WO2023235925A1 - Compounds with mertk activity - Google Patents

Compounds with mertk activity Download PDF

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WO2023235925A1
WO2023235925A1 PCT/AU2023/050497 AU2023050497W WO2023235925A1 WO 2023235925 A1 WO2023235925 A1 WO 2023235925A1 AU 2023050497 W AU2023050497 W AU 2023050497W WO 2023235925 A1 WO2023235925 A1 WO 2023235925A1
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Prior art keywords
optionally substituted
group
compound
alkyl
amino
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PCT/AU2023/050497
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French (fr)
Inventor
Jonathan Baell
Ramesh MUDUDUDDLA
Siu Wai WONG
Lian XUE
Uwe Ackerman
Lucy VIVASH
Trevor Kilpatrick
Michele BINDER
Ylva Elisabet Bergman BOZIKIS
Original Assignee
Monash University
The Florey Institute Of Neuroscience And Mental Health
Olivia Newton-John Cancer Research Institute
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Priority claimed from AU2022901554A external-priority patent/AU2022901554A0/en
Application filed by Monash University, The Florey Institute Of Neuroscience And Mental Health, Olivia Newton-John Cancer Research Institute filed Critical Monash University
Publication of WO2023235925A1 publication Critical patent/WO2023235925A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/041—Heterocyclic compounds
    • A61K51/044—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
    • A61K51/0459—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with two nitrogen atoms as the only ring hetero atoms, e.g. piperazine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/041—Heterocyclic compounds
    • A61K51/044—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
    • A61K51/0463—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/48—Two nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled

Definitions

  • the present invention relates to compounds with activity as inhibitors of MERTK.
  • the present invention also relates to radio-labelled compounds which may be used in diagnostic and imaging compositions.
  • the present invention also relates to methods and uses of the compounds in imaging body tissue, as well as identifying and diagnosing disease states.
  • the present invention also relates to methods and uses of the compounds in the treatment of diseases, particularly conditions and diseases associated with MERTK.
  • MERTK a receptor tyrosine kinase of the TAM (TYRO3, AXL, and MERTK) family
  • MERTK upregulation is associated with M2 polarization of microglia, which plays a vital role in neuroregeneration following damage induced by neuroinflammatory diseases such as multiple sclerosis (MS).
  • MS multiple sclerosis
  • MERTK is also over-expressed in a wide variety of cancers (for example, acute lymphoblastic leukemia (ALL), acute myeloid leukaemia (AML), melanoma, breast cancer, colon cancer, prostate cancer, non-small cell lung cancer (NSCLC), glioblastoma, and others).
  • ALL acute lymphoblastic leukemia
  • AML acute myeloid leukaemia
  • NSCLC non-small cell lung cancer
  • MS Multiple sclerosis
  • MS is the most common immune-mediated disorder affecting the central nervous system, characterized by demyelination of neurons in the brain and spinal cord. This damage disrupts the ability of parts of the nervous system to transmit signals, resulting in a range of signs and symptoms, including physical, mental, and sometimes psychiatric problems.
  • Treatments attempt to improve function after an attack and prevent new attacks. Medications used to treat MS, while modestly effective, can have side effects and be poorly tolerated.
  • about 2.3 million people were affected globally and in the same year, about 18,900 people died from MS. The disease usually begins between the ages of twenty and fifty and is twice as common in women as in men.
  • MERTK is highly expressed in the M2-subclass microglia, which is associated with the reparative mechanisms in response to tissue injury in the brain. It has also been shown that the MERTK gene can significantly impact MS susceptibility and severity.
  • Positron Emission Tomography PET is a non-invasive in vivo imaging technique that allows for both visualization and quantification of a target protein.
  • PET radiotracers are now routinely used in the diagnosis and monitoring of epilepsy, Parkinson’s disease, Alzheimer’s disease and other dementias. Despite these attributes, PET is not routinely used in MS. Numerous radiotracers have been developed for imaging neuroinflammation, which have almost exclusively targeted the TSPO on activated microglia. Despite these advances, various challenges have prevented the routine use of these tracers in MS (and neurology more widely). These challenges include low brain penetrance of existing radiotracers, high background uptake, and rs6971 polymorphism.
  • MERTK is a diagnostic and therapeutic target for a range of diseases and conditions associated with MERTK, such as cancer and MS. Additionally, a specific MERTK radioligand may have wide-ranging and significant applications in the diagnosis, and treatment and theragnostic of a range of diseases and conditions associated with MERTK, such as cancer and MS.
  • the present invention provides a compound of Formula (I): Formula (I) wherein Y is -(-C(O)-NH-)-or -(-C(O)-NH-(CH 2 ))-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl or optionally substituted C 3-8 cycloalkyl; R 2 is independently selected from the group consisting of H, optionally substituted C 1-6 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 2 -C 18 heteroaryl; R 3 is selected from the group consisting of H, OH, optionally substituted C
  • the present invention provides a compound of Formula (Ia): Formula (Ia) wherein Y is -(-C(O)-NH-)-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl or optionally substituted C 3-8 cycloalkyl; R 2 is independently selected from the group consisting of H, optionally substituted C 1-6 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 2 -C 18 heteroaryl; R 3 is selected from the group consisting of H, OH, optionally substituted C 1 -C 12 alkyloxy, NR 9 R 10 , optionally substituted
  • Compounds of Formula (I) have shown potent activity as inhibitors of MERTK. Compounds of the present invention may therefore be used in the treatment of diseases and conditions associated with MERTK. Additionally, compounds of the present invention have shown improved CNS penetrance compared with previously described compounds. Particularly, compounds having a beta-fluoro group as defined in Formula (I) show significant improvements in potency and selectivity for MERTK, as well as improved CNS penetrance. In a further aspect the present invention, the compounds of Formula (I) are radio- labelled, preferably with an 18 F group.
  • the present invention provides a method of imaging body tissue comprising: a) applying an imaging composition to a subject, wherein said composition comprises a radio- labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom.
  • the present invention provides a method of identifying, selecting, or diagnosing a disease state in a subject, comprising the steps of: a) applying an imaging composition to a body tissue of the subject, wherein said composition comprises a radio-labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom, wherein the image indicates a level of microglial subtype present in the body tissue; c) comparing the level with a reference level to determine an increased or decreased level of microglial subtype present in the body tissue, wherein the increased or decreased level of microglial subtype present in the body tissue compared to the reference level is indicative of a disease state or a stage of development of a disease state.
  • the disease state is a condition associated with MERTK. More preferably, the disease state is a neuroinflammatory central-nervous system disease. Even more preferably, the disease state is multiple sclerosis (MS).
  • the imaging is imaging is Positron Emission Tomography imaging.
  • a PET radiotracer specific for MERTK would provide the ability to distinguish pathogenic and reparative processes that underpin conditions such as MS, when compared with compared with currently available radiotracers target the translocator protein (TSPO). This advance has the potential to transform current practice in managing MS.
  • the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt of a of the invention, and a pharmaceutically acceptable excipient.
  • the present invention provides compounds and methods for the identification, treatment, prevention, or amelioration of a condition associated with MERTK.
  • the condition is MS.
  • Figure 1 is a plot of plasma and brain (total) concentrations versus time (Compound 5-02) in male Swiss outbred mice following IV administration (3.7 mg/kg).
  • Figure 3 is a set of images of summed 0-10 uptake (standard uptake value) images in a control (Left image) and cuprizone challenged (Right image, MS model) mouse.
  • Figure 4 is a time activity curve (whole brain) in a control (black) and cuprizone- exposed (grey) mouse.
  • the word “comprise” and variations of the word, such as “comprising” and “comprises”, is not intended to exclude other additives, components, integers or steps.
  • the term “and/or” as used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other.
  • a and/or B is to be taken as specific disclosure of each of (i) A, (ii) B and (iii) A and B, just as if each is set out individually herein.
  • the term "subject” shall be taken to mean any mammalian animal, preferably a human.
  • the term ‘inhibit” and variations thereof such as “inhibiting” means to prevent, block or reduce the function of the thing being inhibited. The term does not require complete inhibition with a reduction of activity at least 50% being considered inhibition.
  • the group may be a terminal group or a bridging group”. This is intended to signify that the use of the term is intended to encompass the situation where the group is a linker between two other portions of the molecule as well as where it is a terminal moiety.
  • alkyl As an example, some publications would use the term “alkylene” for a bridging group and hence in these other publications there is a distinction between the terms “alkyl” (terminal group) and “alkylene” (bridging group). In the present application no such distinction is made and most groups may be either a bridging group or a terminal group.
  • unsubstituted means that there is no substituent or that the only substituents are hydrogen.
  • optionally substituted as used throughout the specification denotes that the group may or may not be further substituted or fused (so as to form a condensed polycyclic system), with one or more non-hydrogen substituent groups.
  • acyl include acetyl and benzoyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the carbonyl carbon.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom.
  • Alkenyl as a group or part of a group denotes an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched preferably having 2-12 carbon atoms, more preferably 2-10 carbon atoms, most preferably 2- 6 carbon atoms, in the normal chain.
  • the group may contain a plurality of double bonds in the normal chain and the orientation about each is independently E or Z.
  • the alkenyl group is preferably a 1-alkenyl group.
  • Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl and nonenyl.
  • the group may be a terminal group or a bridging group.
  • Alkenyloxy refers to an alkenyl-O- group in which alkenyl is as defined herein. Preferred alkenyloxy groups are C 1 -C 6 alkenyloxy groups.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom.
  • "Alkyl" as a group or part of a group refers to a straight or branched aliphatic hydrocarbon group, preferably a C 1 –C 12 alkyl, more preferably a C 1 -C 10 alkyl, most preferably C 1 -C 6 unless otherwise noted.
  • suitable straight and branched C 1 -C 6 alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, t-butyl, hexyl, and the like.
  • the group may be a terminal group or a bridging group.
  • Alkylamino includes both mono-alkylamino and dialkylamino, unless specified.
  • Mono-alkylamino means an Alkyl-NH- group, in which alkyl is as defined herein.
  • Dialkylamino means a (alkyl) 2 N- group, in which each alkyl may be the same or different and are each as defined herein for alkyl.
  • the alkyl group is preferably a C 1 -C 6 alkyl group.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the carbonyl carbon.
  • Alkyloxy refers to an alkyl-O- group in which alkyl is as defined herein. Preferably the alkyloxy is a C 1 -C 6 alkyloxy. Examples include, but are not limited to, methoxy and ethoxy.
  • the group may be a terminal group or a bridging group.
  • Alkyloxyalkyl refers to an alkyloxy-alkyl- group in which the alkyloxy and alkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group.
  • Alkyloxyaryl refers to an alkyloxy-aryl- group in which the alkyloxy and aryl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the aryl group.
  • the alkyl group is preferably a C 1 -C 6 alkyl group. Examples include, but are not limited to, methoxycarbonyl and ethoxycarbonyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the carbonyl carbon.
  • Alkyloxycycloalkyl refers to an alkyloxy-cycloalkyl- group in which the alkyloxy and cycloalkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the cycloalkyl group.
  • Alkyloxyheteroaryl refers to an alkyloxy-heteroaryl- group in which the alkyloxy and heteroaryl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroaryl group.
  • Alkyloxyheterocycloalkyl refers to an alkyloxy-heterocycloalkyl- group in which the alkyloxy and heterocycloalkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heterocycloalkyl group.
  • the alkyl group is preferably a C 1 -C 6 alkyl group.
  • alkylsulfinyl groups include, but not limited to, methylsulfinyl and ethylsulfinyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom.
  • the alkyl group is preferably a C 1 -C 6 alkyl group. Examples include, but not limited to methylsulfonyl and ethylsulfonyl.
  • the group may be a terminal group or a bridging group.
  • Alkynyl as a group or part of a group means an aliphatic hydrocarbon group containing a carbon-carbon triple bond and which may be straight or branched preferably having from 2-12 carbon atoms, more preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms in the normal chain. Exemplary structures include, but are not limited to, ethynyl and propynyl. The group may be a terminal group or a bridging group.
  • Alkynyloxy refers to an alkynyl-O- group in which alkynyl is as defined herein.
  • Preferred alkynyloxy groups are C 1 -C 6 alkynyloxy groups.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom.
  • Aminoalkyl means an NH 2 -alkyl- group in which the alkyl group is as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group.
  • Aryl as a group or part of a group denotes (i) an optionally substituted monocyclic, or fused polycyclic, aromatic carbocycle (ring structure having ring atoms that are all carbon) preferably having from 5 to 12 atoms per ring.
  • aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety in which a phenyl and a C 5-7 cycloalkyl or C 5-7 cycloalkenyl group are fused together to form a cyclic structure, such as tetrahydronaphthyl, indenyl or indanyl.
  • the group may be a terminal group or a bridging group.
  • an aryl group is a C 6 -C 18 aryl group.
  • Arylalkenyl means an aryl-alkenyl- group in which the aryl and alkenyl are as defined herein.
  • Exemplary arylalkenyl groups include phenylallyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkenyl group;
  • Arylalkyl means an aryl-alkyl- group in which the aryl and alkyl moieties are as defined herein.
  • Preferred arylalkyl groups contain a C 1-5 alkyl moiety.
  • Exemplary arylalkyl groups include benzyl, phenethyl, 1-naphthalenemethyl and 2-naphthalenemethyl.
  • the group may be a terminal group or a bridging group.
  • Arylalkyloxy refers to an aryl-alkyl-O- group in which the alkyl and aryl are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom.
  • Arylamino includes both mono-arylamino and di-arylamino unless specified. Mono-arylamino means a group of formula arylNH-, in which aryl is as defined herein.
  • Di-arylamino means a group of formula (aryl) 2 N- where each aryl may be the same or different and are each as defined herein for aryl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom.
  • Arylheteroalkyl means an aryl-heteroalkyl- group in which the aryl and heteroalkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group.
  • Aryloxy refers to an aryl–O- group in which the aryl is as defined herein.
  • the aryloxy is a C 6 -C 18 aryloxy, more preferably a C 6 -C 10 aryloxy.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom.
  • a “bond” is a linkage between atoms in a compound or molecule.
  • the bond may be a single bond, a double bond, or a triple bond.
  • Cycloalkenyl means a non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and preferably having from 5-10 carbon atoms per ring.
  • Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl or cycloheptenyl.
  • the cycloalkenyl group may be substituted by one or more substituent groups.
  • a cycloalkenyl group typically is a C 3 -C 12 alkenyl group.
  • the group may be a terminal group or a bridging group.
  • Cycloalkyl refers to a saturated monocyclic or fused or spiro polycyclic, carbocycle preferably containing from 3 to 9 carbons per ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like, unless otherwise specified. It includes monocyclic systems such as cyclopropyl and cyclohexyl, bicyclic systems such as decalin, and polycyclic systems such as adamantane.
  • a cycloalkyl group typically is a C 3 -C 12 alkyl group.
  • the group may be a terminal group or a bridging group.
  • Cycloalkylalkyl means a cycloalkyl-alkyl- group in which the cycloalkyl and alkyl moieties are as defined herein.
  • Exemplary monocycloalkylalkyl groups include cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl and cycloheptylmethyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group.
  • Cycloalkylalkenyl means a cycloalkyl-alkenyl- group in which the cycloalkyl and alkenyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkenyl group.
  • Cycloalkylheteroalkyl means a cycloalkyl-heteroalkyl- group in which the cycloalkyl and heteroalkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group.
  • Cycloalkyloxy refers to a cycloalkyl-O- group in which cycloalkyl is as defined herein.
  • the cycloalkyloxy is a C 1 -C 6 cycloalkyloxy.
  • Examples include, but are not limited to, cyclopropanoxy and cyclobutanoxy.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom.
  • Cycloalkenyloxy refers to a cycloalkenyl-O- group in which the cycloalkenyl is as defined herein.
  • the cycloalkenyloxy is a C 1 -C 6 cycloalkenyloxy.
  • the group may be a terminal group or a bridging group.
  • Haloalkyl refers to an alkyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine.
  • a haloalkyl group typically has the formula CnH(2n+1- m)Xm wherein each X is independently selected from the group consisting of F, Cl, Br and I. In groups of this type n is typically from 1 to 10, more preferably from 1 to 6, most preferably 1 to 3. m is typically 1 to 6, more preferably 1 to 3.
  • haloalkyl examples include fluoromethyl, difluoromethyl and trifluoromethyl.
  • Haloalkenyl refers to an alkenyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom independently selected from the group consisting of F, Cl, Br and I.
  • Haloalkynyl refers to an alkynyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom independently selected from the group consisting of F, Cl, Br and I.
  • Halogen represents chlorine, fluorine, bromine or iodine.
  • Heteroalkyl refers to a straight- or branched-chain alkyl group preferably having from 2 to 12 carbons, more preferably 2 to 6 carbons in the chain, in which one or more of the carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatomic group selected from S, O, P and NR’ where R’ is selected from the group consisting of H, optionally substituted C 1 -C 12 alkyl, optionally substituted C 3 -C 12 cycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 1 -C 18 heteroaryl.
  • heteroalkyls include alkyl ethers, secondary and tertiary alkyl amines, amides, alkyl sulfides, and the like.
  • heteroalkyl also include hydroxyC 1 -C 6 alkyl, C 1 -C 6 alkyloxyC 1 - C 6 alkyl, aminoC 1 -C 6 alkyl, C 1 -C 6 alkylaminoC 1 -C 6 alkyl, and di(C 1 -C 6 alkyl)aminoC 1 -C 6 alkyl.
  • the group may be a terminal group or a bridging group.
  • Heteroalkyloxy refers to a heteroalkyl-O- group in which heteroalkyl is as defined herein. Preferably the heteroalkyloxy is a C 2 -C 6 heteroalkyloxy.
  • the group may be a terminal group or a bridging group.
  • Heteroaryl either alone or part of a group refers to groups containing an aromatic ring (preferably a 5 or 6 membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring with the remainder of the ring atoms being carbon atoms. Suitable heteroatoms include nitrogen, oxygen and sulphur.
  • the group may be a monocyclic or bicyclic heteroaryl group.
  • heteroaryl examples include thiophene, benzothiophene, benzofuran, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, naphtho[2,3- b]thiophene, furan, isoindolizine, xantholene, phenoxatine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, tetrazole, indole, isoindole, 1H-indazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, cinnoline, carbazole, phenanthridine, acridine, phenazine, thiazole, isothiazole, phenothiazine, oxazole, isooxazole, furazane, phen
  • a heteroaryl group is typically a C 1 - C 18 heteroaryl group.
  • the group may be a terminal group or a bridging group.
  • Heteroarylalkyl means a heteroaryl-alkyl group in which the heteroaryl and alkyl moieties are as defined herein.
  • Preferred heteroarylalkyl groups contain a lower alkyl moiety.
  • Exemplary heteroarylalkyl groups include pyridylmethyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group.
  • Heteroarylalkenyl means a heteroaryl-alkenyl- group in which the heteroaryl and alkenyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkenyl group.
  • Heteroarylheteroalkyl means a heteroaryl-heteroalkyl- group in which the heteroaryl and heteroalkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group.
  • Heteroaryloxy refers to a heteroaryl-O- group in which the heteroaryl is as defined herein. Preferably the heteroaryloxy is a C 1 -C 18 heteroaryloxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. “Heterocyclic” refers to saturated, partially unsaturated or fully unsaturated monocyclic, bicyclic or polycyclic ring system containing at least one heteroatom selected from the group consisting of nitrogen, sulfur and oxygen as a ring atom. Examples of heterocyclic moieties include heterocycloalkyl, heterocycloalkenyl and heteroaryl.
  • Heterocycloalkenyl refers to a heterocycloalkyl group as defined herein but containing at least one double bond.
  • a heterocycloalkenyl group typically is a C 2 - C 12 heterocycloalkenyl group.
  • the group may be a terminal group or a bridging group.
  • Heterocycloalkyl refers to a saturated monocyclic, bicyclic, or polycyclic ring containing at least one heteroatom selected from nitrogen, sulfur, oxygen, preferably from 1 to 3 heteroatoms in at least one ring. Each ring is preferably from 3 to 10 membered, more preferably 4 to 7 membered.
  • heterocycloalkyl substituents include pyrrolidyl, tetrahydrofuryl, tetrahydrothiofuranyl, piperidyl, piperazyl, tetrahydropyranyl, morphilino, 1,3-diazapane, 1,4-diazapane, 1,4-oxazepane, and 1,4-oxathiapane.
  • a heterocycloalkyl group typically is a C 2 -C 12 heterocycloalkyl group. The group may be a terminal group or a bridging group.
  • Heterocycloalkylalkyl refers to a heterocycloalkyl-alkyl- group in which the heterocycloalkyl and alkyl moieties are as defined herein.
  • exemplary heterocycloalkylalkyl groups include (2-tetrahydrofuryl)methyl, (2-tetrahydrothiofuranyl) methyl.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group.
  • Heterocycloalkylalkenyl refers to a heterocycloalkyl-alkenyl- group in which the heterocycloalkyl and alkenyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkenyl group.
  • "Heterocycloalkylheteroalkyl” means a heterocycloalkyl-heteroalkyl- group in which the heterocycloalkyl and heteroalkyl moieties are as defined herein.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group.
  • Heterocycloalkyloxy refers to a heterocycloalkyl-O- group in which the heterocycloalkyl is as defined herein.
  • the heterocycloalkyloxy is a C 1 - C 6 heterocycloalkyloxy.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom.
  • Heterocycloalkenyloxy refers to a heterocycloalkenyl-O- group in which heterocycloalkenyl is as defined herein.
  • the Heterocycloalkenyloxy is a C 1 -C 6 Heterocycloalkenyloxy.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom.
  • “Hydroxyalkyl” refers to an alkyl group as defined herein in which one or more of the hydrogen atoms has been replaced with an OH group.
  • a hydroxyalkyl group typically has the formula C n H (2n+1-x) (OH) x.
  • n is typically from 1 to 10, more preferably from 1 to 6, most preferably 1 to 3.
  • x is typically 1 to 6, more preferably 1 to 3.
  • the group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom.
  • each formula includes compounds having the indicated structure, including the hydrated as well as the non-hydrated forms.
  • pharmaceutically acceptable salts refers to salts that retain the desired biological activity of the above-identified compounds and include pharmaceutically acceptable acid addition salts and base addition salts.
  • Suitable pharmaceutically acceptable acid addition salts of compounds of Formula (I) may be prepared from an inorganic acid or from an organic acid.
  • inorganic acids are hydrochloric, sulfuric, and phosphoric acid.
  • Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic and sulfonic classes of organic acids, examples of which are formic, acetic, propanoic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, fumaric, maleic, alkyl sulfonic, arylsulfonic.
  • base addition salts may be prepared by ways well known in the art using organic or inorganic bases.
  • Suitable organic bases include simple amines such as methylamine, ethylamine, triethylamine and the like.
  • suitable inorganic bases include NaOH, KOH, and the like. Additional information on pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Co., Easton, PA 1995. In the case of agents that are solids, it is understood by those skilled in the art that the inventive compounds, agents and salts may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulae.
  • the term "therapeutically effective amount” or “effective amount” is an amount sufficient to effect beneficial or desired clinical results. An effective amount can be administered in one or more administrations. An effective amount is typically sufficient to palliate, ameliorate, stabilize, reverse, slow or delay the progression of the disease state.
  • Compounds of the Invention As outlined above, the compounds of the present invention have the Formula (I):
  • R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl or optionally substituted C 3-8 cycloalkyl
  • R 2 is independently selected from the group consisting of H, optionally substituted C 1-6 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 2 -C 18 heteroaryl;
  • R 3 is selected from the group consisting of H, OH, optionally substituted C 1 -C 12 alkyloxy, NR 9 R 10 , optionally substituted C
  • R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl or optionally substituted C 3-8 cycloalkyl
  • R 2 is independently selected from the group consisting of H, optionally substituted C 1-6 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3-8 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 2 -C 18 heteroaryl;
  • R 3 is selected from the group consisting of H, OH, optionally substituted C 1 -C 12 alkyloxy, NR 9 R 10 , optionally substituted C 1 -C 12 alkylamino, and halogen;
  • R 4 is selected from the group consisting of H, OH, optionally substituted C 1 -C 12 alkyloxy, NR 9 R 10 , optionally substitute
  • Y is -(-C(O)-NH-)- and n is 1.
  • This provides compounds of Formula (IIb): Formula (IIb) wherein dashed lines indicate an optional methylene bridge; and R 1 , R 2 , R 3 , and R 4 , are defined as above; or a pharmaceutically acceptable salt thereof.
  • R 4 is an optionally substituted 5-7-membered heterocyclic group of the formula: wherein X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; m is 0 to 2; where valency allows, R 7 is selected from the group consisting of H, optionally substituted C 1 - C 12 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3 -C 12 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 1 -C 18 heteroaryl; each R 5 is independently defined as above.
  • the compound of Formula (I) has a structure of Formula (III): Formula (III) wherein R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl or optionally substituted C 3-8 cycloalkyl; X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; m is 0 to 2; where valency allows, R 7 is selected from the group consisting of H, optionally substituted C 1 - C 12 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3 -C 12 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 1 -C 18 heteroaryl; R 2 is selected from the group consisting of H, optionally substituted C 1-6 alkyl, optionally substituted C 1 -C
  • m is 0. In some embodiments of the compound of Formula (III), m is 1. In some embodiments of the compound of Formula (III), m is 2.
  • R 4 is an optionally substituted heterocyclic group of the formula: wherein X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; where valency allows, R 7 is selected from the group consisting of H, optionally substituted C 1 - C 12 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 -C 12 haloalkyl, optionally substituted C 3 -C 12 cycloalkyl, optionally substituted C 2 -C 12 heterocycloalkyl, optionally substituted C 6 -C 18 aryl, and optionally substituted C 1 -C 18 heteroaryl; each R 5 is independently defined as above.
  • the beta-fluoro group has S-chirality.
  • An (S)-beta-fluoro group particularly shows unexpected improvements in potency and selectivity for MERTK, as well as improved CNS penetrance over previously described compounds.
  • the beta-fluoro group has S-chirality, R 2 is H, R 3 is OH, R 4 is attached at the 5-position, m is 1, X is N, and the dashed lines represent double bonds.
  • R 1 is selected from the group consisting of optionally substituted C 1-6 alkyl; and each R 5 is independently defined as above; or a pharmaceutically acceptable salt thereof.
  • R 1 is optionally substituted methyl or optionally substituted ethyl.
  • R 3 is OH or halogen.
  • R 2 is H.
  • R 5 is independently selected from the group consisting of H, CN, halogen, optionally substituted C 1- C 6 alkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 3 -C 12 cycloalkyl, an optionally substituted C 1 -C 12 heterocycloalkyl, optionally substituted C 1 -C 12 alkylsulfonyl, optionally substituted C 1 -C 12 alkylamino, and optionally substituted C 1 -C 12 haloalkyl.
  • R 6 is independently selected from the group consisting of H, halogen, optionally substituted C 1- C 6 alkyl, optionally substituted C 3 -C 12 cycloalkyl, an optionally substituted C 1 - C 12 heterocycloalkyl, optionally substituted C 1 -C 12 alkyloxy, optionally substituted C 1 - C 12 alkylsulfonyl, optionally substituted C 1 -C 12 alkylamino, and optionally substituted C 1 - C 12 haloalkyl.
  • R 4 is selected from the from the group consisting of: , wherein R 5 is as defined herein.
  • R 4 has a structure selected from the group consisting of: , , , , , , , , and .
  • the compound has a structure as shown in Table 1, or a pharmaceutically acceptable salt thereof. Table 1
  • the compound of Formula (I) is CNS penetrant.
  • CNS penetrant means that the compound can penetrate the blood-brain barrier (BBB) and exhibit activity in the central nervous system. Penetration of the BBB may be by any mechanism such as passive transport into the CNS, active transport, efflux, and metabolism. CNS penetrance can be indicated by various parameters described in the art such as Blood Brain Barrier (BBB) Score (M. Gupta, H.J. Lee, C.J. Barden, D.F. Weaver, The Blood-Brain Barrier (BBB) Score, J Med Chem, 62 (2019) 9824-9836) or the CNS PET multi- parameter optimization (MPO) tool (L.
  • BBB Blood Brain Barrier
  • MPO CNS PET multi- parameter optimization
  • the compound of Formula (I) is radio-labelled to provide a radio-tracer compound.
  • the compound of Formula (I) is radio-labelled with 18 F.
  • the radio-label may be attached to any part of the molecule.
  • the beta-fluoro group may be 18 F.
  • in the compound of Formula (I) at least one of R 3 is radio-labelled, preferably with 18 F. Radiolabelling of the compound of Formula (I) can be performed particularly by the synthesis methods described in the accompanying examples.
  • the present invention provides an imaging composition comprising a radio-labelled compound of the invention.
  • the present invention provides a method of imaging body tissue comprising: a) applying an imaging composition to a subject, wherein said composition comprises a radio-labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom.
  • the body tissue is selected from the group consisting of the brain and spinal cord.
  • the imaging method is Positron Emission Tomography imaging.
  • the present invention provides a method of identifying, selecting, or diagnosing a disease state in a subject, comprising the steps of: a) applying an imaging composition to a body tissue of the subject, wherein said composition comprises a radio-labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom, wherein the image indicates a level of microglial subtype present in the body tissue; c) comparing the level with a reference level to determine an increased or decreased level of microglial subtype present in the body tissue, wherein the increased or decreased level of microglial subtype present in the body tissue compared to the reference level is indicative of a disease state or a stage of development of a disease state.
  • the reference level of microglial subtype present in the body tissue is the level of expression of microglial subtype present in the body tissue from a normal subject.
  • the microglial subtype present in the body tissue is M2 microglia.
  • imaging method is Positron Emission Tomography imaging.
  • the disease state is a condition associated with MERTK.
  • the disease state is a neuroinflammatory central- nervous system disease.
  • the disease state is multiple sclerosis (MS).
  • the disease state is cancer.
  • the method of identifying, selecting, or diagnosing a disease state in a subject further comprises administering to the subject a therapeutically effective amount of an anti-disease state therapeutic.
  • the compounds of the invention are ligands of MERTK and therefore have the ability to inhibit these enzymes.
  • the ability to inhibit the enzymes may be a result of the compounds acting directly and solely on the enzyme to modulate/potentiate biological activity. However, it is understood that the compounds may also act at least partially on other factors associated with the activity of the enzyme.
  • the inhibition of MERTK may be carried out in any of a number of ways known in the art. For example if inhibition in vitro is desired an appropriate amount of the compound may be added to a solution containing the MERTK.
  • the inhibition of the MERTK typically involves administering the compound to a mammal containing the compound of the invention.
  • the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt of a compound of the invention, and a pharmaceutically acceptable excipient.
  • the present invention provides a compound of the invention for use in the identification, treatment, prevention, or amelioration of a condition in a mammal.
  • the condition is associated with MERTK.
  • the present invention provides a method of identifying, preventing, treating, or ameliorating a condition in a mammal, said method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of the invention.
  • the condition is associated with MERTK.
  • the present invention provides a use of a compound of the invention in the manufacture of a medicament for the identification, treatment, prevention, or amelioration of a condition in a mammal.
  • the condition is associated with MERTK.
  • the condition is selected from the group consisting of neuroinflammatory central-nervous system disease and cancer.
  • the condition is neuroinflammatory central-nervous system disease.
