 (d, 2H), 7.29 (d, 2H), 3.21 (s, 3H), 3.11 (s, 3H), 3.03 (d, 2H), 2.62 (d, 2H).
Step 2: 1-(4-bromophenyl)-3,3-dimethoxycyclobutanecarboxamide
Hydrogen peroxide (2.05 ml, 20.07 mmol) was added over 2h to a stirred mixture of 1-(4- bromophenyl)-3,3-dimethoxycyclobutanecarbonitrile (3.18g, 10.74 mmol) and Potassium carbonate (0.297 g, 2.15 mmol) in DMSO (11 ml) at 40 degree under a nitrogen atmosphere. After the addition, the reaction mixture was heated to 80 degree for 3h. The reaction mixture was allowed to cool down to room temperature and water (30ml) was added. The reaction mixture was extracted with ethyl acetate (3 X 20ml). The combined organic phase was washed with water (2 x 30ml), brine (30ml), concentrated to give light yellow oil (3.3g). 1H NMR (500 MHz, CDCI3) 7.40 (d, 2H), 7.11 (d, 2H), 5.89 (br, 1 H), 5.58 (br, 1 H), 3.12 (s, 3H), 3.02 (s, 3H), 2.97 (d, 2H), 2,52 (d, 2H).
Step 3 1-(4-bromophenyl)-3,3-dimethoxycvclobutanamine
To a solution of 1-(4-bromophenyl)-3,3-dimethoxycyclobutanecarboxamide (2.3 g, 7.32 mmol) in acetonitrile (9 ml) and Water (9.00 ml) was added phenyliodine
bis(trifluoroacetate) (4.73 g, 11.00 mmol) and the resulting mixture was stirred at room temperature for 18h. The reaction mixture was slowly poured into sodium bicarnate sat. solution (~45ml) and extracted with ethyl acetate (3X30ml). The combined organic phase was washed with water, brine and concentrated to give crude product (3.4g). The crude product was purified by column (biotage, 50g) eluted with 0-5% DCM/Methanol (mixture of 500ml methanol and 60ml 2M NH40H in methanol) to give product 1-(4- bromophenyl)-3,3-dimethoxycyclobutanamine (1.06g), 1H NMR (500 MHz, CDCI3): 7.48 (d, 2H), 7.32 (d, 2H), 3.27 (d, 3H), 3.17 (d, 3H), 2.66 (d, 2H), 2.44 (d, 2H).
Step 4: 2-(1-(4-bromophenyl)-3,3-dimethoxycyclobutyl)isoindoline-1 ,3-dione
A mixture of 1-(4-bromophenyl)-3,3-dimethoxycyclobutanamine (0.66g, 2.306 mmol), ethyl 1 ,3-dioxoisoindoline-2-carboxylate (0.56 g, 2.55 mmol) and triethylamine (1.3 ml, 9.33 mmol) in CHCI3 (15ml) was heated at 70 °C for 16h. The reaction mixture was concentrated to as dry as possible. Methanol (~5ml) was added. The solution was concentrated to dryness. Methanol (~5ml) was added again and the resulted solid was collected by filtration to afford product as a white solid (0.39g). 1H NMR (500 MHz, CDCI3): 7.70 (dd, 2H), 7.60 (dd, 2H), 7.44 (dd, 2H), 7.37 (dd, 2H), 3.18 (m, 4H), 3.10 (s, 3H), 3.02 (s, 3H).
Step 5: 2-(1-(4-bromophenyl)-3-oxocvclobutyl)isoindoline-1 ,3-dione
A solution of 2-(1-(4-bromophenyl)-3,3-dimethoxycyclobutyl)isoindoline-1 ,3-dione (0.3 g, 0.72 mmol) in Acetone (30 ml) was added 6M HCI (3ml) and the resulting solution was stirred at 50 °C for 2h. The reaction mixture was concentrated to give product 0.26g. ). 1H N R (500 MHz, CDCI3): 7.76 (dd, 2H), 7.67 (dd, 2H), 7.41 (m, 4H), 4.21 (m, 2H), 3.87 (m, 2H).
Step 6: 2-((1rl3r)-1-(4-bromophenyl)-3-hvdroxycvclobutyl)isoindoline-1 ,3-dione
A solution of 2-(1-(4-bromophenyl)-3-oxocyclobutyl)isoindoline-1 ,3-dione (194mg, 0.524 mmol) in THF (5 ml) was added L-Selectride (0.524 ml, 0.524 mmol)(1 M THF solution) at -78 °C to give a yellow solution and stirred at -78 °C for 1 h. The reaction mixture was quenched with saturated sodium bicarbonate solution at -78 °C and extracted with ethyl acetate (3X10ml). The combined organic phase was concentrated and purified by column (biotage 25g) to give product (99mg). 1H NMR (500 MHz, CDCI3): 7.76-7.79 (m, 2H), 7.67-7.71 (m, 2H), 7.42-7.50 (m, 4H), 4.5-4.6 (m, 1 H), 3.73-3.84 (m, 2H), 2.76-2.83 (m, 2H).
Step 7: 2-((1 r.3r)-3-hydroxy-1-(4-(4.4,5.5-tetramethyl-1 ,3.2-dioxaborolan-2- yl)phenyl)cvclobutyl)isoindoline-1 ,3-dione
A mixture of 2-((1 r,3r)-1-(4-bromophenyl)-3-hydroxycyclobutyl)isoindoline-1 ,3-dione (99mg, 0.266 mmol), 4)4,4',4,,5,5,5',5'-octamethyl-2,2'-bi(1 ,3,2-dioxaborolane) (67.5 mg, 0.266 mmol) and potassium acetate (131 mg, 1.330 mmol) in Dioxane (10 ml) was evacuated and flushed with nitrogen three times. Pd(dppf)CI2.CH2Cl2 (21.72 mg, 0.027 mmol) was added and the resulting mixture was evacuated and flushed with nitrogen 3 times. The mixture was then heated at 90 °C for 22h. The reaction mixture was concentrated and the residue was purified by chromatography (biotage 25g) eluted with ethyl acetate/cyclohexane 0-70% to give product (45mg). 1H NMR (500 MHz, CDCI3): 7.73-7.78 (m, 4H), 7.65 (m, 2H), 7.57-7.59 (m, 2H), 4.45-4.55 (m, 1 H), 3.81-3.85 (m, 2H), 2.79-2.82(m, 2H), 1.30 (s, 12H). Step 8: 2-(nr,3r)-3-hvdroxy-1-(4-(1-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2.3- b]f 1 ,4loxazin-6-yl)phenyl)cvclobutyl)isoindoline-1 ,3-dione
A mixture of 6-bromo-1-methyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (45mg, 0.141 mmol), 2-((1r,3r)-3-hydroxy-1-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (49.3 mg, 0.117 mmol) and cesium carbonate (191 mg, 0.587 mmol) in Dioxane (3 ml) and Water (1 ml) was degassed. Then
PdCI2(dppf).CH2CI2 (19.19 mg, 0.023 mmol) was added. The reaction mixture was degassed again and heated at 50 °C under stirring for 16h.
The reaction mixture was cooled down to room temperature and diluted with water (15ml) and extracted with ethyl acetate (3X15ml). The combined organic phase was washed with water, dried over sodium sulfate and concentrated to give crude product which was purified by column chromatography (biotage 10g) eluted with ethyl acetate/cyclohexane 0-80% to afford product (26mg) ). 1H NMR (500 MHz, CDCI3): 7.74-7.76 (m, 2H), 7.66- 7.68 (m, 2H), 7.40-7.42 (m, 2H), 7.23-7.30 (m, 6H), 7.17-7.19 (m, 2H), 4.87 (s, 2H), 4.45- 4.55 (m, 1 H), 3.79-3.82 (m, 2H), 3.38 (s, 3H), 2.74-2.78 (m, 2H), LCMS (Method D): RT = 1.22 min, M+H+ = 533.0.
Step 9: 6-(4-((1r,3r)-1-amino-3-hydroxycvclobutyl)phenyl)-1-methyl-7-phenyl-1 H- pyridor2,3-biri,41oxazin-2(3H)-one
A mixture of 2-((1r,3r)-3-hydroxy-1-(4-(1-methyl-2-oxo-7-phenyl-2,3-dihydro-1H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (26mg, 0.049 mmol) in Dioxane (1 ml) and MeOH (1.0 ml) was added hydrazine (0.25ml, 7.97 mmol) monohydrate. The reaction mixture was heated at 120 °C for 30 min under microwave condition. The reaction mixuture was diluted with saturated bicarbonate solution (10ml) and extracted with DCM (3X10ml). The combined organic phase was washed with water, brine and dried over sodium sulfate, filtered and concentrated to give crude product which was purified by prepHPLC (method F) to give product (5mg). 1H NMR (500 MHz, CD3OD): 7.44 (s, 1H), 7.28-7.29 (m, 2H), 7.18-7.20 (m, 5H), 7. 1-7.13 (m, 2H), 4.85 (s, 2H), 4.52-4.54 (m, 1 H), 3.32 (s, 3H), 32.83-2.87 (m, 2H), 2.46-2.51 (m, 2H), LCMS (Method D): RT = 0.67 min, M+H+ = 403.2. Example 138: (1 r.3r)-3-amino-3-(4-(1-ethyl-8-phenyl-4H-pyridor2.3- b][1 ,2,41triazolor4,3-dl[1 ,41oxazin-7-yl)phenylM -methylcyclobutanol
Figure imgf000197_0001
Step 1: 2-(nr.3r)-1-M-M-ethyl-8-phenyl-4H-pyridof2,3-blf1,2.41triazolof4,3-d1f1,41oxazin- 7-yl)phenyl)-3-hvdroxy-3-methylcvclobutyl)isoindoline-1 ,3-dione
In a 15 ml reaction tube was 7-bromo-1-ethyl-8-phenyl-4H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazine (62mg, 0.174 mmol), 2-((1r,3r)-3-hydroxy-3-methyl-1- (4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (62.7 mg, 0.145 mmol), and cesium carbonate (236 mg, 0.723 mmol) in 1 ,4-dioxane (2.5 ml) to give a yellow solution. The reaction mixture was degassed by bubbling nitrogen for 15 minutes, following by addition of PdCl2(dppf)-CH2Cl2 adduct. The reaction mixture was heated to 50°C for 1 hr.
The reaction mixture was allowed to cool to RT and water was added (7ml). The reaction mixture was extracted using ethyl acetate (10ml x 3). The combined organic layers were concentrated under reduced pressure.
The crude was purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n-hexanes, then 0% to 3% methanol in dichloromethane) to give the title compound (65mg, 77%). LCMS (Method D) RT 1.212 min, M+1= 584. 1H NMR (500 MHz, CDCI3): 7.79 (1H, s), 7.77- 7.72 (2H, m), 7.71-7.64 (2H, m), 7.58- 7.51 (2H, m), 7.38- 7.28 (5H, m), 7.22 7.16 (1H, m), 5.54 (2H, s), 3.39-3.29 (2H, m), 3.16-3.06 (2H + 2H, m), 1.52-1.50 (3H, t), 1.42 (3H, s). Step 2: (1 r,3r)-3-amino-3-(4-(1-ethyl-8-phenyl-4H-pyridor2,3-biri ,2.41triazolor4,3- dlf 1.41oxazin-7-yl)phenyl)-1 -methylcyclobutanol
In a 10ml microwave vial was 2-((1r,3r)-1-(4-(1-ethyl-8-phenyl-4H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline-
1.3- dione (30mg, 0.051 mmol) and hydrazine monohydrate (0.016 ml, 0.514 mmol) in
1.4- dioxane (1.5 ml) and ethanol (0.5ml) to give a yellow solution. The reaction mixture was heated to 140°C for 3 hours under microwave conditions.
The reaction mixture was cooled to room temperature and filtered to remove the solid, which has formed during the reaction. This was washed thoroughly with ethanol. The filtrates were combined and concentrated under reduced pressure.
The crude residue (20mg) was dissolved in 1 ml of a solution of methanol:
dichloromethane (1 :1) and added to a scx-2 catch-and-release cartridge (5g). After allowing to binding for 15 minutes, the column was washed with methanol (4 x 2ml) then the product eluted with 2M ammonia in methanol (4x 2ml). The combined product fractions were concentrated to dryness under reduced pressure to give an off-white solid (15mg). This was re-dissolved in methanol/water and freeze dried overnight to give the title compound (15mg, 64%). LCMS (Method D) RT = 0.630 min, M+1= 454.
1H-NMR (500 MHz, CD3OD) δ 7.95(1 H, s), 7.26-7.23 (3H, m), 7.22- 7.20 (3H, m), 7.18- 7.13 (3H, m), 5.52 (2H, s), 3.15-3.09 (2H, q), 2.59-2.52 (2H, m), 2.33-2.24 (2H, m), 1.43 (3H, s), 1.41-1.36 (3H, t).
Example 139: 6-(4-(f1s.3s)-1-amino-3-hvdroxycvclobutyl)phenyl)-1-ethyl-7-phenyl- 1 H-pyridor2.3-bl[1.41oxazin-2(3H)-one
Figure imgf000198_0001
Step 1 : terf-butyl ((1 s.3s)-1 -(4-(1-ethyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridor2.3- b1f1 ,41oxazin-6-yl)phenyl)-3-hvdroxycyclobutyl)carbamate
In a 15 mL reaction tube was added 6-bromo-1-ethyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (50 mg, 0.150 mmol), tert-butyl ((1s,3s)-3-hydroxy-1-(4-(4,4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (49 mg, 0.125 mmol) and cesium carbonate (204 mg, 0.625 mmol) in a mixture of 1,4-dioxane (2.3 ml) and water (0.8 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of [1 ,1 - bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (20 mg, 0.025 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for one hour then allowed to cool to room
temperature, diluted with water (5 ml) and extracted into ethyl acetate (3 x 5 ml). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography
(cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (45 mg, 70% yield). Ή-NMR (500 MHz, CDCI3) δ 7.29-7.35 (5H, m), 7.28 (1 H, s), 7.18-7.24 (4H, m), 4.96 (1 H, br s), 4.88 (2H, s), 4.05 (1 H, br s), 4.01 (2H, q), 2.98 (2H, br s), 2.75 (2H, br s), 1.20-1.51 (9H, br m), 1.32 (3H, t). LCMS (Method D) RT = 1.25 min, M+H+ = 516.20.
Step 2: 6-(4-((1 s,3s)-1-amino-3-hvdroxycyclobutyl)phenyl)-1-ethyl-7-phenyl-1 H- pyridor2,3-bin .41oxazin-2(3H)-one
/erf-butyl ((1s,3s)-1 -(4-(1-ethyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)-3-hydroxycyclobutyl)carbamate (45 mg, 0.087 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as an off-white solid (33 mg, 71% yield). 1H-NMR (500 MHz, MeOD) δ 7.55 (1 H, s), 7.39-7.42 (4H, m), 7.27-7.31 (3H, m), 7.20- 7.24 (2H, m), 4.93 (2H, s), 4.01-4.11 (3H, m), 3.03-3.1 1 (2H, m), 2.42-2.50 (2H, m), 1.28 (3H, t). LCMS (Method D) RT = 0.74 min, M+H+ = 416.20. Example 140: 6-(4-((1s.3s)-1-amino-3-hvdroxycvclobutyl)phenv0-1- (cvclopropylmethyl)-7-phenyl-1H-pyridor2.3-b1M.41oxazin-2(3H)-one
Figure imgf000200_0001
Step 1 : ferf-butyl ((1s,3s)-1 -(4-(1-(cvclopropylmethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H- Pyridof213-biri.41oxazin-6-yl)phenyl)-3-hvdroxycvclobutyl)carbamate
In a 15 mL reaction tube was added 6-bromo-1-(cyclopropylmethyl)-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (50 mg, 0.139 mmol), terf-butyl ((1s,3s)-3-hydroxy-1- (4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (45 mg, 0.1 16 mmol) and cesium carbonate (189 mg, 0.580 mmol) in a mixture of 1 ,4-dioxane (2.2 ml) and water (0.7 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of [1 ,1 - bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (19 mg, 0.023 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for one hour then allowed to cool to room
temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography
(cyclohexane:ethyl acetate, gradient elution from 90:10 to 0: 100) to give the desired product as an off-white solid (43 mg, 68% yield). H-NMR (500 MHz, CDCI3) δ 7.40 (1 H, s), 7.28-7.34 (5H, m), 7.18-7.24 (4H, m), 4.96 (1H, br s), 4.90 (2H, s), 4.05 (1 H, br s), 3.87 (2H, d), 2.99 (2H, br s), 2.75 (2H, br s), 1.14-1.54 (9H, br m), 1.1 1-1.22 (1 H, m), 0.54-0.62 (2H, m), 0.43-0.50 (2H, m). LCMS (Method D) RT = 1.36 min, M+H+ = 542.20.
Step 2: 6-(4-((1s,3s)-1-amino-3-hvdroxycyclobutyl)phenyl)-1-(cvclopropylmethyl)-7- phenyl-1 H-pyridof2.3-b1M ,41oxazin-2(3H)-one
fert-butyl ((1 s,3s)-1-(4-(1-(cyclopropylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)-3-hydroxycyclobutyl)carbamate (43 mg, 0.079 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as a yellow solid (31 mg, 70% yield). 1H-NMR (500 MHz, MeOD) δ 7.65 (1 H, s), 3.60 (4H, s), 7.28-7.31 (3H, m), 7.20- 7.24 (2H, m), 4.94 (2H, s), 4.06 (1 H, quin), 3.96 (2H, d), 3.02-3.12 (2H, m), 2.40-2.51 (2H, m), 1.14-1.25 (1 H, m), 0.53-0.60 (2H, m), 0.40-0.47 (2H, m). LCMS (Method D) RT = 0.83 min, M+H+ = 442.20.
Example 141: 7-(4-((1s,3s)-1-amino-3-hvdroxycvclobutyl)phenyl)-8-phenyl-2,4- dihvdro-1 H-pyridor2,3-b1M ,2.41triazolor4,3-d1M .4loxazin-1-one
Figure imgf000201_0001
Step 1 : 7-bromo-8-phenyl-2.4-dihvdro-1 H-pyridof2,3-b1f 1 ,2,41triazolof4.3-d1f 1 ,4loxazin-1 - one
In a 10 ml microwave vial was 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazine-2(3H)- thione (1g, 3. 1 mmol) and ethylhydrazine carboxylate (1.2g, 12.44 mmol) in p-xylene (10 ml) to give a yellow suspension. The reaction mixture was heated to 180°C for 45 minutes under microwave conditions. The crude residue was concentrated under reduced pressure and purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n-hexane) to give the title compound (293mg, 27%). LCMS (Method D) RT 1.069 min, M+1 = 346.
Step 2: tert-butyl ((1 s,3s)-3-hvdroxy-1-(4-(1-oxo-8-phenyl-2.4-dihvdro-1 H-pyridof2.3- b]ri12,41triazolor4.3-diri.41oxazin-7-yl)phenyl)cvclobutyl)carbamate
In a 15 ml reaction tube was 7-bromo-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-1 -one (70mg, 0.203 mmol), tert-butyl ((1s,3s)-3- hydroxy-1-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (65.8 mg, 0.169 mmol), and cesium carbonate (275 mg, 0.845 mmol) in 1 ,4-dioxane (3ml) and water (0.733ml) to give a yellow solution.
The reaction mixture was degassed by bubbling nitrogen for 15 minutes, following by the addition of PdCl2(dppf)-CH2Cl2 adduct (20mg) and degassing for further 15 minutes.
The reaction mixture was heated to 70OC for 96 hours under nitrogen atmosphere. Reaction mixture was allowed to cool to room temperature and water was added (7ml). The reaction mixture was extracted using ethyl acetate (10ml x 3). The combined organic layers were concentrated under reduced pressure.
The crude was purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate in n-hexanes) to give the title compound (15mg, 16%). LCMS (Method D) RT 1.034 min, M+1= 528. Ή NMR (500 MHz, CDCI3): 8.57 (1 H, s), 7.48-J.27 (5H, m), 7.25- 7.19 (4H, m), 5.30 (2H, s), 3.09-2.94 (2H, m), 2.84-2.70 (2H, m), 2.04 (1 H, s), 1.47-1.32 (9H, bs).
Step 3: 7 -(4-((1s,3s)-1-amino-3-hvdroxycvclobutyl)phenyl)-8-phenyl-2,4-dihvdro-1 H- Pyridor2.3-b1f1.2.4ltriazolor4.3-diri.41oxazin-1-one
Tert-butyl ((1s,3s)-3-hydroxy-1-(4-(1-oxo-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)cyclobutyl)carbamate (15mg, 0.028 mmol) was dissolved in dichloromethane (1 ml). TFA (1 ml) was added at room temperature and the reaction mixture was stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~1 ml) and concentrated to dryness under reduced pressure. This was repeated three times. The residue was then slurred in diethyl ether (1 ml) and after settling the supernatant solvent was removed by pipette. This was repeated twice. The residue was then slurred in n-hexanes (1 ml) and after settling the supernatant solvent was removed by pipette. This was repeated twice. The remaining solvent was removed under reduced pressure and the residue was dried until constant weigh. It was further purified by preparatory HPLC to give the desired product as an off-white solid (4 mg, 33% yield). LCMS (Method D) RT = 0.613 min, M-NH2= 410. 1H-NMR (500 MHz, CD3OD) δ 8.56 (1 H, s), 7.42-7.37 (2H, m), 7.36- 7.28 (5H, m), 7.25- 7.20 (2H, m), 5.41 (2H, s), 3.96- 3.87 (1 H, m), 3.00-2.88 (2H, m), 2.28-2.13 (2H, m). Example 142: (4-(1-ethyl-8-phenyl-4H-pyridof2,3-biri.2.4ltriazolor4,3-dirt4loxazin- 7-yl)phenyl)methanamine
Figure imgf000203_0001
Step l : 2-((1r,3r)-1-(4-(1-ethyl-8-phenyl-4H-pyridor2,3-biri,2.41triazolor4,3-diri,41oxazin- 7-yl)phenyl)-3-hvdroxy-3-methylcyclobutyl)isoindoline-1 ,3-dione
In a 15 ml reaction tube was 7-bromo-1-ethyl-8-phenyl-4H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazine (75mg, 0.210 mmol), tert-butyl 4-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)benzylcarbamate (84 mg, 0.252 mmol), and cesium carbonate (342 mg, 1.050 mmol) in 1 ,4-dioxane (3ml) and water (1 ml) to give a yellow solution. The reaction mixture was degassed by bubbling nitrogen for 15 minutes, following by addition of PdCl2(dppf)-CH2Cl2 adduct (34mg). The reaction mixture was heated to 50°C for 1 hour.
The reaction mixture was allowed to cool to RT and water was added (7ml). The reaction mixture was extracted using ethyl acetate (10ml x 3). The combined organic layers were concentrated under reduced pressure.
The crude was purified by Biotage silica gel chromatography (gradient 0% to 100% ethyl acetate cyclohexane) to give the title compound (51 mg, 50%). LCMS (Method B) RT 1.292 min, M+1= 484.1H NMR (500 MHz, CDCI3): 7.81 (1 H, s), 7.38- 7.30 (5H, m), 7.23- 7.18 (2H, m), 7.16- 7.12 (2H, m), 5.57 (2H, s), 4.29 (2H, bs), 3.19-3.09 (2H, q), 1.58- 1.50 (3H, t), 1.48-1.39 (9H, bs).
Step 2: (4-(1-ethyl-8-phenyl-4H-pyridor2.3-biri .2,4ltriazolof4.3-d1f1 ,41oxazin-7- vDphenvQmethanamine
Following the procedure for 1-((1 H-imidazol-2-yl)methyl)-6-(4-(1- aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one, 2-((1 r,3r)-1 -(4- (1 -ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)phenyl)-3-hydroxy- 3-methylcyclobutyl)isoindoline-1 ,3-dione (21 mg, 0.04 mmol) was reacted to afford the title compound (26 mg, quantitative). LC S: RT = 0.669 min, +1 = 384. 1H N R (500 MHz, MeOD): 7.99 (1H, s), 7.36- 7.32 (2H, m), 7.29-7.20 (5H, m), 7.19 -7.15 (2H, m), 5.55 (2H, s), 4.00 (2H, s), 3.25-3.16 (2H, q), 1.69-1.43 (3H, q).
Example 143: 6-(4-((1 r,3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1-ethyl-7- phenyl-1 H-pyridor2.3-bin .41oxazin-2(3H)-one
Figure imgf000204_0001
Step 1 : 2-((1 r,3r)-1 -(4-(1 -ethyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyrido[2.3-bin ,41oxazin-6- yl)phenyl)-3-hydroxy-3-methylcvclobutyl)isoindoline-1 ,3-dione
In a 15 mL reaction tube was added 6-bromo-1-ethyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (40 mg, 0.120 mmol), 2-((1r,3r)-3-hydroxy-3-methyl-1-(4- (4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (43 mg, 0.100 mmol) and cesium carbonate (163 mg, 0.500 mmol) in a mixture of 1 ,4- dioxane (1.9 ml) and water (0.6 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of [1 ,1'- bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (16 mg, 0.020 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for one hour. The reaction mixture was allowed to cool to room temperature, diluted with water (5 ml) and extracted into ethyl acetate (3 5 ml). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (40 mg, 71% yield). 1H-NMR (500 MHz, CDCI3) δ 7.71-7.77 (2H, m), 7.63-7.69 (2H, m), 7.50 (2H,d), 7.23-7.33 (6H, m), 7.15-7.20 (2H, m), 4.84 (2H, s), 3.99 (2H, q), 3.32 (2H, d), 3.08 (2H, d), 1.40 (3H, s), 1.30 (3H, q). LCMS (Method D) RT = 1.36 min, M+H+ = 560.20. Step 2: 6-(4-((1r.3r)-1-amino-3-hvdroxy-3-rnethylcvclobutyl)phenyl)-1-ethyl-7-phenyl-1 H- pyridor2.3-b1[1 ,4k>xazin-2(3H)-one
To a solution of 2-((1r,3r)-1-(4-(1-ethyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline-1 ,3-dione (40 mg, 0.071 mmol) in a mixture of methanol (1 mL) and 1 ,4-dioxane (1 ml) was added hydrazine monohydrate (0.25 ml, 5.15 mmol) and heated to 120°C under microwave irradiation for 30 minutes. The reaction mixture was diluted with dichloromethane (8 ml) and washed with saturated sodium bicarbonate solution (8 ml), brine (8 ml) and water (8 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was redissoved in 1 ,4-dioxane (1 mL) followed by the dropwise addition of 4M HCI in 1 ,4-dioxane (0.054 mL, 0.214 mmol). The resulting suspension was stirred at room temperature for 15 minutes, then diluted with diethyl ether (6 mL) and slurried for 10 minutes. After settling, the supernatant solvent was removed by pipette. The solid was slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated two times. The remaining solvent was removed by
concentration under reduced pressure and freeze drying overnight to give the desired compound as a white solid (27 mg, 81% yield). 1H-NMR (500 MHz, MeOD) δ 7.56 (1H, s), 7.37-7.44 (4H, m), 7.27-7.31 (3H, m), 7.20-7.25 (2H, m), 4.93 (2H, s), 4.07 (2H, q), 2.85 (2H, d), 2.67 (2H, d), 1.49 (3H, s), 1.28 (3H, q). LCMS (Method D) RT = 0.72 min, M+H+ = 430.20.
Example 144: 6-(4-((1r.3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1- (cvclopropylmethyl)-7-phenyl-1H-pyridor2.3-biri.41oxazin-2(3H)-one
Figure imgf000205_0001
Step 1 : 2-((1r.3r)-1-(4-(1-(cvclopropylmethvn-2-oxo-7-phenyl-2.3-dihvdro-1H-pyridor2,3- biri .4loxazin-6-yl)phenyl)-3-hvdroxy-3-methylcvclobutyl)isoindoline-1 ,3-dione
In a 15 mL reaction tube was added 6-bromo-1-(cyclopropylmethyl)-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (40 mg, 0.11 1 mmol), 2-((1 r,3r)-3-hydroxy-3-methyl-1- (4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1 ,3-dione (40 mg, 0.093 mmol) and cesium carbonate (151 mg, 0.464 mmol) in a mixture of 1 ,4- dioxane (1.7 ml) and water (0.6 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of [1 ,1 - bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (15 mg, 0.019 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for one hour. The reaction mixture was allowed to cool to room temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) then Biotage chromatography (dichloromethane:methanol, gradient elution from 100:0 to 90:10) to give the desired product as an off-white solid (20 mg, 37% yield). 1H-NMR (500 MHz, CDCI3) δ 7.71-7.76 (2H, m), 7.63-7.69 (2H, m), 7.50 (2H,d), 7.36 (1H, s), 7.24-7.33 (5H, m), 7.15-7.19 (2H, m), 4.85 (2H, s), 3.84 (2H, d), 3.32 (2H, d), 3.08 (2H, d), 1.40 (3H, s), 1.09-1.19 (1 H, m), 0.52-0.59 (2H, m), 0.42-0.48 (2H, m). LCMS (Method D) RT = 1.45 min, M+H+ = 586.20.