  • the neuroinflammatory central-nervous system disease is selected from the list consisting of multiple sclerosis (MS), traumatic brain injury, epilepsy, Alzheimer’s disease (AD), Parkinson's disease (PD), Huntington's disease (HD), Multiple system atrophy (MSA), Amyotrophic lateral sclerosis (ALS), stroke, neuromyelitis optica spectrum disorder (NMOSD), acute disseminated encephalomyelitis (ADEM) and myelin oligodendrocyte glycoprotein (MOG) encephalomyelitis, and the leukodystrophies (including Aicardi-Goutines syndrome, adrenoleukodystrophy (ALD), Alexander disease, Canavan disease, cerebrotendinous xanthomatosis, Krabbé disease, metachromatic leukodystrophy (MLD), Niemann-Pick disease, Pelizaeus-Merzbacher disease (PMD), and childhood ataxia with central nervous system hypomyelination).
  • MS multiple sclerosis
  • the condition is multiple sclerosis (MS).
  • the condition is cancer.
  • the cancer is selected from the list consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukaemia (AML), melanoma, breast cancer, lung cancer, colon cancer, prostate cancer, gastric cancer, non-small cell lung cancer (NSCLC), glioblastoma, liver cancer, kidney cancer, ovarian cancer, uterine cancer, and brain cancer.
  • Administration of compounds within Formula (I) to humans can be by any of the accepted modes for enteral administration such as oral or rectal, or by parenteral administration such as subcutaneous, intramuscular, intravenous and intradermal routes. Injection can be bolus or via constant or intermittent infusion.
  • the active compound is typically included in a pharmaceutically acceptable carrier or diluent and in an amount sufficient to deliver to the patient a therapeutically effective dose.
  • they can be administered in any form or mode which makes the compound bioavailable.
  • One skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the compound selected, the condition to be treated, the stage of the condition to be treated and other relevant circumstances. We refer the reader to Remingtons Pharmaceutical Sciences, 19 th edition, Mack Publishing Co. (1995) for further information.
  • the compounds of the present invention can be administered alone or in the form of a pharmaceutical composition in combination with a pharmaceutically acceptable carrier, diluent or excipient.
  • the compounds of the invention while effective themselves, are typically formulated and administered in the form of their pharmaceutically acceptable salts as these forms are typically more stable, more easily crystallised and have increased solubility.
  • the compounds are, however, typically used in the form of pharmaceutical compositions which are formulated depending on the desired mode of administration.
  • the present invention provides a pharmaceutical composition including a compound of Formula (I) and a pharmaceutically acceptable carrier, diluent or excipient.
  • the compositions are prepared in manners well known in the art.
  • the invention in other embodiments provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. In such a pack or kit can be found a container having a unit dosage of the agent(s).
  • kits can include a composition comprising an effective agent either as concentrates (including lyophilized compositions), which can be diluted further prior to use or they can be provided at the concentration of use, where the vials may include one or more dosages.
  • an effective agent either as concentrates (including lyophilized compositions), which can be diluted further prior to use or they can be provided at the concentration of use, where the vials may include one or more dosages.
  • single dosages can be provided in sterile vials so that the physician can employ the vials directly, where the vials will have the desired amount and concentration of agent(s).
  • Associated with such container(s) can be various written materials such as instructions for use, or a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.
  • the compounds of the invention may be used or administered in combination with one or more additional drug(s) for the treatment of the disorder/diseases mentioned.
  • the components can be administered in the same formulation or in separate formulations. If administered in separate formulations the compounds of the invention may be administered sequentially or simultaneously with the other drug(s).
  • the compounds of the invention may be used in a combination therapy. When this is done the compounds are typically administered in combination with each other. Thus one or more of the compounds of the invention may be administered either simultaneously (as a combined preparation) or sequentially in order to achieve a desired effect. This is especially desirable where the therapeutic profile of each compound is different such that the combined effect of the two drugs provides an improved therapeutic result.
  • compositions of this invention for parenteral injection comprise pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use.
  • suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
  • These compositions may also contain adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of micro- organisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like.
  • Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminium monostearate and gelatin.
  • the compounds can be incorporated into slow release or targeted delivery systems such as polymer matrices, liposomes, and microspheres.
  • the injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
  • the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay
  • the dosage form may also comprise buffering agents.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
  • embedding compositions which can be used include polymeric substances and waxes.
  • the active compounds can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
  • inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzy
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • Suspensions in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.
  • compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.
  • Dosage forms for topical administration of a compound of this invention include powders, patches, sprays, ointments and inhalants.
  • the active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives, buffers, or propellants which may be required.
  • the amount of compound administered will preferably treat and reduce or alleviate the condition.
  • a therapeutically effective amount can be readily determined by an attending diagnostician by the use of conventional techniques and by observing results obtained under analogous circumstances. In determining the therapeutically effective amount a number of factors are to be considered including but not limited to, the species of animal, its size, age and general health, the specific condition involved, the severity of the condition, the response of the patient to treatment, the particular compound administered, the mode of administration, the bioavailability of the preparation administered, the dose regime selected, the use of other medications and other relevant circumstances.
  • a preferred dosage will be a range from about 0.01 to 300 mg per kilogram of body weight per day.
  • a more preferred dosage will be in the range from 0.1 to 100 mg per kilogram of body weight per day, more preferably from 0.2 to 80 mg per kilogram of body weight per day, even more preferably 0.2 to 50 mg per kilogram of body weight per day.
  • a suitable dose can be administered in multiple sub-doses per day. Examples The invention will now be further explained and illustrated by reference to the following non-limiting examples. Additional compounds, other than those described below, may be prepared using methods and synthetic protocols or appropriate variations or modifications thereof, as described herein. The agents of the various embodiments may be prepared using the reaction routes and synthesis schemes as described below, employing the techniques available in the art using starting materials that are readily available.
  • reactions can be monitored according to any suitable method known in the art.
  • product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), or mass spectrometry, or by chromatography such as high-performance liquid chromatography (HPLC) or thin layer chromatography.
  • spectroscopic means such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), or mass spectrometry
  • HPLC high-performance liquid chromatography
  • ambient temperature e.g., a reaction temperature
  • room temperature e.g., a temperature from about 20 oC to about 30 oC.
  • Reagents useful for synthesizing compounds may be obtained from commercial suppliers or prepared according to techniques known in the art. Commercial solvents and reagents were used as supplied. Analytical thin-layer chromatography (TLC) was performed on silica gel 60 pre- coated with fluorescent indicator F254 aluminium sheets (0.25 mm, Merck).
  • TLC plates were visualised using UV254 or by chemical staining with a solution of potassium permanganate followed by mild heating. Flash column chromatography was carried out using Davisil silica gel 60, 40-63 ⁇ m (LC60A 40-63, Davisil). NMR spectra were obtained on a Bruker UltraShield 400 spectrometer at 400 MHz for 1 H and 100.6 MHz for 13 C.
  • LC-MS Liquid chromatography mass spectrometry
  • Agilent Technologies 6120 Single Quad LC/MS System coupled with an Agilent 1260 HPLC consisting of G1312B Quaternary pump, 1260 Infinity G1367E 1260 HiP ALS Autosampler and 1290 Infinity G4212A 1290 DAD wavelength detector (monitored at 214nm and 254nm).
  • Software used for LC-MS was LC/MSD Chemstation Rev.B.04.03 coupled with Masshunter Easy Access Software.
  • LC conditions reverse phase HPLC analysis.
  • HPLC was acquired on an Agilent 1260 Infinity Analytical HPLC equipped with a 1260 Degasser G1322A (JPAAJ81416), 1260 Bin Pump G1312B (DEACB04606), 1260 HiP ALS G1367E (DEACO03069), 1260 TCC: G1316A (DEACN16726), 1260 DAD: G4212B (DEAA304233) and a Zorbax Eclipse Plus C18 Rapid Resolution 4.6 X 100mm 3.5 ⁇ Micron Column. Solvent A: water + 0.1% TFA. Solvent B: MeCN + 0.1% TFA.
  • LTBS salts (30-80%).
  • Lithium (pyridin-2-yl)trihydroxyborate (LTBS) According to the general procedure, using 2-bromopyridine to afford the title compound as a light brown solid (665 mg, 71%).
  • Example 5-21 5-((2-(((S)-2-fluorobutyl)amino)-5-(6-fluoropyridin-2-yl)pyrimidin-4- yl)amino)bicyclo[2.2.1]heptan-2-ol: Off-white solid (39%).
  • Example 5-22 (1S,4r)-4-((5-(6-chloropyridin-2-yl)-2-(((S)-2-fluorobutyl)amino)pyrimidin- 4-yl)amino)cyclohexan-1-ol
  • General Procedure Route A using lithium (6-chloropyridin-2- yl)trihydroxyborate, to afford the title compound as pale yellow solid (43 mg, 42% yield);
  • Example 5-23 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)-N-methylpyridine-3-sulfonamide
  • Example 5-24 (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-(3-fluoropropyl)pyridin-2- yl)pyrimidin-4-yl)amino) cyclohexan-1-ol trifluoroacetate salt
  • Example 5-26 (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(methylsulfonyl)pyridin-2- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol trifluoroacetate salt
  • Example 5-28 (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-methoxypyridin-2- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol According to General Procedure Route B, using pyridine 4-methoxypyridine, to afford the title compound as an off-white solid (45 mg, 0.116 mmol, 62%).
  • Example 5-29 (1S,4r)-4-((5-(5-((dimethylamino)methyl)pyridin-2-yl)-2-(((S)-2- fluorobutyl)amino) pyrimidin-4-yl)amino) cyclohexan-1-ol
  • Example 5-31 (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(morpholinomethyl)pyridin-2- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol
  • HPLC tR 3.45 min, >99 % purity at 254 nm
  • 1 H NMR 400 MHz, MeOD
  • Example 5-32 (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(methyl(tetrahydrofuran-3- yl)amino) pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol
  • White amorphous solid (18 % yield, obtained as a TFA salt);
  • Example 5-33 N-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl) amino) pyrimidin-5-yl)pyridin-3-yl)-1-methylpiperidine-4-carboxamide
  • Example 5-35 1-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)-3-methylimidazolidin-2-one 2,2,2- trifluoroacetate According to General Procedure outlined in Scheme 3, using 1- methylimidazolidin-2-one, purification by reverse phase column to afford the trifluoroacetate salt of the title compound as an off-white solid.
  • Example 5-36 1-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)-4-methylpiperazin-2-one 2,2,2- trifluoroacetate According to General Procedure outlined in Scheme 3, using 4-methylpiperazin-2- one, purification by reverse phase column to afford the trifluoroacetate salt of the title compound as an off-white solid.
  • Example 5-40 Scheme 6 (a) (1r,4r)-4-((5-(1-methyl-1H-pyrazol-3-yl)-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol A suspension of (4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin- 5-yl)boronic acid 3 (0.5 g, 1.77 mmol, 1.0 equiv.), 3-bromo-1-methyl-1H-pyrazole (0.312 g, 1.94 mmol, 1.1 equiv.), K 2 CO 3 (0.488 g, 3.53 mmol, 2.0 equiv.), tetrakis(triphenylphosphine)palladium(0) (0.102 g, 0.0885 mmol, 0.05 equiv.) in 1,4-dioxane (8 mL) and water (2 mL) was purged with
  • the mixture was stirred at 100 °C for 6 hours at which point it was concentrated to a dark brown/black residue and filtered through pad of Celite® and adjusted pH to 4 using 10% aqueous citric acid solution.
  • the crude mixture was diluted with ethyl acetate and extracted (3 x 100 mL), the combined organic layer was dried over MgSO 4 and concentrated.
  • Example 5-46 (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-(((2- fluoroethyl)(methyl)amino)methyl)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2-trifluoroacetate
  • a solution of Example 5-45 from step (d)
  • 4 M aqueous HCl solution 0.2 mL, 0.8 mmol
  • 50% aqueous NaOH was used to adjust the pH to 7.
  • Example 5-50 Scheme 10 (a) 6-bromo-N-(2-fluoroethyl)pyridin-3-amine To a solution of 2-bromo-5-iodopyridine (4.1 g, 14.44 mmol, 1.0 equiv.) in 1,4- dioxane were added 2-fluoroethan-1-amine hydrochloride (1.58 g, 15.89 mmol, 1.1 equiv.), Cs 2 CO 3 (14.12 g, 43.33 mmol, 3 equiv.). and xantphos (1.25 g, 2.17 mmol, 0.15 equiv.). The reaction mixture was degassed with nitrogen for 30 min.
  • the reaction mixture was stirred at 150 °C in a sealed tube for 3 days.
  • the solvent was evaporated under reduced pressure and the resulting residue was partitioned between brine and DCM.
  • the aqueous layer was further extracted with DCM (3 ⁇ 20 mL), the combined organic exacts were dried over anhydrous magnesium sulfate (MgSO 4 ), filtered and concentrated under reduced pressure.
  • the residue was purified by silica gel flash chromatography (0-50% EtOAc in petroleum spirits) to afford the title compound as an off- white solid (382.5 mg, 1.06 mmol, 46%).
  • Example 11-03 (a) 2,4-dichloro-N-(1-cyclobutylpiperidin-4-yl)pyrimidine-5-carboxamide To a solution of 2,4-dichloropyrimidine-5-carbonyl chloride (500mg, 2.36 mmol, 1.0 equiv.) in dichloromethane (10 mL) were added 4-(N-cyclobutyl)piperidinyl amine (401.2 mg, 2.60 mmol, 1.1 equiv.) and DIPEA (617.9 ⁇ L, 3.55 mmol, 1.5 equiv.) at 0 °C. The resulting mixture was stirred at 0 °C for 1 h.
  • Example 11-04 Scheme 13 (a) Ethyl 2-chloro-4-(((1r,4r)-4-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate To a solution of ethyl 2,4-dichloropyrimidine- 5-carboxylate (2.0 g, 9.1 mmol), trans-4-aminocyclohexanol (1.2 g, 10 mmol) in i-PrOH (50 mL) was added DIPEA (1.8 g, 14 mmol, 2.7 mL) at 0 °C. The resultant mixture was slowly warmed to room temperature and stirred for 14 h.
  • reaction mixture was concentrated in vacuo before partitioned between ethyl acetate and water.
  • the mixture was further extracted with ethyl acetate (x2).
  • the organic layers were combined and dried with magnesium sulphate.
  • the volatiles were removed in vacuo and the residue was purified by flash silica gel column chromatography, eluting with 10–20% ethyl acetate/petroleum spirits to afford the title as white solid (2.32 g, 86%).
  • the resultant mixture was stirred at room temperature for 16 h.
  • the reaction mixture was concentrated in vacuo and the residue was partitioned between ethyl acetate and water.
  • the mixture was further extracted with ethyl acetate (x2).
  • the organic layers were combined and dried over magnesium sulfate, filtered and concentrated.
  • the residue was purified by flash silica gel column chromatography, eluting with 10% ethyl acetate/petroleum spirits to afford the title compound as a white solid (3.7 g, 67%).
  • the resultant mixture was heated to 50 °C for 16 h.
  • the THF was removed in vacuo and the aqueous layer was acidified with 10% aqueous citric acid to pH 3–4.
  • the white precipitate formed was collected by filtration to afford the title compound as an off-white solid (44.7 mg, 69%).
  • reaction mixture was stirred at 50 °C overnight, the solvent was removed in vacuo and the reaction was mixture was partitioned between ethyl acetate and water. The mixture was further extracted with ethyl acetate (2x). The combined organic layers were dried over magnesium sulfate, filtered and the volatiles were removed in vacuo. The residue was purified by flash silica gel column chromatography, eluting with 5% MeOH in DCM to give the title compound as an off-white solid (18.4 mg, 64%).
  • Example 11-04 N-(1-(Cyclopropylmethyl)piperidin-4-yl)-2-((2-fluorobutyl)amino)-4- (((1r,4r)-4- hydroxycyclohexyl)amino)pyrimidine-5-carboxamide
  • 4-(((1r,4r)-4-((tert-butyldimethylsilyl)oxy)cyclohexyl)amino)-N-(1- (cyclopropylmethyl)piperidin-4-yl)-2-((2-fluorobutyl)amino)pyrimidine-5-carboxamide 15 mg, 0.026 mmol) in DCM (3 mL) was added 4M hydrochloric acid in 1,4-dioxane (1.9 mg , 0.052 mmol, 13 ⁇ L).
  • Example 11-05 Scheme 14 (a) ethyl 4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidine-5-carboxylate To a solution of ethyl 4-chloro-2-(methylthio)pyrimidine-5-carboxylate (5g, 21.5 mmol, 1.0 equiv.) and trans-4-aminocyclohexanol (2.6g, 22.56 mmol, 1.05 equiv.) in i-PrOH (100 mL) was added DIPEA (13.1 mL, 75.21 mmol, 3.5 equiv.) and the reaction mixture was stirred at RT overnight.
  • the stationary phase was a Phenomenex Gemini C-18, 5 ⁇ RP column, 150 ⁇ 4.6 mm.
  • Acetonitrile (A) and water (B) with 0.1% formic acid were used as the mobile phase and a gradient elution technique was used for analysis: 0-18 min: 15-40% A, 18-30 min: isocratic 40% A, 30-34 min: 40-90% A, 34-37 min: isocratic 90% A, 37-40 min: 90- 15% A at a flow rate of 0.5 mL/min.
  • the Bioscan FC-4000 dual BGO coincidence detector was used for the detection of radioactive compounds.
  • the HPLC system used for semi-preparative workup of radiolabelled compounds was a Knauer Smartline 1050 pump equipped with a Smartline Manager 5050 for quaternary gradient capability.
  • the Knauer UV detector 2520 (254 nm) and a CsI(TI) crystal PIN diode radioactivity detector were used for the detection of compounds.
  • the gradient used for the purification of [ 18 F]5-02 was 10-30% acetonitrile (0.1% formic acid) for 50 min at a flow rate of 4 mL/min.
  • the stationary phase was a Phenomenex Gemini C-18, 10 ⁇ RP column, 250 ⁇ 10 mm.
  • [ 18 F]fluoride was transferred from the cyclotron, trapped on a QMA ion exchange cartridge and eluted using 20 mg kryptofix 2.2.2 and 3.5 mg K 2 CO 3 in 0.2 mL of water plus 0.4 mL of acetonitrile as eluent mixture. After the addition of 1 mL of dry acetonitrile, the K[ 18 F]F/kryptofix mixture was dried at 90 °C for 8 min under vacuum with argon flow. After drying, the reactor was cooled to 50 °C and 4 mg of the precursor in 0.5 mL of DMF added. Radiolabelling was achieved by heating to 120 °C for 15 min in a closed reactor.
  • Activity assays were performed in a 384 well, polypropylene microplate in a final volume of 50 ⁇ l of 50 mM Hepes, pH 7.4 containing 10 mM MgCl2, 1.0 mM DTT, 0.01% Triton X-100, 0.1% Bovine Serum Albumin (BSA), containing 1.0 ⁇ M fluorescent substrate and ATP at the Km for each enzyme. All reactions were terminated by addition of 20 ⁇ l of 70 mM EDTA. After a 180 min incubation, phosphorylated and unphosphorylated substrate peptides were separated in buffer supplemented with 1 x CR-8 on a LabChip EZ Reader equipped with a 12-sipper chip. Data were analysed using EZ Reader software.
  • the peptide substrates of each kinase and their concentrations used in the assay are shown in Table 2.
  • Kinase peptide substrates and concentrations used in MCE assay The results of the assay are shown in Table 3.
  • the compounds of the invention exhibit low nanomolar potency activity against MERTK.
  • the compounds of the invention have good selectivity for MERTK over AXL and TYRO3.
  • most of the compounds of the invention are not appreciably active against FLT3, which is known to also be expressed in the CNS.
  • Table 5 Pharmacokinetic parameters for Compound 5-02in male Swiss outbred mice following IV administration at 3.7 mg/kg Plasma and brain (total) concentration versus time profiles are presented in Figure 1 and the unbound plasma and brain concentration versus time profiles are shown in Figure 2.
  • Compound 5-02 was stable in both assay matrices (brain homogenate and plasma) under the conditions of the RED assay, and fraction unbound values (fu) in mouse plasma and brain were 0.018 ( ⁇ 0.002) and 0.0072 ( ⁇ 0.0001), respectively.
  • values for Kp,uu were close to unity at all sample times suggesting that Compound 5-02is not subject to efflux at the blood brain barrier.
  • the absence of notable time-dependence in B:P and Kp,uu suggests that that distributional equilibrium between brain and plasma was achieved soon after dosing.
  • PET dynamic series were reconstructed with the Terratomo 3D ordered subset estimate maximization (OSEM) algorithm (four iterations and six subsets). Voxel size was 0.6mm 3 .
  • CT was used attenuation correction as well as correction for randoms and scatter.
  • Each frame of the dynamic series was corrected for radioactive decay and calibrated in SUV.
  • Analysis and interpretation were performed using PMOD (version 4.1, PMOD Technologies, Zurich, Switzerland). Regions of interest were drawn for the whole brain to derive the time-activity curves for each scan performed. The results of this analysis are shown in Figures 3 and 4.
  • Figure 3 shows the brain uptake summed over 10 minutes (standard uptake value) in both a control (left image) and cuprizone challenged (right image, MS model) mouse.
  • FIG. 3 shows, [ 18 F]5-02 is CNS-penetrant.
  • Figure 4 is a time activity curve (whole brain) in a control (black) and cuprizone-exposed (grey) mouse. Rapid uptake and washout are seen with minimal change in tracer concentration in the brain beyond 15 minutes (900 sec).
  • Radiometric protein kinase assay Reaction Biology A radiometric protein kinase assay (33PanQinaseTM Activity Assay) was used for measuring the kinase activity of the MERTK.
  • MERTK inhibition assay was performed in 96- well ScintiPlates TM from PerkinElmer (Boston, MA, USA) in a 50 ⁇ l reaction volume.
  • reaction cocktail was pipetted in four steps in the following order: 25 ⁇ l of assay buffer (standard buffer/ ⁇ -33P]-ATP), 10 ⁇ l of ATP solution (in H 2 O), 5 ⁇ l of test compound (in 10 % DMSO) and 10 ⁇ l of enzyme/substrate mixture.
  • the assay contained 70 mM HEPES-NaOH pH 7.5, 3 mM MgCl 2 , 3 mM MnCl 2 , 3 ⁇ M Na-orthovanadate, 1.2 mM DTT, 50 ⁇ g/ml PEG20000, ATP (corresponding to the apparent ATP-Km of the kinase, see Table 6), [ ⁇ - 33P]-ATP (approx.9 x 1005 cpm per well), protein kinase (see Table 6), and substrate (see Table 6). Table 6.
  • Plasma and brain concentrations of Compound 11-02 were detected for the duration of the 7.5 h sampling period, with an apparent half-life of ⁇ 2.4 h (Table 8).
  • the apparent blood volume of distribution and blood clearance were both high.
  • the apparent in vivo blood clearance (173 mL/min/kg) is higher than the nominal mouse hepatic blood flow (120 mL/min/kg) suggesting that Compound 11-02 may be subject to extrahepatic clearance processes.
  • a The terminal elimination phase has been estimated on the basis of the last two time points therefore values based on extrapolation to infinity are approximations only. Table 8.

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Abstract

The present invention provides compounds of Formula (I), which have activity as inhibitors of MERTK. The present invention also provides radio-labelled compounds which may be used in diagnostic and imaging compositions. The present invention also provides methods and uses of the compounds in imaging body tissue, as well as identifying and diagnosing disease states. The present invention further provides methods and uses of the compounds in the treatment of diseases, particularly conditions and diseases associated with MERTK.