Step 2: 6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcvclobutyl)phenyl)-1- (cvclopropylmethyl)-7-phenyl-1H-pyridof2,3-biri ,4]oxazin-2(3H)-one
To a solution of 2-((1r,3r)-1-(4-(1-(cyclopropylmethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline-1 ,3-dione (20 mg, 0.034 mmol) in a mixture of methanol (1 ml) and 1 ,4-dioxane (1 ml) was added hydrazine monohydrate (0.25 mL, 5.15 mmol) and heated to 120°C under microwave irradiation for 30 minutes. The reaction mixture was diluted with dichloromethane (8 ml) and washed with saturated sodium bicarbonate solution (8 ml), brine (8 ml) and water (8 ml), dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was redissoved in 1 ,4-dioxane (1 ml) followed by the dropwise addition of 4M HCI in 1 ,4-dioxane (0.026 mL, 0.102 mmol). The resulting suspension was stirred at room temperature for 15 minutes, then diluted with diethyl ether (6 ml) and slurried for 10 minutes. After settling, the supernatant solvent was removed by pipette. The solid was slurried in diethyl ether (3 ml) and after settling the supernatant solvent removed by pipette. This was repeated two times. The remaining solvent was removed by
concentration under reduced pressure and freeze drying overnight to give the desired compound as a white solid (18 mg, quantitative yield). 1H-NMR (500 MHz, MeOD) δ 7.65 (1 H, s), 7.39-7.45 (4H, m), 7.27-7.31 (3H, m), 7.20-7.25 (2H, m), 4.94 (2H, s), 3.96 (2H, d), 2.85 (2H, d), 2.68 (2H, d), 1.49 (3H, s), 1.14-1.26 (1H, m), 0.54-0.61 (2H, m), 0.40- 0.46 (2H, m). LCMS (Method D) RT = 0.81 min, M+H+ = 456.20.
Example 145: 6-(4-((1r,3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-1-(2- fluoroethyl)-7-phenyl-1H-pyridor2.3-bin,41oxazin-2(3H)-one
Figure imgf000207_0001
Step 1 : 6-bromo-1 -(2-fluoroethyl)-7-phenyl-1 H-pyridor2.3-blH ,41oxazin-2(3H)-one
A mixture of 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (400mg, 1.311 mmol), 1-fluoro-2-iodoethane (684 mg, 3.93 mmol) and potassium carbonate (544 mg, 3.93 mmol) in DMF (4ml) was heated under stirring for 3h. The reaction mixture was cooled down and diluted with water (20ml), the resulted mixture was extracted with DCM (3X20ml). The combined organic phase was washed with water, brine, dried over sodium sulfate, concentrated to give the crude material which was purified by column
chromatography eluted with ethyl acetate/cyclohexane 0-40% to give product 0.21g. H NMR (500 MHz, CDCI3): 7.39-7.48 (m, 5H), 4.89 (s, 2H), 4.77 (t, 1 H), 4.67 (t, 1 H), 4.21 (t, 1 H), 4.16 (t, 1 H), LCMS (Method D): RT = 1.24 min, M+H+ = 353.0.
Step 2: 2-((1r.3r)-1-(4-(1-(2-fluoroethyl)-2-oxo-7-phenyl-2.3-dihvdro-1H-pyridof2,3- biri.41oxazin-6-yl)phenyl)-3-hvdroxy-3-methylcyclobutyl)isoindoline-1 ,3-dione
A mixture of 6-bromo-1-(2-fluoroethyl)-7-phenyl-1H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (38.9 mg, 0.1 mmol), 2-((1r,3r)-3-hydroxy-3-methyl-1-(4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)phenyl)cyclobutyl)isoindoline-1,3-dione (40mg, 0.092 mmol) and cesium carbonate (150 mg, 0.462 mmol) in Dioxane (3 ml) and Water (1 ml) was degased. Pd(dppf)CI2.CH2Cl2 (15.08 mg, 0.018 mmol) was added, after degassed, The reaction mixture was heating at 50 °C under stirring for 2h. The reaction mixture was cooled down to room temperature and diluted with water (20ml) and the resulted mixture was extracted with ethyl acetate (3X20ml). The combined organic phase was washed with water (30ml) and dried with sodium sulfate, filtered and concentrated to give crude product which was purified by chromatography (biotage 10g) eluted with ethyl acetate/cyclohexane 0-60% to afford product (31 mg). LCMS (Method D): RT = 1.31 min, M+H+ = 578.2.
Step 3: 6-(4-((1 r.3r)-1-amino-3-hvdroxy-3-methylcvclobutvnphenyl)-1-(2-fluoroethyl)-7- phenyl-1 H-pyridof2.3-bin ,41oxazin-2(3H)-one
A mixture of 2-((1 r,3r)-1-(4-(1-(2-fluoroethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline-1 ,3-dione (31 mg, 0.054 mmol) in Dioxane (1 ml) and Methanol (1.000 ml) was added HYDRAZINE monohydrate (0.25ml, 7.97 mmol). The reaction mixture was heated at 120 °C for 30 min under microwave condition. The reaction mixture was diluted with saturated bicarbonate solution (10ml) and extracted with DCM (3X10ml). The combined organic phase was washed with water, brine and dried over sodium sulfate, filtered and concentrated to give product (13mg) 1H NMR (500 MHz, CD3OD) 7.54 (s, 1 H), 7.20-7.22 (m, 2H), 7.15-7.17 (m, 5H), 7.07-7.09 (m, 2H), 4.86 (s, 2H), 4.66 (t, 1 H), 4.57 (t, 1 H), 4.27 (t, 1 H), 4.22 (t, 1H), 2.53 (d, 2H), 2.56 (d, 2H). LCMS (Method D): RT = 0.63 min, M+H+ = 448.2.
Example 146: 6-(4-(Aminomethyl)phenyl)-7-phenyl-1-(2,2,2-trifluoroethyl)-1H- pyridor2.3-bin.41oxazin-2(3H)-one
Figure imgf000208_0001
Step 1 : tert-Butyl 4-(2-oxo-7-phenyl-1 -(2.2.2-trifluoroethyl)-2,3-dihvdro-1 H-pyridof2.3- blf1 ,4]oxazin-6-yl)benzylcarbamate
In a 15 mL reaction tube was added 6-bromo-7-phenyl-1-(2,2,2-trifluoroethyl)-1 H- pyhdo[2,3-b][1 ,4]oxazin-2(3H)-one (40mg, 0.103 mmol), tert-butyl 4-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)benzylcarbamate (41.3 mg, 0.124 mmol), and cesium carbonate (168 mg, 0.517 mmol) in a mixture of 1 ,4-Dioxane (1937 μΙ) and water (646 μΙ) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of PdCI2(dppf)-CH2CI2Adduct (16.87 mg, 0.021 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50 °C under a nitrogen atmosphere for one hour. LC-MS analysis showed reaction completion. The reaction mixture was allowed to cool to room temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a brown solid. The resulting residue was purified via Biotage (0 - 45 % cyclohexane:EtOAc; 10g column) to give the title compound (45 mg, 85%) as off-white solid. Ή NMR (500 MHz,CDCI3): 7.36 (1 H, s), 7.32-7.26 (5H, m), 7.17-7.13 (4H,m), 4.96 (2H, s), 4.77 (1 H, br), 4.63-4.58 (2H, q), 4.28 (2H, s), 1.48 -1.45 (9H, br).
Step 2: 6-(4-(Aminomethvnphenyl)-7-phenyl-1 -(2,2,2-trifluoroethyl)-1 H-pyridof2,3- bin ,41oxazin-2(3H)-one
tert-Butyl 4-(2-oxo-7-phenyl-1 -(2,2,2-trifluoroethyl)-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)benzylcarbamate (45 mg, 0.088 mmol) was dissolved in TFA (1 mL). The resulting mixture was stirred for 30 seconds at room temperature and evaporated under reduced pressure. The deprotected compound was taken back twice into diethyl ether and the solid formed was washed twice with diethylether. The remaining solvent was removed under reduced pressure and dried to afford the title compound (34 mg, 94%) as off-white solid. LCMS (Method D): RT = 0.82 min, M+H+ =414. H NMR (500 MHz, CDCI3): 8.25 (2H, br), 7.47 (1 H, s), 7.28-7.26 (4H, m), 7.20-7.18 (3H,m), 7.10-7.08 (2H, m), 4.88 (2H, s), 4.64-4.59 (2H, q), 3.92 (2H, s).
Example 147: 2-(6-(4-((1s,3s)-1 -amino-3-hvdroxycvclobutyl)phenyl)-2-oxo-7-phenyl- 2,3-dihydro-1 H-pyridor2,3-biri.41oxazin-1-yl)acetonitrile
Figure imgf000209_0001
Step 1 : 2-(6-bromo-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2.3-b1f 1 ,4loxazin-1 - vDacetonitrile
A mixture of 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (0.4 g, .311 mmol), 2-bromoacetonitrile (0.472 g, 3.93 mmol) and potassium carbonate (0.544 g, 3.93 mmol) in DMF (4 ml) was stirred at 50 °C for 2h. The reaction mixture was diluted with water (20ml) and extracted with dichloromethane (3X20ml). The combined organic phase was washed with water, brine, dried over sodium sulfate and concentrated to give crude product which was purified by column chromatography (biotage 25g) eluted with 0-50% ethyl acetate/cyclohexane to afford product (0.19g) 1H N R (500 MHz, CDCI3) 7.45-7.50 (m, 3H), 7.4-7.43 (m, 2H), 7.31 (s, 1 H), 4.93 (s, 2H), 4.83 (s, 2H).
Step 2: tert-butyl ((1s,3s)-1-(4-(1-(cvanomethvn-2-oxo-7-phenyl-2,3-dihydro-1 H- Pyridof2,3-b1[1 ,4loxazin-6-yl)phenyl)-3-hydroxycvclobutyl)carbamate
A mixture of 2-(6-bromo-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1- yl)acetonitrile (42.4 mg, 0.123 mmol), tert-butyl ((1s,3s)-3-hydroxy-1 -(4-(4,4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (40mg, 0.103 mmol) and CESIUM CARBONATE (167 mg, 0.514 mmol) in Dioxane (3 ml) and Water (1 ml) was degassed. Pd(dppf)Cl2.CH2Cl2 (16.78 mg, 0.021 mmol) was added, after degassed, The reaction mixture was heating at 50 °C under stirring for 1 h.
The reaction mixture was cooled down to room temperature and diluted with water (20ml) and the resulted mixture was extracted with ethyl acetate (3X20ml). The combined organic phase was washed with water (30ml) and dried with sodium sulfate, filtered and concentrated to give crude product which was purified by chromatography (biotage 10g) eluted with ethyl acetate/cyclohexane 0-70% to afford product (21 mg). LCMS (Method D): RT = 1.15 min, M+H+ = 527.2.
Step 3: 2-(6-(4-(aminomethyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyridof2,3- b1[1 ,4loxazin-1 -vDacetonitrile
To o a round bottomed flask containing tert-butyl ((1s,3s)-1 -(4-(1 -(cyanomethyl)-2-oxo-7- phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)-3- hydroxycyclobutyl)carbamate (18mg, 0.034 mmol) was added TRIFLUOROACETIC ACID (0.5ml, 6.49 mmol). The resulting solution was stirred for 60 seconds then concentrated to dryness under reduced pressure. The residue was suspended in diethyl ether (1 ml) and concentrated to dryness three times to give the trifluoroacetic acid salt of the product (8.5mg) H NMR (500 MHz, CD3OD) 7.58 (s, 1 H), 7.31-7.35 (m, 4H), 7.19- 7.22 (m, 3H), 7.13-7.15 (m, 2H), 5.02 (s, 2H), 4.92 (s, 2H), 3.92-3.96 (m, 1 H), 2.95-2.99 (m, 2H), 2.35-2.39 (m, 2H). LCMS (Method A): RT = 3.53 min, M+H+ = 428.11. Example 148: 6-(4-((1s.3s)-1-amino-3-hvdroxycyclobutyl)phenyl)-7-phenyl-1 -(2.2.2- trifluoroethvn-1 H-pyridor2.3-bin ,41oxazin-2(3H)-one
Figure imgf000211_0001
Step 1 : 6-bromo-7-phenyl-1-(2,2,2-trifluoroethyl)-1 H-pyridor2,3-blf1,41oxazin-2(3H)-one A mixture of 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (0.4g, 1.31 1 mmol), potassium carbonate (0.544 g, 3.93 mmol) and 1 ,1 ,1-trifluoro-2-iodoethane (0.826 g, 3.93 mmol) in DMF (2ml) was heated at 70 °C under stirring for 18h
The reaction mixture was cooled down and diluted with water(15ml) and extracted with DCM (15mlX3). The combined organic phase was washed with water (25ml) and dried with Na2S0 and concentrated. The residue was purified by column chromatography (biotage 25g) eluted with ethyl acetate/cyclohexane 0-60% to afford product 6-bromo-7- phenyl-1-(2,2,2-trifluoroethyl)-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (0.17g) 1H NMR (500 MHz, CDCI3) 7.45-7.50 (m, 3H), 7.38-7.40 (m, 2H), 7.28 (s, 1 H), 4.94 (s, 2H), 4.55
(q, 2H).
Step 2: tert-butyl ((1s.3s)-3-hvdroxy-1-(4-(2-oxo-7-phenyl-1-(2.2,2-trifluoroethvn-2.3- dihvdro-1 H-pyridof2,3-b1[1 ,41oxazin-6-yl)phenyl)cvclobutyl)carbamate
A mixture of 6-bromo-7-phenyl-1-(2,2,2-trifluoroethyl)-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one (47.7 mg, 0.123 mmol), tert-butyl ((1s,3s)-3-hydroxy-1-(4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (40mg, 0.103 mmol) and cesium carbonate (167 mg, 0.514 mmol) in Dioxane (3ml) and Water (1 ml) was degassed. Pd(dppf)Cl2.CH2CI2 (16.78 mg, 0.021 mmol) was added, after degassed, The reaction mixture was heating at 50 °C under stirring for 1h.
The reaction mixture was cooled down to room temperature and diluted with water (20ml) and the resulted mixture was extracted with ethyl acetate (3X20ml). The combined organic phase was washed with water (30ml) and dried with sodium sulfate, filtered and concentrated to give crude product which was purified by chromatography (biotage 10g) eluted with ethyl acetate/cyclohexane 0-70% to afford product (32mg). 1H NMR (500 MHz, CDCI3) 7.31-7.37 (m, 5H), 7.16-7.22 (m, 5H), 4.96 (s, 2H), 4.95 (br, 1 H), 4.61 (q, 2H), 4.09 (m, 1 H), 2.95-3.05 (m, 2H), 2.7-2.8 (m, 2H), 1.40 (br, 9H). LCMS (Method D): RT = 1.31 min, M+H+ = 571.0
Step 3: 6-(4-((1s.3s)-1-amino-3-hvdroxycvclobutyl)phenyl)-7-phenyl-1-(2.2.2- trifluoroethylH H-pyridor2.3-blM .41oxazin-2(3H)-one
to a round bottomed flask containing to a round bottomed flask containing tert-butyl ((1 s,3s)-3-hydroxy-1-(4-(2-oxo-7-phenyl-1 -(2,2,2-trifluoroethyl)-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (30mg, 0.053 mmol) was added
TRIFLUOROACETIC ACID (0.5ml, 6.49 mmol). The resulting solution was stirred for 60 seconds then concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (1 ml) and concentrated to dryness three times to give the trifluoroacetic acid salt of the product (21 mg) 1H NMR (500 MHz, CD3OD) 7.76 (s, 1 H), 7.45 (m, 4H), 7.31-7.32 (m, 3H), 7.21-7.23 (m, 2H), 5.03 (s, 2H), 4.92 (q, 2H), 4.06-4.09 (m, 1 H), 3.08-3.1 1 (m, 2H), 2.46-2.50 (m, 2H). LCMS (Method A): RT = 3.94 min, M+H+ = 471.04.
Example 149: 6-(4-(1-aminocvclopropyl)phenyl)-1-methyl-7-phenyl-1 H-pyridor2,3- bin,41oxazin-2(3H)-one
Figure imgf000212_0001
Step 1 : 1-(4-(4,4.5,5-tetramethyl-1 ,3.2-dioxaborolan-2-yl)phenyl)cyclopropanamine
In a 40 mL reaction tube was added 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)benzonitrile (500 mg, 2.18 mmol) and titanium(IV) isopropoxide (710 μΙ, 2.40 mmol) in anhydrous tetrahydrofuran (11 ml) to give a colourless solution. The reaction mixture was cooled to -70°C under a nitrogen atmosphere, followed by the dropwise addition of ethylmagnesium bromide (1 M in THF, 4.80 ml, 4.80 mmol) over a period of 30 minutes. The resulting orange solution was stirred at -78°C for 10 minutes, followed by warming to room temperature and stirring for 60 minutes. After this period, boron trifluoride diethyl etherate (540 μΙ, 4.37 mmol) was added and the mixture stirred at room temperature for 60 minutes. To the reaction was then added 1M HCI solution (6 ml) and stirred for 15 minutes before basification with 2M NaOH solution. The organic phase was separated and the aqueous washed with ethyl acetate (3 x 4 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane.ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (70 mg, 12% yield). 1H-NMR (500 MHz, CDCI3) δ 7.76 (2H, d), 7.29 (2H, d), 2.32 (2H, br s), 1.34 (12H, s), 1.12 (2H, dd), 1.01 (2H, dd). LCMS RT = 0.68 min, +H+ = 260.20.
Step 2: terf-butyl (1 -(4-(4,4.5,5-tetramethyl-1 ,3.2-dioxaborolan-2- yl)phenyl)cvclopropyl)carbamate
To a solution of 1-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl)cyclopropanamine (70 mg, 0.270 mmol) in anhydrous THF (1.35 ml) was added triethylamine (83 μΙ, 0.594 mmol) and di-terf-butyl dicarbonate (69 μΙ, 0.297 mmol). The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 3 hours then concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 40:60) to give the desired product as a white solid (54 mg, 56% yield). H-NMR (500 MHz, CDCI3) δ 7.73 (2H, d), 7.19 (2H, d), 1.39-1.59 (9H, br m), 1.33 (12H, s), 1.30 (2H, br s), 1.25 (2H, br s).
Step 3: te/f-butyl (1-(4-(1-methyl-2-oxo-7-phenyl-2.3-dihvdro-1 H-pyridof2.3-b1[1 ,41oxazin- 6-yl)phenvDcvclopropyl)carbamate
In a 15 mL reaction tube was added 6-bromo-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (40 mg, 0.125 mmol) and ferf-butyl (1-(4-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)phenyl)cyclopropyl)carbamate (54 mg, 0.150 mmol) in 1 ,4- dioxane (1 ml), followed by a solution of sodium carbonate (40 mg, 0.3 mmol) in water (0.25 ml) to give a white suspension. This was degassed by bubbling nitrogen for 10 minutes, followed by the addition of [1 ,1 - bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (9 mg, 0.013 mmol). The reaction mixture was heated to 80°C overnight. The reaction mixture was allowed to cool to room temperature, diluted with brine (4 ml) and extracted into ethyl acetate (3 x 4 ml). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the desired product as a white solid (60 mg, quantitative yield). 1H-NMR (500 MHz, CDCI3) δ 7.14-7.26 (6H, m), 7.08-7.14 (2H, m), 6.93 (2H, d), 4.99-5.27 ( H, br m), 4.80 (2H, s), 3.31 (3H, s), 1.35 (9H, br s), 1.25 (2H, br s), 1.03 (2H, br s). LCMS (Method D): RT = 1.44 min, M+H+ = 472.20.
Step 4: 6-(4-(1-arninocvclopropyl)phenyl)-1-methyl-7-phenyl-1 H-pyrido[2,3-biri.4loxazin- 2(3H)-one
To a solution of feri-butyl (1-(4-(1-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)phenyl)cyclopropyl)carbamate (60 mg, 0.127 mmol) in 1 ,4-dioxane (1 mL) was added dropwise 4M HCI in 1 ,4-dioxane (0.5 mL, 2.00 mmol) and the resulting solution stirred at room temperature for 1 hour. The reaction mixture was diluted by the dropwise addition of diethyl ether (6 mL) and stirred for 15 minutes to give a white precipitate. The layers were allowed to settle and the supernatant solution removed. The solid was washed twice more with diethyl ether (6 mL), allowing to settle and removing the supernatant solvent each time. The remaining solvent was removed by concentration to dryness under reduced pressure, then freeze-drying over the weekend to give the desired product as an off-white solid (27 mg, 52% yield). 1H-NMR (500 MHz, MeOD) δ 7.53 (1 H, s), 7.34 (2H, d), 7.31 (2H, d), 7.25-7.29 (3H, m), 7.18-7.23 (2H, m), 4.94 (2H, s), 3.41 (3H, s), 1.29-1.35 (2H, m), 1.24-1.29 (2H, m). LCMS (Method D): RT = 0.75 min, M+H+ = 372.10.
Example 150: 2-(6-(4-((1r,3r)-1-amino-3-hvdroxy-3-methylcyclobutyl)phenyl)-2-oxo- 7-phenyl-2,3-dihydro-1 H-pyridor2,3-biri ,41oxazin-1 -vDacetonitrile
Figure imgf000214_0001
Step 1 : terf-butyl ((1r.3r)-1-(4-(1-(cvanomethvn-2-oxo-7-phenyl-2.3-dihvdro- H- pyridor2,3-biri,41oxazin-6-yl)phenyl)-3-hydroxy-3-methylcvclobutyl)carbamate
In a 15 mL reaction tube was added 2-(6-bromo-2-oxo-7-phenyl-2,3-dihydro-1H- pyrido[2,3-b][1 ,4]oxazin-1-yl)acetonitrile (40 mg, 0.116 mmol), fert-butyl ((1r,3r)-3- hydroxy-3-methyl-1-(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2- yl)phenyl)cyclobutyl)carbamate (39 mg, 0.097 mmol) and cesium carbonate (158 mg, 0.484 mmol) in a mixture of 1 ,4-dioxane (1.8 ml) and water (0.6 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of [1 ,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (16 mg, 0.019 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for one hour. The reaction mixture was allowed to cool to room temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified twice by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (30 mg, 57% yield). 1H-NMR (500 MHz, de-Acetone) δ 7.67 (1 H, s), 7.23-7.35 (9H, m), 6.55 (1 H, br s), 5.21 (2H, s), 5.04 (2H, s), 2.70-2.78 (2H, m), 2.45-2.60 (2H, m), 1.49 (3, s), 1.15-1.40 (br m, 9H). LCMS (Method D) RT = 1.22 min, M+H+ = 541.15.
Step 2: 2-(6-(4-((1r,3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-2-oxo-7-phenyl- 2,3-dihydro-1 H-pyridor2,3-blf1 ,4]oxazin-1-yl)acetonitrile
fert-Butyl ((1r,3r)-1-(4-(1-(cyanomethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1,4]oxazin-6-yl)phenyl)-3-hydroxy-3-methylcyclobutyl)carbamate (30 mg, 0.055 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as a white solid (18 mg, 59% yield). 1H-NMR (500 MHz, MeOD) δ 7.68 (1 H, s), 7.40-7.45 (4H, m), 7.28-7.31 (3H, m), 7.22-7.26 (2H, m), 5.12 (2H, s), 5.00 (2H, s), 2.85 (2H, d), 2.68 (2H, d), 1.48 (3H, s). LCMS (Method D) RT = 0.64 min, M+H+ = 441.10.
Example 151: 6-(4-(Aminomethyl)phenyl)-1-ethyl-7-phenyl-1 H-pyridor2.3- biri.41oxazin-2(3H)-one
Figure imgf000215_0001
Step 1 : tert-Butyl 4-n-ethyl-2-oxo-7-phenyl-2.3-dihydro-1 H-pyriclof2.3-blf1.41oxazin-6- vDbenzylcarbamate
In a 15 mL reaction tube was added 6-bromo-1-ethyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one (50 mg, 0.150 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1 ,3,2- dioxaborolan-2-yl)benzylcarbamate (60.0 mg, 0.180 mmol), and CESIUM CARBONATE (244 mg, 0.750 mmol) in a mixture of 1 ,4-Dioxane (2814 μΙ) and water (938 μΙ) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of PdCI2(dppf)-CH2CI2Adduct (24.51 mg, 0.030 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50 °C under a nitrogen
atmosphere for one hour. LC-MS analysis showed reaction completion. The reaction mixture was allowed to cool to room temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give an off-white solid. The resulting residue was purified via Biotage (15 - 85 % cyclohexane:EtOAc; 10g column) to give the title compound (45 mg, 65.3%) as off-white solid. Ή NMR (500 MHz,CDCI3): 7.31-7.27 (6H, m), 7.19-7.18 (2H,d), 7.1 -7.10 (2H, d), 4.87 (2H, s), 4.84 (1 H, br), 4.27 (2H, s), 4.03-3.98 (2H, q), 1.45 (9H, br), 1.32-1.30 (3H, t).
Step 2: 6-(4-(Aminomethyl)phenyl)-1 -ethyl-7-phenyl-1 H-pyridor2,3-b1f 1 ,41oxazin-2(3H)- one
tert-Butyl 4-(1-ethyl-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3-b][1 ,4]oxazin-6- yl)benzylcarbamate (45 mg, 0.098 mmol) was dissolved in TFA (1 mL). The resulting mixture was stirred for 30 seconds at room temperature and evaporated under reduced pressure. The deprotected compound was taken back twice into diethyl ether and the solid formed was washed twice with diethylether. The remaining solvent was removed under reduced pressure and dried to afford the title compound (35 mg, 99%) as white solid. LCMS: RT = 0.75 min, M+H+ =360. 1H NMR (500 MHz, CDCI3): 8.34 (2H, br), 7.30- 7.27 (5H, m), 7.18 (3H, m), 7.13-7.1 1 (2H,m), 4.80 (2H, s), 4.03-3.99 (2H, q), 3.96 (2H, s), 1.32-1.30 93H, t). Example 152: 6-(4-((1r,3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenyl)-7-phenyl- 1 -(2.2,2-trifluoroethyl)-1 H-pyridor2.3-biri .41oxazin-2f 3H)-one
Figure imgf000217_0001
Step 1 : tert-butyl ((1 r,3r)-3-hvdroxy-3-methyl-1-(4-(2-oxo-7-phenyl-1-(2,2,2-trifluoroethyl)- 2,3-dihvdro-1 H-pyridor2,3-biri,4loxazin-6-yl)phenyl)cyclobutyl)carbamate
In a 15 mL reaction tube was added 6-bromo-7-phenyl-1-(2,2,2-trifluoroethyl)-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (45 mg, 0.116 mmol), ferf-butyl ((1 r,3r)-3-hydroxy-3- methyl-1 -(4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)cyclobutyl)carbamate (39 mg, 0.097 mmol) and cesium carbonate (158 mg, 0.484 mmol) in a mixture of 1 ,4- dioxane (1.8 ml) and water (0.6 ml) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of [1,1 - bis(diphenylphosphino)ferrocene]dichloropalladium(ll) dichloromethane adduct (16 mg, 0.019 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50°C under a nitrogen atmosphere for one hour. The reaction mixture was allowed to cool to room temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure. The residue was purified twice by Biotage chromatography (cyclohexane:ethyl acetate, gradient elution from 90:10 to 0:100) to give the desired product as an off-white solid (25 mg, 44% yield). 1H-NMR (500 MHz, CDCI3) δ 7.35 (1 H, s), 7.26-7.32 (5H, m), 7.21-7.25 (2H, m), 7.14-7.18 (2H, m), 4.94 (2H, s), 2.93 (1 H, br s), 4.60 (2H, q), 2.40-2.68 (4H, m), 1.55 (3H, s), 1.20-1.40 (9H, br m). LCMS (Method D) RT = 1.38 min, M+H+ = 584.10.
Step 2: 6-(4-((1r.3r)-1-amino-3-hvdroxy-3-methylcvclobutyl)phenvn-7-phenyl-1 -(2,2.2- trifluoroethyl)-1 H-pyhdor2.3-blM ,41oxazin-2(3H one
terf-Butyl ((1r,3r)-3-hydroxy-3-methyl-1-(4-(2-oxo-7-phenyl-1-(2,2,2-trifluoroethyl)-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutyl)carbamate (25 mg, 0.043 mmol) was dissolved in TFA (1 mL) and stirred for 30 seconds. The solution was immediately concentrated to dryness under reduced pressure. The residue was dissolved in diethyl ether (~3 mL) and concentrated to dryness under reduced pressure three times. The residue was then slurried in diethyl ether (3 mL) and after settling the supernatant solvent removed by pipette. This was repeated three times. The remaining solvent was removed by freeze drying overnight to give the desired compound as a white solid (8 mg, 31 % yield). 1H-NMR (500 MHz, MeOD) δ 7.73 (1 H, s), 7.39-7.44 (4H, m), 7.26-7.32 (3H, m), 7.17-7.23 (2H, m), 5.01 (2H, s), 4.89 (2H, q), 2.85 (2H, d), 2.66 (2H, d), 1.49 (3H, s). LCMS (Method D) RT = 0.80 min, M+H+ = 484.05.