Description

COMPOUNDS WITH MERTK ACTIVITY This application claims priority from Australian Patent Application No. 2022901554 filed on 7 June 2022, the contents of which are to be taken as incorporated herein by this reference. Technical Field The present invention relates to compounds with activity as inhibitors of MERTK. The present invention also relates to radio-labelled compounds which may be used in diagnostic and imaging compositions. The present invention also relates to methods and uses of the compounds in imaging body tissue, as well as identifying and diagnosing disease states. The present invention also relates to methods and uses of the compounds in the treatment of diseases, particularly conditions and diseases associated with MERTK. Background of Invention MER tyrosine kinase (MERTK), a receptor tyrosine kinase of the TAM (TYRO3, AXL, and MERTK) family, is associated with many chronic inflammatory and autoimmune diseases and disorders. MERTK upregulation is associated with M2 polarization of microglia, which plays a vital role in neuroregeneration following damage induced by neuroinflammatory diseases such as multiple sclerosis (MS). MERTK is also over-expressed in a wide variety of cancers (for example, acute lymphoblastic leukemia (ALL), acute myeloid leukaemia (AML), melanoma, breast cancer, colon cancer, prostate cancer, non-small cell lung cancer (NSCLC), glioblastoma, and others). Multiple sclerosis (MS) is the most common immune-mediated disorder affecting the central nervous system, characterized by demyelination of neurons in the brain and spinal cord. This damage disrupts the ability of parts of the nervous system to transmit signals, resulting in a range of signs and symptoms, including physical, mental, and sometimes psychiatric problems. There is no known cure for multiple sclerosis. Treatments attempt to improve function after an attack and prevent new attacks. Medications used to treat MS, while modestly effective, can have side effects and be poorly tolerated. In 2015, about 2.3 million people were affected globally and in the same year, about 18,900 people died from MS. The disease usually begins between the ages of twenty and fifty and is twice as common in women as in men. Life expectancy is typically 5 to 10 years lower than that of the unaffected population. Microglial activation facilitates the proliferation and the release of both cytotoxic and pro-inflammatory substances, in response to neural insults in the CNS. Both microglial activation and oligodendrocyte apoptosis have been revealed to be some of the earliest events during lesion formation in MS. It has recently been shown that MERTK is highly expressed in the M2-subclass microglia, which is associated with the reparative mechanisms in response to tissue injury in the brain. It has also been shown that the MERTK gene can significantly impact MS susceptibility and severity. Positron Emission Tomography (PET) is a non-invasive in vivo imaging technique that allows for both visualization and quantification of a target protein. Many advances have been made in the use of PET in neurology in the last ~20 years. PET radiotracers are now routinely used in the diagnosis and monitoring of epilepsy, Parkinson’s disease, Alzheimer’s disease and other dementias. Despite these attributes, PET is not routinely used in MS. Numerous radiotracers have been developed for imaging neuroinflammation, which have almost exclusively targeted the TSPO on activated microglia. Despite these advances, various challenges have prevented the routine use of these tracers in MS (and neurology more widely). These challenges include low brain penetrance of existing radiotracers, high background uptake, and rs6971 polymorphism. Previous work by the present inventors has shown that a suitable MERTK radiotracer could be of high clinical value for phenotyping different MS types and for monitoring disease progression by assessing M2-microglial activity. Early studies have demonstrated MERTK ligands with promising activity but with a CNS penetrance which is sub-optimal for clinical use. Therefore, MERTK is a diagnostic and therapeutic target for a range of diseases and conditions associated with MERTK, such as cancer and MS. Additionally, a specific MERTK radioligand may have wide-ranging and significant applications in the diagnosis, and treatment and theragnostic of a range of diseases and conditions associated with MERTK, such as cancer and MS. There is therefore an ongoing need for improved diagnostic and therapeutic approaches to management of MS, which at least partially addresses one or more of the above-mentioned short-comings or provides a useful alternative. There is also an ongoing need for new compounds which inhibit MERTK, which at least partially addresses one or more of the above-mentioned short-comings or provides a useful alternative. There is also an ongoing need for new specific MERTK radioligands, which at least partially addresses one or more of the above-mentioned short-comings or provides a useful alternative. A reference herein to a patent document or other matter which is given as prior art is not to be taken as an admission that the document or matter was known or that the information it contains was part of the common general knowledge as at the priority date of any of the claims. Summary of Invention In a first aspect the present invention provides a compound of Formula (I):
Figure imgf000004_0001
Formula (I) wherein Y is -(-C(O)-NH-)-or -(-C(O)-NH-(CH2))-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R1 is selected from the group consisting of optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; R2 is independently selected from the group consisting of H, optionally substituted C1-6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of H, OH, optionally substituted C1-C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; R4 is a cyclic group selected from C6-C18aryl, C2-C18heteroaryl, C3-C12cycloalkyl or C1- C12heterocycloalkyl; and is optionally substituted up to four times with R5 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2- C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3- C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2- C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1- C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; wherein each R5 is optionally further substituted with R6 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3-C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; and wherein R9 and R10 are independently selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C1-C12heterocycloalkyl, optionally substituted C1-C12alkylamino, optionally substituted C5-C7lactone, optionally substituted C4-C7lactam, and optionally substituted C1-C12haloalkyl; or wherein R9 and R10 when taken together with the atoms to which they are attached form an optionally substituted C3-C12cycloalkyl or an optionally substituted C1-C12heterocycloalkyl; or a pharmaceutically acceptable salt thereof. In a further aspect the present invention provides a compound of Formula (Ia):
Figure imgf000006_0001
Formula (Ia) wherein Y is -(-C(O)-NH-)-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R1 is selected from the group consisting of optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; R2 is independently selected from the group consisting of H, optionally substituted C1-6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of H, OH, optionally substituted C1-C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; R4 is a cyclic group selected from C6-C18aryl, C2-C18heteroaryl, C3-C12cycloalkyl or C1- C12heterocycloalkyl; and is optionally substituted up to four times with R5 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2- C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3- C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2- C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; wherein each R5 is optionally further substituted with R6 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3-C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; and wherein R9 and R10 are independently selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C1- C12heterocycloalkyl, optionally substituted C1-C12alkylamino, optionally substituted C5- C7lactone, optionally substituted C4-C7lactam, and optionally substituted C1-C12haloalkyl; or wherein R9 and R10 when taken together with the atoms to which they are attached form an optionally substituted C3-C12cycloalkyl or an optionally substituted C1-C12heterocycloalkyl; or a pharmaceutically acceptable salt thereof. Compounds of Formula (I) have shown potent activity as inhibitors of MERTK. Compounds of the present invention may therefore be used in the treatment of diseases and conditions associated with MERTK. Additionally, compounds of the present invention have shown improved CNS penetrance compared with previously described compounds. Particularly, compounds having a beta-fluoro group as defined in Formula (I) show significant improvements in potency and selectivity for MERTK, as well as improved CNS penetrance. In a further aspect the present invention, the compounds of Formula (I) are radio- labelled, preferably with an 18F group. Compounds of the present invention may therefore be used as radio-labelled ligands of MERTK, which can be used in radio imaging methods as well as methods for the identification, diagnosis and management of diseases and conditions associated with MERTK. Therefore, in a further aspect the present invention provides a method of imaging body tissue comprising: a) applying an imaging composition to a subject, wherein said composition comprises a radio- labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom. In a yet further aspect, the present invention provides a method of identifying, selecting, or diagnosing a disease state in a subject, comprising the steps of: a) applying an imaging composition to a body tissue of the subject, wherein said composition comprises a radio-labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom, wherein the image indicates a level of microglial subtype present in the body tissue; c) comparing the level with a reference level to determine an increased or decreased level of microglial subtype present in the body tissue, wherein the increased or decreased level of microglial subtype present in the body tissue compared to the reference level is indicative of a disease state or a stage of development of a disease state. Preferably, the disease state is a condition associated with MERTK. More preferably, the disease state is a neuroinflammatory central-nervous system disease. Even more preferably, the disease state is multiple sclerosis (MS). Preferably, the imaging is imaging is Positron Emission Tomography imaging. A PET radiotracer specific for MERTK would provide the ability to distinguish pathogenic and reparative processes that underpin conditions such as MS, when compared with compared with currently available radiotracers target the translocator protein (TSPO). This advance has the potential to transform current practice in managing MS. In a still further aspect, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt of a of the invention, and a pharmaceutically acceptable excipient. In yet a further aspect the present invention provides compounds and methods for the identification, treatment, prevention, or amelioration of a condition associated with MERTK. Preferably, the condition is MS. Further aspects of the invention appear below in the detailed description of the invention. Brief Description of Drawings Embodiments of the invention will herein be illustrated by way of example only with reference to the accompanying drawings in which: Figure 1 is a plot of plasma and brain (total) concentrations versus time (Compound 5-02) in male Swiss outbred mice following IV administration (3.7 mg/kg). Figure 2 is a plot of unbound concentrations of Compound 5-02 in plasma and brain of male Swiss outbred mice following IV administration (3.7 mg/kg). Data are presented as the mean ± SD of n=3 mice at each time point. Figure 3 is a set of images of summed 0-10 uptake (standard uptake value) images in a control (Left image) and cuprizone challenged (Right image, MS model) mouse. Figure 4 is a time activity curve (whole brain) in a control (black) and cuprizone- exposed (grey) mouse. Figure 5 is a plot of plasma concentrations of Compound 11-02 in male Swiss outbred mice following IV administration at 4.6 mg/kg. Data are presented as the mean ± SD of n=3 mice at each time point. Figure 6 is a plot of unbound concentrations of Compound 11-02 in plasma and brain of male Swiss outbred mice following IV administration (4.6 mg/kg). Data are presented as the mean ± SD of n=3 mice at each time point. Detailed Description Before describing the present invention in detail, it is to be understood that the terminology used herein is for the purpose of describing embodiments only and is not intended to be limiting. Definitions In this specification, a number of terms are used that are well known to a skilled addressee. Nevertheless, for the purposes of clarity a number of terms will be defined. Unless specifically defined otherwise, all technical and scientific terms shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art. As used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the content clearly dictates otherwise. . Throughout the description and claims of the specification the word “comprise” and variations of the word, such as “comprising” and “comprises”, is not intended to exclude other additives, components, integers or steps. The term "and/or" as used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. For example "A and/or B" is to be taken as specific disclosure of each of (i) A, (ii) B and (iii) A and B, just as if each is set out individually herein. As used herein, the term "subject" shall be taken to mean any mammalian animal, preferably a human. As used herein, "disease", "disorder", "condition" and the like, as they relate to a subject's health, are used interchangeably and have meanings ascribed to each and all such terms. The term ‘inhibit” and variations thereof such as “inhibiting” means to prevent, block or reduce the function of the thing being inhibited. The term does not require complete inhibition with a reduction of activity at least 50% being considered inhibition. In the definitions of a number of substituents below it is stated that “the group may be a terminal group or a bridging group”. This is intended to signify that the use of the term is intended to encompass the situation where the group is a linker between two other portions of the molecule as well as where it is a terminal moiety. Using the term alkyl as an example, some publications would use the term “alkylene” for a bridging group and hence in these other publications there is a distinction between the terms “alkyl” (terminal group) and “alkylene” (bridging group). In the present application no such distinction is made and most groups may be either a bridging group or a terminal group. As used herein, the term “unsubstituted” means that there is no substituent or that the only substituents are hydrogen. The term "optionally substituted" as used throughout the specification denotes that the group may or may not be further substituted or fused (so as to form a condensed polycyclic system), with one or more non-hydrogen substituent groups. In certain embodiments the substituent groups are one or more groups independently selected from the group consisting of halogen, =O, =S, -CN, -NO2, -CF3, -OCF3, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyloxycycloalkyl, alkyloxyheterocycloalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkyloxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, phenoxy, benzyloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, -C(=O)OH, -C(=O)Re, -C(=O)ORe, C(=O)NReRf, C(=NOH)Re, C(=NRe)NRfRg, NReRf, NReC(=O)Rf, NReC(=O)ORf, NReC(=O)NRfRg, NReC(=NRf)NRgRh, NReSO2Rf, -SRe, SO2NReRf, -ORe , OC(=O)NReRf, OC(=O)Re and acyl, wherein Re, Rf, Rg and Rh are each independently selected from the group consisting of H, C1-C12alkyl, C1-C12haloalkyl, C2-C12alkenyl, C2-C12alkynyl, C1- C10heteroalkyl, C3-C12cycloalkyl, C3-C12cycloalkenyl, C1-C12heterocycloalkyl, C1- C12heterocycloalkenyl, C6-C18aryl, C1-C18heteroaryl, and acyl, or any two or more of Ra, Rb, Rc and Rd, when taken together with the atoms to which they are attached form a heterocyclic ring system with 3 to 12 ring atoms. In some embodiments each optional substituent is independently selected from the group consisting of: halogen, =O, =S, -CN, -NO2, -CF3, -OCF3, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, aminoalkyl, -COOH, -SH, and acyl. Examples of particularly suitable optional substituents include F, Cl, Br, I, CH3, CH2CH3, OH, OCH3, CF3, OCF3, NO2, NH2, and CN. "Acyl" means an R-C(=O)- group in which the R group may be an alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group as defined herein. Examples of acyl include acetyl and benzoyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the carbonyl carbon. "Acylamino" means an R-C(=O)-NH- group in which the R group may be an alkyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom. "Alkenyl" as a group or part of a group denotes an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched preferably having 2-12 carbon atoms, more preferably 2-10 carbon atoms, most preferably 2- 6 carbon atoms, in the normal chain. The group may contain a plurality of double bonds in the normal chain and the orientation about each is independently E or Z. The alkenyl group is preferably a 1-alkenyl group. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl and nonenyl. The group may be a terminal group or a bridging group. "Alkenyloxy" refers to an alkenyl-O- group in which alkenyl is as defined herein. Preferred alkenyloxy groups are C1-C6 alkenyloxy groups. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. "Alkyl" as a group or part of a group refers to a straight or branched aliphatic hydrocarbon group, preferably a C1–C12 alkyl, more preferably a C1-C10 alkyl, most preferably C1-C6 unless otherwise noted. Examples of suitable straight and branched C1-C6 alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, t-butyl, hexyl, and the like. The group may be a terminal group or a bridging group. "Alkylamino" includes both mono-alkylamino and dialkylamino, unless specified. "Mono-alkylamino" means an Alkyl-NH- group, in which alkyl is as defined herein. "Dialkylamino" means a (alkyl)2N- group, in which each alkyl may be the same or different and are each as defined herein for alkyl. The alkyl group is preferably a C1-C6alkyl group. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom. "Alkylaminocarbonyl" refers to a group of the formula (Alkyl)x(H)yNC(=O)- in which alkyl is as defined herein, x is 1 or 2, and the sum of X+Y =2. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the carbonyl carbon. "Alkyloxy" refers to an alkyl-O- group in which alkyl is as defined herein. Preferably the alkyloxy is a C1-C6alkyloxy. Examples include, but are not limited to, methoxy and ethoxy. The group may be a terminal group or a bridging group. "Alkyloxyalkyl" refers to an alkyloxy-alkyl- group in which the alkyloxy and alkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group. "Alkyloxyaryl" refers to an alkyloxy-aryl- group in which the alkyloxy and aryl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the aryl group. "Alkyloxycarbonyl" refers to an alkyl-O-C(=O)- group in which alkyl is as defined herein. The alkyl group is preferably a C1-C6 alkyl group. Examples include, but are not limited to, methoxycarbonyl and ethoxycarbonyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the carbonyl carbon. "Alkyloxycycloalkyl" refers to an alkyloxy-cycloalkyl- group in which the alkyloxy and cycloalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the cycloalkyl group. "Alkyloxyheteroaryl" refers to an alkyloxy-heteroaryl- group in which the alkyloxy and heteroaryl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroaryl group. "Alkyloxyheterocycloalkyl" refers to an alkyloxy-heterocycloalkyl- group in which the alkyloxy and heterocycloalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heterocycloalkyl group. "Alkylsulfinyl" means an alkyl-S-(=O)- group in which alkyl is as defined herein. The alkyl group is preferably a C1-C6 alkyl group. Exemplary alkylsulfinyl groups include, but not limited to, methylsulfinyl and ethylsulfinyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom. "Alkylsulfonyl" refers to an alkyl-S(=O)2- group in which alkyl is as defined above. The alkyl group is preferably a C1-C6alkyl group. Examples include, but not limited to methylsulfonyl and ethylsulfonyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom. "Alkynyl” as a group or part of a group means an aliphatic hydrocarbon group containing a carbon-carbon triple bond and which may be straight or branched preferably having from 2-12 carbon atoms, more preferably 2-10 carbon atoms, more preferably 2-6 carbon atoms in the normal chain. Exemplary structures include, but are not limited to, ethynyl and propynyl. The group may be a terminal group or a bridging group. "Alkynyloxy" refers to an alkynyl-O- group in which alkynyl is as defined herein. Preferred alkynyloxy groups are C1-C6alkynyloxy groups. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. "Aminoalkyl" means an NH2-alkyl- group in which the alkyl group is as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group. "Aminosulfonyl" means an NH2-S(=O)2- group. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom. "Aryl" as a group or part of a group denotes (i) an optionally substituted monocyclic, or fused polycyclic, aromatic carbocycle (ring structure having ring atoms that are all carbon) preferably having from 5 to 12 atoms per ring. Examples of aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety in which a phenyl and a C5-7cycloalkyl or C5-7cycloalkenyl group are fused together to form a cyclic structure, such as tetrahydronaphthyl, indenyl or indanyl. The group may be a terminal group or a bridging group. Typically, an aryl group is a C6-C18 aryl group. "Arylalkenyl" means an aryl-alkenyl- group in which the aryl and alkenyl are as defined herein. Exemplary arylalkenyl groups include phenylallyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkenyl group; "Arylalkyl" means an aryl-alkyl- group in which the aryl and alkyl moieties are as defined herein. Preferred arylalkyl groups contain a C1-5alkyl moiety. Exemplary arylalkyl groups include benzyl, phenethyl, 1-naphthalenemethyl and 2-naphthalenemethyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group. “Arylalkyloxy" refers to an aryl-alkyl-O- group in which the alkyl and aryl are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the oxygen atom. "Arylamino" includes both mono-arylamino and di-arylamino unless specified. Mono-arylamino means a group of formula arylNH-, in which aryl is as defined herein. Di-arylamino means a group of formula (aryl)2N- where each aryl may be the same or different and are each as defined herein for aryl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom. "Arylheteroalkyl" means an aryl-heteroalkyl- group in which the aryl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group. "Aryloxy" refers to an aryl–O- group in which the aryl is as defined herein. Preferably the aryloxy is a C6-C18aryloxy, more preferably a C6-C10aryloxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. "Arylsulfonyl" means an aryl-S(=O)2- group in which the aryl group is as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom. A “bond” is a linkage between atoms in a compound or molecule. The bond may be a single bond, a double bond, or a triple bond. "Cycloalkenyl" means a non-aromatic monocyclic or multicyclic ring system containing at least one carbon-carbon double bond and preferably having from 5-10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl or cycloheptenyl. The cycloalkenyl group may be substituted by one or more substituent groups. A cycloalkenyl group typically is a C3-C12 alkenyl group. The group may be a terminal group or a bridging group. "Cycloalkyl" refers to a saturated monocyclic or fused or spiro polycyclic, carbocycle preferably containing from 3 to 9 carbons per ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like, unless otherwise specified. It includes monocyclic systems such as cyclopropyl and cyclohexyl, bicyclic systems such as decalin, and polycyclic systems such as adamantane. A cycloalkyl group typically is a C3-C12 alkyl group. The group may be a terminal group or a bridging group. "Cycloalkylalkyl" means a cycloalkyl-alkyl- group in which the cycloalkyl and alkyl moieties are as defined herein. Exemplary monocycloalkylalkyl groups include cyclopropylmethyl, cyclopentylmethyl, cyclohexylmethyl and cycloheptylmethyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group. "Cycloalkylalkenyl" means a cycloalkyl-alkenyl- group in which the cycloalkyl and alkenyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkenyl group. "Cycloalkylheteroalkyl" means a cycloalkyl-heteroalkyl- group in which the cycloalkyl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group. "Cycloalkyloxy" refers to a cycloalkyl-O- group in which cycloalkyl is as defined herein. Preferably the cycloalkyloxy is a C1-C6cycloalkyloxy. Examples include, but are not limited to, cyclopropanoxy and cyclobutanoxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. "Cycloalkenyloxy" refers to a cycloalkenyl-O- group in which the cycloalkenyl is as defined herein. Preferably the cycloalkenyloxy is a C1-C6cycloalkenyloxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. “Haloalkyl” refers to an alkyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom selected from the group consisting of fluorine, chlorine, bromine and iodine. A haloalkyl group typically has the formula CnH(2n+1- m)Xm wherein each X is independently selected from the group consisting of F, Cl, Br and I. In groups of this type n is typically from 1 to 10, more preferably from 1 to 6, most preferably 1 to 3. m is typically 1 to 6, more preferably 1 to 3. Examples of haloalkyl include fluoromethyl, difluoromethyl and trifluoromethyl. “Haloalkenyl” refers to an alkenyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom independently selected from the group consisting of F, Cl, Br and I. “Haloalkynyl” refers to an alkynyl group as defined herein in which one or more of the hydrogen atoms has been replaced with a halogen atom independently selected from the group consisting of F, Cl, Br and I. "Halogen" represents chlorine, fluorine, bromine or iodine. “Heteroalkyl" refers to a straight- or branched-chain alkyl group preferably having from 2 to 12 carbons, more preferably 2 to 6 carbons in the chain, in which one or more of the carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatomic group selected from S, O, P and NR’ where R’ is selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C1-C18heteroaryl. Exemplary heteroalkyls include alkyl ethers, secondary and tertiary alkyl amines, amides, alkyl sulfides, and the like. Examples of heteroalkyl also include hydroxyC1-C6alkyl, C1-C6alkyloxyC1- C6alkyl, aminoC1-C6alkyl, C1-C6alkylaminoC1-C6alkyl, and di(C1-C6alkyl)aminoC1-C6alkyl. The group may be a terminal group or a bridging group. "Heteroalkyloxy" refers to a heteroalkyl-O- group in which heteroalkyl is as defined herein. Preferably the heteroalkyloxy is a C2-C6heteroalkyloxy. The group may be a terminal group or a bridging group. "Heteroaryl" either alone or part of a group refers to groups containing an aromatic ring (preferably a 5 or 6 membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring with the remainder of the ring atoms being carbon atoms. Suitable heteroatoms include nitrogen, oxygen and sulphur. The group may be a monocyclic or bicyclic heteroaryl group. Examples of heteroaryl include thiophene, benzothiophene, benzofuran, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, naphtho[2,3- b]thiophene, furan, isoindolizine, xantholene, phenoxatine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, tetrazole, indole, isoindole, 1H-indazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, cinnoline, carbazole, phenanthridine, acridine, phenazine, thiazole, isothiazole, phenothiazine, oxazole, isooxazole, furazane, phenoxazine, 2-, 3- or 4- pyridyl, 2-, 3-, 4-, 5-, or 8- quinolyl, 1-, 3-, 4-, or 5- isoquinolinyl 1-, 2-, or 3- indolyl, and 2-, or 3-thienyl. A heteroaryl group is typically a C1- C18heteroaryl group. The group may be a terminal group or a bridging group. "Heteroarylalkyl" means a heteroaryl-alkyl group in which the heteroaryl and alkyl moieties are as defined herein. Preferred heteroarylalkyl groups contain a lower alkyl moiety. Exemplary heteroarylalkyl groups include pyridylmethyl. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkyl group. "Heteroarylalkenyl" means a heteroaryl-alkenyl- group in which the heteroaryl and alkenyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group it is bonded to the remainder of the molecule through the alkenyl group. "Heteroarylheteroalkyl" means a heteroaryl-heteroalkyl- group in which the heteroaryl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group. "Heteroaryloxy" refers to a heteroaryl-O- group in which the heteroaryl is as defined herein. Preferably the heteroaryloxy is a C1-C18heteroaryloxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. “Heterocyclic” refers to saturated, partially unsaturated or fully unsaturated monocyclic, bicyclic or polycyclic ring system containing at least one heteroatom selected from the group consisting of nitrogen, sulfur and oxygen as a ring atom. Examples of heterocyclic moieties include heterocycloalkyl, heterocycloalkenyl and heteroaryl. "Heterocycloalkenyl" refers to a heterocycloalkyl group as defined herein but containing at least one double bond. A heterocycloalkenyl group typically is a C2- C12heterocycloalkenyl group. The group may be a terminal group or a bridging group. "Heterocycloalkyl" refers to a saturated monocyclic, bicyclic, or polycyclic ring containing at least one heteroatom selected from nitrogen, sulfur, oxygen, preferably from 1 to 3 heteroatoms in at least one ring. Each ring is preferably from 3 to 10 membered, more preferably 4 to 7 membered. Examples of suitable heterocycloalkyl substituents include pyrrolidyl, tetrahydrofuryl, tetrahydrothiofuranyl, piperidyl, piperazyl, tetrahydropyranyl, morphilino, 1,3-diazapane, 1,4-diazapane, 1,4-oxazepane, and 1,4-oxathiapane. A heterocycloalkyl group typically is a C2-C12heterocycloalkyl group. The group may be a terminal group or a bridging group. "Heterocycloalkylalkyl" refers to a heterocycloalkyl-alkyl- group in which the heterocycloalkyl and alkyl moieties are as defined herein. Exemplary heterocycloalkylalkyl groups include (2-tetrahydrofuryl)methyl, (2-tetrahydrothiofuranyl) methyl. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkyl group. "Heterocycloalkylalkenyl" refers to a heterocycloalkyl-alkenyl- group in which the heterocycloalkyl and alkenyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the alkenyl group. "Heterocycloalkylheteroalkyl" means a heterocycloalkyl-heteroalkyl- group in which the heterocycloalkyl and heteroalkyl moieties are as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the heteroalkyl group. "Heterocycloalkyloxy" refers to a heterocycloalkyl-O- group in which the heterocycloalkyl is as defined herein. Preferably the heterocycloalkyloxy is a C1- C6heterocycloalkyloxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. "Heterocycloalkenyloxy" refers to a heterocycloalkenyl-O- group in which heterocycloalkenyl is as defined herein. Preferably the Heterocycloalkenyloxy is a C1-C6 Heterocycloalkenyloxy. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the oxygen atom. “Hydroxyalkyl” refers to an alkyl group as defined herein in which one or more of the hydrogen atoms has been replaced with an OH group. A hydroxyalkyl group typically has the formula CnH(2n+1-x)(OH)x. In groups of this type n is typically from 1 to 10, more preferably from 1 to 6, most preferably 1 to 3. x is typically 1 to 6, more preferably 1 to 3. "Sulfinyl" means an R-S(=O)- group in which the R group may be OH, alkyl, cycloalkyl, heterocycloalkyl; aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom. "Sulfinylamino" means an R-S(=O)-NH- group in which the R group may be OH, alkyl, cycloalkyl, heterocycloalkyl; aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom. "Sulfonyl" means an R-S(=O)2- group in which the R group may be OH, alkyl, cycloalkyl, heterocycloalkyl; aryl or heteroaryl group as defined herein. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the sulfur atom. "Sulfonylamino" means an R-S(=O)2-NH- group. The group may be a terminal group or a bridging group. If the group is a terminal group, it is bonded to the remainder of the molecule through the nitrogen atom. It is understood that included in the family of compounds of Formula (I) are isomeric forms including diastereoisomers, enantiomers, tautomers, and geometrical isomers in "E" or "Z" configurational isomer or a mixture of E and Z isomers. It is also understood that some isomeric forms such as diastereomers, enantiomers, and geometrical isomers can be separated by physical and/or chemical methods and by those skilled in the art. For those compounds where there is the possibility of geometric isomerism the applicant has drawn the isomer that the compound is thought to be although it will be appreciated that the other isomer may be the correct structural assignment. Some of the compounds of the disclosed embodiments may exist as single stereoisomers, racemates, and/or mixtures of enantiomers and /or diastereomers. All such single stereoisomers, racemates and mixtures thereof, are intended to be within the scope of the subject matter described and claimed. Additionally, Formula (I) is intended to cover, where applicable, solvated as well as unsolvated forms of the compounds. Thus, each formula includes compounds having the indicated structure, including the hydrated as well as the non-hydrated forms. The term "pharmaceutically acceptable salts" refers to salts that retain the desired biological activity of the above-identified compounds and include pharmaceutically acceptable acid addition salts and base addition salts. Suitable pharmaceutically acceptable acid addition salts of compounds of Formula (I) may be prepared from an inorganic acid or from an organic acid. Examples of such inorganic acids are hydrochloric, sulfuric, and phosphoric acid. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic and sulfonic classes of organic acids, examples of which are formic, acetic, propanoic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, fumaric, maleic, alkyl sulfonic, arylsulfonic. In a similar vein base addition salts may be prepared by ways well known in the art using organic or inorganic bases. Example of suitable organic bases include simple amines such as methylamine, ethylamine, triethylamine and the like. Examples of suitable inorganic bases include NaOH, KOH, and the like. Additional information on pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Co., Easton, PA 1995. In the case of agents that are solids, it is understood by those skilled in the art that the inventive compounds, agents and salts may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulae. The term "therapeutically effective amount" or "effective amount" is an amount sufficient to effect beneficial or desired clinical results. An effective amount can be administered in one or more administrations. An effective amount is typically sufficient to palliate, ameliorate, stabilize, reverse, slow or delay the progression of the disease state. Compounds of the Invention As outlined above, the compounds of the present invention have the Formula (I):
Figure imgf000021_0001
Formula (I) wherein Y is -(-C(O)-NH-)- or -(-C(O)-NH-(CH2))-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R1 is selected from the group consisting of optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; R2 is independently selected from the group consisting of H, optionally substituted C1-6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of H, OH, optionally substituted C1-C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; R4 is a cyclic group selected from C6-C18aryl, C2-C18heteroaryl, C3-C12cycloalkyl or C1- C12heterocycloalkyl; and is optionally substituted up to four times with R5 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2- C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3- C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2- C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1- C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; wherein each R5 is optionally further substituted with R6 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3-C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; and wherein R9 and R10 are independently selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C1- C12heterocycloalkyl, optionally substituted C1-C12alkylamino, optionally substituted C5- C7lactone, optionally substituted C4-C7lactam, and optionally substituted C1-C12haloalkyl; or wherein R9 and R10 when taken together with the atoms to which they are attached form an optionally substituted C3-C12cycloalkyl or an optionally substituted C1-C12heterocycloalkyl; or a pharmaceutically acceptable salt thereof. In some embodiments the compound of Formula (I) is the compound of Formula (Ia):
Figure imgf000023_0001
Formula (Ia) wherein Y is -(-C(O)-NH-)-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R1 is selected from the group consisting of optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; R2 is independently selected from the group consisting of H, optionally substituted C1-6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of H, OH, optionally substituted C1-C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; R4 is a cyclic group selected from C6-C18aryl, C2-C18heteroaryl, C3-C12cycloalkyl or C1- C12heterocycloalkyl; and is optionally substituted up to four times with R5 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2- C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3- C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2- C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1- C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; wherein each R5 is optionally further substituted with R6 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3-C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; and wherein R9 and R10 are independently selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C1- C12heterocycloalkyl, optionally substituted C1-C12alkylamino, optionally substituted C5- C7lactone, optionally substituted C4-C7lactam, and optionally substituted C1-C12haloalkyl; or wherein R9 and R10 when taken together with the atoms to which they are attached form an optionally substituted C3-C12cycloalkyl or an optionally substituted C1-C12heterocycloalkyl; or a pharmaceutically acceptable salt thereof. In some embodiments of the compound of Formula (I), n is 0. This provides compounds of Formula (II):
Figure imgf000025_0001
Formula (II) wherein dashed lines indicate an optional methylene bridge; and R1, R2, R3, and R4, are defined as above; or a pharmaceutically acceptable salt thereof. In some embodiments of the compound of Formula (I), Y is -(-C(O)-NH-)- and n is 1. This provides compounds of Formula (IIb):
Figure imgf000025_0002
Formula (IIb) wherein dashed lines indicate an optional methylene bridge; and R1, R2, R3, and R4, are defined as above; or a pharmaceutically acceptable salt thereof. In some embodiments of the compound of Formula (I), R4 is an optionally substituted 5-7-membered heterocyclic group of the formula:
Figure imgf000026_0001
wherein X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; m is 0 to 2; where valency allows, R7 is selected from the group consisting of H, optionally substituted C1- C12alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C1-C18heteroaryl; each R5 is independently defined as above. In some embodiments, the compound of Formula (I) has a structure of Formula (III):
Figure imgf000026_0002
Formula (III) wherein R1 is selected from the group consisting of optionally substituted C1-6alkyl or optionally substituted C3-8cycloalkyl; X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; m is 0 to 2; where valency allows, R7 is selected from the group consisting of H, optionally substituted C1- C12alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C1-C18heteroaryl; R2 is selected from the group consisting of H, optionally substituted C1-6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3- 8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of OH, optionally substituted C1-C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; each R5 is independently defined as above. In some embodiments of the compound of Formula (III), m is 0. In some embodiments of the compound of Formula (III), m is 1. In some embodiments of the compound of Formula (III), m is 2. This provides compounds of Formula (I) where R4 is an optionally substituted heterocyclic group of the formula:
Figure imgf000027_0001
wherein X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; where valency allows, R7 is selected from the group consisting of H, optionally substituted C1- C12alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C1-C18heteroaryl; each R5 is independently defined as above. In some preferred embodiments of the compound of Formula (I), the beta-fluoro group has S-chirality. An (S)-beta-fluoro group particularly shows unexpected improvements in potency and selectivity for MERTK, as well as improved CNS penetrance over previously described compounds. In some embodiments of the compound of Formula (III), the beta-fluoro group has S-chirality, R2 is H, R3 is OH, R4 is attached at the 5-position, m is 1, X is N, and the dashed lines represent double bonds. This provides compounds of Formula (I) having a structure of Formula (IV):
Figure imgf000028_0001
Formula (IV) wherein R1 is selected from the group consisting of optionally substituted C1-6alkyl; and each R5 is independently defined as above; or a pharmaceutically acceptable salt thereof. In some preferred embodiments of the compound of Formula (I), R1 is optionally substituted methyl or optionally substituted ethyl. In some preferred embodiments of the compound of Formula (I), R3 is OH or halogen. In some preferred embodiments of the compound of Formula (I), R2 is H. In some preferred embodiments of the compound of Formula (I), R5 is independently selected from the group consisting of H, CN, halogen, optionally substituted C1- C6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C3-C12cycloalkyl, an optionally substituted C1-C12heterocycloalkyl, optionally substituted C1-C12alkylsulfonyl, optionally substituted C1-C12alkylamino, and optionally substituted C1-C12haloalkyl. In some further preferred embodiments of the compound of Formula (I), R6 is independently selected from the group consisting of H, halogen, optionally substituted C1- C6alkyl, optionally substituted C3-C12cycloalkyl, an optionally substituted C1- C12heterocycloalkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1- C12alkylsulfonyl, optionally substituted C1-C12alkylamino, and optionally substituted C1- C12haloalkyl. In some embodiments of the compound of Formula (I), R4 is selected from the from the group consisting of: ,
Figure imgf000029_0001
wherein R5 is as defined herein. In some preferred embodiments of the compound of Formula (I), R4 has a structure selected from the group consisting of: ,
Figure imgf000029_0002
Figure imgf000030_0001
, , , ,
Figure imgf000031_0001
, , , , , and . In some embodiments of the compound of Formula (I), the compound has a structure as shown in Table 1, or a pharmaceutically acceptable salt thereof. Table 1
Figure imgf000031_0002
Figure imgf000032_0001
Figure imgf000033_0001
Figure imgf000034_0001
Figure imgf000035_0001
Radiochemical and Pharmaceutical Methods In some embodiments of the present invention, the compound of Formula (I) is CNS penetrant. “CNS penetrant” means that the compound can penetrate the blood-brain barrier (BBB) and exhibit activity in the central nervous system. Penetration of the BBB may be by any mechanism such as passive transport into the CNS, active transport, efflux, and metabolism. CNS penetrance can be indicated by various parameters described in the art such as Blood Brain Barrier (BBB) Score (M. Gupta, H.J. Lee, C.J. Barden, D.F. Weaver, The Blood-Brain Barrier (BBB) Score, J Med Chem, 62 (2019) 9824-9836) or the CNS PET multi- parameter optimization (MPO) tool (L. Zhang, A. Villalobos, Strategies to facilitate the discovery of novel CNS PET ligands, EJNMMI Radiopharm Chem, 1 (2017) 13). In some embodiments of the present invention, the compound of Formula (I) is radio-labelled to provide a radio-tracer compound. In preferred embodiments of the present invention, the compound of Formula (I) is radio-labelled with 18F. The skilled person will appreciate that the radio-label may be attached to any part of the molecule. For example, the beta-fluoro group may be 18F. In further preferred embodiments of the present invention, in the compound of Formula (I) at least one of R3 is radio-labelled, preferably with 18F. Radiolabelling of the compound of Formula (I) can be performed particularly by the synthesis methods described in the accompanying examples. The skilled person will appreciate that other radio-labelling methods are available and within the scope of the present invention. In an aspect, the present invention provides an imaging composition comprising a radio-labelled compound of the invention. In a further aspect, the present invention provides a method of imaging body tissue comprising: a) applying an imaging composition to a subject, wherein said composition comprises a radio-labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom. In a preferred embodiment, the body tissue is selected from the group consisting of the brain and spinal cord. In another preferred embodiment, the imaging method is Positron Emission Tomography imaging. In a further aspect, the present invention provides a method of identifying, selecting, or diagnosing a disease state in a subject, comprising the steps of: a) applying an imaging composition to a body tissue of the subject, wherein said composition comprises a radio-labelled compound of the invention; b) detecting radiation emitted by said composition and forming an image therefrom, wherein the image indicates a level of microglial subtype present in the body tissue; c) comparing the level with a reference level to determine an increased or decreased level of microglial subtype present in the body tissue, wherein the increased or decreased level of microglial subtype present in the body tissue compared to the reference level is indicative of a disease state or a stage of development of a disease state. In a preferred embodiment, the reference level of microglial subtype present in the body tissue is the level of expression of microglial subtype present in the body tissue from a normal subject. Preferably, the microglial subtype present in the body tissue is M2 microglia. In another preferred embodiment, imaging method is Positron Emission Tomography imaging. In a preferred embodiment, the disease state is a condition associated with MERTK. In a further preferred embodiment, the disease state is a neuroinflammatory central- nervous system disease. In a more preferred embodiment, the disease state is multiple sclerosis (MS). In another preferred embodiment, the disease state is cancer. In some embodiments, the method of identifying, selecting, or diagnosing a disease state in a subject, further comprises administering to the subject a therapeutically effective amount of an anti-disease state therapeutic. The compounds of the invention are ligands of MERTK and therefore have the ability to inhibit these enzymes. The ability to inhibit the enzymes may be a result of the compounds acting directly and solely on the enzyme to modulate/potentiate biological activity. However, it is understood that the compounds may also act at least partially on other factors associated with the activity of the enzyme. The inhibition of MERTK may be carried out in any of a number of ways known in the art. For example if inhibition in vitro is desired an appropriate amount of the compound may be added to a solution containing the MERTK. In circumstances where it is desired to inhibit MERTK in a mammal, the inhibition of the MERTK typically involves administering the compound to a mammal containing the compound of the invention. In a further aspect, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt of a compound of the invention, and a pharmaceutically acceptable excipient. In a further aspect, the present invention provides a compound of the invention for use in the identification, treatment, prevention, or amelioration of a condition in a mammal. In a preferred embodiment, the condition is associated with MERTK. In a yet further aspect, the present invention provides a method of identifying, preventing, treating, or ameliorating a condition in a mammal, said method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of the invention. In a preferred embodiment, the condition is associated with MERTK. In a yet further aspect, the present invention provides a use of a compound of the invention in the manufacture of a medicament for the identification, treatment, prevention, or amelioration of a condition in a mammal. In a preferred embodiment, the condition is associated with MERTK. In some embodiments the condition is selected from the group consisting of neuroinflammatory central-nervous system disease and cancer. In some embodiments, the condition is neuroinflammatory central-nervous system disease. In some embodiments, the neuroinflammatory central-nervous system disease is selected from the list consisting of multiple sclerosis (MS), traumatic brain injury, epilepsy, Alzheimer’s disease (AD), Parkinson's disease (PD), Huntington's disease (HD), Multiple system atrophy (MSA), Amyotrophic lateral sclerosis (ALS), stroke, neuromyelitis optica spectrum disorder (NMOSD), acute disseminated encephalomyelitis (ADEM) and myelin oligodendrocyte glycoprotein (MOG) encephalomyelitis, and the leukodystrophies (including Aicardi-Goutières syndrome, adrenoleukodystrophy (ALD), Alexander disease, Canavan disease, cerebrotendinous xanthomatosis, Krabbé disease, metachromatic leukodystrophy (MLD), Niemann-Pick disease, Pelizaeus-Merzbacher disease (PMD), and childhood ataxia with central nervous system hypomyelination). Preferably, the condition is multiple sclerosis (MS). In some embodiments, the condition is cancer. In some embodiments, the cancer is selected from the list consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukaemia (AML), melanoma, breast cancer, lung cancer, colon cancer, prostate cancer, gastric cancer, non-small cell lung cancer (NSCLC), glioblastoma, liver cancer, kidney cancer, ovarian cancer, uterine cancer, and brain cancer. Administration of compounds within Formula (I) to humans can be by any of the accepted modes for enteral administration such as oral or rectal, or by parenteral administration such as subcutaneous, intramuscular, intravenous and intradermal routes. Injection can be bolus or via constant or intermittent infusion. The active compound is typically included in a pharmaceutically acceptable carrier or diluent and in an amount sufficient to deliver to the patient a therapeutically effective dose. In using the compounds of the invention, they can be administered in any form or mode which makes the compound bioavailable. One skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the compound selected, the condition to be treated, the stage of the condition to be treated and other relevant circumstances. We refer the reader to Remingtons Pharmaceutical Sciences, 19th edition, Mack Publishing Co. (1995) for further information. The compounds of the present invention can be administered alone or in the form of a pharmaceutical composition in combination with a pharmaceutically acceptable carrier, diluent or excipient. The compounds of the invention, while effective themselves, are typically formulated and administered in the form of their pharmaceutically acceptable salts as these forms are typically more stable, more easily crystallised and have increased solubility. The compounds are, however, typically used in the form of pharmaceutical compositions which are formulated depending on the desired mode of administration. As such in some embodiments the present invention provides a pharmaceutical composition including a compound of Formula (I) and a pharmaceutically acceptable carrier, diluent or excipient. The compositions are prepared in manners well known in the art. The invention in other embodiments provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the invention. In such a pack or kit can be found a container having a unit dosage of the agent(s). The kits can include a composition comprising an effective agent either as concentrates (including lyophilized compositions), which can be diluted further prior to use or they can be provided at the concentration of use, where the vials may include one or more dosages. Conveniently, in the kits, single dosages can be provided in sterile vials so that the physician can employ the vials directly, where the vials will have the desired amount and concentration of agent(s). Associated with such container(s) can be various written materials such as instructions for use, or a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. The compounds of the invention may be used or administered in combination with one or more additional drug(s) for the treatment of the disorder/diseases mentioned. The components can be administered in the same formulation or in separate formulations. If administered in separate formulations the compounds of the invention may be administered sequentially or simultaneously with the other drug(s). In addition to being able to be administered in combination with one or more additional drugs, the compounds of the invention may be used in a combination therapy. When this is done the compounds are typically administered in combination with each other. Thus one or more of the compounds of the invention may be administered either simultaneously (as a combined preparation) or sequentially in order to achieve a desired effect. This is especially desirable where the therapeutic profile of each compound is different such that the combined effect of the two drugs provides an improved therapeutic result. Pharmaceutical compositions of this invention for parenteral injection comprise pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils (such as olive oil), and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. These compositions may also contain adjuvants such as preservative, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of micro- organisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminium monostearate and gelatin. If desired, and for more effective distribution, the compounds can be incorporated into slow release or targeted delivery systems such as polymer matrices, liposomes, and microspheres. The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium just prior to use. Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active compounds can also be in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof. Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound. Dosage forms for topical administration of a compound of this invention include powders, patches, sprays, ointments and inhalants. The active compound is mixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives, buffers, or propellants which may be required. The amount of compound administered will preferably treat and reduce or alleviate the condition. A therapeutically effective amount can be readily determined by an attending diagnostician by the use of conventional techniques and by observing results obtained under analogous circumstances. In determining the therapeutically effective amount a number of factors are to be considered including but not limited to, the species of animal, its size, age and general health, the specific condition involved, the severity of the condition, the response of the patient to treatment, the particular compound administered, the mode of administration, the bioavailability of the preparation administered, the dose regime selected, the use of other medications and other relevant circumstances. A preferred dosage will be a range from about 0.01 to 300 mg per kilogram of body weight per day. A more preferred dosage will be in the range from 0.1 to 100 mg per kilogram of body weight per day, more preferably from 0.2 to 80 mg per kilogram of body weight per day, even more preferably 0.2 to 50 mg per kilogram of body weight per day. A suitable dose can be administered in multiple sub-doses per day. Examples The invention will now be further explained and illustrated by reference to the following non-limiting examples. Additional compounds, other than those described below, may be prepared using methods and synthetic protocols or appropriate variations or modifications thereof, as described herein. The agents of the various embodiments may be prepared using the reaction routes and synthesis schemes as described below, employing the techniques available in the art using starting materials that are readily available. The preparation of particular compounds of the invention is described in detail in the following examples, but the skilled addressee will recognize that the chemical reactions described may be readily adapted to prepare a number of other agents of the various embodiments. For example, the synthesis of non-exemplified compounds may be successfully performed by modifications apparent to those skilled in the art, e.g., by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications of reaction conditions. A list of suitable protecting groups in organic synthesis can be found in T.W. Greene's Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, 1991. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the various embodiments. Reactions can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1H or 13C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), or mass spectrometry, or by chromatography such as high-performance liquid chromatography (HPLC) or thin layer chromatography. The symbols, abbreviations and conventions in the processes, schemes, and examples are consistent with those used in the contemporary scientific literature. Unless otherwise indicated, all temperatures are expressed in °C (degree centigrade). All reactions conducted at room temperature unless otherwise mentioned. The expressions, “ambient temperature,” “room temperature,” and “r.t.”, as used herein, are understood in the art, and refer generally to a temperature, e.g., a reaction temperature, that is about the temperature of the room in which the reaction is carried out, for example, a temperature from about 20 ºC to about 30 ºC. Materials and Methods Reagents useful for synthesizing compounds may be obtained from commercial suppliers or prepared according to techniques known in the art. Commercial solvents and reagents were used as supplied. Analytical thin-layer chromatography (TLC) was performed on silica gel 60 pre- coated with fluorescent indicator F254 aluminium sheets (0.25 mm, Merck). TLC plates were visualised using UV254 or by chemical staining with a solution of potassium permanganate followed by mild heating. Flash column chromatography was carried out using Davisil silica gel 60, 40-63 µm (LC60A 40-63, Davisil). NMR spectra were obtained on a Bruker UltraShield 400 spectrometer at 400 MHz for 1H and 100.6 MHz for 13C. Liquid chromatography mass spectrometry (LC-MS) was carried out on an Agilent Technologies 6120 Single Quad LC/MS System coupled with an Agilent 1260 HPLC consisting of G1312B Quaternary pump, 1260 Infinity G1367E 1260 HiP ALS Autosampler and 1290 Infinity G4212A 1290 DAD wavelength detector (monitored at 214nm and 254nm). Software used for LC-MS was LC/MSD Chemstation Rev.B.04.03 coupled with Masshunter Easy Access Software. LC conditions: reverse phase HPLC analysis. Column: Poroshell 120 EC-C18 (3.0 X 50mm 2.7-Micron); column temperature: 35°C; injection volume: 1 μl; sample was eluted out using a binary gradient: solvent A: 0.1% formic acid in water, solvent B: 0.1 % formic acid in MeCN. MS conditions: quadrupole ion source with multimode-ESI; drying gas temperature: 350°C; capillary voltage: 3000 V (positive or negative mode); scan range: 100– 1000; step size: 0.1 second; acquisition time: 5 min; typical gradient run: 95% solvent A + 5% solvent B gradually increased to 100% solvent B over 2.5 min and 100% solvent B for 1.3 min then was back to 95% solvent A + 5% solvent B. HPLC was acquired on an Agilent 1260 Infinity Analytical HPLC equipped with a 1260 Degasser G1322A (JPAAJ81416), 1260 Bin Pump G1312B (DEACB04606), 1260 HiP ALS G1367E (DEACO03069), 1260 TCC: G1316A (DEACN16726), 1260 DAD: G4212B (DEAA304233) and a Zorbax Eclipse Plus C18 Rapid Resolution 4.6 X 100mm 3.5‐Micron Column. Solvent A: water + 0.1% TFA. Solvent B: MeCN + 0.1% TFA. Typical gradient run: 95% solvent A + 5% solvent B gradually increased to 100% solvent B over 9 min then 100% solvent B for 1 min, with a flow rate of 1 ml/min. HRMS was acquired on an Agilent 6224 TOF LC/MS Mass Spectrometer coupled to an Agilent 1290 Infinity (Agilent, Palo Alto, CA). Acquisition was performed using the Agilent Mass Hunter Data Acquisition software version B.05.00 Build 5.0.5042.2 and analysis was performed using Mass Hunter Qualitative Analysis (version B.05.00 Build 5.0.519.13). Mass spectrometer conditions: ionisation mode: electrospray ionisation; drying gas flow: 11 L/min; nebuliser: 45 psi; drying gas temperature: 325°C; capillary voltage: 4000 V; fragmentor: 160 V; Skimmer: 65 V; OCT RFV: 750 V; scan range acquired: 100–1500 m/z; internal reference ions: positive ion mode = m/z = 121.050873 & 922.009798. Synthetic Schemes As stated above there are a number of ways in which the compounds of the invention can be synthesized as would be appreciated by a person skilled in the art. Nevertheless, we provide reaction schemes for making certain compounds of the invention in Schemes 1 to 15. Synthesis of 2-fluorobutan-1-amine hydrochloride intermediates The intermediates 2-fluorobutan-1-amine hydrochloride and its chiral counterparts, (R)-2-fluorobutan-1-amine hydrochloride and (S)-2-fluorobutan-1-amine hydrochloride were synthesised using previous published methods such as, for example, those found in Org. Lett.2011, 13, 20, 5684–5687. 2-Fluorobutan-1-amine hydrochloride:
Figure imgf000044_0001
A white solid (44.5 mg, 91%).1H NMR (400 MHz, DMSO-d6) δ 8.17 (s, 3H), 4.78 – 4.59 (m, 1H), 3.04 (dddd, J = 22.5, 16.0, 13.9, 5.6 Hz, 2H), 1.71 – 1.58 (m, 2H), 0.93 (t, J = 7.5 Hz, 3H). (R)-2-fluorobutan-1-amine hydrochloride:
Figure imgf000044_0002
A white solid (62.1 mg, 95%).1H NMR (400 MHz, MeOD) δ 4.78 – 4.59 (m, 1H), 3.18 – 3.05 (m, 2H), 1.77 – 1.66 (m, 2H), 1.05 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, MeOD) δ 92.91 (d, J C-F = 170.8 Hz), 44.03 (d, J C-F = 21.0 Hz), 26.52 (d, J C-F = 20.2 Hz), 9.20 (d, J C- F = 5.6 Hz). (S)-2-Fluorobutan-1-amine hydrochloride:
Figure imgf000045_0001
A white solid (55.9 mg, 93%).1H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 3H), 4.81 – 4.60 (m, 1H), 3.17 – 2.92 (m, 2H), 1.72 – 1.57 (m, 2H), 0.93 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, DMSO-d6) δ –187.28 (s); 13C NMR (101 MHz, DMSO-d6) δ 91.87 (d, 1JC-F = 169.2 Hz), 41.93 (d, 2JC-F = 21.1 Hz), 24.85 (d, 2JC-F = 20.0 Hz), 8.78 (d). Examples 5-01 to 5-28:
Figure imgf000045_0002
Scheme 1 (a) General procedure for preparation of LTBS salts To a solution of 2-bromopyridine (1.0 equiv.) in anhydrous toluene and THF (4:1, v/v) solvent mixture was added tri-isopropyl borate (1.1 equiv.). The mixture was cooled to - 78°C and then added n-BuLi (1.05 equiv; 2.5 M solution in hexane) in a dropwise manner over 15-20 min time. The resulting mixture was stirred at the same temperature for 3 h, then the cooling bath was removed and the mixture was slowly warmed to 0 °C (around 1.5 h), the reaction was quenched with isopropyl alcohol. The resulting suspension was stirred overnight. The solvent was then removed under reduced pressure, and the resulting residue was treated with acetone and water (9:1, v/v) until solid was precipitated. The resultant precipitate was filtered, further washed with acetone and water (9:1, v/v) and then dried under reduced pressure to afford the desired LTBS salts (30-80%). Lithium (pyridin-2-yl)trihydroxyborate (LTBS)
Figure imgf000046_0001
According to the general procedure, using 2-bromopyridine to afford the title compound as a light brown solid (665 mg, 71%).1H NMR (400 MHz, D2O) δ 8.50 (d, J = 6.0 Hz, 1H), 8.34 – 8.27 (m, 1H), 8.01 (d, J = 7.9 Hz, 1H), 7.73 (t, J = 6.9 Hz, 1H). (b) General procedure for preparation of compound (4) (Route A) To a solution of (1r,4r)-4-((5-bromo-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol (2) (1.0 equiv.) and LTBS (3.0 equiv.) in DMF and H2O (4:1, v/v) was purged with nitrogen for 30 mins. Pd(dppf)Cl2 (0.05eq), CuBr (0.2 equiv.) and K2CO3 (3.0 equiv.) were added and the resulting reaction mixture heated to 110°C and stirred for 30 min- 1 hour. The reaction was monitored by TLC. Upon completion, the mixture was diluted with EtOAc, and the resulting suspension was filtered through the Celite pad, it was washed with EtOAc. The filtrate was further washed with ice cold water (3 × 20 mL) and brine. The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (10-70% EtOAc in Pet. Sp.) to afford the desired compound 4 in 18-74% yield. (c) Synthesis of (1r,4r)-4-((5-bromo-2-(methylthio)pyrimidin-4-yl)amino)cyclohexan-1-ol (2) To a solution of 5-bromo-4-chloro-2-(methylthio)pyrimidine 1 (1.0 equiv.) and trans-4-aminocyclohexanol (1.1 equiv.) in i-PrOH was added DIPEA (2.0 equiv.). The reaction mixture was stirred at 70°C overnight. The reaction was monitored by TLC. Upon completion, the solvent was evaporated under reduced pressure. The resulting residue was partitioned between DCM and brine. The aqueous layer was further extracted with DCM (3 × times). All the combined organic exacts were dried over anhydrous magnesium sulphate, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography using 0-50% ethyl acetate in petroleum spirits to afford the desired compound as an off white solid, 98% yield. (d) Preparation of (4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin-5- yl)boronic acid (3) To a solution of 2 (1.0 equiv.) and bis(pinacolato)diboron (2.0 equiv.) in dioxane was added KOAc (3.0 equiv.). The reaction mixture was degassed with nitrogen for 30 min, and then Pd(dppf)Cl2 (0.1 equiv.) was added. The reaction mixture was heated to 90°C in microwave for 90 min. Upon completion, the mixture was concentrated under reduced pressure and then was purified by silica gel flash chromatography using DCM in MeOH (0- 100%) to afford the desired compound 3 as a half white solid 72%. (e) General procedure for preparation of compound (4) (Route B): To a solution of 3 (1.0 equiv.) in a mixture of dioxane and water (4:1, v/v) was added substituted 2-bromo pyridine (1.5 equiv.) and K2CO3 (3.0 equiv.). The reaction mixture was degassed with nitrogen for 30 min. Then, Pd(PPh3)4 (0.1 eq) was added and the resulting reaction mixture was heated to 90 °C and stirred overnight. The reaction was monitored by TLC. Upon completion, the mixture was diluted with EtOAc, and the resulting suspension was passed through a pad of Celite. The filtrate was further washed with water (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography using 0-50% EtOAc in Pet. Spirits to afford the desired compound 4 in 20-68% yields. (f) Synthesis of pyridyl pyrimidine compounds (Example Compounds 5-01-5-28): To a solution of 4 (1.0 equiv.) in THF (2.0 mL) was added m-CPBA (2.05 equiv.). The mixture was stirred at room temperature for 1.0 h. The reaction was monitored by TLC. Upon all starting material was consumed, the desired 2-fluoroalkyl amine hydrochloride (4.0 equiv.) and DIPEA (4.0 equiv.) were added. The reaction mixture was heated under microwave irradiation at 140 °C for 30 min-1 h. Then the solvent was removed under reduced pressure. The resulting residue was purified by silica gel flash chromatography (20-100% EtOAc in Pet. Sp.) to afford the desired pyridylpyrimidyl compound 5 in 40-89% yields. Example 5-01: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(pyridin-2-yl)pyrimidin-4- yl)amino)cyclohexan-1-ol: White solid, (58% yield); 1H NMR (400 MHz, MeOD) δ 8.48 (dt, J = 4.8, 1.6 Hz, 1H), 8.33 (s, 1H), 7.77 – 7.75 (m, 2H), 7.19 – 7.13 (m, 1H), 4.70 – 4.65 (m, 1H), 4.58 – 4.52 (m, 1H), 4.07 – 4.00 (m, 1H), 3.75 – 3.61 (m, 2H), 3.52 – 3.43 (m, 1H), 2.17 – 2.15 (m, 2H), 2.01 – 1.99 (m, 2H), 1.77 – 1.63 (m, 2H), 1.48 – 1.38
Figure imgf000048_0001
(m, 4H), 1.04 (t, J = 7.6 Hz, 3H) ppm; 19F NMR (376 MHz, MeOD) δ −188.64 ppm; HPLC: tR 4.53 min > 95% purity at 254 nm; LCMS (m/z): 360.2 [M+H]+; HRMS (m/z): [M + H]+ calcd for C19H26FN5O 360.2194 found 360.2202. Example 5-02: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(6-fluoropyridin-2-yl)pyrimidin- 4-yl)amino)cyclo hexan-1-ol: White solid (65% yield, obtained as a TFA salt); 1H NMR (400 MHz, MeOD) δ 8.41 (s, 1H), 8.09 (q, J = 8.1 Hz, 1H), 7.80 (dd, J = 7.8, 2.0 Hz, 1H), 7.11 (dd, J = 8.2, 2.1 Hz, 1H), 4.79 – 4.53 (m, 1H), 4.16 (dd, J = 11.7, 8.0 Hz, 1H), 3.90 – 3.50 (m, 3H), 2.20 (d,
Figure imgf000048_0002
J = 7.2 Hz, 2H), 2.12 – 1.95 (m, 2H), 1.85 – 1.63 (m, 2H), 1.51 (tt, J = 23.7, 11.8 Hz, 4H), 1.08 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -70.21, -188.38; HPLC: tR 6.67 min > 98 % purity at 254 nm; LCMS: m/z 378.2 [M+H]+; HRMS [M+H]+: m/z calculated 378.2100 found 378.2109. Example 5-03: (1S,4r)-4-((5-(5-chloro-6-fluoropyridin-2-yl)-2-(((S)-2- fluorobutyl)amino)pyrimidin-4-yl)amino) cyclohexan-1-ol: Off-white solid (38% yield); 1H NMR (400 MHz, MeOD) δ 8.39 (s, 1H), 8.04 – 7.84 (m, 1H), 7.68 (d, J = 8.4 Hz, 1H), 4.60 (d, J = 51.4 Hz, 1H), 4.04 (dd, J = 8.9, 5.1 Hz, 1H), 3.79 – 3.58 (m, 2H), 3.52 (d, J = 7.4 Hz, 1H), 2.17 (d, J = 8.9 Hz, 2H), 2.01 (d, J =
Figure imgf000048_0003
9.4 Hz, 2H), 1.87 – 1.58 (m, 2H), 1.52 – 1.33 (m, 4H), 1.04 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -74.96, -188.50; HPLC: tR 4.87 min, > 98 % purity at 254 nm; LCMS: m/z 412.2 [M+H]+;; HRMS [M+H]+: m/z calculated 412.1710 found 412.1718. Example 5-04: (1S,4r)-4-((5-(5,6-difluoropyridin-2-yl)-2-(((S)-2- fluorobutyl)amino)pyrimidin-4-yl)amino) cyclohexan-1-ol: Off-white solid (42% yield); 1H NMR (400 MHz, MeOD) δ 8.31 (s, 1H), 8.04 – 7.85 (m, 1H), 7.78 (dd, J = 8.5, 2.7 Hz, 1H), 4.76 – 4.51 (m, 1H), 4.24 – 4.03 (m, 1H), 3.90 – 3.53 (m, 3H), 2.16 (d, J = 4.5 Hz, 2H), 2.11 – 1.94 (m, 2H), 1.86 – 1.61 (m, 2H), 1.61 – 1.37 (m,
Figure imgf000049_0001
4H), 1.06 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -90.02 (d), -143.41 (d), -188.32; HPLC: tR 4.54 min, >98 % purity at 254 nm; LCMS: m/z 396.2 [M+H]+; HRMS [M+H]+: m/z calculated 396.20 found 396.2021. Example 5-05: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-fluoropyridin-2-yl)pyrimidin- 4-yl)amino)cyclo hexan-1-ol: Off-white solid (64%, yield over two steps); 1H NMR (400 MHz, CDCl3) δ 8.34 (d, J = 2.8 Hz, 1H), 8.28 (s, 1H), 7.62 (dd, J = 9.1, 4.1 Hz, 1H), 7.45 (td, J = 8.6, 3.0 Hz, 1H), 5.82 (s, 1H), 4.68 -4.56 (dm, 1H), 4.02 (dd, J = 12.1, 7.9 Hz, 1H), 3.92 – 3.60 (m, 2H), 3.89 – 3.62
Figure imgf000049_0002
(m, 2H), 3.60 – 3.39 (m, 1H), 2.52 (s, 1H), 2.24 – 2.10 (m, 2H), 2.04 (d, J = 9.7 Hz, 2H), 1.82 – 1.54 (m, 2H), 1.54 – 1.29 (m, 4H), 1.03 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, CDCl3) δ -130.71, -186.69; HPLC: tR 4.47 min, 97 % purity at 254 nm; LRMS: m/z 378.2 [M+H]+; HRMS [M+H]+: m/z calculated 378.2100 found 378.2115. Example 5-06: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-fluoropyridin-2-yl)pyrimidin- 4-yl)amino)cyclo hexan-1-ol: Off-white solid (68% yield, obtained as a TFA salt); 1H NMR (400 MHz, MeOD) δ 8.63 (dd, J = 8.5, 5.8 Hz, 1H), 8.41 (s, 1H), 7.77 (dd, J = 10.7, 2.0 Hz, 1H), 7.23 (ddd, J = 8.2, 5.8, 2.3 Hz, 1H), 4.76 –
Figure imgf000049_0003
4.54 (m, 1H), 4.13 (td, J = 10.3, 5.2 Hz, 1H), 3.92 – 3.54 (m, 3H), 2.16 (dd, J = 7.4, 2.4 Hz, 2H), 2.09 – 1.95 (m, 2H), 1.83 – 1.62 (m, 2H), 1.62 – 1.36 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -77.18, -101.37, -188.33; HPLC: tR 4.46 min, > 97 % purity at 254 nm; LCMS: m/z 378.2 [M+H]+; HRMS [M+H]+: m/z calculated 378.2100 found 378.2116. Example 5-07: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(fluoromethyl)pyridin-2- yl)pyrimidin-4-yl)amino) cyclohexan-1-ol: Off-white solid (68%, yield over two steps); HPLC: tR 4.64 min, >98 % purity at 254 nm; 1H NMR (400 MHz, CDCl3) δ 9.95 (s, 1H), 8.44 (s, 1H), 8.29 (s, 1H), 7.68 (d, J = 8.3 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.19 (s, 1H), 5.45 – 5.18 (m, 2H), 4.56
Figure imgf000050_0003
(ddt, J = 15.2, 10.6, 7.6 Hz, 1H), 3.98 (dd, J = 7.0, 3.5 Hz, 1H), 3.85 – 3.56 (m, 2H), 3.45 (t, J = 16.1 Hz, 1H), 2.12 (d, J = 11.5 Hz, 2H), 1.98 (d, J = 9.7 Hz, 2H), 1.75 – 1.55 (m, 2H), 1.48 – 1.23 (m, 4H), 1.03 – 0.91 (m, 3H); 19F NMR (376 MHz, CDCl3) δ -186.46, -207.82; LRMS: m/z 392.2 [M+H]+; HRMS [M+H]+: m/z calculated 392.2256 found 392.2273. Example 5-08: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(2-fluoroethoxy)pyridin-2- yl)pyrimidin-4-yl)amino) cyclohexan-1-ol: White solid (76% yield, obtained as a TFA salt); 1H NMR (400 MHz, Methanol-d4) δ 8.45 – 8.32 (m, 1H), 8.26 (d, J = 11.14 Hz, 1H), 7.84 (d, J = 8.90 Hz, 1H), 7.55 (dd, J = 8.94, 3.03
Figure imgf000050_0002
Hz, 1H), 4.78 – 4.54 (m, 2H), 4.36 (ddd, J = 28.73, 4.79, 2.70 Hz, 2H), 4.12 (td, J = 10.42, 8.52, 4.99 Hz, 1H), 3.91 – 3.50 (m, 2H), 2.16 (d, J = 12.15 Hz, 2H), 2.10 – 1.97 (m, 2H), 1.83 – 1.64 (m, 2H), 1.61 – 1.37 (m, 4H), 1.06 (t, J = 7.45 Hz, 3H); LCMS [M+H]+ 422.3 m/z Example 5-09: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(3-fluoropropyl)pyridin-2- yl)pyrimidin-4-yl) amino)cyclohexan-1-ol: white solid (65% yield, obtained as a TFA salt); HPLC: tR 4.79 min, 100 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.50 (d, J = 6.4 Hz, 1H), 8.33 (s, 1H), 7.86 – 7.77 (m, 2H), 4.64 (m, 1H), 4.53 (t, J = 5.8 Hz, 1H), 4.41 (t, J = 5.8 Hz, 1H), 4.19 – 4.08 (m, 1H), 3.68 (m, 3H), 2.90 – 2.75
Figure imgf000050_0001
(m, 2H), 2.17 (d, J = 9.3 Hz, 2H), 2.12 – 1.93 (m, 4H), 1.83 – 1.61 (m, 2H), 1.61 – 1.35 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.32 (aliphatic fluorine atom is in out of the scale); HPLC: tR 4.79 min, > 99% purity at 254 nm; LCMS: m/z 420.3 [M+H]+; HRMS [M+H]+: m/z calculated 420.2569 found 420.2585. Example 5-10: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-(fluoromethyl)pyridin-2- yl)pyrimidin-4-yl)amino) cyclohexan-1-ol: Off-white solid (62%, yield over two steps); 1H NMR (400 MHz, CDCl3) δ 9.79 (s, 1H), 8.47 (d, J = 5.2 Hz, 1H), 8.38 (s, 1H), 7.57 (s, 1H), 7.05 (d, J = 5.2 Hz, 1H), 5.43 (d, J = 46.7 Hz, 2H), 4.78 – 4.47 (m, 1H), 4.02 (dtd, J = 10.5, 6.9, 3.7 Hz, 1H), 3.90 –
Figure imgf000051_0003
3.64 (m, 2H), 3.48 (dddd, J = 19.6, 14.5, 7.5, 5.3 Hz, 1H), 2.09 (ddd, J = 9.5, 8.9, 5.4 Hz, 5H), 1.86 – 1.55 (m, 2H), 1.55 – 1.29 (m, 4H), 1.03 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, CDCl3) δ 16.35, -186.60; HPLC: tR 4.51 min, > 96% purity at 254 nm; LCMS: m/z 392.2 [M+H]+; HRMS [M+H]+: m/z calculated 392.2256 found 392.2273. Example 5-11: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-(2-fluoroethoxy)pyridin-2- yl)pyrimidin-4-yl) amino)cyclohexan-1-ol: Off-white solid (78%, yield over two steps); 1H NMR (400 MHz, MeOD) δ 8.51 (d, J = 6.1 Hz, 1H), 8.23 (s, 1H), 7.50 (s, 1H), 7.18 (d, J = 2.0 Hz, 1H), 4.88 – 4.80 (m, 1H), 4.78 – 4.65
Figure imgf000051_0002
(m, 1H), 4.63 – 4.38 (m, 3H), 4.08 (d, J = 10.0 Hz, 1H), 3.90 – 3.54 (m, 3H), 2.20 – 1.95 (m, 4H), 1.82 – 1.58 (m, 2H), 1.58 – 1.31 (m, 4H), 1.06 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.22 (aliphatic fluorine atom on RHS is in out of the scale); HPLC: tR 4.20 min, > 99% purity at 254 nm; LRMS: m/z 422.2 [M+H]+; HRMS [M+H]+: m/z calculated 422.2362 found 422.2379. Example 5-12: 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)nicotine nitrile: Off-white solid (58%, yield over two steps); 1H NMR (400 MHz, Chloroform-d) δ 9.89 (s, 1H), 8.73 (dd, J = 2.15, 0.85 Hz, 1H), 8.49 (s, 1H), 7.89 (dd, J = 8.68, 2.24 Hz, 1H), 7.76 (d, J = 8.76 Hz, 1H), 6.34 (s, 1H), 4.62 (dtdd, J = 49.53, 7.69, 5.07, 3.05
Figure imgf000051_0001
Hz, 1H), 4.05 (dh, J = 10.84, 3.11 Hz, 1H), 3.90 – 3.64 (m, 2H), 3.52 (dddd, J = 19.48, 14.53, 7.50, 5.43 Hz, 1H), 3.02 – 2.42 (m, 1H), 2.28 – 2.12 (m, 2H), 2.12 – 1.96 (m, 2H), 1.84 – 1.58 (m, 2H), 1.55 – 1.30 (m, 4H), 1.03 (t, J = 7.46 Hz, 3H); 19F NMR (376 MHz, Chloroform-d) δ -186.44, -187.40; HPLC: tR = 4.3 min > 95% purity at 254 nm; LCMS [M+H]+ 385.2 m/z; HRMS [M+H+: m/z calcd 385.2147 found 385.2157. Example 5-13: 2-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)isonicotinonitrile: Off-white solid (51%, yield over two steps); 1H NMR (400 MHz, Chloroform-d) δ 9.80 (s, 1H), 8.62 (d, J = 5.17 Hz, 1H), 8.39 (s, 1H), 7.86 (s, 1H), 7.29 (d, J = 5.17 Hz, 1H), 6.36 (s, 1H), 4.62 (dtdd, J = 49.50, 7.66, 5.02, 3.04 Hz, 1H), 4.04 (tdt, J =
Figure imgf000052_0003
10.68, 7.26, 3.85 Hz, 1H), 3.76 (dtd, J = 21.57, 14.71, 14.14, 9.73 Hz, 2H), 3.62 – 3.40 (m, 1H), 2.17 (dt, J = 12.03, 3.59 Hz, 2H), 2.09 – 1.90 (m, 2H), 1.84 – 1.58 (m, 2H), 1.54 – 1.31 (m, 4H), 1.03 (t, J = 7.45 Hz, 3H); 19F NMR (376 MHz, Chloroform- d) δ -186.46, -187.39; HPLC - tR = 4.3 min >95% purity at 254 nm; LCMS [M+H]+ 385.2 m/z; HRMS calcd for [M+H]+ 385.2147 m/z, found 385.2161. Example 5-14: (1S,4r)-4-((5-(5-((cyclohexylamino)methyl)pyridin-2-yl)-2-(((S)-2- fluorobutyl)amino)pyrimidin-4-yl)amino)cyclohexan-1-ol: Off-white solid (15 %, yield over two steps); 1H NMR (400 MHz, Methanol-d4) δ 9.78 (d, J = 7.35 Hz, 1H), 8.43 (s, 1H), 8.28 (s, 1H), 7.85 – 7.63 (m, 2H), 4.02 – 3.91 (m, 1H), 3.82 (s, 2H), 3.60 (d, J = 9.30 Hz, 2H), 3.51 – 3.04
Figure imgf000052_0002
(m, 2H), 2.51 (d, J = 10.55 Hz, 1H), 2.09 (d, J = 10.60 Hz, 2H), 1.95 (d, J = 10.31 Hz, 4H), 1.80 – 1.55 (m, 5H), 1.36 (q, J = 11.30 Hz, 4H), 1.17 (dq, J = 22.47, 11.69, 10.17 Hz, 5H), 0.99 (t, J = 7.44 Hz, 3H); HPLC - tR = 4.0 min >95% purity at 254 nm; LCMS [M+H]+ 471.2 m/z. Example 5-15: (1S,4r)-4-((5-(5-((cyclopropylamino)methyl)pyridin-2-yl)-2-(((S)-2- fluorobutyl)amino)pyrimidin -4-yl)amino)cyclohexan-1-ol: Off-white solid (19%, yield over two steps); 1H NMR (400 MHz, Methanol-d4) δ 8.46 (s, 1H), 8.33 (s, 1H), 7.83 – 7.67 (m, 2H), 4.74 – 4.49 (m, 2H), 4.03 (dt, J = 10.53, 5.71 Hz, 1H), 3.82 (s, 2H), 3.76 – 3.59 (m, 2H), 3.47 (ddd, J = 17.17, 14.43,
Figure imgf000052_0001
7.27 Hz, 1H), 2.29 – 2.10 (m, 3H), 2.00 (d, J = 9.49 Hz, 2H), 1.70 (ddq, J = 28.01, 14.31, 7.27 Hz, 2H), 1.54 – 1.35 (m, 4H), 1.04 (t, J = 7.46 Hz, 3H), 0.63 – 0.26 (m, 4H).19F NMR (376 MHz, Methanol-d4) δ -188.49; LCMS [M+H]+ 429.2 m/z. Example 5-16: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(4-(2-fluoroethyl)piperazin-1- yl)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol: Light brown solid (26%, yield over two steps); 1H NMR (400 MHz, Methanol-d4) δ 8.20 (d, J = 2.82 Hz, 2H), 7.60 (d, J = 8.90 Hz, 1H), 7.39 (dd, J = 8.91, 2.95 Hz, 1H), 4.68 (t, J = 4.85 Hz, 1H), 4.56 (t, J = 4.87 Hz, 1H),
Figure imgf000053_0003
4.00 (dt, J = 10.61, 5.28 Hz, 1H), 3.78 – 3.55 (m, 2H), 3.45 (ddd, J = 17.14, 14.36, 7.31 Hz, 1H), 3.28 (d, J = 4.70 Hz, 4H), 2.81 (t, J = 4.82 Hz, 1H), 2.73 (t, J = 4.91 Hz, 5H), 2.15 (d, J = 10.28 Hz, 2H), 1.99 (d, J = 10.00 Hz, 2H), 1.70 (dq, J = 20.80, 7.04, 5.40 Hz, 2H), 1.45 – 1.25 (m, 4H), 1.04 (t, J = 7.46 Hz, 3H).19F NMR (376 MHz, Methanol-d4) δ -188.54, -220.16; HPLC - tR = 3.8 min >95% purity at 254 nm; LCMS [M+H]+ 490.3 m/z. Example 5-17: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(methyl(tetrahydrofuran-3- yl)amino)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol: Pale yellow solid (10%, obtained as a TFA salt); 1H NMR (400 MHz, MeOD) δ 8.21 (s, 1H), 8.11 (s, 1H), 7.69 (d, J = 9.1 Hz, 1H), 7.42 (d, J = 9.1 Hz, 1H), 4.74 – 4.50 (m, 2H), 4.22 – 4.02 (m, 2H), 3.93 (dd, J = 9.7, 2.7 Hz, 1H), 3.89 – 3.50 (m, 5H),
Figure imgf000053_0001
2.96 (s, 3H), 2.35 (td, J = 13.1, 8.1 Hz, 1H), 2.16 (d, J = 8.8 Hz, 2H), 2.08 – 1.88 (m, 3H), 1.84 – 1.59 (m, 3H), 1.48 (tt, J = 22.5, 11.2 Hz, 4H), 1.06 (t, J = 7.4 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.33; HPLC: tR 4.43 min, > 99% purity at 254 nm; LRMS: m/z 459.2 [M+H]+; HRMS [M+H]+: m/z calculated 459.2878 found 459.2887. Example 5-18: 3-((6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)methyl)oxazolidin-2-one: Pale yellow solid (25%, obtained as a TFA salt); 1H NMR (400 MHz, MeOD) δ 8.59 (s, 1H), 8.39 (s, 1H), 7.92 (d, J = 8.4 Hz, 2H), 4.72 (d, J = 5.6 Hz, 1H), 4.60 (d, J = 4.9 Hz, 1H), 4.53
Figure imgf000053_0002
(d, J = 9.9 Hz, 2H), 4.38 (t, J = 8.0 Hz, 2H), 4.14 (t, J = 10.3 Hz, 1H), 3.80 (dd, J = 27.9, 14.5 Hz, 1H), 3.72 – 3.65 (m, 1H), 3.59 (dd, J = 15.7, 7.9 Hz, 2H), 2.16 (s, 2H), 2.03 (d, J = 12.7 Hz, 2H), 1.73 (ddd, J = 28.3, 14.1, 7.0 Hz, 2H), 1.49 (tt, J = 24.5, 12.4 Hz, 4H), 1.07 (t, J = 7.4 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.33; HPLC: tR 4.22 min, > 96% purity at 254 nm; LRMS: m/z 459.2 [M+H]+; HRMS [M+H]+: m/z calculated 459.2514 found 459.2516. Example 5-19: 3-((2-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-4-yl)methyl)dihydrofuran-2(3H)-one: Off-white solid (29%); 1H NMR (401 MHz, CDCl3) δ 9.79 (s, 1H), 8.45 – 8.25 (m, 2H), 7.45 (s, 1H), 6.96 (dd, J = 5.2, 1.0 Hz, 1H), 5.46 (s, 1H), 4.76 – 4.47 (m, 1H), 4.31 (td, J = 8.9, 2.4 Hz, 1H), 4.18 (td, J = 9.6, 6.6 Hz, 1H), 4.08 –
Figure imgf000054_0003
3.93 (m, 1H), 3.92 – 3.60 (m, 2H), 3.58 – 3.36 (m, 1H), 3.24 (dd, J = 13.8, 4.3 Hz, 1H), 2.96 – 2.83 (m, 1H), 2.78 (dd, J = 13.8, 9.1 Hz, 1H), 2.39 – 2.23 (m, 1H), 2.23 – 2.10 (m, 2H), 2.10 – 1.85 (m, 5H), 1.82 – 1.56 (m, 2H), 1.54 – 1.31 (m, 4H), 1.30 – 1.16 (m, 2H), 1.03 (t, J = 7.5 Hz, 3H); 19F NMR (377 MHz, CDCl3) δ -186.64; LCMS: m/z 458.3 [M+H]+; HPLC: tR 4.42 min, 100 % purity at 254 nm; HRMS [M+H]+: m/z calculated 458.256 found 458.2565. Example 5-20: (1S,4r)-4-((5-(5-chloropyridin-2-yl)-2-(((S)-2-fluorobutyl)amino)pyrimidin- 4-yl)amino) cyclohexan-1-ol: According to General Procedure Route B, using 2-bromo-5- chloropyridine, to afford the title compound as a pale yellow amorphous solid; (218 mg, 48%); 1H NMR (400 MHz, CDCl3) δ 9.47 (bs, 1H), 8.44
Figure imgf000054_0001
(d, J = 2.4 Hz, 1H), 8.33 (s, 1H), 7.65 (dd, J = 8.8, 2.5 Hz, 1H), 7.57 (d, J = 8.8 Hz, 1H), 5.39 (bs, 1H), 4.79 – 4.48 (m, 1H), 4.03 (m, 1H), 3.81 (m, 2H), 3.56 – 3.37 (m, 1H), 2.18 (d, J = 10.5 Hz, 2H), 2.03 (d, J = 7.9 Hz, 2H), 1.83 – 1.53 (m, 4H), 1.45 (ddd, J = 37.1, 18.6, 9.3 Hz, 2H), 1.10 – 0.98 (m, 3H); LCMS: m/z 394.1 [M+H]+. Example 5-21: 5-((2-(((S)-2-fluorobutyl)amino)-5-(6-fluoropyridin-2-yl)pyrimidin-4- yl)amino)bicyclo[2.2.1]heptan-2-ol: Off-white solid (39%).1H NMR (400 MHz, Chloroform-d) δ 9.60 (s, 1H), 8.35 (s, 1H), 7.74 (q, J = 7.99 Hz, 1H), 7.45 (dd, J = 7.82, 2.37 Hz, 1H), 6.69 (dd, J = 8.07, 2.76 Hz, 1H), 5.90 (s, 1H), 4.77 – 4.04 (m, 3H), 3.78 (dd, J = 28.70, 14.43 Hz, 1H), 3.60 – 3.33 (m, 1H), 3.05 –