Example 153: 6-(4-(Aminomethyl)phenyl)-1-(cvclopropylmethyl)-7-phenyl-1 H- pyridor2.3-biri.41oxazin-2(3H)-one
Figure imgf000218_0001
Stepl : tert-Butyl 4-(1-(cvclopropylmethyl)-2-oxo-7-phenyl-2,3-dihvdro-1 H-pyridof2,3- bin ,41oxazin-6-yl)benzylcarbamate
In a 15 mL reaction tube was added 6-bromo-1-(cyclopropylmethyl)-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one (80mg, 0.223 mmol), tert-butyl 4-(4,4,5,5-tetramethyl- 1 ,3,2-dioxaborolan-2-yl)benzylcarbamate (89 mg, 0.267 mmol) and CESIUM
CARBONATE (363 mg, 1.1 14 mmol) in a mixture of 1 ,4-Dioxane (3341 μΙ) and water (1 1 14 μΙ) to give a colourless solution. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of PdCI2(dppf)-CH2CI2Adduct (36.4 mg, 0.045 mmol) and degassing for a further 5 minutes. The reaction mixture was heated to 50 °C under a nitrogen atmosphere for one hour. LC-MS analysis showed reaction completion. The reaction mixture was allowed to cool to room temperature, diluted with water (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give an off-white solid. The resulting residue was purified via Biotage (15 - 85 % cyclohexane:EtOAc; 10g column) to give the title compound (80 mg, 74%). Ή NMR (500 MHz,CDCI3): 7.39 (1 H, s), 7.32-7.26 (5H, m), 7.20-7.18 (2H, dd), 7.12-7.10 (2H, d), 4.89 (2H, s), 4.80 (1 H, br), 4.27 (2H, s), 3.87-3.86 (2H, d), 1.48 -1.45 (9H, br), 1.18-1.15 (1 H, m), 0.60-0.56 (2H, m), 0.48-0.45 (2H, m). Step 2: 6-(4-(Aminomethyl)phenyl)-1-(cyclopropylmethyl)-7-phenyl-1 H-pyrido[2,3- b1[1 ,41oxazin-2(3H)-one
tert-Butyl 4-(1-(cyclopropylrnethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-6-yl)benzylcarbamate (80 mg, 0.165 mmol) was dissolved in TFA (1 ml_). The resulting mixture was stirred for 30 seconds at room temperature and evaporated under reduced pressure. The deprotected compound was taken back twice into diethyl ether and the solid formed was washed twice with diethylether. The remaining solvent was removed under reduced pressure and dried to afford the title compound (45 mg, 70.9%) as white solid. LCMS: RT = 0.84 min, M+H+ =386. 1H NMR (500 MHz, CDCI3): 8.33 (2H, br), 7.41 (1 H, s), 7.27-7.26 (4H, m), 7.18 (3H, m), 7.13-7.12 (2H, m), 4.82 (2H, s), 3.94 (2H, s), 3.87-3.85 (2H, d), 1.16-1.14 (1 H, m), 0.60-0.57 (2H, m), 0.48-0.45 (2H, m). tert-butyl 1-(4-(6-chloro-5-nitro-3-phenylpyridin-2-yl)phenyl)cyclobutylcarbamate
Step 1 : tert-butyl (1-(4-(5-nitro-6-oxo-3-phenyl-1 ,6-dihydropyridin-2- yl)phenyl)cyclobutyl)carbamate
A solution of (E)-tert-butyl 1-(4-(3-(dimethylamino)-2-phenylacryloylphenyl) cyclobutyl carbamate (95 g, 214 mmol) and 2-nitroacetamide (20 g, 192 mmol) in acetic acid (600 mL) was stirred at 45°C for 3hours, then at room temperature for 18 hours. The resulting precipitate was filtered, washed with acetic acid (150 mL), diethyl ether (400 mL) and dried at ambient temperature to afford the title compound as a yellow solid (41 g). The filtrate was concentrated under reduced pressure. The residue was taken up in diethyl ether. The resulting precipitate was filtered and dried at ambient temperature to afford a second crop of product (9 g). The overall yield of title compound was 48%.1H NMR (500 MHz, d6-DMSO): 13.02 (1H, brs), 8.42 (1H, s), 7.6-7.1 (9H, m), 2.35 (4H, m), 1.98 ( H, m), 1.76 (1H, m), 1.3-1.1 (9H, m).
Step 2: tert-butyl 1-(4-(5-amino-6-hvdroxy-3-phenylpyridin-2-yl)phenyl)cvclobutyl carbamate A solution of tert-butyl 1 -(4-(6-hydroxy-5-nitro-3-phenylpyridin-2-yl)phenyl)cyclobutyl carbamate (50 g, 2.16 mmol) and 10% Pd/C (0.2 g) in THF (2 L) was hydrogenated in an autoclave (2 bar) for 5hours at room temperature. The solution was then filtered through celite and the celite was washed with THF (0.5 L). The combined filtrates were concentrated under reduced pressure. The residue was triturated in diethyl ether. The resulting white precipitate was filtered and dried at ambient to afford the title compound (50 g, 91%). It was used for the next step without further purification.
Step 3: tert-butyl 1-(4-(6-chloro-5-nitro-3-phenylpyridin-2-yl)phenyl)cyclobutylcarbamate To a Schlenck tube were added tert-butyl 1-(4-(6-chloro-5-nitro-3-phenylpyridin-2- yl)phenyl)cyclobutylcarbamate (10g, 21.67 mmol), triphenylphosphine (17.05 g, 65 mmol), DCE (200 mL) and carbon tetrachloride (2.19 mL, 22.75 mmol). The resulting heterogeneous solution was heated at 100°C for 35 minutes. It was then concentrated under reduced pressure. The residue was purified by Biotage chromatography (gradient 0% to 3% methanol in dichloromethane) to afford the title compound as a white solid (6 g, 57%). H NMR (500 MHz, CDCI3): 8.30 (1 H, s), 7.4-7.2 (9H, m), 2.51 (4H, m), 2.10 (1 H, m), 1.88 (1H, m), 1.3-1.1 (9H, m). tert-butyl (1 -(4-(5-amino-6-oxo-3-phenyl-1 ,6-dihydropyridin-2- vDphenvDcyclobutvDcarbamate
Figure imgf000220_0001
Step 1 : tert-butyl (1 -(4-(5-nitro-6-oxo-3-phenyl-1 ,6-dihvdropyridin-2- vDphenvDcvclobutvDcarbamate
(E)-tert-butyl-1-(4-(3-(dimethylamino)phenyl)cyclobutylcarbamate (10g, 23.78mmol) and 2-nitroacetamide (2.45g, 23.78mmol) were dissolved in acetic acid (125mL) to give a yellow solution. This reaction mixture was heated to 50°C for 2 hours and then it was stirred at room temperature for 16 hours. The obtained precipitate was filtered, washed with diethyl ether and dried until constant weight to give the title compound as a yellow solid (4.1g, 37.4%). Step 2: tert-butyl (1-(4-(5-amino-6-oxo-3-phenyl-1.6-dihydropyridin-2- yl)phenyl)cyclobutyl)carbamate
Tert-butyl (1 -(4-(5-nitro-6-oxo-3-phenyl-1 ,6-dihydropyridin-2- yl)phenyl)cyclobutyl)carbamate (10.95g, 23.77mmol) was dissolved in acetic acid (250mL) at room temperature. Zinc powder (10g, 153mmol) was added at 30°C and the resulting suspension was stirred at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure and the obtained residue was partitioned between water (250mL) and ethyl acetate (250mL). The aqueous phase was extracted with fresh ethyl acetate (3 x 200mL) and the combined organic layers were washed with water (200ml_), brine (200ml_), dried on anhydrous Na2S04, filtered and concentrated to dryness under reduced pressure. The obtained crude was purified by Biotage silica gel chromatography (gradient 0% to 3% methanol in dichloromethane) to give the title compound (4.3g, 43%). It was used for the next step without further purification. LCMS (Method A) RT 5.96 min, M+1 =432. tert-butyl (1-(4-(1-methyl-7-phenyl-2-thioxo-2,3-dihvdro-1 H-pyridor2,3-b1[1,4]oxazin-
6-yl)phenyl)cvclobutyl)carbamate
Figure imgf000221_0001
Tert-butyl (1-(4-(1-methyl-7-phenyl-2-thioxo-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutyl)carbamate (200mg, 0.412mmol) was suspended in toluene (5ml). Lawesson's reagent (125mg, 0.309mmol) was added at room temperature. The resulting mixture was heated under reflux for 5 hours. After it was cooled down to room
temperature, the mixture was concentrated to dryness under reduced pressure. The resulting residue was dissolved in the minimum amount of dichloromethane (1 mL) and treated with diisopropyl ether (7ml_). A yellow solid crushed out. It was filtered and dried until constant weight (200mg, 96%). LCMS (Method A): RT = 7.08 min, M+1 = 502. 6-bromo-1 -methyl-7-phenyl-1 H-pyridor2,3-b1[1 ,4]oxazin-2(3H)-one
Figure imgf000222_0001
In a 15 mL reaction tube was added 6-bromo-7-phenyl- H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one (1.00 g, 3.28 mmol) in anhydrous N,N-dimethylformamide (5 mL), potassium carbonate (1.359 g, 9.83 mmol) and iodomethane (0.246 mL, 3.93 mmol) to give a brown suspension. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. The reaction mixture was diluted with saturated sodium bicarbonate solution (30 mL) and extracted into dichloromethane (3 x 15 mL). The combined organic phases were dried over anhydrous Na2S04, filtered and concentrated to dryness under reduced pressure to give a brown oil. The residue was purified via Biotage chromatography (silica 50 g column, cyclohexane:ethyl acetate, gradient elution from 88:12 to 0:100) to give a pale yellow solid (780 mg, 74.6 % yield). 1H NMR (500 MHz, CDCI3): 7.51-7.44 (5H, m), 7.20 (1 H. s), 4.91 (2H, s), 3.36 (3H, s).
7-bromo-1 H-pyridor2.3-biri .41oxazin-2(3H)-one
Figure imgf000222_0002
In a 500 mL round-bottomed flask was added ethyl 2-(5-bromo-3-nitropyridin-2- yloxy)acetate (5.2 g, 17.04 mmol) in hydrochloric acid, 37% (50 mL) to give a yellow suspension. The mixture was cooled to 0-5°C followed by the portion wise addition of tin powder (10.1168 g, 85 mmol). Caution must be taken during the addition as it is very exothermic. The mixture was stirred at room temperature for a further 30 minutes after the addition. The reaction mixture was heated to 80 °C under a nitrogen atmosphere for 3 hours. The reaction mixture was diluted with water (250 mL). The precipitate was filtered, washed with water (200mL) and diethyl ether (200mL) and dried until constant weight to give the title compound (3.2g, 82%). 1H NMR (500 MHz, d6-DMSO): 10.41 (2H, s), 7.88 (2H, s), 7.34 (2H, s), 4.85 (2H, s). ethyl 2-((5-bromo-3-nitropyridin-2-yl)oxy)acetate
Figure imgf000223_0001
To a suspension of sodium hydride (5.31 g, 133 mmol) in 1 ,4-dioxane (250 mL) was added drop wise ethyl glycolate (12.56 ml, 133 mmol) over a period of 30 minutes ensuring that the temperature was maintained below 30°C. The resulting thick
suspension was stirred at room temperature for 15 minutes. In a separate 1 L round- bottomed flask was added 5-bromo-2-chloro-3-nitropyridine (21 g, 88 mmol) in 1 ,4- dioxane (150 mL) to give a brown solution. The sodium hydride/ethyl glycolate slurry was added drop wise over a period of 30 minutes. The resulting reaction mixture was heated to 80 °C overnight. The reaction mixture was concentrated under reduced pressure and the residue was purified via Biotage chromatography (silica 340 g column, gradient elution from 0% to 10% ethyl acetate in cyclohexane) to give a pale yellow solid (11.8g, 43 % yield). Ή NMR (500 MHz, CDCI3): 8.48 (1 H, d), 8.42 (1 H, d), 5.07 (2H, s), 4.25 (2H, q), 1.30 (3H, s).
6-bromo-1 -ethyl-7-phenyl-1 H-pyridor2,3-b1H ,41oxazin-2(3H)-one
Figure imgf000223_0002
In a 15 mL reaction tube was added 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin- 2(3H)-one (300 mg, 0.983 mmol), iodoethane (0.095 mL, 1.180 mmol) and potassium carbonate (408 mg, 2.95 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give a brown suspension. This was stirred at 50 °C under a nitrogen atmosphere for 60 minutes. The reaction mixture was diluted with saturated sodium bicarbonate solution (5 mL) and extracted into ethyl acetate (3 x 5 mL). The combined organic phases were washed with 50:50 water.brine (3 x 5 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a brown solid. This was purified by Biotage chromatography (25g silica cartridge, cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the title compound as a beige solid (160 mg, 48.8 % yield).
1H NMR (500 MHz, CDCI3): 7.58-7.37 (5H, m), 7.21 (1H, s), 4.86 (2H, s), 3.96 (2H, q), 1.27 (3H, t). LCMS (Method D) RT 1.293 min, M+1= 334.
6-bromo-1 -(cvclopropy lmethyl)-7-phenyM H-pyrido[2,3-b]ri ,4]oxazin-2(3H)-one
Figure imgf000224_0001
In a 15 mL reaction tube was added 6-bromo-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one (300 mg, 0.983 mmol), (bromomethyl)cyclopropane (0.114 mL, 1.180 mmol) and potassium carbonate (408 mg, 2.95 mmol) in anhydrous N,N-dimethylformamide (1 mL) to give a brown suspension. This was stirred at 50°C under a nitrogen atmosphere for 60 minutes. The reaction mixture was diluted with saturated sodium bicarbonate solution (5 mL) and extracted into ethyl aceate (3 x 5 mL). The combined organic phases were washed with 50:50 watenbrine (3 x 5 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a brown solid. This was purified by Biotage chromatography (25g silica cartridge, cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the title compound as a beige solid (160 mg, 0.445 mmol, 45.3 % yield). 1H NMR (500 MHz, CDCI3): 7.58-7.37 (5H, m), 7.32 (1 H, s), 4.88 (2H, s), 3.81 (2H, d), 1.14-1.01 (1 H. m), 0.60-0.53 (2H, m), 0.47-0.40 (2H, m). tert-butyl ((1 r.3r)-1 -(4-(5.5-dimethyl-1.3.2-dioxaborinan-2-yl)phenyl)-3-hvdroxy-3- methylcvclobutvDcarbamate
Figure imgf000224_0002
Step 1 : 2-((1 r,3r)-1-(4-(5,5-dimethyl-1 ,3.2-dioxaborinan-2-yl)phenyl)-3-hvdroxy-3- methylcyclobutvOisoindoline- 1 ,3-dione
In a 100 mL round-bottomed flask was added 2-((1 r,3r)-1-(4-bromophenyl)-3-hydroxy-3- methylcyclobutyl)isoindoline-1 ,3-dione (0.710 g, 1.838 mmol), S.S.S'.S'-tetramethyl^.Z- bi(1 ,3,2-dioxaborinane) (0.700 g, 3.10 mmol) and potassium acetate (0.361 g, 3.68 mmol) in anhydrous DMSO (35.4 ml) to give a colourless suspension. This was degassed by bubbling nitrogen for 15 minutes, followed by the addition of PdCI2(dppf)- CH2CI2 adduct (0.150 g, 0.184 mmol). The resulting suspension was heated to 80 °C under a nitrogen atmosphere overnight. The reaction mixture was allowed to cool to room temperature, diluted with ethyl acetate (180 mL), filtered through celite, washed with water (2 x 180 mL) then brine (180 mL), dried over Na2S04, filtered and
concentrated to dryness under reduced pressure to give a brown oil. This was purified by Biotage chromatography (silica 50g cartridge, cyclohexane:ethyl acetate, gradient elution from 90:10 to 0.Ί 00) to give the title compound as an off-white solid (263 mg, 0.627 mmol, 34.1 % yield). 1H NMR (500 MHz, CDCI3): 7.82-7.72 (5H, m), 7.70-7.63 (4H, m), 3.73 (3H, s), 3.36 (2H, d), 3.19 (2H, d). LCMS (Method D) RT 0.890 min, M+1 = 420.
Step 2: tert-butyl ((1 r,3r)-1-(4-(5.5-dimethyl-1 ,3.2-dioxaborinan-2-yl)phenyl)-3-hvdroxy-3- methylcyclobutvDcarbamate
In a 15 mL reaction tube was added 2-((1 r,3r)-1-(4-(5,5-dimethyl-1 ,3,2-dioxaborinan-2- yl)phenyl)-3-hydroxy-3-methylcyclobutyl)isoindoline-1 ,3-dione (220 mg, 0.525 mmol) and hydrazine monohydrate (0.2 mL, 4.12 mmol) in ethanol (4 mL) to give a pale yellow solution. This was heated to 80 °C overnight giving a white suspension. The reaction mixture was filtered and the filtrates concentrated to dryness under reduced pressure. The residue was redissolved in methanol (4 mL) followed by the addition of di-tert-butyl dicarbonate (0.244 mL, 1.049 mmol) and triethylamine (0.183 mL, 1.312 mmol). The reaction mixture was heated to 50°C overnight. The reaction mixture was diluted with ethyl acetate (16 mL) and washed with water (8 mL), dried over Na2S04, filtered and concentrated to dryness under reduced pressure to give a yellow solid. This was purified by Biotage chromatography (25g silica cartridge, cyclohexane:ethyl acetate, gradient elution from 90:10 to 20:80) to give the title compound as an off-white solid (88 mg, 0.226 mmol, 43.1 % yield). 1H-NMR (500 MHz, CD3CI3) δ 7.75 (2H, m), 7.38 (2H, s), 3.76 (3H, s), 2.80-2.63 (4H, m), 2.60-2.45 (2H, m), 2.16-2.03 (2H, m), 1.49-1.29 (9H, m), 1.01 (6H, s). LCMS (Method D) RT = 0.814 min, M+1 = 390. AKT Kinase Assay testing
Testing of the compounds was performed using an AKT Kinase Assay:
Activated AKT isoforms 1 , 2 and 3 were assayed utilising a 5' FAM Crosstide (Seq. GRPRTSSFAEG-OH). The extent of kinase phosphorylation was determined by fluorescent polarisation using IMAP progressive binding reagent, which introduces binding beads which allow the reagent to specifically bind to phosphate residues via covalent co-ordination complex bonds. iMAP binding solution stops Crosstide / kinase interaction and specifically binds phosphorylated substrates. The degree of phosphorylation is determined by fluorescent polarisation (excitation 485nm; emission 528nm) or the reduction in speed of rotation of the excited substrate.
The following materials were used in the assay:
a) Activated AKT isoforms (SignalChem.) dissolved in Complete Reaction buffer at a pre-determined concentration selected so that the assay was carried out in the linear range.
b) AKT substrate peptide: FAM Crosstide (R7110) Molecular Devices, diluted in complete reaction buffer.
c) iMAP Progressive Screening Express Kit (R8127) Molecular Devices
d) Complete Reaction Buffer containing 0.1% BSA, 10mM Tris-HCI, 10mM MgCI2, 0.05% NaN3 and 0.01% phosphate free BSA, 1mM DTT
e) Progressive Binding Solution containing 75% Buffer A, 25% Buffer B and low volume Binding Reagent which contains the binding entity for the assay f) ATP diluted in complete reaction buffer
g) Black polystyrene 384 well assay plates (Nunc).
h) Biotek Synergy 4 Hybrid Plate reader.
5μΙ of test compound was dissolved in DMSO (Sigma Aldrich) and serially diluted in complete reaction buffer to give a fourteen point half log dose response and plated into 384 well black plates. The compound was incubated at room temperature with activated AKT isoform (5μΙ) at the pre-determined concentration, for 45 minutes. 2.5μΙ of ATP solution mixed with 2.5μΙ of AKT substrate peptide (FAM Crosstide (R7110) Molecular Devices) were dispensed into each well and the plate centrifuged at lOOOrpm for 20 seconds to ensure homogenous mixing of reagents. The reaction mix was incubated in the dark for one hour at room temperature.
The kinase reaction was stopped by the addition of Progressive Binding Solution and the mixture allowed to equilibrate for one hour in the dark, at room temperature.
The fluorescent polarisation generated in each well was determined using a Biomek Synergy 4 Hybrid plate reader. In brief, each reaction solution was excited at 485nm with the emission measured at 528nm in both the parallel and perpendicular pathway.
The polarisation value generated in each well was calculated by Gen5 software (Biotek) and the % inhibition of kinase activity compared to vehicle control was calculated via GraphPad Prism. IC50 values for each compound were calculated by non-linear regression analysis using Prism software.
All plates were internally controlled by two methods. Firstly, by calculating the
signaknoise ratio; based on kinase polarisation without inhibitor and polarisation generated by complete reaction buffer in the absence of activated kinase. Secondly by determining IC50 values generated by known inhibitors of the AKT isoforms.
Testing of the compounds was also performed using in vitro cell proliferation assays:
Cell Titre Glo (Promega) is a highly sensitive homogeneous reagent used to determine the viability of cells. The reagent uses a stable form of luciferase to measure ATP as an output of viability. The luminescent values generated in the assay are directly
proportional to the number of viable cells in your assay.
The following materials were used: White, clear bottomed 96 well assay plates (Costar); Cell titre Glo reagents; LnCaP (ECACC) cells grown in RPMI medium (Invitrogen) supplemented with 10mM HEPES (Invitrogen), 1mM Sodium pyruvate (Invitrogen), 2mM L-Glutamine (Invitrogen) and 10% Foetal calf serum (Invitrogen); PC3 (ECACC) cells grown in RPMI medium supplemented with 10% Foetal calf serum (Invitrogen); Trypsin (Invitrogen); PBS (Invitrogen); Biotek Synergy 4 Hybrid Plate reader; 96 well plate shaker (Stuart SSL5); Eppendorf 5414 desk-top centrifuge; Beckman Coulter cell counter Z1 single threshold system.
Prostate cell lines, PC3 and LnCaP, were washed, detached and re-suspended in their respective fresh media. The cells were pelleted by centrifugation (Eppendorf 5414 ) and spent supernatant discarded. The cells were re-suspended by vortex mixing, counted and seeded into clear bottom white 96 well plates at a density of 5000 cells per well. The cells were incubated (Sanyo) overnight at 37°C (95% 02 / 5% C02), and next day treated with increasing concentrations of test compound formulated in fresh medium. The plates were returned to the incubator for 72 hours.
Cell Titre Glo (Promega) was prepared by mixing the supplied reagents as per manufacturer's instructions and left to stand at room temperature. The cell plates were removed from the incubator and 80DI of the Cell Titre Glo solution added to each well. The plate was shaken for five minutes to ensure homogenous mixing of reagents and cells, then left to stand for ten minutes at room temperature.
The cell viability post compound treatment was determined by the luminescent intensity emitted from the drug treated wells in the plate. In brief, the assay plate was placed in the Biotex Synergy 4 Hybrid plate reader and the luminescence read in each well. The compound treated wells were compared to vehicle treated wells and the % inhibition of cell viability calculated.
The data was analysed using GraphPad Prism, with IC50 values generated using nonlinear regression of the data set.
Analysis of compound effects on AKT signalling pathways
Phosphorylation status of various members of the AKT/PI3K pathway were investigated via western blotting.
Materials Required for this Assay: 20x Running buffer (Invitrogen); Rainbow marker ladder (GE Healthcare); Reducing buffer (Invitrogen); 20x Transfer buffer (Invitrogen); 4- 12% Bis-Tris Gels (Invitrogen); Filter paper (Whatman); Nitrocellulose (Amersham); ECL plus detection reagents (GE Healthcare); Radiographic film (Kodak); Biorad Protein determination reagent (Biorad); AKT pathway signalling antibodies (Cell Signalling) LnCaP and PC3 cell lines were washed, detached and re-suspended in fresh medium. They were seeded in 90mm2 dishes and incubated overnight (95% 02 / 5% C02) to allow adherence. When the cells had reached 60% confluence, the medium was removed and replaced with compound or vehicle supplemented medium. The plates were incubated for a range of time points.
The medium was removed and the cells placed on ice and washed in PBS. 300μΙ of lysis buffer was added to the dish and left for a few minutes, before the cells were scraped into the solution and pipetted into a centrifuge tube. The tube was placed on ice for 10 minutes and then vortexed to aid cellular lysis. The sample was centrifuged (Eppendorf bench-top ultra-fuge) at 13.2k rpm for 10 minutes at 4°C. The resultant supernatant was assayed for protein content using the Bradford method (Biorad) and equal quantities of protein calculated and heated in sample reducing buffer to 95°C for ten minutes.
The samples were run on 4-12% Bis-Tris gels (Invitrogen), transferred onto
nitrocellulose membrane (Amersham) and blocked with a 5% non-fat milk solution.
The membranes were used to determine difference in total AKT, pSer473 AKT, pGSK3p and total GSK3p (all sourced from Cell Signalling~)The respective primary antibodies were diluted in 1% non-fat milk blocking solution and incubated on the membranes overnight at 4°C. The membranes were washed three times in PBS and incubated with the respective secondary antibodies (Sigma Aldrich) for two hours and the proteins were detected using ECL plus detection reagents (GE Healthcare).