Figure imgf000054_0002
2.71 (m, 2H), 2.50 (dt, J = 7.96, 4.54 Hz, 1H), 2.35 (d, J = 4.48 Hz, 1H), 2.05 – 1.39 (m, 6H), 1.02 (t, J = 7.41 Hz, 3H).13C NMR (101 MHz, Chloroform-d) δ 161.7 (d, J = 270.9 Hz), 160.7, 154.8 (d, J = 12.1 Hz), 141.6 (d, J = 7.9 Hz), 127.6, 115.4 (d, J = 4.3 Hz), 104.9 (d, J = 35.7 Hz), 104.2, 94.4 (d, J = 169.6 Hz), 74.4, 72.1, 51.8, 45.1 (d, J = 21.8 Hz), 44.9, 43.0, 41.3, 39.6, 37.6, 35.2, 34.2 (d, J = 3.6 Hz), 34.1 (d, J = 4.2 Hz), 33.0, 29.8, 28.7 (d, J = 4.4 Hz), 25.9 (d, J = 20.9 Hz), 9.4 (d, J = 6.0 Hz).19F NMR (376 MHz, Chloroform-d) δ -68.32, -68.52, -186.41. LCMS [M+H]+ 390.2 m/z; HPLC - tR = 4.7, 4.8 min >95% purity at 254 nm; HRMS calcd for [M+H]+ 390.2100 m/z, found 390.2114. Example 5-22: (1S,4r)-4-((5-(6-chloropyridin-2-yl)-2-(((S)-2-fluorobutyl)amino)pyrimidin- 4-yl)amino)cyclohexan-1-ol
Figure imgf000055_0001
According to General Procedure Route A, using lithium (6-chloropyridin-2- yl)trihydroxyborate, to afford the title compound as pale yellow solid (43 mg, 42% yield); HPLC: tR 4.78 min, > 97 % purity at 254 nm; 1H NMR (400 MHz, CDCl3) δ 9.48 (bs, 1H), 8.40 (s, 1H), 7.62 (t, J = 7.9 Hz, 1H), 7.54 (d, J = 8.0 Hz, 1H), 7.09 (d, J = 7.6 Hz, 1H), 5.48 (bs, 1H), 4.78 – 4.48 (m, 1H), 4.04 (d, J = 6.6 Hz, 1H), 3.92 – 3.69 (m, 2H), 3.48 (m, 1H), 2.20 (m, 2H), 2.06 (m, 2H), 1.96 – 1.57 (m, 3H), 1.57 – 1.34 (m, 4H), 1.03 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 159.81, 156.74, 155.40, 148.72, 139.03, 119.65, 116.33, 104.35, 95.27- 93.58 (d, 1JCF= 170.7 Hz), 69.56, 48.23, 45.14-44.93 (d, 2JCF= 21.2 Hz), 33.41, 29.77, 25.86- 25.66 (d, 2JCF= 20.2 Hz), 9.31, 9.24; 19F NMR (376 MHz, CDCl3) δ -186.55; LCMS: m/z 394.1 [M+H]+; HRMS [M+H]+: m/z calculated 394.1804 found 394.1820. Example 5-23: 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)-N-methylpyridine-3-sulfonamide
Figure imgf000055_0002
According to General Procedure Route B, using tert-butyl ((6-chloropyridin-3- yl)sulfonyl)(methyl)carbamate, purification by reverse-phase column chromatography (KP- C18-HS Flash Cartridge) using ACN and water to afford to afford title compound as off-white solid; (22 mg, 33%); HPLC: tR 4.96 min, >95 % purity at 254 nm; 1H NMR (400 MHz, DMSO) δ 9.81 (s, 1H), 8.80 (s, 1H), 8.63 (s, 1H), 8.10 (d, J = 8.8 Hz, 1H), 8.01 (d, J = 8.7 Hz, 1H), 7.65 – 7.45 (m, 1H), 4.73-4.48 (m, 2H), 3.94 (s, 1H), 3.48 (s, 1H), 2.45 (d, J = 5.0 Hz, 3H), 2.00 (s, 2H), 1.84 (d, J = 10.2 Hz, 2H), 1.72-1.52 (m, 2H), 1.43 – 1.18 (m, 4H), 0.95 (t, J = 7.0 Hz, 3H); 13C NMR (101 MHz, CDCl3 + MeOD (1:1)) δ 159.51, 158.73, 145.59, 134.77, 131.02, 129.13, 127.28, 117.88, 94.42 – 92.74 (d, 1JCF = 169.7 Hz), 68.66, 44.60 – 44.38 (d, 2JCF = 22.2 Hz), 32.87, 29.77, 29.12, 28.01, 25.42 – 25.21 (d, 2JCF = 20.9 Hz), 8.50 (d) (one carbon is missing i.e. pyrimidine C5); 19F NMR (377 MHz, CDCl3) δ -183.58; LCMS: m/z 453.1 [M+H]+; HRMS [M+H]+: m/z calculated 453.2079 found 453.2094. Example 5-24: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-(3-fluoropropyl)pyridin-2- yl)pyrimidin-4-yl)amino) cyclohexan-1-ol trifluoroacetate salt
Figure imgf000056_0001
According to General Procedure Route B, using 2-bromo-4-(3- fluoropropyl)pyridine, purification by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN (0.1% TFA) and water (0.1% TFA) to afford the trifluoroacetate salt of the title compound as an off-white solid (28 mg, 33%); HPLC: tR 4.72 min, 100 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.50 (d, J = 5.2 Hz, 1H), 8.37 (s, 1H), 7.78 (s, 1H), 7.30 (d, J = 4.0 Hz, 1H), 4.66 (dd, J = 49.0, 7.3 Hz, 1H), 4.54 (t, J = 5.8 Hz, 1H), 4.42 (t, J = 5.8 Hz, 1H), 4.13 (dd, J = 12.2, 8.2 Hz, 1H), 3.91 – 3.53 (m, 4H), 2.92 – 2.81 (m, 2H), 2.24 – 1.97 (m, 6H), 1.73 (tdd, J = 14.8, 10.0, 4.8 Hz, 2H), 1.49 (tt, J = 25.3, 12.6 Hz, 4H), 1.07 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 160.10, 153.72, 152.94, 152.62, 147.43, 140.51, 123.04, 119.63, 105.42, 93.99-92.27 (d, 1JCF = 173.7 Hz), 83.54-81.89 (d, 1JCF = 166.6 Hz), 69.40, 53.55, 49.76, 44.38 (d, 2JCF = 23.2 Hz), 33.47, 31.46, 31.07, 30.88, 29.54, 26.05 (d, 2JCF = 21.2 Hz), 9.31; 19F NMR (376 MHz, CDCl3) δ -186.80, -220.42; LCMS: m/z 420.3 [M+H]+; HRMS [M+H]+: m/z calculated 420.2569 found 420.2585. Example 5-25: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(6-nitropyridin-2-yl)pyrimidin-4- yl)amino)cyclohexan-1-ol
Figure imgf000056_0002
According to General Procedure Route B, using 2-bromo-6-nitropyridine, to afford the title compound as pale yellow solid (43 mg, 42% yield); HPLC: tR 4.50 min, 100 % purity at 254 nm; 1H NMR (400 MHz, CDCl3) δ 10.03 (bs, 1H), 8.55 (bs, 1H), 8.03 – 7.96 (m, 2H), 7.74 – 7.61 (m, 1H), 7.58 – 7.42 (m, 2H), 4.75 – 4.49 (m, 1H), 4.05 (dt, J = 13.8, 5.3 Hz, 1H), 3.93 – 3.68 (m, 2H), 3.62 – 3.40 (m, 1H), 2.23 (m, 2H), 2.16 – 2.01 (m, 2H), 1.84 – 1.60 (m, 2H), 1.58 – 1.42 (m, 3H), 1.27 (d, J = 14.5 Hz, 1H), 1.04 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 159.90, 154.19, 139.94, 133.22, 132.31, 132.21, 132.10, 132.07, 128.71, 128.59, 123.01, 113.16, 95.23 – 93.56 (d, 1JCF = 168.0 Hz), 77.36, 70.02, 45.28 – 45.07 (d, 2JCF = 21.2 Hz), 34.13, 34.09, 30.48, 26.03 – 25.82 (d, 2JCF = 21.2 Hz), 9.44 (d); 19F NMR (376 MHz, CDCl3) δ -186.56; LCMS: m/z 405.2 [M+H]+; HRMS [M+H]+: m/z calculated 405.2045 found 405.2060. Example 5-26: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(methylsulfonyl)pyridin-2- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol trifluoroacetate salt
Figure imgf000057_0001
According to General Procedure Route B, using 2-bromo-5- (methylsulfonyl)pyridine, purification by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN (0.1% TFA) and water (0.1% TFA) to afford the title compound as white solid (30 mg, 36% yield); HPLC: tR 4.83 min, 96.3 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 9.11 (d, J = 2.0 Hz, 1H), 8.56 (s, 1H), 8.39 (dd, J = 8.7, 2.4 Hz, 1H), 8.12 (d, J = 8.7 Hz, 1H), 4.79 – 4.55 (m, 1H), 4.23 – 4.04 (m, 1H), 3.92 – 3.52 (m, 3H), 3.24 (s, 3H), 2.17 (d, J = 9.7 Hz, 2H), 2.04 (d, J = 11.1 Hz, 2H), 1.82 – 1.64 (m, 2H), 1.64 – 1.35 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, MeOD) δ 160.80, 158.35, 154.10, 147.92, 144.33, 137.98, 136.86, 121.51, 106.25, 95.31 – 93.60 (d, 1JCF = 172.7 Hz), 69.85, 51.41, 46.13 – 45.91 (d, 2JCF = 22.2 Hz), 44.49, 34.30, 30.54, 26.91 – 26.70 (d, 2JCF = 21.2 Hz), 9.57 (d); 19F NMR (376 MHz, MeOD) δ -188.29; LCMS: m/z 438.1 [M+H]+; HRMS [M+H]+: m/z calculated 438.1970 found 438.1981. Example 5-27: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(morpholinosulfonyl)pyridin- 2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol trifluoroacetate salt
Figure imgf000057_0002
According to General Procedure Route B, using 4-((6-chloropyridin-3- yl)sulfonyl)morpholine, purification by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN (0.1% TFA) and water (0.1% TFA) to afford the trifluoroacetate salt of the title compound as white solid (31 mg, 38% yield); HPLC: tR 5.26 min, 97.4 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.97 (d, J = 2.0 Hz, 1H), 8.55 (s, 1H), 8.23 (dd, J = 8.7, 2.3 Hz, 1H), 8.12 (d, J = 8.7 Hz, 1H), 4.79 – 4.51 (m, 1H), 4.22 – 4.06 (m, 1H), 3.92 – 3.56 (m, 7H), 3.15 – 2.98 (m, 4H), 2.17 (d, J = 10.9 Hz, 2H), 2.04 (d, J = 11.4 Hz, 2H), 1.83 – 1.63 (m, 2H), 1.57 (dd, J = 23.6, 12.3 Hz, 2H), 1.51 – 1.33 (m, 2H), 1.07 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, MeOD) δ 160.81, 157.83, 154.07, 147.91, 144.07, 138.25, 132.10, 121.54, 106.32, 95.36 – 93.63 (d, 1JCF = 174.7Hz), 69.90, 67.19, 51.46, 47.27, 46.13 – 45.92 (d, 2JCF = 21.2 Hz), 34.37, 30.54, 26.91 – 26.70 (d, 2JCF = 21.2 Hz), 9.56 (d); 19F NMR (376 MHz, MeOD) δ -188.34; LCMS: m/z 509.3 [M+H]+; HRMS [M+H]+: m/z calculated 509.2341 found 509.2359. Example 5-28: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-methoxypyridin-2- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol
Figure imgf000058_0001
According to General Procedure Route B, using pyridine 4-methoxypyridine, to afford the title compound as an off-white solid (45 mg, 0.116 mmol, 62%).1H NMR (400 MHz, Chloroform-d) δ 9.88 (s, 1H), 8.29 (d, J = 5.55 Hz, 2H), 7.06 (d, J = 2.44 Hz, 1H), 6.66 (dd, J = 5.88, 2.33 Hz, 1H), 5.67 (s, 1H), 4.62 (dqd, J = 49.67, 4.61, 2.09 Hz, 1H), 3.99 (dp, J = 14.63, 4.92, 4.23 Hz, 1H), 3.86 (s, 3H), 3.80 (dt, J = 14.99, 3.71 Hz, 1H), 3.71 (ddt, J = 13.69, 9.45, 3.91 Hz, 1H), 3.47 (dddd, J = 18.42, 14.36, 7.47, 4.85 Hz, 1H), 2.54 (s, 1H), 2.16 (dd, J = 9.89, 5.20 Hz, 2H), 2.07 – 1.94 (m, 2H), 1.68 (dddd, J = 34.17, 12.18, 9.02, 6.07 Hz, 2H), 1.55 – 1.28 (m, 4H), 1.02 (t, J = 7.46 Hz, 3H).13C NMR (101 MHz, Chloroform-d) δ 166.3, 160.2, 160.2, 157.8, 153.7, 148.7 (d, J = 5.7 Hz), 107.6, 105.0, 104.0 (d, J = 4.3 Hz), 94.5 (d, J = 169.4 Hz), 69.9, 55.2, 48.5, 45.2 (d, J = 21.6 Hz), 34.0, 30.3, 25.9 (d, J = 20.9 Hz), 9.4 (d, J = 6.0 Hz).19F NMR (376 MHz, Chloroform-d) δ -186.70. HPLC - tR = 3.8 min >95% purity at 254 nm; LCMS [M+H]+ 390.3 m/z. Examples 5-29 to 5-32:
Figure imgf000058_0002
Scheme 2 General reductive amination procedure for preparation of pyridyl pyrimidine compounds A mixture of 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)nicotinaldehyde and amine ( 1.0 equiv.) in CH2Cl2 (1 mL) was added DIPEA (1.0 equiv.) and the resulting mixture refluxed for 30 mins. After cooling the reaction to room temp, NaBH(OAc)3 (1.0 equiv.) added and the reaction mixture was stirred at room temperature overnight. The solvent was removed and the residue was dissolved in CH2Cl2 and washed with a sat. solution of NaHCO3 and brine. The organic layer was dried (MgSO4) and concentrated. The residue was purified by prep-HPLC to afford a TFA salt of the title compound. Example 5-29: (1S,4r)-4-((5-(5-((dimethylamino)methyl)pyridin-2-yl)-2-(((S)-2- fluorobutyl)amino) pyrimidin-4-yl)amino) cyclohexan-1-ol White amorphous solid (55% yield, obtained as a TFA salt); HPLC: tR 3.62 min, >99 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.74 (d, J = 1.6 Hz, 1H), 8.49 (s, 1H), 8.07 (dt, J = 20.5, 5.3 Hz, 2H), 4.78 – 4.53 (m, 1H), 4.43 (s, 2H), 4.15 (dd,
Figure imgf000059_0001
J = 12.1, 8.4 Hz, 1H), 3.94 – 3.54 (m, 3H), 2.92 (s, 6H), 2.17 (d, J = 9.3 Hz, 2H), 2.10 – 1.93 (m, 2H), 1.83 – 1.61 (m, 2H), 1.61 – 1.29 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.31; LCMS: m/z 417.2 [M+H]+. Example 5-30: (1S,4r)-4-((5-(5-((4,4-difluoropiperidin-1-yl)methyl)pyridin-2-yl)-2-(((S)-2- fluorobutyl) amino)pyrimidin-4-yl)amino)cyclohexan-1-ol White amorphous solid; (61% yield, obtained as a TFA salt); HPLC: tR 3.62 min, >99 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.75 (s, 1H), 8.48 (s, 1H), 8.09 (d, J = 8.3 Hz, 1H), 8.05 – 7.98 (m, 1H), 4.70 (m, 1H), 4.44 (m, 2H), 4.15 (s,
Figure imgf000059_0002
2H), 3.90 – 3.56 (m, 3H), 3.50 (s, 3H), 3.36 (d, J = 6.8 Hz, 1H), 2.47 – 2.29 (m, 3H), 2.16 (s, 2H), 2.03 (m, 2H), 1.82 – 1.62 (m, 3H), 1.49 (m, 4H), 1.07 (t, J = 7.4 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -76.82, -188.31; LCMS: m/z 493.4 [M+H]+; HRMS [M+H]+: m/z calculated 493.2897 found 493.2903. Example 5-31: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(morpholinomethyl)pyridin-2- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol White amorphous solid; (68% yield, obtained as a TFA salt); HPLC: tR 3.45 min, >99 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.75 (s, 1H), 8.49 (s, 1H), 8.10 (d, J = 8.3 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H), 4.67 (dm, 1H), 4.47 (s, 2H), 4.13
Figure imgf000060_0003
(bs, 1H), 4.09 – 3.74 (m, 4H), 3.74 – 3.53 (m, 2H), 3.32 (s, 6H), 2.17 (bs, 2H), 2.04 (m, 2H), 1.84 – 1.62 (m, 2H), 1.62 – 1.36 (m, 4H), 1.08 (t, J = 7.4 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.37; LCMS: m/z 454.3 [M+H]+. Example 5-32: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(methyl(tetrahydrofuran-3- yl)amino) pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol White amorphous solid (18 % yield, obtained as a TFA salt); HPLC: tR 3.66 min, 100 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.60 (s, 1H), 8.40 (d, J = 10.4 Hz, 1H), 8.00 – 7.85 (m, 2H), 5.10 (s, 1H), 4.72 (m, 1H), 4.59 (s,
Figure imgf000060_0001
1H), 4.21 (m, 1H), 3.79 (s, 3H), 3.73 – 3.53 (m, 2H), 3.35 (bs, 2H), 2.90 (s, 6H), 2.16 (s, 2H), 2.03 (m, 2H), 1.86 – 1.62 (m, 3H), 1.62 – 1.36 (m, 3H), 1.07 (t, J = 7.4 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.35; LCMS: m/z 472.3 [M+H]+; HRMS [M+H]+: m/z calculated 472.3195 found 472.3203. Examples 5-33 to 5-36
Figure imgf000060_0002
(a) General Procedure for preparation of N-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl) amino) pyrimidin-5-yl)pyridin-3-yl)-1-methylpiperidine-4- carboxamide (5) A mixture of 4-((5-(5-chloropyridin-2-yl)-2-(((S)-2-fluorobutyl)amino)pyrimidin-4- yl)amino) cyclohexan-1-ol (1.0 equiv.), 1-methylpiperidine-4-carboxamide (1.2eq), Pd2dba3 (0.05 equiv.), di-tert-butyl(2',4',6'-triisopropyl-3,4,5,6-tetramethyl-[1,1'-biphenyl]-2- yl)phosphine (0.1 equiv.), K3PO4 in t-BuOH (2.0 mL) was heated at 110°C overnight. The solvent was removed under reduced pressure. The residue was purified by pre-HPLC to afford a TFA salt of the title compound. Example 5-33: N-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl) amino) pyrimidin-5-yl)pyridin-3-yl)-1-methylpiperidine-4-carboxamide According to General Procedure outlined in Scheme 3, using -methylpiperidine-4-carboxamide to afford the title compound as a white amorphous solid (65% yield,
Figure imgf000061_0002
obtained as a TFA salt); HPLC: tR 3.94 min, >95 % pure at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.86 (d, J = 2.3 Hz, 1H), 8.31 (s, 1H), 8.16 (dd, J = 8.8, 2.5 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 4.77 – 4.49 (m, 1H), 4.13 (t, J = 10.2 Hz, 1H), 3.91 – 3.50 (m, 4H), 3.44 (s, 1H), 3.09 (t, J = 11.9 Hz, 2H), 2.90 (s, 3H), 2.75 (m, 1H), 2.21-2.16 (m, 4H), 2.10 – 1.93 (m, 4H), 1.82 – 1.61 (m, 2H), 1.61 – 1.35 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.36; LCMS: m/z 500.3 [M+H]+. Example 5-34: 1-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)pyrrolidin-2-one
Figure imgf000061_0001
According to General Procedure outlined in Scheme 3, using pyrrolidin-2-one, to afford the title compound as a white amorphous solid (28 mg, 38% yield); HPLC: tR 5.06 min, >95 % purity at 254 nm; 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.74 (d, J = 2.7 Hz, 1H), 8.44 (s, 1H), 8.10 (dd, J = 9.0, 2.8 Hz, 1H), 7.87 (d, J = 9.1 Hz, 1H), 7.06 (s, 1H), 4.60 (d, J = 49.5 Hz, 2H), 3.86 (q, J = 10.1, 7ss.1 Hz, 3H), 3.65 – 3.41 (m, 3H), 2.05 (dq, J = 23.9, 9.2, 8.3 Hz, 4H), 1.83 (dt, J = 10.4, 4.4 Hz, 2H), 1.71 – 1.43 (m, 2H), 1.40 – 1.10 (m, 4H), 0.94 (t, J = 7.4 Hz, 3H); 13C NMR (101 MHz, DMSO) δ 174.30, 161.15, 159.21, 155.19, 150.96, 137.55, 132.85, 127.52, 118.23, 114.86, 94.56, 92.88 (d, 1JCF = 169.8 Hz), 67.92, 47.34, 33.71, 31.85, 30.16, 25.49, 25.29, 17.48, 9.10; 19F NMR (376 MHz, DMSO) δ -184.87; LCMS: m/z 500.3 [M+H]+; HRMS [M+H]+: m/z calculated 500.3144 found 500.3151. Example 5-35: 1-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)-3-methylimidazolidin-2-one 2,2,2- trifluoroacetate
Figure imgf000062_0001
According to General Procedure outlined in Scheme 3, using 1- methylimidazolidin-2-one, purification by reverse phase column to afford the trifluoroacetate salt of the title compound as an off-white solid.1H NMR (400 MHz, Methanol-d4) δ 8.81 (d, J = 2.7 Hz, 1H), 8.26 (s, 1H), 8.16 (dd, J = 9.0, 2.8 Hz, 1H), 7.83 (d, J = 9.0 Hz, 1H), 4.77 – 4.52 (m, 1H), 4.24 – 4.07 (m, 1H), 3.99 – 3.87 (m, 2H), 3.87 – 3.53 (m, 5H), 2.90 (s, 3H), 2.30 – 2.12 (m, 2H), 2.03 (dd, J = 10.9, 4.1 Hz, 2H), 1.85 – 1.62 (m, 2H), 1.60 – 1.38 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H). HPLC - tR = 4.0 min >95% purity at 254 nm; LCMS [M+H]+ 458.2 m/z; HRMS calcd for [M+H]+ 458.2674 m/z, found 458.2680. Example 5-36: 1-(6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)-4-methylpiperazin-2-one 2,2,2- trifluoroacetate
Figure imgf000062_0002
According to General Procedure outlined in Scheme 3, using 4-methylpiperazin-2- one, purification by reverse phase column to afford the trifluoroacetate salt of the title compound as an off-white solid.1H NMR (400 MHz, Methanol-d4) δ 8.73 – 8.66 (m, 1H), 8.40 (s, 1H), 7.98 (d, J = 2.7 Hz, 2H), 4.78 – 4.52 (m, 1H), 4.21 – 4.02 (m, 5H), 3.93 – 3.55 (m, 5H), 3.03 (s, 3H), 2.22 – 2.07 (m, 2H), 2.08 – 1.97 (m, 2H), 1.84 – 1.61 (m, 2H), 1.61 – 1.37 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H). HPLC - tR = 4.0 min >95% purity at 254 nm; LCMS [M+H]+ 472.2 m/z; HRMS calcd for [M+H]+ 472.2831 m/z, found 472.2830. Examples 5-37 and 5-38:
Figure imgf000063_0001
Scheme 4 (a) Example 5-37: (1R,4r)-4-((2-(((R)-2-Fluorobutyl)amino)-5-(pyridin-2-yl)pyrimidin-4- yl)amino)cyclohexan-1-ol To a solution of (1r,4r)-4-((2-(methylthio)-5-(pyridin-2-yl)pyrimidin-4- yl)amino)cyclohexan-1-ol (22 mg, 0.07 mmol) in DCM (4 mL) was added m-CPBA (75% purity; 18 mg, 0.10 mmol) and the mixture was stirred at room temperature for 2 h to give a mixture of sulfoxide and sulfone. The mixture was diluted with water and extracted with DCM (x2). The volatiles were removed in vacuo and the residue was then dissolved in THF (3 mL) and (R)-2-fluorobutan-1-amine hydrochloride (36 mg, 0.28 mmol) and DIPEA (45 mg, 0.35 mmol, 60 μl) were added. The resultant mixture was heated to 150 °C under microwave irradiation for 20 min. The volatiles were removed, the reaction was diluted with EtOAc and water, the layers were separated and the aqueous layer was extracted with EtOAc (x2). The combined organic layers were dried over magnesium sulfate, filtered and the volatiles were removed in vacuo. The residue was purified by silica gel flash column chromatography, eluting with 3% MeOH in DCM to yield the title compound as an off-white solid (18 mg, 72%). HPLC – tR = 4.516 min >97% purity at 254 mm; LRMS [M+H]+ 360.2 m/z; HRMS calcd for [M+H]+ 360.2194 m/z, found 360.2201 m/z; 1H NMR (400 MHz, MeOD) δ 8.58 (d, J = 4.8 Hz, 1H), 8.39 (s, 1H), 7.90 – 7.83 (m, 2H), 7.33 – 7.28 (m, 1H), 4.79 – 4.58 (m, 1H), 4.16 – 4.08 (m, 1H), 3.84 – 3.75 (m, 1H), 3.74 – 3.66 (m, 1H), 3.59 (ddd, J = 17.7, 14.5, 7.4 Hz, 1H), 2.28 – 2.16 (m, 2H), 2.10 – 2.02 (m, 2H), 1.82 – 1.70 (m, 2H), 1.56 – 1.43 (m, 4H), 1.10 (t, J = 7.5 Hz, 3H). (a) Example 5-38: (1r,4r)-4-((2-((2-Fluorobutyl)amino)-5-(pyridin-2-yl)pyrimidin-4- yl)amino)cyclohexan-1-ol dihydrochloride Using the same procedure outlined in Scheme 5, with 2-fluorobutan-1-amine hydrochloride. After chromatography, the crude solid was treated with 4M HCl in 1,4-dioxane (0.1 mL) followed by trituration with diethyl ether to give the title compound as a dihydrochloride salt (18.1 mg, 76%). HPLC – tR = 4.965 min >97% purity at 254 mm; LRMS [M+H]+ 360.0 m/z; HRMS [M+H]+ 360.2194 m/z, found 360.2200 m/z; 1H NMR (400 MHz, MeOD) δ 8.57 – 8.51 (m, 1H), 8.36 (s, 1H), 7.87 – 7.80 (m, 2H), 7.27 (ddd, J = 6.0, 5.1, 2.3 Hz, 1H), 4.65 (dtd, J = 12.8, 10.1, 7.1 Hz, 1H), 4.13 – 4.04 (m, 1H), 3.81 – 3.72 (m, 1H), 3.71 – 3.63 (m, 1H), 3.59 – 3.50 (m, 1H), 2.22 – 2.15 (m, 2H), 2.05 – 1.99 (m, 2H), 1.80 – 1.67 (m, 2H), 1.51 – 1.41 (m, 4H), 1.07 (t, J = 7.5 Hz, 3H). Example 5-39:
Figure imgf000064_0001
Scheme 5 (a) (1r,4r)-4-((5-(1,4-dimethyl-1H-pyrazol-3-yl)-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol A suspension of (4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin- 5-yl)boronic acid 3 (0.5g, 1.77 mmol, 1.0 equiv.), 3-bromo-1,4-dimethyl-1H-pyrazole (0.34g, 1.94 mmol, 1.1equiv.), K2CO3 (0.488g, 3.53 mmol, 2.0 equiv.), tetrakis(triphenylphosphine)palladium(0) (0.102 g, 0.0885, 0.05 equiv.) in 1,4-dioxane (8 mL) and water (2 mL) was purged with nitrogen for 10 min. The mixture was stirred at 90 °C for 4 hours. The mixture was diluted with EtOAc, and the resulting suspension was filtered through the Celite® pad which was washed with EtOAc. The filtrate was washed with ice cold water (3 × 20 mL) and brine, dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (5-70% EtOAc in Pet. Sp.) to afford the title compound (0.318 g, 54% yield) as a white solid; 1H NMR (400 MHz, Chloroform-d) δ 8.24 (s, 1H), 7.25 – 7.18 (m, 1H), 4.20 – 4.01 (m, 1H), 3.87 (s, 3H), 3.72 (dq, J = 9.8, 4.8 Hz, 1H), 2.57 (s, 3H), 2.21 (d, J = 0.8 Hz, 5H), 2.04 (t, J = 6.6 Hz, 2H), 1.56 – 1.29 (m, 4H); LCMS: m/z 334.2 [M+H]+ (b) Example 5-39: (1S,4r)-4-((5-(1,4-dimethyl-1H-pyrazol-3-yl)-2-(((S)-2- fluorobutyl)amino)pyrimidin-4-yl)amino)cyclohexan-1-ol To a solution of (1r,4r)-4-((5-(1,4-dimethyl-1H-pyrazol-3-yl)-2- (methylthio)pyrimidin-4-yl)amino)cyclohexan-1-ol (from step (a), 75 mg, 0.22 mmol, 1.0 equiv.) in THF (2.0 mL) was added m-CPBA (77%) (108.6 mg, 0.45 mmol, 2.01 equiv.). The reaction mixture was stirred at room temperature for 1 h. Then (S)-2-fluorobutan-1-amine hydrochloride (119.4 mg, 1.12 mmol) was added and the resulting reaction mixture was heated under microwave irradiation at 150 °C for 1h. The solvent was removed under reduced pressure, the residue was washed with saturated aqueous NaHCO3 solution and extracted with EtOAc (2x 50 mL). The combined organic fractions were dried over MgSO4 and concentrated. The residue was purified by silica gel flash chromatography (10-80% EtOAc in Pet. Sp.) to afford the title compound as white amorphous solid (32 mg, 37%); HPLC: tR 5.16 min, >95 % purity at 254 nm; 1H NMR (400 MHz, CDCl3) δ 8.02 (s, 1H), 7.92 – 7.72 (m, 1H), 7.16 (s, 1H), 5.59 (bs, 1H), 4.75 – 4.46 (m, 1H), 3.95 (m, 1H), 3.83 (s, 3H), 3.78 – 3.61 (m, 2H), 3.45 (m, 1H), 2.74 (s, 2H), 2.25 – 2.08 (m, 5H), 2.08 – 1.91 (m, 2H), 1.81 – 1.54 (m, 2H), 1.53 – 1.37 (m, 2H), 1.37 – 1.15 (m, 3H), 1.01 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, CDCl3) δ -186.90; 13C NMR (101 MHz, CDCl3) δ 160.21, 159.48, 152.91, 145.64, 130.73, 113.91, 102.05, 95.50 – 93.81 (d, 1JCF = 170.7 Hz), 69.95, 48.60, 45.19 (d, 2JCF = 21.9 Hz), 38.87, 34.00, 30.41, 27.04, 26.92 (d, 2JCF = 21.2 Hz), 10.77, 9.43; 19F NMR (376 MHz, CDCl3) δ - 186.90; LCMS: m/z 377.2 [M+H]+; HRMS [M+H]+: m/z calculated 377.2460 found 377.2462. Example 5-40:
Figure imgf000065_0001
Scheme 6 (a) (1r,4r)-4-((5-(1-methyl-1H-pyrazol-3-yl)-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol A suspension of (4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin- 5-yl)boronic acid 3 (0.5 g, 1.77 mmol, 1.0 equiv.), 3-bromo-1-methyl-1H-pyrazole (0.312 g, 1.94 mmol, 1.1 equiv.), K2CO3 (0.488 g, 3.53 mmol, 2.0 equiv.), tetrakis(triphenylphosphine)palladium(0) (0.102 g, 0.0885 mmol, 0.05 equiv.) in 1,4-dioxane (8 mL) and water (2 mL) was purged with nitrogen for 10 min. The mixture was stirred at 90 °C for 4 h. The mixture was diluted with EtOAc and the resulting suspension was filtered through the Celite® pad, which was washed with EtOAc. The filtrate was washed with ice cold water (3 × 20 mL) and brine. The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (5-70% EtOAc in Pet. Sp.) to afford the title compound (0.272 g, 48% yield) as a white solid; 1H NMR (400 MHz, Chloroform-d) δ 8.67 (s, 1H), 8.31 (s, 1H), 7.39 (d, J = 2.6 Hz, 1H), 6.56 (d, J = 2.6 Hz, 1H), 4.13 (ddt, J = 10.0, 7.1, 3.4 Hz, 1H), 3.93 (d, J = 2.7 Hz, 3H), 3.76 (d, J = 10.7 Hz, 1H), 2.57 (s, 3H), 2.33 – 2.13 (m, 2H), 2.13 – 1.92 (m, 2H), 1.46 (m, 4H); LCMS: m/z 320.1 [M+H]+ (b) Example 5-40: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(1-methyl-1H-pyrazol-3- yl)pyrimidin-4-yl)amino)cyclohexan-1-ol To a solution of (1r,4r)-4-((5-(1-methyl-1H-pyrazol-3-yl)-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol (from step (a), 70 mg, 0.219 mmol, 1.0 equiv.) in THF (2.0 mL) was added m-CPBA (77%) (108.6 mg, 0.45 mmol, 2.01 equiv.). The reaction mixture was stirred at room temperature for 1 h. Then (S)-2-fluorobutan-1-amine hydrochloride (116.3 mg, 1.10 mmol, 5.0 equiv.) and DIPEA (133.6 µL, 0.766 mmol, 3.5 equiv.) were added and the resulting reaction mixture was heated under microwave irradiation at 150 °C for 1h. The solvent was removed under reduced pressure, washed with saturated aqueous NaHCO3 solution and extracted with EtOAc (2x 50 mL). The combined organic fractions were dried over MgSO4, filtered and concentrated. The residue was purified by silica gel chromatography (10-80% EtOAc in Pet. Sp.) followed by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN and water to afford the title compound as white amorphous solid (29 mg, 36%); 1H NMR (400 MHz, CDCl3) δ 8.50 – 8.22 (m, 1H), 8.14 (s, 1H), 7.32 (d, J = 2.4 Hz, 1H), 6.43 (d, J = 2.4 Hz, 1H), 5.52 (s, 1H), 4.62 (m, 1H), 4.01 (m, 1H), 3.89 (s, 3H), 3.77 (m, 2H), 3.57 – 3.27 (m, 1H), 2.25 – 2.13 (m, 2H), 2.04 (dt, J = 13.0, 3.7 Hz, 2H), 1.79 – 1.59 (m, 2H), 1.55 – 1.32 (m, 4H), 1.27 (d, J = 13.0 Hz, 2H), 1.02 (t, J = 7.4 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 158.81, 152.27, 148.74, 130.82, 125.24, 101.27, 95.48 – 93.80 (d, 1JCF = 169.7 Hz), 70.00, 48.58, 45.34 – 45.12 (d, 2JCF = 22.27 Hz), 39.02, 34.00, 33.96, 30.42, 29.83, 26.04 – 25.83 (d, 1JCF = 21.2 Hz), 9.45; 19F NMR (376 MHz, CDCl3) δ -186.87; LCMS: m/z 363.2 [M+H]+; HRMS [M+H]+: m/z calculated 363.2303 found 363.2308.
Figure imgf000067_0001