Claims

CLAIMS:
1. A compound according to Formula (I):
Figure imgf000230_0001
(I)
wherein:
R and R2 are each independently selected from hydrogen, aryl, C1-C10 alkyl, CN, CHO, C02H, CONH2, CONHR3, CONR3aR3b, COR3, C02R3, NH2, NHR3, NR3aR3b, NHCOR3, NHS02R3, NR3aCOR3b, NR3aS02R3 , OH, OR3, SH, SR3, SOR3, S02R3, S02NHR3, S02NR3aR3b, F, CI, Br and I, wherein each R3, R3a and R3b is independently selected from C1-C10 alkyl, including wherein R3a and R3 are joined to one another to form a heterocycle that includes the nitrogen to which they are attached . wherein separate R1 and R2 may be joined to one another to form an optionally substituted and optionally saturated heterocycle or carbocycle that includes the C atom to which they are attached; or R1 and R2 together are oxo or optionally C1-C10 alkyl O- substituted oxime;
W is O, S, SO, S02, NR', or CRaRb where R' is either hydrogen or C1-C10 alkyl, and where Ra and R are each independently selected from the members of the group from which R1 and R2 are selected above;
X is either absent or is CR R5 where R4 and R5 are each independently selected from the members of the group from which R1 and R2 are selected above; Y and Z are independently either substituted or unsubstituted nitrogen or carbon and where carbon is substituted by substituents independently selected from the members of the group from which R1 and R2 are selected above or when nitrogen then the substituent is selected from aryl, C1-C10 alkyl, S02R3, CONHR3, CONR3aR3b, COR3 and C02R3 where R3, R3a and R3b are as defined above, or where Y and Z together form an optionally substituted heterocyclyl or carbocyclic group, or where Y is S02;
R7a and R7b are independently selected from H and alkyl, including wherein R7a and R7 are joined to one another to form a heterocycle that includes the nitrogen to which they are attached; and
R63 and R6b are independently selected from: H, (C C6)alkyl, (d-C6)alkenyl, and (C
C6)alkynyl, wherein said alkyl is optionally substituted with up to three substituents selected from: OH and halo; or R^and R6 can be taken together to form a monocyclic or bicyclic carbo- or heterocycle with 3-7 members in each ring, said heterocycle having one or more heteroatoms selected from N, O and S, and said carbo- or heterocycle is optionally substituted with one or more substituents selected from: (Ci-C6)alkyl, (C3-C6)cycloalkyl, (C C6)alkoxy, C02H, halo, OH, CN and NR3aR3b, said alkyl, cycloalkyl and alkoxy is optionally substituted with one or more substituents selected from halo, CN, OH and NR3aR3b; and ring Cy is selected from (C3 to C8)cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaryl and aryl, wherein m is 0, 1 , 2, 3, 4 or 5, and each R8 is independently selected from alkyl, CN, CHO, C02H, CONH2, CONHR9, CONHR9aR9b, COR9, C02R9, NH2, NHR9, NR9aR9b, NHCOR9, NHS02R9, NR9aCOR9 , NR9aS02R9b, OH, OR9, SH, SR9, F, CI, Br and I, wherein each R9, R9a and R9b is independently selected from alkyl, including wherein R9a and R9b form a heterocycle that includes the nitrogen to which they are attached or Cy may be iodine; or a compound selected from the group:
Figure imgf000232_0001
and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
2. The compound of claim 1 , wherein X is absent and is W is CH2 or O.
3. The compound of claim 1 or claim 2, wherein R63 and R6b together form
Figure imgf000232_0002
that is they preferably form cyclobutane.
4. The compound of claim 3, wherein the group bound to the phenyl ring in structure I is 1-aminocyclobutyl.
5. The compound of any one of the preceding claims, wherein the bond between Y and Z is a single bond.
6 The compound of claim 2 where Y is carbonyl and Z is optionally substituted amino, or Z is carbonyl and Y is optionally substituted amino.
7. The compound of claim 6 wherein W is O and R and R2 are hydrogen.
8. The compound of claim 6 or claim 7 wherein, when amino is substituted, it is preferably methyl or acetamido substituted.
9 The compound of any preceding claim wherein the substituents bound to Y and Z together with Y and Z themselves form a 5 or 6 membered optionally substituted carbocyclic or heterocyclic ring.
10. The compound of any one of the preceding claims wherein the substituents bound to Y and Z together with Y and Z themselves form a moiety selected from the group:
Figure imgf000233_0001
11. The compound of any one of the preceding claims, wherein Cy is phenyl and m is 0.
12. The compound of claim 1 being:
2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin-5(6H)-one
2-(4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin-5(6H)-one oxime
2-(4-(1 -aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydroquinolin-5(6H)-one O-methyl oxime
1-(4-(8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7-yl)phenyl)cyclobutanamine
1-(4-(2-methyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
1 -(4-(1 -methyl-8-phenyl-4,5-dihydro-1 H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
7-(4-(1-aminocyclobutyl)phenyl)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-3-ol
7-(4-(1-aminocyclobutyl)phenyl)-3a-methyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-3(3aH)-one
1-(4-(3-(methylthio)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutanamine 1 - (4-(3-(methylsulfonyl)-8-phenyl-4,5-dihydro-2H-pyra2olo[3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
7-(4-(1 -aminocyclobutyl)phenyl)-N-methyl-8-p enyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-3-amine
2- ((7-(4-(1 -aminocyclobutyl)phenyl)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-3- yl)amino)ethanol
1-(4-(3-morpholino-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
7- (4-(1 -aminocyclobutyl)phenyl)-2-methyl-8-phenyl-4,5-dihydro-3H-imidazo[4,5-f]quinolin-
3- ol
1 -(4-(2-methyl-8-phenyl-4,5-dihydro-3H-imidazo[4,5-f]quinolin-7- yl)phenyl)cyclobutanamine
1 - (4-(9-phenyl-5,6-dihydropyrido[2,3-h]quinazolin-8-yl)phenyl)-cyclobutanamine
8- (4-(9-phenyl-5,6-dihydropyrido[2,3-h]quinazolin-2-amine
2- (4-(1 -aminocyclobutyl)phenyl)-7,7-dimethyl-3-phenyl-7,8-dihydroquinolin-5(6H)-one
1 - (4-(4,4-dimethyl-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-7- yl)phenyl)cyclobutanamine
2- (4-(1 -aminocyclobutyl)phenyl)-3-phenyl-6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-5- one
l^-ig-phenyl^^.S.e-tetrahydropyrazolotS'^'iS^lcycloheptafl ^-blpyridin-e- yl)phenyl)cyclobutanamine
2-(4-(1 -aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydro-1 ,6-naphthyridin-5(6H)-one
2-(4-(1 -aminocyclobutyl)phenyl)-6-methyl-3-phenyl-7,8-dihydro-1 ,6-naphthyridin-5(6H)- one
2-(2-(4-(1 -aminocyclobutyl)phenyl)-5-oxo-3-phenyl-7,8-dihydro-1 ,6-naphthyridin-6(5H)- yl)acetonitrile
2-(4-(1 -aminocyclobutyl)phenyl)-6-(2-dimethylamino)ethyl)-3-phenyl-7,8-dihydro-1 ,6- naphthyridin-5(6H)-one
8-(4-(1 -aminocyclobutyl)phenyl)-9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4- f][1 ,6]naphthyridin-3(2H)-one
l^-ie-ethyl-S-phenyl-S.ej.e-tetrahydro-I.e-naphthyridin^-y pheny cyclobutanamine
1 -(4-(9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4-f][1 ,6]naphthyridin-8- yl)phenyl)cyclobutanamine
1-(4-(3-(1 -methyl-1 H-imidazol-4-yl)-9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4- f][1 ,6]naphthyridin-8-yl)phenyl)cyclobutanamine
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one 6-(4-(1-aminocyclobutyl)phenyl)-1-methyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(prop-2-yn-1-yl)-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-1-yl)acetonitrile
6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-(dimethylamino)ethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-1 -yl)acetamide
6- (4-(1-aminocyclobutyl)phenyl)-1-(2-oxopropyl)-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin- 2(3H)-one
1 -(4-(7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6-yl)phenyl)cyclobutanamine
7- (4-(1-arninocyclobutyl)phenyl)-8-phenyl-2l4-dihydro-1 H-pyrido[2,3-b][1 ,2I4]triazolo[4,3- d][1 ,4]oxazin-1-one
1 -(4-(8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
1 -(4-(1 -methyl-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutanamine
1 -(6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1 - yl)ethanone
1- ((1H-imidazol-2-yl)methyl)-6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3- b][1,4]oxazin-2(3H)-one
2- (6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1- yl)acetonitrile
2-(6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1- yl)acetamide
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl- 1 -(pyridin-4-ylmethyl)- 1 H-pyrido[2, 3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(pyridin-3-ylmethyl)-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
1-(4-(1-ethyl-8-phenyl-4H-pyrido[2I3-b][1 ,2l4]triazolo[4I3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
6-(4-(1-aminocyclobutyl)phenyl)-1-(cyclobutylmethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one ethyl 2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-1-yl)acetate
7-(4-(1-aminocyclobutyl)phenyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-1 -amine
6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-methoxyethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-1-ethyl-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin-2(3H)-one
1- ((1 H-imidazol-5-yl)methyl)-6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
2- (6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-1-yl)propanenitrile
6-(4-(1 -aminocyclobutyl)phenyl)-1 -(cyclopropylmethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(pyridin-2-ylmethyl)-1H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-1 -isopropyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one
6-(4-(1-aminocyclobutyl)phenyl)-1-cyclopentyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(thiazol-4-ylmethyl)-1H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 -(2,2,2-trifluoroethyl)-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-1 -cyclobutyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one
3- (6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-1-yl)propanenitrile
1-(4-(1-methyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
4- (6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-1-yl)butanenitrile
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(tetrahydro-2H-pyran-4-yl)-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one 5- (6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-1 -yl)pentanenitrile
1-allyl-6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6- (4-(1-aminocyclobutyl)phenyl)-1-(2-hydroxyethyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
1- (4-(8-phenyl-1-propyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
1 -(4-(1 -(methylsulfonyl)-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutanamine
6-(4-(1 -aminocyclobutyl)phenyl)-3-(2-hydroxyethyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-4-methyl-7-phenyl-3,4-dihydropyrido[2,3-b]pyrazin- 2(1 H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-1 -methyl-7-phenyl-1 H-pyrido[3,2-e][1 ,3,4]oxathiazine 2,2-dioxide
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 -(pyridin-3-yl)-1 H-pyrido[3,2- e][1 ,3,4]oxathiazine 2,2-dioxide
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-(pyridin-3-yl)-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
2- (6-(4-(1-aminocyclobutyl)phenyl)-2,2-dioxido-7-phenyl-1H-pyrido[3,2- e][1 ,3,4]oxathiazin-1 -yl)acetonitrile
6-(4-(1 -aminocyclobutyl)phenyl)-3-(2-hydroxyethyl)-1 -methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
2-(6-(4-(1-aminocyclobutyl)phenyl)-3-(2-hydroxyethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-1-yl)acetonitrile
6-(4-(1-aminocyclobutyl)phenyl)-3-methyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one
2-(6-(4-(1-aminocyclobutyl)phenyl)-3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-1 -yl)acetonitrile
6-(4-(1-aminocyclobutyl)phenyl)-1-(methylsulfonylmethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-1-(1-methyl-1 H-pyrazol-4-yl)-7-phenyl-1H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one e-i^il-aminocyclobuty pheny -I .S-dimethyl^-phenyl-I H-pyrido^.S-blll ^Joxazin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-1 -(2,2-difluoroethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-fluoroethyl)-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-3,4-dihydropyrido[3,2-d]pyrimidin-2(1 H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-N-ethyl-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazine-1 -carboxamide
6-(4-(1 -aminocyclobutyl)phenyl)-1 ,3-dimethyl-7-phenyl-3,4-dihydropyrido[3,2-d]pyrimidi 2(1 H)-one
6-(4-(1 -Aminocyclobutyl)phenyl)-1 -methyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one O-methyl oxime
1 - (4-(7-phenyl-1-(pyridin-3-yl)-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-6- yl)phenyl)cyclobutanamine
1 -(4-(1 -bromo-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
(R)-2-(6-(4-(1-aminocyclobutyl)phenyl)-3-methyl-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-1 -yl)acetonitrile
2- (6-(4-(1-aminocyclobutyl)phenyl)-3-(methoxymethyl)-2-oxo-7-phenyl-2,3-dihydro-1 H- pyrido[2,3-b][1 ,4]oxazin-1-yl)acetonitrile
6-(4-(1 -aminocyclobutyl)phenyl)-3-(methoxymethyl)-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6- (4-((1 s,3s)-1 -amino-3-hydroxycyclobutyl)phenyl)-1 -methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
1-(4-(1 -(difluoromethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cylobutanamine
7- (4-(1-aminocyclobutyl)phenyl)-2-methyl-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-1 -one
1 - (4-(8-phenyl-1-(trifluoromethyl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin^ yl)phenyl)cyclobutanamine
2- (7-(4-(1-aminocyclobutyl)phenyl)-1-oxo-8-phenyl-1 H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-2(4H)-yl)acetonitrile 6- (4-(1 -aminocyclobutyl)phenyl)-1 -isobutyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one
7- (4-(1-aminocyclobutyl)phenyl)-8-phenyl-2-(2,2,2-trifluoroethyl)-2^-dihydro-1 H- pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-1-one
7-(4-(1-aminocyclobutyl)phenyl)-8-phenyl-4,5-dihydro-1 H-pyrido[2,3-b][1 ,4]diazepin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-7-phenyl-1-propyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6- (4-(1-aminocyclobutyl)phenyl)-1-(3-hydroxy-2-(hydroxymethyl)-2-methylpropyl)-7- phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
7- (4-(1-Aminocyclobutyl)phenyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-1-yl)phenyl)methanol
6-(4-(1-aminocyclobutyl)phenyl)-1-methyl-7-phenyl-3,4-dihydropyrido[3,2-d]pyrimidin- 2(1 H)-one
6- (4-(1-aminocyclobutyl)phenyl)-1 ,3-bis(2-fluoroethyl)-7-phenyl-3,4-dihydropyrido[3,2- d]pyrimidin-2(1 H)-one
1-(4-(1-cyclopropyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
7- (4-(1 -aminocyclobutyl)phenyl)-1 ,5-dimethyl-8-phenyl-4,5-dihydro-1 H-pyrido[2,3- b][1 ,4]diazepin-2(3H)-one
1-(4-(1-isopropyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
6- (4-((1 r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1-rnethyl-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
7- (4-(1 -aminocyclobutyl)phenyl)-1 -methyl-8-phenyl-4,5-dihydro-1 H-pyrido[2,3- b][1 ,4]diazepin-2(3H)-one
1 -(4-(1 -isobutyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
7-(4-(1-aminocyclobutyl)phenyl)-N,N-dimethyl-8-phenyl-4H-pyrido[2,3- b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-1 -amine
7-(4-(1-aminocyclobutyl)phenyl)-1-(2-fluoroethyl)-8-phenyl-4,5-dihydro-1 H-pyrido[2,3- b][1 ,4]diazepin-2(3H)-one
1 -(4-(8-phenyl-1 -(2,2,2-trifluoroethyl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine 7-(4-(1-aminocyclobutyl)phenyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazine-1 -carboxamide
1-(4-(1-(fluoromethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
1-(4-(1-(2-methoxyethyl)-8-phenyl-4H-pyrido[2l3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
1-(4-(8-phenyl-1-(pyridin-3-yl)-4H-pyrido[2)3-b][1 )2,4]triazolo[4l3-d)[1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
1- (4-(8-phenyl-1-(pyridin-4-yl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
6-(4-(1-aminocyclobutyl)phenyl)-1-methyl-7-(pyridin-3-yl)-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-1-methyl-7-(thiophen-3-yl)-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-1 -methyl-7-(thiophen-2-yl)-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
2- (4-(1-aminocyclobutyl)phenyl)-3-phenyl-6,8-dihydro-5H-pyrano[3,4-b]pyridin-5-one (1r,3r)-3-amino-3-(4-(1-(difluoromethyl)-8-phenyl-4H-pyndo[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)-1 -methylcyclobutanol
1 -(4-(1 ,8-diphenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
1- (4-(8-phenyl-1-(pyridin-2-yl)-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
6-(4-(aminomethyl)phenyl)-1-methyl-7-phenyl-1 H-pyrido[2,3-b][1,4]oxazin-2(3H)-one 6-(4-(1 -aminocyclobutyl)phenyl)-7-(4-fluorophenyl)-1 -methyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-7-(cyclopropylmethyl)-1 -methyl-1 H-pyrido[2, 3- b][1 ,4]oxazin-2(3H)-one
6-(4-((1s,3s)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1-methyl-7-phenyl-1H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
2- (6-(4-(aminomethyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1- yl)acetonitrile
(1 s,3s)-3-Amino-3-(4-(1 -ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanol 6-(4-((1r,3r)-1-amino-3-hydroxycyclobutyl)phenyl)-1-methyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
(1 r,3r)-3-amino-3-(4-(1-ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)-1 -methylcyclobutanol
6-(4-((1s,3s)-1-amino-3-hydroxycyclobutyl)phenyl)-1-ethyl-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6- (4-((1 s,3s)-1 -amino-3-hydroxycyclobutyl)phenyl)-1 -(cyclopropylmethyl)-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
7- (4-((1 s,3s)-1 -amino-3-hydroxycyclobutyl)phenyl)-8-phenyl-2,4-dihydro-1 H-pyrido[2,3- b][1,2,4]triazo)o[4,3-d][1,4]oxazin-1-one
(4-(1-ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)methanamine
6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1-ethyl-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1-(cyclopropylmethyl)-7- phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1-(2-fluoroethyl)-7-phenyl- 1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-(Aminomethyl)phenyl)-7-phenyl-1-(2,2,2-trifluoroethyl)-1H-pyrido[2,3-b][1 ,4]oxazi 2(3H)-one
2-(6-(4-((1s,3s)-1-amino-3-hydroxycyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H- pyrido[2,3-b][1 ,4]oxazin-1 -yl)acetonitrile
6-(4-((1s,3s)-1-amino-3-hydroxycyclobutyl)phenyl)-7-phenyl-1-(2,2,2-trifluoroethyl)-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclopropyl)phenyl)-1 -methyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)- one
2-(6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-2-oxo-7-phenyl-2,3- dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)acetonitrile
6-(4-(Aminomethyl)phenyl)-1-ethyl-7-phenyl-1H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one 6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-7-phenyl-1 -(2,2,2- trifluoroethyl)-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-(Aminomethyl)phenyl)-1-(cyclopropylmethyl)-7-phenyl-1H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one 7-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-8-phenyl-2,4-dihydro-1 H- pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-1 -one and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
13. The compound of claim 1 being:
1- (4-(8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-flquinolin-7- yl)phenyl)cyclobutanamine
7- (4-(1-aminocyclobutyl)phenyl)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4-f]quinolin-
3-ol
2- ((7-(4-(1-aminocyclobutyl)phenyl)-8-phenyl-4,5-dihydro-2H-pyrazolo[3,4- f]quinolin-3-yl)amino)ethanol
1- (4-(9-phenyl-5,6-dihydropyrido[2,3-h]quinazolin-8-yl)phenyl)-cyclobutanamine
2- (4-(1-aminocyclobutyl)phenyl)-3-phenyl-7,8-dihydro-1 ,6-naphthyridin-5(6H)-one
8- (4-(1-aminocyclobutyl)phenyl)-9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4- f][1 ,6]naphthyridin-3(2H)-one
1 -(4-(3-(1 -methyl-1 H-imidazol-4-yl)-9-phenyl-5,6-dihydro-[1 ,2,4]triazolo[3,4- f][1 ,6]naphthyridin-8-yl)phenyl)cyclobutanamine
6-(4-(1-aminocyclobutyl)phenyl)-1-methyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1H-pyrido[2,3- b][1 ,4]oxazin-1-yl)acetonitrile and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
14. The compound of claim 1 being:
2-(6-(4-(1-aminocyclobutyl)phenyl)-2-oxo-7-phenyl-2,3-dihydro-1 H-pyrido[2,3- b][1 ,4]oxazin-1-yl)acetonitrile and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
15. The compound of claim 1 being:
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 -(prop-2-yn-1 -yl)-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
1 -(4-(1 -ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine 6-(4-(1 -aminocyclobutyl)phenyl)-1 -ethyl-7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin- 2(3H)-one
6-(4-(1-aminocyclobutyl)phenyl)-1-(cyclopropylmethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-7-phenyl-1 -(2,2,2-trifluoroethyl)-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
6-(4-(1 -aminocyclobutyl)phenyl)-1 -(2-fluoroethyl)-7-phenyl-1 H-pyrido[2,3- b][1 ,4]oxazin-2(3H)-one
l^-il -idifluoromethylJ-S-phenyl^H-pyrido^.S-blll ^.^triazolo^.S-dltl .^oxazin- 7-yl)phenyl)cylobutanamine
1- (4-(1-(fluoromethyl)-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3-d][1 ,4]oxazin-7- yl)phenyl)cyclobutanamine
6-(4-((1 r,3r)-1-amino-3-hydroxycyclobutyl)phenyl)-1-methyl-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
(1 rl3r)-3-amino-3-(4-(1-ethyl-8-phenyl-4H-pyrido[2,3-b][1 ,2,4]triazolo[4,3- d][1 ,4]oxazin-7-yl)phenyl)-1 -methylcyclobutanol
6-(4-((1 r,3r)-1 -amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1 -ethyl-7-phenyl-1 H- pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-((1r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-1-(cyclopropylmethyl)- 7-phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
6-(4-((1r,3r)-1-amino-3- ydroxy-3-iTiethylcyclobutyl)phenyl)-1-(2-fluoroethyl)-7- phenyl-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one
2- (6-(4-((1 r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-2-oxo-7-phenyl- 2,3-dihydro-1 H-pyrido[2,3-b][1 ,4]oxazin-1-yl)acetonitrile
6-(4-((1 r,3r)-1-amino-3-hydroxy-3-methylcyclobutyl)phenyl)-7-phenyl-1-(2,2,2- trifluoroethyl)-1 H-pyrido[2,3-b][1 ,4]oxazin-2(3H)-one and pharmaceutically acceptable salts, stereoisomers and tautomers thereof.
16. A pharmaceutical composition comprising a pharmaceutical carrier and, dispersed therein, a compound of any one of the previous claims.
17. The compound of any one of claims 1 to 15 for use in therapy.
18. The compound of any one of claims 1 to 15 for use in the treatment or prevention of cancer.
19. A method of treating or preventing cancer comprising administering a compound according to any one of claims 1 to 15 to a subject in need thereof, optionally with radiotherapy.
PCT/GB2010/002329 2009-12-23 2010-12-23 Inhibitors of akt activity WO2011077098A1 (en)

Priority Applications (18)

Application Number Priority Date Filing Date Title
JP2012545434A JP5923043B2 (en) 2009-12-23 2010-12-23 Inhibitor of AKT activity
PL10800980T PL2516435T3 (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
CA2783340A CA2783340C (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
ES10800980.4T ES2503794T3 (en) 2009-12-23 2010-12-23 Akt activity inhibitors
EP10800980.4A EP2516435B8 (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
KR1020127019033A KR20140015157A (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
AU2010334591A AU2010334591C1 (en) 2009-12-23 2010-12-23 Inhibitors of AKT activity
BRBR112012015677-2A BR112012015677A2 (en) 2009-12-23 2010-12-23 Pharmaceutical compounds.
DK10800980.4T DK2516435T3 (en) 2009-12-23 2010-12-23 Inhibitors of AKT activity
SG2012041232A SG181517A1 (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
NZ600453A NZ600453A (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
MX2012007274A MX2012007274A (en) 2009-12-23 2010-12-23 Inhibitors of akt activity.
RU2012131115/04A RU2579513C2 (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
CN201080059321.9A CN102791708B (en) 2009-12-23 2010-12-23 The inhibitor of AKT activity
US13/517,219 US9221838B2 (en) 2009-12-23 2010-12-23 Inhibitors of AKT activity
SI201030774T SI2516435T1 (en) 2009-12-23 2010-12-23 Inhibitors of akt activity
IL220153A IL220153A (en) 2009-12-23 2012-06-04 Inhibitors of akt activity
HK12111698.8A HK1171013A1 (en) 2009-12-23 2012-11-16 Inhibitors of akt activity akt

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0922589.7 2009-12-23
GBGB0922589.7A GB0922589D0 (en) 2009-12-23 2009-12-23 Pharmaceutical compounds

Publications (2)

Publication Number Publication Date
WO2011077098A1 true WO2011077098A1 (en) 2011-06-30
WO2011077098A9 WO2011077098A9 (en) 2012-04-19

Family

ID=41716932

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2010/002329 WO2011077098A1 (en) 2009-12-23 2010-12-23 Inhibitors of akt activity

Country Status (22)

Country Link
US (1) US9221838B2 (en)
EP (1) EP2516435B8 (en)
JP (1) JP5923043B2 (en)
KR (1) KR20140015157A (en)
CN (1) CN102791708B (en)
AU (1) AU2010334591C1 (en)
BR (1) BR112012015677A2 (en)
CA (1) CA2783340C (en)
CY (1) CY1115686T1 (en)
DK (1) DK2516435T3 (en)
ES (1) ES2503794T3 (en)
GB (1) GB0922589D0 (en)
HK (1) HK1171013A1 (en)
IL (1) IL220153A (en)
MX (1) MX2012007274A (en)
NZ (1) NZ600453A (en)
PL (1) PL2516435T3 (en)
PT (1) PT2516435E (en)
RU (1) RU2579513C2 (en)
SG (1) SG181517A1 (en)
SI (1) SI2516435T1 (en)
WO (1) WO2011077098A1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013140189A1 (en) 2012-03-23 2013-09-26 Almac Discovery Limited 6- (4- (1 -amino- 3 -hydroxycyclobutyl) phenyl) - 5 - phenyl (furo, thieno or pyrrolo) [2, 3-d] pyrimidin- 4 - one derivatives for the treatment of cancer
EP3493818A4 (en) * 2016-08-07 2020-04-29 The Wistar Institute Of Anatomy And Biology Methods of detecting and treating a tumor expressing pt346 pdk1
WO2022069552A1 (en) 2020-09-30 2022-04-07 Vaderis Therapeutics Ag Allosteric akt inhibitors for use in the treatment of hereditary hemorrhagic telangiectasia

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2971763A1 (en) * 2014-12-23 2016-06-30 Bergenbio Asa Inhibitors of akt kinase
US10323044B2 (en) * 2015-02-27 2019-06-18 Taiho Pharmaceutical Co., Ltd. Crystal of imidazo-oxazine, pharmaceutical composition containing said crystal, and method for producing said crystal
GB201603779D0 (en) 2016-03-04 2016-04-20 Mission Therapeutics Ltd Novel compounds
UA125730C2 (en) * 2017-06-22 2022-05-25 Курадев Фарма Лімітед Small molecule modulators of human sting
JP2020533317A (en) 2017-09-07 2020-11-19 オーガスタ ユニバーシティ リサーチ インスティテュート，インコーポレーテッド Specific AKT3 activator and its use

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006065601A2 (en) * 2004-12-15 2006-06-22 Merck & Co., Inc. Inhibitors of akt activity
WO2008070041A2 (en) 2006-12-06 2008-06-12 Merck & Co., Inc. Inhibitors of akt activity
WO2008070134A1 (en) 2006-12-06 2008-06-12 Merck & Co., Inc. Inhibitors of akt activity
WO2009148916A1 (en) 2008-06-03 2009-12-10 Merck & Co., Inc. Inhibitors of akt activity
WO2009148887A1 (en) 2008-06-03 2009-12-10 Merck & Co., Inc. Inhibitors of akt activity

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1085666C (en) * 1994-11-14 2002-05-29 沃尼尔·朗伯公司 6-aryl pyrido (2,3-d) pyrimidines and naphthyridines for inhibiting protein tyrosine kinase medicated cellular proliferation
TWI301760B (en) * 2004-02-27 2008-10-11 Merz Pharma Gmbh & Co Kgaa Tetrahydroquinolinones and their use as antagonists of metabotropic glutamate receptors
US8008317B2 (en) * 2005-06-10 2011-08-30 Merck Sharp & Dohme Corp. Inhibitors of akt activtiy
WO2008011032A1 (en) * 2006-07-17 2008-01-24 Amgen Inc. Quinazoline and pyridopyrimidine derivatives as p38 kinase inhibitors
US7999107B2 (en) * 2007-01-31 2011-08-16 Merck Sharp & Dohme Corp. Substituted pyrano[2,3-B]pyridine derivatives as cannabinoid-1 receptor modulators

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006065601A2 (en) * 2004-12-15 2006-06-22 Merck & Co., Inc. Inhibitors of akt activity
WO2008070041A2 (en) 2006-12-06 2008-06-12 Merck & Co., Inc. Inhibitors of akt activity
WO2008070134A1 (en) 2006-12-06 2008-06-12 Merck & Co., Inc. Inhibitors of akt activity
WO2008070016A2 (en) 2006-12-06 2008-06-12 Merck & Co., Inc. Inhibitors of akt activity
WO2009148916A1 (en) 2008-06-03 2009-12-10 Merck & Co., Inc. Inhibitors of akt activity
WO2009148887A1 (en) 2008-06-03 2009-12-10 Merck & Co., Inc. Inhibitors of akt activity

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
WU Z ET AL: "Rapid assembly of diverse and potent allosteric Akt inhibitors", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, PERGAMON, ELSEVIER SCIENCE, GB, vol. 18, no. 6, 15 March 2008 (2008-03-15), pages 2211 - 2214, XP025695048, ISSN: 0960-894X, [retrieved on 20071017], DOI: DOI:10.1016/J.BMCL.2007.10.023 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013140189A1 (en) 2012-03-23 2013-09-26 Almac Discovery Limited 6- (4- (1 -amino- 3 -hydroxycyclobutyl) phenyl) - 5 - phenyl (furo, thieno or pyrrolo) [2, 3-d] pyrimidin- 4 - one derivatives for the treatment of cancer
CN104245705A (en) * 2012-03-23 2014-12-24 阿尔麦克探索有限公司 6- (4- (1 -amino- 3 -hydroxycyclobutyl) phenyl) - 5 - phenyl (furo, thieno or pyrrolo) [2, 3-d] pyrimidin- 4 - one derivatives for the treatment of cancer
JP2015510923A (en) * 2012-03-23 2015-04-13 アルマック・ディスカバリー・リミテッドＡｌｍａｃ Ｄｉｓｃｏｖｅｒｙ Ｌｉｍｉｔｅｄ 6- (4- (1-Amino-3-hydroxycyclobutyl) phenyl) -5-phenyl- (furo, thieno or pyrrolo) [2,3-d] pyrimidin-4-one derivatives for the treatment of cancer
US9156853B2 (en) 2012-03-23 2015-10-13 Almac Discovery Limited AKT inhibitor compounds for treatment of cancer
EP3493818A4 (en) * 2016-08-07 2020-04-29 The Wistar Institute Of Anatomy And Biology Methods of detecting and treating a tumor expressing pt346 pdk1
US11208478B2 (en) 2016-08-07 2021-12-28 The Wistar Institute Of Anatomy And Biology Methods of detecting and treating a tumor expressing pT346 PDK1
WO2022069552A1 (en) 2020-09-30 2022-04-07 Vaderis Therapeutics Ag Allosteric akt inhibitors for use in the treatment of hereditary hemorrhagic telangiectasia
GB2600384A (en) * 2020-09-30 2022-05-04 Vaderis Therapeutics Ag Allosteric AKT inhibitors for use in the treatment of hereditary hemorrhagic telangiectasia

Also Published As

Publication number Publication date
JP2013515710A (en) 2013-05-09
ES2503794T3 (en) 2014-10-07
CN102791708B (en) 2016-03-02
RU2579513C2 (en) 2016-04-10
EP2516435A1 (en) 2012-10-31
CA2783340C (en) 2015-11-24
EP2516435B8 (en) 2014-10-15
NZ600453A (en) 2013-10-25
GB0922589D0 (en) 2010-02-10
BR112012015677A2 (en) 2015-06-30
PT2516435E (en) 2014-11-13
HK1171013A1 (en) 2013-03-15
CN102791708A (en) 2012-11-21
AU2010334591C1 (en) 2014-07-31
AU2010334591B2 (en) 2014-04-10
EP2516435B1 (en) 2014-08-06
AU2010334591A1 (en) 2012-06-21
KR20140015157A (en) 2014-02-06
IL220153A (en) 2015-05-31
WO2011077098A9 (en) 2012-04-19
US20130116243A1 (en) 2013-05-09
MX2012007274A (en) 2012-10-03
CY1115686T1 (en) 2017-01-25
US9221838B2 (en) 2015-12-29
SI2516435T1 (en) 2014-12-31
CA2783340A1 (en) 2011-06-30
RU2012131115A (en) 2014-01-27
JP5923043B2 (en) 2016-05-24
DK2516435T3 (en) 2014-10-13
SG181517A1 (en) 2012-07-30
IL220153A0 (en) 2012-07-31
PL2516435T3 (en) 2015-01-30

Similar Documents

Publication Publication Date Title
EP2516435B1 (en) Inhibitors of akt activity
US8546376B2 (en) Pharmaceutical compounds
AU2015203480B2 (en) Certain Triazolopyridines and Triazolopyrazines, Compositions Thereof and Methods of Use Therefor
AU2014241152B2 (en) Benzimidazolone derivatives as bromodomain inhibitors
US20120309739A1 (en) Akt / pkb inhibitors
CN106928219B (en) Nitrogen-containing fused heterocyclic compound, preparation method, intermediate, composition and application
WO2016007736A1 (en) Imidazopyrazines as lsd1 inhibitors
AU2016243939A1 (en) Heterocyclic compounds as LSD1 inhibitors
WO2012019426A1 (en) Phthalazinone derivative, and preparation method and pharmaceutical use thereof
US9156853B2 (en) AKT inhibitor compounds for treatment of cancer
TW200936141A (en) Substituted heteroarylamide oxazepinopyrimidone derivatives
WO2018121766A1 (en) Nitrogen-containing fused heterocyclic compound, preparation method therefor, and intermediate, composition, and application thereof
WO2021032004A9 (en) Azaheteroaryl compound and application thereof
CN114907350A (en) Nitrogen-containing condensed ring compounds, preparation method and application
EP4055013A1 (en) Wdr5 inhibitors and modulators

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 201080059321.9

Country of ref document: CN

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

Ref document number: 10800980

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 220153

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: 2010334591

Country of ref document: AU

ENP Entry into the national phase

Ref document number: 2783340

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2010334591

Country of ref document: AU

Date of ref document: 20101223

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: MX/A/2012/007274

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 2012545434

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 20127019033

Country of ref document: KR

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 6394/CHENP/2012

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2012131115

Country of ref document: RU

Ref document number: 2010800980

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 13517219

Country of ref document: US

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112012015677

Country of ref document: BR

REG Reference to national code

Ref country code: BR

Ref legal event code: B01E

Ref document number: 112012015677

Country of ref document: BR

Free format text: COM BASE NA RESOLUCAO 228/09, SOLICITA-SE QUE SEJAM APRESENTADOS NOVOS CDS/DVDS, COM AS RESPECTIVAS DECLARACOES E CODIGO ALFANUMERICO, POIS O TITULO DA LISTAGEM DE SEQUENCIA NAO ESTA EM LINGUA VERNACULA.

ENP Entry into the national phase

Ref document number: 112012015677

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20120625