Scheme 7 (a) 6-(4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin-5-yl)nicotinic acid A suspension of (4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin- 5-yl)boronic acid 3 (1.0 g, 3.53 mmol, 1.0 equiv.), methyl 6-bromonicotinate (0.762 g, 3.53 mmol, 1.0 equiv.), K2CO3 (1.46 g, 10.59 mmol, 3.0 equiv.), tetrakis(triphenylphosphine)palladium(0) (204.0 mg, 0.176 mmol, 0.05 equiv.) in 1,4-dioxane (16 mL) and water (4 mL) was purged with nitrogen for 10 min. The mixture was stirred at 100 °C for 6 hours at which point it was concentrated to a dark brown/black residue and filtered through pad of Celite® and adjusted pH to 4 using 10% aqueous citric acid solution. The crude mixture was diluted with ethyl acetate and extracted (3 x 100 mL), the combined organic layer was dried over MgSO4 and concentrated. The crude material was purified by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN and water to afford the title product (720 mg, 56%) as a pale yellow solid; 1H NMR (400 MHz, Methanol- d4) δ 9.06 (s, 1H), 8.53 (s, 1H), 8.31 (dd, J = 8.4, 2.2 Hz, 1H), 7.92 (dd, J = 8.7, 0.9 Hz, 1H), 4.23 – 4.07 (m, 1H), 3.78 – 3.60 (m, 1H), 2.56 (s, 3H), 2.26 – 2.12 (m, 2H), 2.03 (d, J = 5.6 Hz, 2H), 1.61 – 1.38 (m, 4H); LCMS: m/z 361.0 [M+H]+ (b) 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl)amino)pyrimidin-5- yl)nicotinic acid To a solution of 6-(4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidin- 5-yl)nicotinic acid (from step (a), 0.3 mg, 0.832 mmol, 1.0 equiv.) in NMP (2.0 mL) was added m-CPBA (77%) (374.93 mg, 1.67 mmol, 2.01 equiv.). The reaction mixture was stirred at room temperature for 1.0 h. Then (S)-2-fluorobutan-1-amine hydrochloride (414.9 mg, 4.16 mmol, 5.0 equiv.) and DIPEA (507.4 µL, 2.91 mmol, 3.5 equiv.) were added and the resulting reaction mixture was heated under microwave irradiation at 150 °C for 1h. The solvent was removed under reduced pressure, washed with saturated aqueous NaHCO3 solution and extracted with EtOAc (2x 50 mL). The combined organic fractions were dried over MgSO4, filtered and concentrated. The residue was purified by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN and water to afford title compound as a light yellow solid (210 mg, 62%); 1H NMR (400 MHz, Acetone-D6) δ 7.99 (d, J = 1.9 Hz, 1H), 7.66 – 7.59 (m, 1H), 7.59 – 7.46 (m, 2H), 4.75 – 4.46 (m, 1H), 3.82 – 3.15 (m, 4H), 2.08 (d, J = 4.6 Hz, 2H), 2.05 (dd, J = 4.4, 2.2 Hz, 2H), 1.80 – 1.50 (m, 4H), 1.42 (dd, J = 6.6, 3.3 Hz, 2H), 0.97 (dd, J = 14.7, 7.4 Hz, 3H). LCMS: m/z 404.3 [M+H]+ (c) Example 5-41: (6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl) (isoxazolidin-2-yl)methanone trifluoroacetate salt To a solution of 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)nicotinic acid (from step (b), 30 mg, 0.074 mmol, 1.0 equiv.), O-(benzotriazol-1-yl)-N,N,N′,N′- tetramethyluronium tetrafluoroborate (TBTU) (36.7 mg, 0.096 mmol, 1.3 equiv.) and DIPEA (36.86 µL, 0.223 mmol, 3.0 equiv.) in anhydrous DMF (3.0 mL) was added isoxazolidine (6.52 mg, 0.089 mmol, 1.2 equiv.) in DMF (1.0 mL) drop-wise at room temperature. The resulting mixture was stirred overnight. The reaction was then treated with saturated aqueous NaHCO3 solution, extracted with DCM and the organic layer was dried (MgSO4) and concentrated. The residue was purified by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN (0.1% TFA) and water (0.1% TFA) to afford the trifluoroacetate salt of the title compound as a white solid (13.0 mg, 38%); 1H NMR (400 MHz, DMSO) δ 10.88 (s, 1H), 8.91 (s, 1H), 8.71 (s, 1H), 8.19 (d, J = 8.4 Hz, 1H), 8.07 (d, J = 8.4 Hz, 1H), 4.66 (bd, 1H), 3.98 (t, J = 6.7 Hz, 2H), 3.82 (t, J = 7.2 Hz, 2H), 2.38 – 2.23 (m, 2H), 1.99 (d, J = 5.3 Hz, 2H), 1.87 (d, J = 10.1 Hz, 2H), 1.78 – 1.55 (m, 3H), 1.46 (m, 2H), 1.35 – 1.11 (m, 3H), 0.98 (t, J = 7.3 Hz, 3H) (Two protons hiding under residual water peak); 19F NMR (376 MHz, DMSO) δ -184.96; LCMS: m/z 459.4 [M+H]+; HRMS [M+H]+: m/z calculated 459.2528 found 459.2514. (c) Example 5-42: (6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)pyridin-3-yl)(4-fluoropiperidin-1-yl)methanone trifluoroacetate salt Using the same procedure as for Example 5-41, with 4-fluoropiperidine hydrochloride, to afford the trifluoroacetate salt of the title compound as a brown amorphous solid (21.0 mg, 34%); HPLC: tR 5.13 min, >99 % purity at 254 nm; 1H NMR (400 MHz, CDCl3) δ 10.82 (d, J = 7.0 Hz, 1H), 9.70 (bs, 1H), 8.61 (s, 1H), 8.20 (s, 1H), 7.84 (d, J = 7.1 Hz, 1H), 7.65 (d, J = 7.8 Hz, 1H), 4.94 (d, J = 47.6 Hz, 1H), 4.63 (m, 1H), 4.08 (bs, 2H), 3.86 – 3.36 (m, 6H), 3.24 (m, 2H), 2.18 (s, 2H), 1.99 (d, J = 42.1 Hz, 6H), 1.80 – 1.56 (m, 2H), 1.47 (s, 3H), 1.04 (t, J = 7.4 Hz, 3H); 19F NMR (376 MHz, CDCl3) δ -184.22, -186.76; 13C NMR (101 MHz, CDCl3) δ 166.86, 159.63, 153.85, 153.69, 146.06, 141.32, 136.49, 130.02, 118.83, 104.61, 93.78 - 92.06 (d, 1JCF = 173.7 Hz), 87.83 - 86.12 (d, 1JCF = 172.7 Hz), 69.10, 66.99, 49.72, 44.90 - 44.66 (d, 2JCF = 24.2 Hz), 33.24, 29.42, 29.37, 25.98 - 25.77 (d, 2JCF = 21.2 Hz), 9.10; LCMS: m/z 463.1 [M+H]+; HRMS [M+H]+: m/z calculated 489.2784 found 489.2802. (c) Example 5-43: 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)-N-(2-fluoroethyl)-N-methylnicotinamide trifluoroacetate salt Using the same procedure as for Example 5-41, with 2-fluoro-N-methylethan-1- amine hydrochloride salt, to afford the trifluoroacetate salt of the title compound as a light brown amorphous solid (12.0 mg, 52%); HPLC: tR 4.85 min, >95 % purity at 254 nm; 1H NMR (400 MHz, CDCl3) δ 10.82 (d, J = 7.0 Hz, 1H), 9.70 (s, 1H), 8.61 (s, 1H), 8.20 (s, 1H), 7.84 (d, J = 7.1 Hz, 1H), 7.65 (d, J = 7.8 Hz, 1H), 4.94 (bd, 1H), 4.81 – 4.46 (m, 1H), 4.08 (bs, 2H), 3.86 – 3.36 (m, 6H), 3.24 (d, J = 27.7 Hz, 2H), 2.18 (s, 2H), 1.99 (d, J = 42.1 Hz, 6H), 1.80 – 1.56 (m, 2H), 1.04 (t, J = 7.4 Hz, 3H); 13C NMR (101 MHz, CDCl3) δ 167.01, 159.78, 154.00, 153.84, 146.21, 141.47, 136.64, 130.17, 118.98, 104.76, 93.93 – 92.21 (d, 1JCF = 173.7 Hz), 87.98 -86.27 (d, 1JCF = 172.7 Hz), 77.36, 69.25, 67.14, 49.87, 45.05 – 44.81 (d, 2JCF = 24.2 Hz), 33.39, 29.57, 26.13 – 25.92 (d, 2JCF = 24.2 Hz), 9.25 (d); 19F NMR (376 MHz, CDCl3) δ - 184.22, -186.76; LCMS: m/z 463.1 [M+H]+; HRMS [M+H]+: m/z calculated 463.2628 found 463.2651. (c) Example 5-44: 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)-N-(1-methylpiperidin-4-yl)nicotinamide trifluoroacetate salt Using the same procedure as for Example 5-41, with 1-methylpiperidin-4-amine, to afford the trifluoroacetate salt of the title compound as a brown amorphous solid (22.0 mg, 35%); HPLC: tR 4.15 min, >95 % purity at 254 nm; 1H NMR (401 MHz, Acetone) δ 11.19 (s, 1H), 9.98 (s, 1H), 9.12 (s, 1H), 8.s64 (s, 1H), 8.39 (d, J = 6.7 Hz, 1H), 8.20 (s, 1H), 4.70 (dm, 1H), 4.22 (m, 2H), 3.91 – 3.46 (m, 4H), 3.20 (m, 8H), 2.92 (s, 3H), 2.21 (s, 2H), 2.11 – 1.94 (m, 2H), 1.86 – 1.65 (m, 2H), 1.65 – 1.37 (m, 3H), 1.04 (t, J = 7.4 Hz, 3H); 13C NMR (101 MHz, Acetone) δ 163.81, 159.61, 155.31, 146.72, 136.41, 130.06, 128.06, 119.29, 115.55, 104.43, 93.87 – 92.17 (d, 1JCF = 171.7 Hz), 68.13, 53.19, 49.65, 49.48, 44.68, 44.60, 44.37, 42.60, 33.28, 25.69 – 25.48 (d, 2JCF = 21.2 Hz), 8.63 (d); 19F NMR (377 MHz, Acetone) δ - 187.00; LCMS: m/z 500.2 [M+H]+; HRMS [M+H]+: m/z calculated 500.3144 found 500.3153. Examples 5-45 and 5-46:
Figure imgf000070_0001
Scheme 8 (a) 2-bromo-4-(1,3-dioxolan-2-yl)pyridine To a solution of 2-bromoisonicotinaldehyde (5.0 g, 26.88 mmol, 1.0 equiv.) in toluene (300 mL) was added ethylene glycol (6.01 mL, 0.108 mmol, 4.0 equiv.) and PTSA (463 mg, 2.69 mmol, 0.1 equiv.), and the reaction mixture was heated to reflux overnight with azeotropic removal of the water using a Dean-Stark trap. The mixture was cooled down and washed with saturated NaHCO3 solution (3 × 100 mL) and brine (1 × 100 mL). The organic phase was dried over anhydrous MgSO4 and concentrated to afford title compound as a yellow oil (5.30 g, 23.04 mmol, 86%).1H NMR (400 MHz, Chloroform-d) δ 8.37 (d, J = 5.02 Hz, 1H), 7.59 (s, 1H), 7.33 (dd, J = 4.97, 1.41 Hz, 1H), 5.78 (s, 1H), 4.04 (s, 4H).13C NMR (101 MHz, Chloroform-d) δ 150.3, 142.3, 139.8, 125.7, 120.4, 101.1, 65.5. HPLC - tR = 3.5 min >95% purity at 254 nm; LCMS [M+H]+ 229.9 m/z (Br79), 231.9 m/z (Br81). (b) Lithium (4-(1,3-dioxolan-2-yl)pyridin-2-yl)trihydroxyborate To a solution of 2-bromo-4-(1,3-dioxolan-2-yl)pyridine (from step (a), 5.25 g, 22.83 mmol, 1.0 equiv.) in a mixture of toluene (200 mL) and tetrahydrofuran (50 mL) and under an atmosphere of nitrogen was added B(i-OPr)3 (5.27 mL, 22.83 mmol, 1.0 equiv.). The mixture was cooled to -78 °C and stirred for 30 min. Then 2.0 M solution of n-BuLi in hexane (10.84 mL, 21.69 mmol, 1.0 equiv.) was slowly added over 30 min. The reaction mixture was stirred at the same temperature for 3 h, then the cooling bath was removed. Upon the mixture slowly warmed to 0 °C (around 1.5 h), the reaction was quenched with isopropyl alcohol. The resulting suspension was stirred overnight. The solvent was then removed under reduced pressure, and the resulting residue was treated with acetone and water (9:1, v/v) until solid was precipitated. The precipitate was filtered, further washed with acetone and water (9:1, v/v) and then dried under reduced pressure to afford the title compound as a brown solid (3.46 g, 15.78 mmol, 69%).1H NMR (400 MHz, Deuterium Oxide) δ 8.56 (s, 1H), 7.88 – 7.72 (m, 1H), 7.67 (d, J = 8.00 Hz, 1H), 5.88 (s, 1H), 4.22 (d, J = 6.66 Hz, 2H), 4.12 (d, J = 6.94 Hz, 2H). (c) (1r,4r)-4-((5-(4-(1,3-dioxolan-2-yl)pyridin-2-yl)-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol To a solution of (1r,4r)-4-((5-bromo-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol 2 (1.6 g, 5.03 mmol, 1.0 equiv.) in a mixture of DMF (48 mL) and H2O (12 mL) were added lithium (4-(1,3-dioxolan-2-yl)pyridin-2-yl)trihydroxyborate (from step (b), 3.30 g, 15.08 mmol, 3.0 equiv.), CuBr (144 mg, 1.01 mmol, 0.2 equiv.) and K2CO3 (2.08 g, 15.08 mmol, 3.0 equiv.). The reaction mixture was degassed with nitrogen for 30 min. Then Pd(dppf)Cl2 (368 mg, 0.503 mmol, 10 mol%) was added under nitrogen flushing. The reaction mixture was heated to 110 °C and stirred for 1 h. The mixture was diluted with EtOAc, and the resulting suspension was filtered through a pad of Celite with the aid of EtOAc. The filtrate was further washed with water (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (0-50% EtOAc in Pet. Sp.) to afford the title compound as an off-white solid (445 mg, 1.15 mmol, 23%). HPLC - tR = 4.1 min >95% purity at 254 nm; LCMS [M+H]+ 389.2 m/z. (d) Example 5-45: (1S,4r)-4-((5-(4-(1,3-dioxolan-2-yl)pyridin-2-yl)-2-(((S)-2- fluorobutyl)amino)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2-trifluoroacetate To a solution of (1r,4r)-4-((5-(4-(1,3-dioxolan-2-yl)pyridin-2-yl)-2- (methylthio)pyrimidin-4-yl)amino)cyclohexan-1-ol (from step (c), 435 mg, 1.12 mmol, 1.0 equiv.) in THF (5.0 mL) was added m-CPBA (75% purity, 518 mg, 2.25 mmol, 2.0 equiv.). The mixture was stirred at room temperature for 1 h. The reaction was monitored by TLC. Upon all starting material was consumed, (S)-2-fluorobutan-1-amine hydrochloride (381 mg, 2.99 mmol, 4.0 equiv.) and DIPEA (520 µL, 2.99 mmol, 4.0 equiv.) were added. The reaction mixture was heated under microwave irradiation at 140 °C for 1 h. Then the solvent was removed under reduced pressure. The resulting residue was purified by a reverse phase C18 column to afford the trifluoroacetate salt of the title compound as a yellow solid (63 mg, 0.146 mmol, 10%).1H NMR (400 MHz, Methanol-d4) δ 8.62 (d, J = 5.20 Hz, 1H), 8.41 (s, 1H), 7.95 (s, 1H), 7.47 (dd, J = 5.22, 1.32 Hz, 1H), 4.76 – 4.54 (m, 1H), 4.22 – 3.99 (m, 5H), 3.92 – 3.52 (m, 3H), 2.26 – 2.13 (m, 2H), 2.03 (dd, J = 11.40, 4.54 Hz, 2H), 1.84 – 1.64 (m, 2H), 1.61 – 1.38 (m, 4H), 1.07 (t, J = 7.46 Hz, 3H).13C NMR (101 MHz, Methanol-d4) δ 161.1, 153.9, 150.7, 148.9, 144.7, 141.8, 121.4, 118.7, 109.9, 102.9, 94.5 (d, J = 171.4 Hz), 69.8, 66.6, 51.2, 45.9 (d, J = 22.1 Hz), 34.2, 30.5, 26.8 (d, J = 20.9 Hz), 9.6 (d, J = 5.8 Hz).19F NMR (376 MHz, Methanol-d4) δ -76.93 (TFA), -188.32. HPLC - tR = 4.5 min >95% purity at 254 nm; LCMS [M+H]+ 432.2 m/z. (e) Example 5-46: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(4-(((2- fluoroethyl)(methyl)amino)methyl)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2-trifluoroacetate To a solution of Example 5-45 (from step (d), 33 mg, 85 µmol, 1.0 equiv.) in water was added 4 M aqueous HCl solution (0.2 mL, 0.8 mmol). The mixture was stirred under reflux for 30 min. Then 50% aqueous NaOH was used to adjust the pH to 7. The mixture was again diluted and extracted with DCM. The organic layer was collected and evaporated to give a yellow residue. The residue was diluted in DCE (1 mL), 2-fluoro-N-methylethan-1- amine hydrochloride (97 mg, 0.852 mmol, 10.0 equiv.) and DIPEA (0.148 mL, 0.852 mmol, 10.0 equiv.) were added. The mixture was stirred under reflux for 30 min before NaBH(OAc)3 (27 mg, 0.128 mmol, 1.5 equiv.) was added. Then the reaction was stirred at room temperature overnight. The mixture was diluted with DCM and further washed with saturated aqueous NaHCO3 (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase C18 column to afford to afford the trifluoroacetate salt of the title compound as an off-white solid (34.4 mg, 0.061 mmol, 79%).1H NMR (400 MHz, Methanol-d4) δ 8.73 (d, J = 5.17 Hz, 1H), 8.44 (s, 1H), 8.09 (s, 1H), 7.65 – 7.48 (m, 1H), 5.03 – 4.95 (m, 1H), 4.76 – 4.58 (m, 1H), 4.54 (s, 2H), 4.15 (td, J = 10.44, 8.50, 5.00 Hz, 1H), 3.92 – 3.46 (m, 5H), 2.96 (s, 3H), 2.25 – 2.14 (m, 2H), 2.09 – 1.95 (m, 2H), 1.86 – 1.64 (m, 2H), 1.62 – 1.37 (m, 4H), 1.07 (t, J = 7.45 Hz, 3H).13C NMR (101 MHz, Methanol-d4) δ 163.9, 161.0, 154.9, 149.7, 142.3, 141.6, 125.3, 123.3, 106.8, 94.5 (d, J = 171.4 Hz), 79.4 (d, J = 168.2 Hz), 69.8, 59.9, 57.4 (d, J = 19.5 Hz), 51.2, 45.9 (d, J = 22.3 Hz), 41.3 (d, J = 2.4 Hz), 34.2, 30.5, 26.8 (d, J = 20.5 Hz), 9.6 (d, J = 6.0 Hz).19F NMR (376 MHz, Methanol-d4) δ -77.09 (TFA), -188.32, -222.18. HPLC pending; LCMS [M+H]+ 449.2 m/z. Examples 5-47 - 5-49:
Figure imgf000073_0001
Scheme 9 (a) 2-bromo-5-(1,3-dioxolan-2-yl)pyridine To a solution of 6-bromonicotinaldehyde (1.23 g, 6.61 mmol, 1.0 equiv.) in toluene (300 mL) were added ethylene glycol (1.48 mL, 26.45 mmol, 4.0 equiv.) and PTSA (114 mg, 0.661 mmol, 0.1 equiv.) and the reaction mixture was heated to reflux overnight with azeotropic removal of the water using a Dean-Stark trap. The mixture was cooled down and washed with saturated aqueous NaHCO3 solution (3 × 100 mL) and brine (1 × 100 mL). The organic phase was dried over anhydrous MgSO4 and concentrated to afford title compound as a yellow oil (1.23 g, 5.34 mmol, 81%).1H NMR (400 MHz, Chloroform-d) δ 8.46 (d, J = 2.30 Hz, 1H), 7.66 (dd, J = 8.15, 2.42 Hz, 1H), 7.51 (d, J = 8.20 Hz, 1H), 5.82 (s, 1H), 4.16 – 3.95 (m, 4H).13C NMR (101 MHz, Chloroform-d) δ 148.4, 142.3, 137.4, 133.6, 128.2, 101.2, 65.6. HPLC - tR = 3.4 min >95% purity at 254 nm; LCMS [M+H]+ 229.9 m/z (Br79), 231.9 m/z (Br81). (b) Lithium (5-(1,3-dioxolan-2-yl)pyridin-2-yl)trihydroxyborate To a solution of 2-bromo-5-(1,3-dioxolan-2-yl)pyridine (from step (a), 1.23 g, 5.35 mmol, 1.0 equiv.) in a mixture of toluene (48 mL) and tetrahydrofuran (12 mL) and under an atmosphere of nitrogen was added B(i-OPr)3 (1.23 mL, 5.35 mmol, 1.0 equiv.). The mixture was cooled to -78 °C and stirred for 30 min. Then 2.0 M solution of n-BuLi in hexane (2.54 mL, 5.08 mmol, 1.0 equiv.) was slowly added over 30 min. The reaction mixture was stirred at the same temperature for 3 h, then the cooling bath was removed. Upon the mixture slowly warmed to 0 °C (around 1.5 h), the reaction was quenched with isopropyl alcohol. The resulting suspension was stirred overnight. The solvent was then removed under reduced pressure and the resulting residue was treated with acetone and water (9:1, v/v) until solid was precipitated. The precipitate was filtered, further washed with acetone and water (9:1, v/v) and then dried under reduced pressure to afford the title compound as a brown solid (711 mg, 3.25 mmol, 61%).1H NMR (400 MHz, Deuterium Oxide) δ 8.56 (s, 1H), 7.88 – 7.72 (m, 1H), 7.67 (d, J = 8.00 Hz, 1H), 5.88 (s, 1H), 4.22 (d, J = 6.66 Hz, 2H), 4.12 (d, J = 6.94 Hz, 2H). (c) (1r,4r)-4-((5-(5-(1,3-dioxolan-2-yl)pyridin-2-yl)-2-(methylthio)pyrimidin-4- yl)amino)cyclohexan-1-ol To a solution of 2 (1.91 g, 5.99 mmol, 1.0 equiv.) in a mixture of DMF (60 mL) and H2O (15 mL) were added lithium (5-(1,3-dioxolan-2-yl)pyridin-2-yl)trihydroxyborate (from step (b), 3.94 g, 17.98 mmol, 3.0 equiv.), CuBr (172 mg, 1.20 mmol, 0.2 equiv.) and K2CO3 (2.48 g, 17.98 mmol, 3.0 equiv.). The reaction mixture was degassed with nitrogen for 30 min. Then Pd(dppf)Cl2 (438 mg, 0.599 mmol, 10 mol%) was added under nitrogen flushing. The reaction mixture was heated to 110 °C and stirred for 1 h. The mixture was diluted with EtOAc and the resulting suspension was filtered through a pad of Celite with the aid of EtOAc. The filtrate was further washed with water (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (0-50% EtOAc in Pet. Sp.) to afford the title compound as an off-white solid (1.62 g, 4.18 mmol, 70%).1H NMR (400 MHz, Methanol-d4) δ 10.12 (d, J = 7.16 Hz, 1H), 8.61 (s, 1H), 8.45 (s, 1H), 7.88 (s, 2H), 5.83 (s, 1H), 4.21 – 3.89 (m, 5H), 3.65 (dt, J = 10.04, 4.86 Hz, 1H), 2.53 (s, 3H), 2.26 – 2.12 (m, 2H), 2.00 (dt, J = 11.80, 4.22 Hz, 2H), 1.43 (p, J = 6.95, 6.37 Hz, 4H).13C NMR (101 MHz, Methanol-d4) δ 172.7, 159.9, 156.8, 154.0, 147.2, 136.8, 133.3, 120.7, 109.6, 102.9, 70.3, 66.6, 50.2, 34.5, 31.2, 14.1. HPLC - tR = 4.1 min >95% purity at 254 nm; LCMS [M+H]+ 389.1 m/z. (d) 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl)amino)pyrimidin-5- yl)nicotinaldehyde To a solution of (1r,4r)-4-((5-(5-(1,3-dioxolan-2-yl)pyridin-2-yl)-2- (methylthio)pyrimidin-4-yl)amino)cyclohexan-1-ol (from step (c), 535 mg, 1.38 mmol, 1.0 equiv.) in THF (5.0 mL) was added m-CPBA (75% purity, 637.84 mg, 2.77 mmol, 2.01 equiv.). The mixture was stirred at room temperature for 1 h and (S)-2-fluorobutan-1-amine hydrochloride (502 mg, 3.93 mmol, 2.85 equiv.) and DIPEA (961 µL, 5.52 mmol, 4.0 equiv.) were added. The reaction mixture was heated under microwave irradiation at 140 °C for 1 h. Then the solvent was removed under reduced pressure and the resulting residue was diluted in water and added 4 M aqueous HCl solution (2 mL, 8 mmol). The mixture was stirred under reflux for 30 min. Then 50% aqueous NaOH was used to adjust the pH to 7. The mixture was again diluted and extracted with DCM. The organic layer was collected and evaporated to give the crude title compound as a yellow solid (162 mg, 0.418 mmol, 33%).1H NMR (400 MHz, Chloroform-d) δ 10.04 (s, 1H), 8.92 (d, J = 2.1 Hz, 1H), 8.51 (d, J = 7.6 Hz, 1H), 8.15 (d, J = 8.6 Hz, 1H), 7.81 (d, J = 8.5 Hz, 1H), 4.78 – 4.48 (m, 1H), 4.07 (ddt, J = 15.3, 11.8, 5.8 Hz, 1H), 3.76 (td, J = 9.6, 4.8 Hz, 2H), 3.56 (ddd, J = 20.1, 13.9, 6.9 Hz, 1H), 2.19 (d, J = 11.1 Hz, 2H), 2.06 (d, J = 11.0 Hz, 2H), 1.86 – 1.60 (m, 2H), 1.59 – 1.36 (m, 4H), 1.04 (t, J = 7.5 Hz, 3H). (e) Example 5-47: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-((4-fluoropiperidin-1- yl)methyl)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2-trifluoroacetate To a solution of 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)nicotinaldehyde (from step (d), 30 mg, 77 µmol, 1.0 equiv.) in DCE (1 mL) were added 4-fluoropiperidine hydrochloride (108 mg, 0.774 mmol, 10.0 equiv.) and DIPEA (0.17 mL, 1.55 mmol, 20.0 equiv.). The mixture was stirred under reflux for 30 min before NaBH(OAc)3 (33 mg, 0.0.155 mmol, 2 equiv.) was added. Then the reaction was stirred at room temperature overnight. The mixture was diluted with DCM and further washed with saturated aqueous NaHCO3 (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase C18 column to afford to afford the trifluoroacetate salt of the title compound as an off-white solid (25.5 mg, 0.043 mmol, 56%).1H NMR (400 MHz, Methanol-d4) δ 8.74 (s, 1H), 8.48 (s, 1H), 8.09 (dd, J = 8.48, 2.20 Hz, 1H), 8.02 (d, J = 8.43 Hz, 1H), 4.78 – 4.54 (m, 1H), 4.46 (s, 2H), 4.16 (ddt, J = 14.46, 10.23, 3.98 Hz, 1H), 3.91 – 3.55 (m, 3H), 3.48 – 3.34 (m, 4H), 2.45 – 1.94 (m, 8H), 1.86 – 1.64 (m, 2H), 1.61 – 1.37 (m, 4H), 1.07 (t, J = 7.45 Hz, 3H).13C NMR (101 MHz, Methanol-d4) δ 161.0, 155.3, 151.1, 142.8, 141.9, 132.8 – 131.1 (m), 125.4, 121.4, 115.2, 94.5 (d, J = 170.9 Hz), 69.8, 51.2, 46.0 (d, J = 22.2 Hz), 34.2, 30.5, 26.8 (d, J = 20.1 Hz), 9.6 (d, J = 5.9 Hz). Five aliphatic carbon missing. 19F NMR (376 MHz, Methanol-d4) δ -76.86 (TFA), -77.09 (TFA), -188.34, -190.21. HPLC - tR = 3.7 min >95% purity at 254 nm; LCMS [M+H]+ 475.3 m/z. (e) Example 5-48: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-((4-(2- fluoroethyl)piperazin-1-yl)methyl)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2-trifluoroacetate To a solution of 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)nicotinaldehyde (from step (d), 114 mg, 0.294 mmol, 1.0 equiv.) in DCE (2 mL) were added 1-(2-fluoroethyl)piperazine dihydrochloride (603 mg, 2.94 mmol, 10.0 equiv.) and DIPEA (1.03 mL, 5.88 mmol, 20.0 equiv.). The mixture was stirred under reflux for 30 min before NaBH(OAc)3 (112 mg, 0.530 mmol, 1.8 equiv.) was added. Then the reaction was stirred under r.t. overnight. The mixture was diluted with DCM and further washed with saturated aqueous NaHCO3 (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase C18 column to afford to afford the trifluoroacetate salt of the title compound as an off-white solid (24 mg, 0.039 mmol, 13%).1H NMR (400 MHz, Methanol-d4) δ 8.64 (d, J = 2.07 Hz, 1H), 8.41 (s, 1H), 7.98 (dd, J = 8.43, 2.25 Hz, 1H), 7.93 (d, J = 8.39 Hz, 1H), 4.80 – 4.50 (m, 2H), 4.21 – 4.08 (m, 1H), 3.97 (s, 2H), 3.90 – 3.53 (m, 3H), 3.52 – 3.44 (m, 1H), 3.40 (dt, J = 9.58, 4.57 Hz, 5H), 3.05 (t, J = 4.91 Hz, 4H), 2.24 – 2.12 (m, 2H), 2.07 – 1.96 (m, 2H), 1.84 – 1.62 (m, 2H), 1.49 (ddddd, J = 24.45, 17.18, 14.55, 9.90, 3.57 Hz, 4H), 1.06 (t, J = 7.45 Hz, 3H).13C NMR (101 MHz, Methanol-d4) δ 161.0, 154.0, 153.6, 149.5, 141.8, 140.4, 131.3, 121.1, 106.9, 94.5 (d, J = 171.8 Hz), 79.8 (d, J = 167.9 Hz), 69.8, 58.9, 57.8 (d, J = 19.6 Hz), 53.0, 51.1, 50.9, 45.9 (d, J = 22.4 Hz), 34.2, 30.5 (d, J = 4.7 Hz), 26.8 (d, J = 20.8 Hz), 9.6 (d, J = 5.9 Hz).19F NMR (376 MHz, Methanol-d4) δ - 76.83 (TFA), -77.18 (TFA), -188.26, -222.00. HPLC - tR = 4.2 min >95% purity at 254 nm; LCMS [M+H]+ 504.3 m/z; HRMS calcd for [M+H]+ 504.3257 m/z, found 504.3268. (e) Example 5-49: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-(((2- fluoroethyl)(methyl)amino)methyl)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2-trifluoroacetate To a solution of 6-(2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4- hydroxycyclohexyl)amino)pyrimidin-5-yl)nicotinaldehyde (from step (d), 55 mg, 0.142 mmol, 1.0 equiv.) in DCE (1 mL) were added 2-fluoro-N-methylethan-1-amine hydrochloride (129 mg, 1.14 mmol, 8 equiv.) and DIPEA (0.198 mL, 1.14 mmol, 8 equiv.). The mixture was stirred under reflux for 30 min before NaBH(OAc)3 (48mg, 0.227 mmol, 1.6equiv.) was added. Then the reaction was stirred at room temperature overnight. The mixture was diluted with DCM and further washed with saturated aqueous NaHCO3 (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase C18 column to afford to afford the trifluoroacetate salt of the title compound as an off-white solid (40.1 mg, 0.071 mmol, 50%).1H NMR (400 MHz, Methanol-d4) δ 8.76 (d, J = 2.21 Hz, 1H), 8.50 (d, J = 10.05 Hz, 1H), 8.11 (dd, J = 8.51, 2.39 Hz, 1H), 8.03 (d, J = 8.48 Hz, 1H), 5.03 – 4.93 (m, 1H), 4.78 – 4.56 (m, 1H), 4.54 (s, 2H), 4.16 (td, J = 10.29, 8.34, 4.91 Hz, 1H), 3.95 – 3.49 (m, 5H), 2.95 (s, 3H), 2.29 – 2.11 (m, 2H), 2.10 – 1.98 (m, 2H), 1.89 – 1.64 (m, 2H), 1.63 – 1.35 (m, 4H), 1.07 (t, J = 7.47 Hz, 3H).13C NMR (101 MHz, Methanol-d4) δ 161.0, 155.4, 154.0, 151.1, 142.8, 141.8, 125.6, 121.4, 102.3, 94.5 (d, J = 171.6 Hz), 79.4 (d, J = 168.3 Hz), 69.8, 58.2, 56.9 (d, J = 19.4 Hz), 51.2, 45.9 (d, J = 22.2 Hz), 40.7 (d, J = 2.3 Hz), 34.2, 30.5 (d, J = 4.7 Hz), 26.8 (d, J = 20.9 Hz), 9.6 (d, J = 5.9 Hz).19F NMR (376 MHz, Methanol-d4) δ -76.83, -77.06, -188.27, -222.45. HPLC - tR = 4.2 min >95% purity at 254 nm; LCMS [M+H]+ 449.2 m/z; HRMS calcd for [M+H]+ 449.2835 m/z, found 449.2841. Example 5-50:
Figure imgf000077_0001
Scheme 10 (a) 6-bromo-N-(2-fluoroethyl)pyridin-3-amine To a solution of 2-bromo-5-iodopyridine (4.1 g, 14.44 mmol, 1.0 equiv.) in 1,4- dioxane were added 2-fluoroethan-1-amine hydrochloride (1.58 g, 15.89 mmol, 1.1 equiv.), Cs2CO3 (14.12 g, 43.33 mmol, 3 equiv.). and xantphos (1.25 g, 2.17 mmol, 0.15 equiv.). The reaction mixture was degassed with nitrogen for 30 min. Then Pd(OAc)2 (259 mg, 1.16 mmol, 8.0 mol%) was added under nitrogen flushing. The reaction mixture was heated to 90 °C and stirred for 1 h. The mixture was diluted with EtOAc and the resulting suspension was filtered through a pad of Celite with the aid of EtOAc. The filtrate was further washed with water (3 × 20 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (0-50% EtOAc in Pet. Sp.) to afford the title compound as a yellow oil (1.09 g, 4.96 mmol, 34%).1H NMR (400 MHz, Chloroform-d) δ 7.80 (d, J = 2.97 Hz, 1H), 7.23 (d, J = 8.51 Hz, 1H), 6.83 (dd, J = 8.65, 3.01 Hz, 1H), 4.67 (t, J = 4.83 Hz, 1H), 4.56 (t, J = 4.78 Hz, 1H), 3.92 (s, 1H), 3.46 (t, J = 4.83 Hz, 1H), 3.39 (t, J = 4.80 Hz, 1H).13C NMR (101 MHz, Chloroform-d) δ 143.4, 135.3, 128.8, 128.0, 122.7, 82.2 (d, J = 168.4 Hz), 44.0 (d, J = 20.4 Hz). HPLC - tR = 3.1 min >95% purity at 254 nm; LCMS [M+H]+ 219.0 m/z (Br79), 220.9 m/z (Br81). (b) 6-bromo-N-(2-fluoroethyl)-N-methylpyridin-3-amine To a solution of 6-bromo-N-(2-fluoroethyl)pyridin-3-amine (from step (a), 244 mg, 1.11 mmol, 1.0 equiv.) in DMF (6 mL) was added NaH (60% dispersion in mineral oil, 71 mg, 1.78 mmol, 1.6 equiv.) at 0 °C. The mixture was stirred for 15 min at room temperature before iodomethane (0.139 mL, 2.23 mmol, 2.0 equiv.) was added and the reaction was then stirred for 1 h at room temperature. The reaction was quenched by saturated NaHCO3 aqueous solution (5 mL) and was further washed by saturated aqueous NaHCO3 solution (3 × 5 mL). The organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (0-50% EtOAc in Pet. Sp.) to afford the title compound as a clear oil (200 mg, 0.858 mmol, 77%).1H NMR (400 MHz, Chloroform-d) δ 7.78 (s, 1H), 7.19 (dd, J = 8.88, 1.93 Hz, 1H), 6.84 (dt, J = 8.95, 2.42 Hz, 1H), 4.58 (td, J = 4.91, 1.72 Hz, 1H), 4.46 (td, J = 4.95, 1.77 Hz, 1H), 3.56 (dtd, J = 25.31, 4.97, 1.74 Hz, 2H), 2.94 (d, J = 1.83 Hz, 3H).13C NMR (101 MHz, Chloroform-d) δ 144.6, 134.0, 127.9, 127.3, 122.4, 81.5 (d, J = 170.8 Hz), 52.6 (d, J = 20.7 Hz), 38.8.19F NMR (376 MHz, Chloroform-d) δ -222.14. HPLC - tR = 3.7 min >95% purity at 254 nm; LCMS [M+H]+ 233.0 m/z (Br79), 235.0 m/z (Br81). (c) Lithium (5-((2-fluoroethyl)(methyl)amino)pyridin-2-yl)trihydroxyborate To a solution of 6-bromo-N-(2-fluoroethyl)-N-methylpyridin-3-amine (from step (b), 691 mg, 2.96 mmol, 1.0 equiv.) in a mixture of toluene (20 mL) and tetrahydrofuran (THF) (5 mL) under an atmosphere of nitrogen was added B(i-OPr)3 (0.82 mL, 3.56 mmol, 1.2 equiv.). The mixture was cooled to -78 °C and stirred for 30 min. Then 2.2 M solution of n-BuLi in hexane (2.7 mL, 5.93 mmol, 2.0 equiv.) was slowly added over 30 min. The reaction mixture was stirred at the same temperature for 3 h, then the cooling bath was removed. Upon the mixture slowly warmed to 0 °C (around 1.5 h), the reaction was quenched with isopropyl alcohol and the suspension was stirred overnight. The solvent was then removed under reduced pressure and the resulting residue was treated with acetone and water (9:1, v/v) until solid was precipitated. The precipitate was collected by filtration, further washed with acetone and water (9:1, v/v) and then dried under reduced pressure to afford the title compound as a brown solid (545 mg, 2.46 mmol, 83%).1H NMR (400 MHz, Deuterium Oxide) δ 7.99 (s, 1H), 7.61 (d, J = 8.88 Hz, 1H), 7.56 – 7.48 (m, 1H), 4.63 (d, J = 4.93 Hz, 1H), 3.71 (s, 1H), 3.64 (s, 1H), 2.97 (s, 3H). (d) (1r,4r)-4-((5-bromo-2-chloropyrimidin-4-yl)amino)cyclohexan-1-ol To a solution of 5-bromo-2,4-dichloropyrimidine (2 g, 8.78 mmol, 1.0 equiv.) and (1r,4r)-4-aminocyclohexan-1-ol (1.01 g, 8.78 mmol, 1.0 equiv.) in i-PrOH was added DIPEA (2.29 mL, 13.17 mmol, 1.5 equiv.). The reaction mixture was stirred at 70 °C overnight. The solvent was evaporated under reduced pressure and the resulting residue was partitioned between brine and DCM. The aqueous layer was further extracted with DCM (3 × 20 mL). The combined organic exacts were dried over anhydrous magnesium sulfate (MgSO4), filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (0-50% EtOAc in petroleum spirits) to afford the title compound as an off- white solid (1.99 g, 6.50 mmol, 74%).1H NMR (400 MHz, CDCl3) δ 8.07 (s, 1H), 5.31 (d, J = 7.7 Hz, 1H), 3.98 (tdt, J = 11.5, 8.0, 4.1 Hz, 1H), 3.65 (tt, J = 10.5, 4.2 Hz, 1H), 2.16 – 2.05 (m, 2H), 2.05 – 1.96 (m, 2H), 1.46 (qd, J = 13.1, 3.4 Hz, 2H), 1.38 – 1.23 (m, 2H); 13C NMR (101 MHz, CDCl3) δ 159.3, 158.7, 156.2, 102.9, 69.5, 49.3, 33.6, 30.4. %). HPLC - tR = 4.2 min >95% purity at 254 nm; LCMS [M+H]+ 306.1 m/z (Br79), 308.0 m/z (Br81). (e) (1S,4r)-4-((5-bromo-2-(((S)-2-fluorobutyl)amino)pyrimidin-4-yl)amino)cyclohexan-1- ol To a solution of (1r,4r)-4-((5-bromo-2-chloropyrimidin-4-yl)amino)cyclohexan-1-ol (from step (d), 700 mg, 2.28 mmol, 1.00 equiv.) and (S)-2-fluorobutan-1-amine hydrochloride (335 mg, 2.63 mmol, 1.15 equiv.) in toluene (3 mL) was added DIPEA (1.19 mL, 6.85 mmol, 3.00 equiv.). The reaction mixture was stirred at 150 °C in a sealed tube for 3 days. The solvent was evaporated under reduced pressure and the resulting residue was partitioned between brine and DCM. The aqueous layer was further extracted with DCM (3 × 20 mL), the combined organic exacts were dried over anhydrous magnesium sulfate (MgSO4), filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography (0-50% EtOAc in petroleum spirits) to afford the title compound as an off- white solid (382.5 mg, 1.06 mmol, 46%). HPLC - tR = 3.6 min >95% purity at 254 nm; LCMS [M+H]+ 361.1 m/z (Br79), 363.1 m/z (Br81). (f) Example 5-50: (1S,4r)-4-((2-(((S)-2-fluorobutyl)amino)-5-(5-((2- fluoroethyl)(methyl)amino)pyridin-2-yl)pyrimidin-4-yl)amino)cyclohexan-1-ol 2,2,2- trifluoroacetate To a solution of (1S,4r)-4-((5-bromo-2-(((S)-2-fluorobutyl)amino)pyrimidin-4- yl)amino)cyclohexan-1-ol (from step (e), 258 mg, 0.713 mmol, 1.0 equiv.) in a mixture of DMF (20 mL) and H2O (5 mL) were added lithium (5-((2-fluoroethyl)(methyl)amino)pyridin-2- yl)trihydroxyborate (from step (c), 522 mg, 2.35 mmol, 3.3 equiv.), CuBr (20 mg, 0.142 mmol, 0.2 equiv.) and K2CO3 (296 mg, 2.14 mmol, 3.0 equiv.). The reaction mixture was degassed with nitrogen for 30 min. Then Pd(dppf)Cl2 (52 mg, 71 µmol, 10 mol%) was added under nitrogen flushing. The reaction mixture was heated to 110 °C and stirred for 1 h then diluted with EtOAc and the resulting suspension was filtered through a pad of Celite with the aid of EtOAc. The filtrate was further washed with water (3 × 20 mL), the organic phase was then dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by a reverse phase C18 column to afford the trifluoroacetate salt of the title compound as a brown solid (20.3 mg, 0.047 mmol, 5%).1H NMR (400 MHz, Methanol-d4) δ 8.18 – 8.03 (m, 2H), 7.66 (d, J = 9.07 Hz, 1H), 7.30 (dd, J = 9.04, 3.11 Hz, 1H), 4.69 (t, J = 4.75 Hz, 1H), 4.57 (t, J = 4.81 Hz, 1H), 4.10 (ddt, J = 14.63, 10.55, 4.27 Hz, 1H), 3.85 – 3.49 (m, 5H), 3.09 (s, 3H), 2.27 – 2.09 (m, 2H), 2.08 – 1.95 (m, 2H), 1.72 (dp, J = 27.73, 6.89 Hz, 2H), 1.49 (dddd, J = 24.76, 14.78, 12.42, 6.36 Hz, 4H), 1.06 (t, J = 7.43 Hz, 3H).13C NMR (101 MHz, Methanol-d4) δ 161.1, 153.7, 145.9, 140.4, 138.0, 132.3, 121.7, 121.6, 108.0, 94.6 (d, J = 171.3 Hz), 83.0 (d, J = 168.4 Hz), 69.9, 52.9 (d, J = 20.1 Hz), 51.0, 45.8 (d, J = 22.1 Hz), 38.6, 34.3, 30.6, 26.8 (d, J = 20.8 Hz), 9.6 (d, J = 6.2 Hz).19F NMR (376 MHz, Methanol- d4) δ -76.95 (TFA), -188.28, -224.13. HPLC - tR = 4.5 min >95% purity at 254 nm. Examples 11-01 to 11-02:
Figure imgf000080_0001
Scheme 11 (a) Synthesis of methyl 2-chloro-4-(((1r,4r)-4-hydroxycyclohexyl)amino)pyrimidine-5- carboxylate (8) To a solution of 2,4-dichloropyrimidine-5-carbonyl chloride 6 (1.0eq) in dichloromethane (30 mL) was added methanol (1.1 eq) and diisopropylethylamine (1.0 eq) at 0 °C. The resulting mixture was stirred for 1 h at 0 °C. Then the solvent was removed. The residue was dissolved in IPA (20 mL) and followed by the addition of trans-4- aminocyclohexanol (1.1 eq) then DIEA (1.5 eq) drop wisely. The resulting mixture was stirred at 0 °C for 90 min. Water was then added, the resulting mixture was extracted with EtOAc (3×). The combined organic layers were dried (Na2SO4), filtered and concentrated. The residue was purified on ISCO to provide methyl 2-Chloro-4-(((trans-4- hydroxycyclohexyl)amino)pyrimidine-5-carboxylate 8 (84% over 2 steps). (b) Synthesis of methyl 2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl) amino)pyrimidine-5-carboxylate (9) To a stirred solution of 8 (1.0 eq) in IPA (5 mL) was added β-fluoro amine hydrochloride (4.0eq) and followed by the addition of DIEA (461.4 mg, 3.57 mmol) drop wisely. The resulting mixture was stirred at 100 °C for 16h. Water was then added. The resulting mixture was extracted with EtOAc (3×). The combined organic layers were dried (Na2SO4), filtered and concentrated. The residue was purified on ISCO to provide title compound 9 in 60% yield. (c) Synthesis of 2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl)amino) pyrimidine-5-carboxylic acid (10) A mixture of pyrimidine-5-carboxylate 9 (1.0eq) and lithium hydroxide monohydrate (10.0eq) in a mixture of THF and water (25 mL, 3:2, v/v) was heated at reflux for 10 h. Then the reaction mixture was cooled to room temperature and acidified by a 4.0N solution of HCl to pH 3. The resulting mixture was extracted with EtOAc (4×). The combined organic layers were dried (Na2SO4) and concentrated to provide the title compound 10 (81%) as a white solid. (d) Synthesis of pyrimidine 5-carboxamide compounds using TBTU coupling A solution of 2-(butylamino)-4-((trans-4-hydroxycyclohexyl)amino) pyrimidine-5- carboxylic acid 10 (1.0eq), O-(benzotriazol-1-yl)-N,N,N′,N′- tetramethyluronium tetrafluoroborate (TBTU, 1.3 eq) and N,N-diisopropylethylamine (DIEA, 3.0 eq) in anhydrous DMF (4.0 mL) and was added right hand side amine (1.2 eq) in DMF (1.0 mL) drop wisely at room temperature. The resulting mixture was stirred for overnight, then diluted with EtOAc (15 mL) and washed with water (3×). The organic layer was dried (Na2SO4) and concentrated. The residue was purified on HPLC to give the title compound (55-58%) as a white solid. Example 11-01: N-(1-(cyclopropylmethyl)piperidin-4-yl)-2-(((S)-2-fluorobutyl)amino)-4- (((1r,4S)-4-hydroxy cyclohexyl)amino)pyrimidine-5-carboxamide: Off-white solid (55% yield); 1H NMR (400 MHz, MeOD) δ 8.30 (s, 1H), 4.72 – 4.53 (m, 1H), 4.13 – 3.99 (m, 2H), 3.78 – 3.70 (m, 2H), 3.67 – 3.50 (m, 3H), 3.16 – 3.06 (m, 2H), 3.03 (d, J = 7.3 Hz, 2H), 2.27 – 2.19 (m, 2H), 2.14 –
Figure imgf000081_0001
2.06 (m, 2H), 2.02 – 1.96 (m, 2H), 1.95 – 1.83 (m, 2H), 1.78 – 1.66 (m, 2H), 1.48 – 1.34 (m, 4H), 1.15 – 1.10 (m, 1H), 1.05 (t, J = 7.5 Hz, 3H), 0.77 (q, J = 6.0 Hz, 2H), 0.43 (q, J = 4.8 Hz, 2H); HPLC: tR 4.007 min > 98% purity at 254 mm; LCMS (m/z): 463.1 [M+H]+; HRMS (m/z): [M+H]+ calcd 463.3197 found 463.3185. Example 11-02: 2-(((S)-2-fluorobutyl)amino)-N-(1-(3-fluoropropyl)piperidin-4-yl)-4- (((1r,4S)-4-hydroxy cyclohexyl)amino)pyrimidine-5-carboxamide: Off-white solid (58% yield); 1H NMR (400 MHz, MeOD) δ 8.24 (s, 1H), 4.69 (d, J = 45.1 Hz, 1H), 4.53 (t, J = 5.8 Hz, 1H), 4.42 (t, J = 5.8 Hz, 1H), 4.02 – 3.85 (m, 1H), 3.82-3.75 (m, 1H), 3.72 – 3.53 (m, 1H), 3.50 (m, 1H),
Figure imgf000082_0001
2.98 (d, J = 11.9 Hz, 2H), 2.58 – 2.40 (m, 2H), 2.23 – 2.03 (m, 4H), 1.99-1.83 (m, 5H), 1.76 – 1.49 (m, 4H), 1.49 – 1.16 (m, 6H), 1.03 (t, J = 7.5 Hz, 3H); 19F NMR (376 MHz, MeOD) δ -188.32, -221.60; HPLC: tR 3.46 min, 100 % purity at 254 nm; LCMS: m/z 469.2 [M+H]+; HRMS [M+H]+: m/z calculated 469.3097 found 469.3098. Example 11-03:
Figure imgf000082_0002
(a) 2,4-dichloro-N-(1-cyclobutylpiperidin-4-yl)pyrimidine-5-carboxamide To a solution of 2,4-dichloropyrimidine-5-carbonyl chloride (500mg, 2.36 mmol, 1.0 equiv.) in dichloromethane (10 mL) were added 4-(N-cyclobutyl)piperidinyl amine (401.2 mg, 2.60 mmol, 1.1 equiv.) and DIPEA (617.9 µL, 3.55 mmol, 1.5 equiv.) at 0 °C. The resulting mixture was stirred at 0 °C for 1 h. Water was added and the resulting mixture was extracted with EtOAc (3×). The combined organic layers were dried over MgSO4, filtered and concentrated. The residue was purified by silica gel flash chromatography (0-10% MeOH in DCM) to afford the title compound as a white solid (701.2 mg, 84%); 1H NMR (400 MHz, MeOD) δ 8.72 (s, 1H), 4.00 – 3.81 (m, 1H), 2.95 (m, 3H), 2.24 – 1.99 (m, 6H), 1.94 (dd, J = 19.3, 10.2 Hz, 2H), 1.82 – 1.71 (m, 2H), 1.66 (dt, J = 21.4, 7.9 Hz, 2H); LCMS: m/z 329.0 [M+H]+ (b) 2-chloro-N-(1-cyclobutylpiperidin-4-yl)-4-(((1r,4r)-4- hydroxycyclohexyl)amino)pyrimidine-5-carboxamide To a solution of 2,4-dichloro-N-(4-(n-cyclobutylpiperidinyl amine) pyrimidine-5- carboxamide (from step (a), 400 mg, 1.21 mmol, 1.0 equiv.) in i-PrOH (15 mL) were added trans-4-aminocyclohexanol (181.9 mg, 1.58 mmol, 1.3 equiv.) and DIPEA (423.27 µL, 2.43 mmol, 2.0 equiv.) at 0 °C. The resulting mixture was stirred at 0 °C for 50 min and warmed to room temperature and stirred for another 50 min. Then the solvent was removed, to the residue was added a mixture of DCM and methanol (20 mL, 3:2, v/v) and the precipitate was collected by filtration to give the title compound as a white solid (410 mg, 82%); 1H NMR (400 MHz, MeOD) δ 8.40 (s, 1H), 4.05 – 3.92 (m, 1H), 3.87 (t, J = 11.2 Hz, 1H), 3.60 (td, J = 9.9, 4.2 Hz, 1H), 3.08 – 2.79 (m, 3H), 2.10 (t, J = 20.1 Hz, 5H), 1.97 (dd, J = 21.0, 13.3 Hz, 6H), 1.81 – 1.50 (m, 4H), 1.49 – 1.28 (m, 5H); LCMS: m/z 408.0 [M+H]+ (c) Example 11-03: N-(1-cyclobutylpiperidin-4-yl)-2-(((S)-2-fluorobutyl)amino)-4- (((1r,4S)-4-hydroxycyclohexyl)amino )pyrimidine-5-carboxamide To a solution of 2-chloro-N-(1-cyclobutylpiperidin-4-yl)-4-(((1r,4r)-4- hydroxycyclohexyl)amino)pyrimidine-5-carboxamide (from step (b), 150 mg, 0.367 mmol, 1.0 equiv.) in THF (3 mL) were added (S)-2-fluorobutan-1-amine hydrochloride (211.1 mg, 1.65 mmol, 5.0 equiv.) and DIPEA (89.72 µL, 1.84 mmol, 5.0 equiv.) and the resulting reaction mixture was heated under microwave irradiation at 150 °C for 6 h. The solvent was removed under reduced pressure, washed with saturated aqueous NaHCO3 solution and extracted with EtOAc (2 x 50 mL). The combined organics were dried over MgSO4, filtered and concentrated. The residue was purified by silica gel column chromatography (0-20% MeOH in DCM) to afford title compound as a white solid (90 mg, 53%); 1H NMR (400 MHz, MeOD) δ 8.31 (s, 1H), 4.80 – 4.58 (m, 2H), 4.11 – 3.89 (m, 2H), 3.76 – 3.55 (m, 2H), 3.49 (m, 2H), 3.27 – 3.04 (m, 2H), 2.94 (dd, J = 8.4, 6.9 Hz, 1H), 2.41 – 2.24 (m, 2H), 2.23 – 2.05 (m, 2H), 2.03 – 1.83 (m, 5H), 1.81 – 1.61 (m, 4H), 1.50 – 1.28 (m, 4H), 1.11 – 1.01 (m, 6H); 13C NMR (101 MHz, MeOD) δ 168.99, 162.14, 157.38, 156.42, 100.31, 93.75 – 92.05 (d, 1JCH = 171.7 Hz), 70.26, 60.33, 46.06, 45.51, 44.15 – 43.94 (d, 2JCH = 20.9 Hz), 34.55, 31.31, 29.48, 26.97, 26.76, 26.60 – 26.40 (d, 2JCH = 20.2 Hz), 14.27, 9.20 (d); 19F NMR (376 MHz, MeOD) δ - 188.44, -191.09 (mixture of diastereomers); LCMS: m/z 463.3 [M+H]+; HRMS [M+H]+: m/z calculated 463.3191 found 463.3197. Example 11-04:
Figure imgf000084_0001
Scheme 13 (a) Ethyl 2-chloro-4-(((1r,4r)-4-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate To a solution of ethyl 2,4-dichloropyrimidine- 5-carboxylate (2.0 g, 9.1 mmol), trans-4-aminocyclohexanol (1.2 g, 10 mmol) in i-PrOH (50 mL) was added DIPEA (1.8 g, 14 mmol, 2.7 mL) at 0 °C. The resultant mixture was slowly warmed to room temperature and stirred for 14 h. The reaction mixture was concentrated in vacuo before partitioned between ethyl acetate and water. The mixture was further extracted with ethyl acetate (x2). The organic layers were combined and dried with magnesium sulphate. The volatiles were removed in vacuo and the residue was purified by flash silica gel column chromatography, eluting with 10–20% ethyl acetate/petroleum spirits to afford the title as white solid (2.32 g, 86%). HPLC – tR = 5.89 min >98% purity at 254 mm; LRMS [M+H]+ 300.9 m/z; 1H NMR (400 MHz, CDCl3) δ 8.65 (s, 1H), 8.30 (s, 1H), 4.34 (q, J = 7.1 Hz, 2H), 4.15 – 4.05 (m, 1H), 3.74 – 3.66 (m, 1H), 2.16 – 2.07 (m, 2H), 2.06 – 1.98 (m, 2H), 1.54 – 1.42 (m, 3H), 1.41 – 1.30 (m, 5H). (b) Ethyl 2-(butylamino)-4-(((1r,4r)-4-((tert- butyldimethylsilyl)oxy)cyclohexyl)amino)pyrimidine-5-carboxylate To a solution of ethyl 2-chloro-4-(((1r,4r)-4-hydroxycyclohexyl)amino)pyrimidine-5- carboxylate (from step (a), 4.0 g, 13 mmol) in DCM (50 ml) were added imidazole (1.4 g, 20.1 mmol) and tert-butyldimethylsilyl chloride (3.0 g, 20 mmol). The resultant mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo and the residue was partitioned between ethyl acetate and water. The mixture was further extracted with ethyl acetate (x2). The organic layers were combined and dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash silica gel column chromatography, eluting with 10% ethyl acetate/petroleum spirits to afford the title compound as a white solid (3.7 g, 67%). HPLC – tR = 9.93 min >98% purity at 254 mm; LRMS [M+H]+ 413.9 m/z; 1H NMR (400 MHz, CDCl3) δ 8.64 (s, 1H), 8.33 (s, 1H), 4.34 (q, J = 7.1 Hz, 2H), 4.13 – 4.05 (m, 1H), 3.70 – 3.63 (m, 1H), 2.11 – 2.05 (m, 2H), 1.90 – 1.83 (m, 2H), 1.58 – 1.47 (m, 4H), 1.38 (t, J = 7.1 Hz, 3H), 0.89 (s, 9H), 0.06 (s, 6H); 13C NMR (101 MHz, CDCl3) δ 166.43, 163.79, 161.38, 160.27, 103.77, 69.92, 61.60, 48.63, 33.55 (2C), 31.07 (2C), 29.91, 26.00 (3C), 14.34, -4.50 (2C). (c) Ethyl 4-(((1r,4r)-4-((tert-butyldimethylsilyl)oxy)cyclohexyl)amino)-2-((2- fluorobutyl)amino)pyrimidine-5-carboxylate To a solution of ethyl 2-(butylamino)-4-(((1r,4r)-4-((tert- butyldimethylsilyl)oxy)cyclohexyl)amino)pyrimidine-5-carboxylate (from step (b), 115 mg, 0.278 mmol) in i-PrOH (15 ml) were added amine 2-fluorobutan-1-amine hydrochloride (37 mg, 0.29 mmol) and DIPEA (72 mg, 0.55 mmol, 97 μL) at room temperature. The reaction was heated to 60 °C and stirred for 20 h. The volatiles were removed in vacuo and the residue was partitioned between ethyl acetate and water and extracted with ethyl acetate (x2). The combined organic layers were dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash silica gel column chromatography, eluting with 20% ethyl acetate/petroleum spirits to give the title compound as a white solid (84.6 mg, 65%). HPLC – tR = 8.605 min >95% purity at 254 mm; LRMS [M+H] + 469.0 m/z; HRMS calcd for [M+H]+ 469.3005 m/z, found 469.3017 m/z; 1H NMR (400 MHz, CDCl3) δ 8.52 (s, 1H), 8.13 (s, 1H), 5.88 (s, 1H), 4.67 – 4.48 (m, 1H), 4.25 (q, J = 7.1 Hz, 2H), 4.00 – 3.91 (m, 1H), 3.88 – 3.72 (m, 1H), 3.68 – 3.61 (m, 1H), 3.56 – 3.41 (m, 1H), 2.10 – 2.04 (m, 2H), 1.90 – 1.83 (m, 2H), 1.74 – 1.63 (m, 2H), 1.50 – 1.38 (m, 3H), 1.36 – 1.31 (m, 4H), 1.02 (t, J = 7.5 Hz, 3H), 0.89 (s, 9H), 0.06 (s, 6H); 19F NMR (376 MHz, CDCl3) δ –186.52 (s); 13C NMR (101 MHz, CDCl3) δ 167.42, 162.75, 161.41, 160.78, 97.00, 94.28 (d, 1JC-F = 170.1 Hz), 70.33, 60.18, 48.33, 45.03 (d, 2JC-F = 21.4 Hz), 34.00 (2C), 32.91, 30.20 (2C), 26.01 (3C), 25.84 (d, 2JC-F = 20.8 Hz), 14.50, 9.42 (d), -4.52 (2C). (d) 4-(((1r,4r)-4-((tert-Butyldimethylsilyl)oxy)cyclohexyl)amino)-2-((2- fluorobutyl)amino)pyrimidine-5-carboxylic acid To a solution of ethyl 4-(((1r,4r)-4-((tert-butyldimethylsilyl)oxy)cyclohexyl)amino)- 2-((2- fluorobutyl)amino)pyrimidine-5-carboxylate (from step (c), 69 mg, 0.15 mmol) in THF/H2O (3:2, v/v, 10 mL) was added LiOH (35 mg, 1.5 mmol) at room temperature. The resultant mixture was heated to 50 °C for 16 h. The THF was removed in vacuo and the aqueous layer was acidified with 10% aqueous citric acid to pH 3–4. The white precipitate formed was collected by filtration to afford the title compound as an off-white solid (44.7 mg, 69%). HPLC – tR = 7.735 min >96% purity at 254 mm; LRMS [M+H]+ 441.0 m/z; HRMS calcd for [M+H]+ 441.2692 m/z, found 441.2705 m/z; 1H NMR (400 MHz, MeOD) δ 8.30 (s, 1H), 4.53 – 4.45 (m, 1H), 4.03 – 3.90 (m, 2H), 3.80 – 3.69 (m, 1H), 3.67333 – 3.58 (m, 1H), 3.54 – 3.45 (m, 1H), 2.14 – 2.06 (m, 2H), 1.97 – 1.88 (m, 2H), 1.70 (ddd, J = 22.2, 14.9, 7.3 Hz, 2H), 1.48 – 1.36 (m, 4H), 1.04 (t, J = 7.5 Hz, 3H), 0.91 (s, 9H), 0.06 (s, 6H); 19F NMR (376 MHz, MeOD) δ –188.24 (s); 13C NMR (101 MHz, DMSO) δ 171.37, 168.71, 161.08, 160.03, 98.21, 93.58 (d, 1JC-F = 169.2 Hz), 69.78, 48.02, 44.48 (d, 2JC-F = 24.0 Hz), 33.80 (2C), 30.13 (2C), 29.68, 25.81 (3C), 25.39 (d, 2JC-F = 21.8 Hz), 9.12, -4.61 (2C). (e) 4-(((1r,4r)-4-((tert-Butyldimethylsilyl)oxy)cyclohexyl)amino)-N-(1- (cyclopropylmethyl)piperidin-4-yl)-2-((2-fluorobutyl)amino)pyrimidine-5-carboxamide To a solution containing 4-(((1r,4r)-4-((tert- Butyldimethylsilyl)oxy)cyclohexyl)amino)-2-((2- fluorobutyl)amino)pyrimidine-5-carboxylic acid (from step (d), 22 mg, 0.05 mmol) in acetonitrile (2.0 mL) was added EDC hydrochloride (10 mg, 0.052 mmol), HOAt (7.0 mg, 0.052 mmol), triethylamine (10 mg, 0.1 mmol, 14 μL) followed by 4-(N-cyclobutyl)piperidinyl amine (10 mg, 0.055 mmol) at room temperature. The reaction mixture was stirred at 50 °C overnight, the solvent was removed in vacuo and the reaction was mixture was partitioned between ethyl acetate and water. The mixture was further extracted with ethyl acetate (2x). The combined organic layers were dried over magnesium sulfate, filtered and the volatiles were removed in vacuo. The residue was purified by flash silica gel column chromatography, eluting with 5% MeOH in DCM to give the title compound as an off-white solid (18.4 mg, 64%). HPLC – tR = 7.517 min >95% purity at 254 mm; LRMS [M/2+H]+ 289.0 [M+H]+ 578.0 m/z; 1H NMR (400 MHz, MeOD) δ 8.26 (s, 1H), 4.70 – 4.48 (m, 1H), 3.99 – 3.89 (m, 1H), 3.83 – 3.73 (m, 2H), 3.72 – 3.62 (m, 1H), 3.45 (ddd, J = 17.7, 14.5, 7.4 Hz, 1H), 3.12 (d, J = 12.0 Hz, 2H), 2.30 (d, J = 6.7 Hz, 2H), 2.21 – 2.13 (m, 2H), 2.12 – 2.05 (m, 2H), 1.96 – 1.86 (m, 4H), 1.75 – 1.59 (m, 4H), 1.49 – 1.28 (m, 4H), 1.03 (t, J = 7.5 Hz, 3H), 0.95 – 0.92 (m, 1H), 0.90 (s, 3349H), 0.56 (q, J = 5.8 Hz, 2H), 0.16 (q, J = 4.6 Hz, 2H), 0.08 (s, 6H); 19F NMR (376 MHz, MeOD) δ –188.18 (s); 13C NMR (101 MHz, MeOD) δ 168.93, 163.25, 162.23, 157.17, 101.00, 95.02 (d, 1JC-F = 170.1 Hz), 71.56, 64.48, 53.70 (2C), 51.2247.94, 45.99 (d, 2JC-F = 23.0 Hz), 35.04 (2C), 32.24 (2C), 31.01 (2C), 30.69, 26.90 (d, 2JC-F = 20.8 Hz), 26.39 (3C), 9.75 (d), 8.84, 4.60 (2C), -4.46 (2C). (f) Example 11-04: N-(1-(Cyclopropylmethyl)piperidin-4-yl)-2-((2-fluorobutyl)amino)-4- (((1r,4r)-4- hydroxycyclohexyl)amino)pyrimidine-5-carboxamide To a solution of 4-(((1r,4r)-4-((tert-butyldimethylsilyl)oxy)cyclohexyl)amino)-N-(1- (cyclopropylmethyl)piperidin-4-yl)-2-((2-fluorobutyl)amino)pyrimidine-5-carboxamide (from step (e), 15 mg, 0.026 mmol) in DCM (3 mL) was added 4M hydrochloric acid in 1,4-dioxane (1.9 mg , 0.052 mmol, 13 μL). The reaction was stirred at room temperature for 4 h, the volatiles were then removed in vacuo and the resultant residue was triturated using diethyl ether (3x). The precipitate was collected by filtration and air-dried to give the title compound as hydrochloride salt (12 mg, 86%). HPLC – tR = 4.181 min >98% purity at 254 mm; LRMS [M+H]+ 463.0 m/z; HRMS calcd for [M+H]+ 463.3191 m/z, found 463.3204 m/z; 1H NMR (400 MHz, MeOD) δ 8.41 (s,1H), 4.66 (dd, J = 49.0, 5.3 Hz, 1H), 4.18 – 4.03 (m, 2H), 3.80 – 3.74 (m, 2H), 3.66 – 3.53 (m, 2H), 3.20 – 3.11 (m, 2H), 3.07 (d, J = 7.3 Hz, 2H), 2.30 – 2.21 (m, 2H), 2.20 – 2.09 (m, 3H), 2.08 – 1.94 (m, 4H), 1.81 – 1.68 (m, 2H), 1.52 – 1.40 (m, 4H), 1.25 – 1.14 (m, 1H), 1.08 (t, J = 7.5 Hz, 3H), 0.81 (q, J = 6.1 Hz, 2H), 0.49 (q, J = 4.8 Hz, 2H); 19F NMR (376 MHz, MeOD) δ –188.35 (s); 13C NMR (101 MHz, MeOD) δ 168.80, 163.65, 162.63, 158.55, 100.73, 96.13 (d, 1JC-F = 166.2 Hz), 69.79, 62.81, 52.72 (2C), 51.08, 46.25, 46.03 (d, 2JC-F = 22.1 Hz), 34.21 (2C), 32.52 (2C), 29.96 (2C), 26.96 (d, 2JC-F = 58.9 Hz), 9.58 (d), 6.56, 4.98 (2C). Example 11-05:
Figure imgf000087_0001
Scheme 14 (a) ethyl 4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidine-5-carboxylate To a solution of ethyl 4-chloro-2-(methylthio)pyrimidine-5-carboxylate (5g, 21.5 mmol, 1.0 equiv.) and trans-4-aminocyclohexanol (2.6g, 22.56 mmol, 1.05 equiv.) in i-PrOH (100 mL) was added DIPEA (13.1 mL, 75.21 mmol, 3.5 equiv.) and the reaction mixture was stirred at RT overnight. The solvent was evaporated under reduced pressure and the resulting residue was partitioned between DCM and brine. The aqueous layer was extracted with DCM (3 × times), the combined organic exacts were dried over anhydrous magnesium sulphate, filtered and concentrated under reduced pressure. The residue was purified by silica gel flash chromatography using 0-60% ethyl acetate in petroleum spirits to afford the title compound as an off white solid (6.1g, 91%); 1H NMR (400 MHz, CDCl3) δ 8.63 (s, 1H), 8.28 (bs, 1H), 4.19 – 4.00 (m, 1H), 3.71 (td, J = 9.9, 5.0 Hz, 1H), 2.54 (s, 3H), 2.22 – 2.08 (m, 2H), 2.08 – 1.97 (m, 2H), 1.53 – 1.39 (m, 4H); LCMS: m/z 312.0[M+H]+ (b) 4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidine-5-carboxylic acid To a stirring solution of ethyl 4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2- (methylthio)pyrimidine-5-carboxylate (from step (a), 5g, 16.06 mmol, 1.0 equiv.) in a mixture of THF/water (60 mL, 3:2, v/v) was added with lithium hydroxide monohydrate (3.85g, 160.5 mmol, 10.0 equiv.) at room temperature. The milky white mixture was heated to 50 °C for 16 h. Volatiles were removed in vacuo and the residual mixture was acidified by 10 % aqueous citric acid to pH 3 to 4. The white precipitate was then collected by filtration and dried to yield the title product as a white amorphous solid (4.05g, 89%); 1H NMR (400 MHz, DMSO) δ 8.50 (s, 1H), 8.27 (d, J = 7.5 Hz, 1H), 4.58 (bs, 1H), 3.93 (dt, J = 10.4, 8.6 Hz, 1H), 3.57 – 3.40 (m, 1H), 2.47 (s, 3H), 2.01 – 1.93 (m, 2H), 1.83 (d, J = 9.9 Hz, 2H), 1.39 – 1.20 (m, 4H); LCMS: m/z 284.1[M+H]+ (c) 4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)-N-(pyridin-3- ylmethyl)pyrimidine-5-carboxamide To a solution of 4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)pyrimidine- 5-carboxylic acid (from step (b), 200 mg, 0.705 mmol, 1.0 equiv.), O-(benzotriazol-1-yl)- N,N,N′,N′- tetramethyluronium tetrafluoroborate (TBTU, 233.8 mg, 0.917 mmol, 1.3 equiv.) and DIPEA (368.85 µL, 2.12 mmol, 3.0 equiv.) in anhydrous DMF (5.0 mL) was added benzylamine (114.5 mg, 1.06 mmol, 1.5 equiv.) in DMF (0.5 mL) drop-wise at room temperature. The resulting mixture was stirred overnight, then diluted with EtOAc (15 mL) and washed with water (3×). The organic layer was dried over MgSO4, filtered and concentrated. The residue was purified by silica gel flash chromatography (10-80% EtOAc in Pet. Sp.) to give the title compound (192.0 mg, 73%) as a white solid; 1H NMR (400 MHz, DMSO) δ 9.12 (t, J = 5.7 Hz, 1H), 8.84 (d, J = 7.5 Hz, 1H), 8.53 (d, J = 3.1 Hz, 2H), 8.46 (dd, J = 4.7, 1.5 Hz, 1H), 7.70 (dt, J = 7.8, 1.9 Hz, 1H), 7.36 (dd, J = 7.6, 4.5 Hz, 1H), 4.56 (d, J = 4.3 Hz, 1H), 4.44 (d, J = 5.7 Hz, 2H), 3.88 (s, 1H), 3.53 – 3.39 (m, 2H), 2.45 (s, 3H), 2.01 – 1.88 (m, 2H), 1.82 (d, J = 3.3 Hz, 2H), 1.39 – 1.18 (m, 4H); LCMS: m/z 374.2 [M+H]+ (d) Example 11-05: 2-(((S)-2-fluorobutyl)amino)-4-(((1r,4S)-4-hydroxycyclohexyl)amino)- N-(pyridin-3-ylmethyl)pyrimidine-5-carboxamide trifluoroacetate salt To a solution of (4-(((1r,4r)-4-hydroxycyclohexyl)amino)-2-(methylthio)-N-(pyridin- 3-ylmethyl)pyrimidine-5-carboxamide (from step (c), 150 mg, 0.401 mmol, 1.0 equiv.) in THF (5.0 mL) was added m-CPBA (77%) (180.1 mg, 0.807 mmol, 2.01 equiv.). The reaction mixture was stirred at room temperature for 1.0 h. Then (S)-2-fluorobutan-1-amine hydrochloride (256.2 mg, 2.01 mmol, 5.0 equiv.) and DIPEA (209.8 µL, 1.2 mmol, 3.5 equiv.) were added and the resulting reaction mixture was heated under microwave irradiation at 150 °C for 1h. The solvent was removed under reduced pressure, washed with saturated aqueous NaHCO3 solution and extracted with EtOAc (2x 50 mL). The combined organics were dried over MgSO4, filtered and concentrated. The residue was purified by reverse-phase column chromatography (KP-C18-HS Flash Cartridge) using ACN (0.1% TFA) and water (0.1% TFA) to afford the trifluoroacetate salt of the title compound as white amorphous solid (75 mg, 45%) (79% yield); HPLC: tR 3.60 min, 94.3 % purity at 254 nm; 1H NMR (400 MHz, MeOD) δ 8.90 (s, 1H), 8.80 (s, 1H), 8.61 (d, J = 8.0 Hz, 1H), 8.39 (s, 1H), 8.12 – 8.01 (m, 1H), 4.84 – 4.51 (m, 3H), 4.24 – 4.01 (m, 1H), 3.89 – 3.53 (m, 3H), 2.26 – 1.94 (m, 5H), 1.84 – 1.61 (m, 3H), 1.54 – 1.33 (m, 2H), 1.06 (t, J = 7.5 Hz, 3H); 13C NMR (101 MHz, MeOD) δ 165.63, 159.83, 153.03, 145.04, 143.51, 141.26, 140.76, 139.40, 126.85, 100.67, 93.77-92.07 (d, 1JCF = 92.8 Hz), 68.42, 49.70, 44.69-44.47 (d, 2JCF = 22.2 Hz), 39.93, 32.83, 29.12, 25.49-25.28 (d, 2JCF = 21.2 Hz), 8.18-8.12 (d, 3JCF = 6 Hz); 19F NMR (376 MHz, MeOD) δ -188.22 (s). LCMS: m/z 417.2 [M+H]+; HRMS [M+H]+: m/z calculated 417.2409 found 417.2419. Radiochemistry As stated above there are a number of ways in which the compounds of the invention can be synthesized as would be appreciated by a person skilled in the art. Nevertheless, we provide a reaction scheme for making certain compounds of the invention in Scheme 15.
Figure imgf000089_0001
Compound 5-02 (left) and [18F]5-02 (right) Scheme 15 No-carrier-added [18F]fluoride was produced by the 18O(p, n)18F nuclear reaction with a 18 MeV proton beam generated by the IBA Cyclone 18/18 cyclotron in a niobium target using [18O]H2O at Austin Health, Centre for PET. The identity and quality control of [18F]5-02 was performed using radio-LCMS measurements using a Shimadzu 2010 LCMS system equipped with a 5 μL injection loop, a SPD-20A UV-Vis detector (254 nm) and two LC-20AD solvent pumps for high pressure mixing of mobile phase. The stationary phase was a Phenomenex Gemini C-18, 5µ RP column, 150×4.6 mm. Acetonitrile (A) and water (B) with 0.1% formic acid were used as the mobile phase and a gradient elution technique was used for analysis: 0-18 min: 15-40% A, 18-30 min: isocratic 40% A, 30-34 min: 40-90% A, 34-37 min: isocratic 90% A, 37-40 min: 90- 15% A at a flow rate of 0.5 mL/min. The Bioscan FC-4000 dual BGO coincidence detector was used for the detection of radioactive compounds. The HPLC system used for semi-preparative workup of radiolabelled compounds was a Knauer Smartline 1050 pump equipped with a Smartline Manager 5050 for quaternary gradient capability. The Knauer UV detector 2520 (254 nm) and a CsI(TI) crystal PIN diode radioactivity detector were used for the detection of compounds. The gradient used for the purification of [18F]5-02 was 10-30% acetonitrile (0.1% formic acid) for 50 min at a flow rate of 4 mL/min. The stationary phase was a Phenomenex Gemini C-18, 10µ RP column, 250×10 mm. [18F]fluoride was transferred from the cyclotron, trapped on a QMA ion exchange cartridge and eluted using 20 mg kryptofix 2.2.2 and 3.5 mg K2CO3 in 0.2 mL of water plus 0.4 mL of acetonitrile as eluent mixture. After the addition of 1 mL of dry acetonitrile, the K[18F]F/kryptofix mixture was dried at 90 °C for 8 min under vacuum with argon flow. After drying, the reactor was cooled to 50 °C and 4 mg of the precursor in 0.5 mL of DMF added. Radiolabelling was achieved by heating to 120 °C for 15 min in a closed reactor. After cooling to 50 °C 8 mL of water was added to the crude mixture. The mixture was then passed through a C-18 SPE cartridge and eluted with 1 mL of acetonitrile into a loop vial. After dilution with 4 mL of 0.1M formic acid, the product was purified using a semi-preparative HPLC system. A gradient elution technique was used to separate the radiolabelled compound from its precursor. The radioactive peak at 24.5 min was collected and reformulated using the solid phase extraction technique by trapping on a C-18 Sep-Pak cartridge and elution with 1 mL of ethanol followed by the addition of 9 mL of saline. A sample was taken for quality control before the tracer was used in the animal experiments. Decay corrected radiochemical yields were 12.6±3%, specific activity was 2.6±0.6 Ci/µmol. The representative quality control of [18F]5-02 was positive. Radio-LCMS analysis of collected product revealed the same tR as the Compound 5-02 standard. The mass-to- charge ratio of [18F]5-02 detected was 377.4 m/z. The radiochemical purity of [18F]5-02 was determined as >99%. Microfluidic capillary electrophoresis (MCE) assay Inhibition of kinase activity was tested using a microfluidic capillary electrophoresis (MCE) assay. MCE assays were performed as previously described in the art (Zhang et al., J Med Chem, 56 (2013) 9693-9700). Activity assays were performed in a 384 well, polypropylene microplate in a final volume of 50 μl of 50 mM Hepes, pH 7.4 containing 10 mM MgCl2, 1.0 mM DTT, 0.01% Triton X-100, 0.1% Bovine Serum Albumin (BSA), containing 1.0 μM fluorescent substrate and ATP at the Km for each enzyme. All reactions were terminated by addition of 20 μl of 70 mM EDTA. After a 180 min incubation, phosphorylated and unphosphorylated substrate peptides were separated in buffer supplemented with 1 x CR-8 on a LabChip EZ Reader equipped with a 12-sipper chip. Data were analysed using EZ Reader software. The peptide substrates of each kinase and their concentrations used in the assay are shown in Table 2.
Figure imgf000091_0002
Table 2. Kinase peptide substrates and concentrations used in MCE assay The results of the assay are shown in Table 3. As can be seen, the compounds of the invention exhibit low nanomolar potency activity against MERTK. In addition, the compounds of the invention have good selectivity for MERTK over AXL and TYRO3. Additionally, most of the compounds of the invention are not appreciably active against FLT3, which is known to also be expressed in the CNS.
Figure imgf000091_0001
Figure imgf000092_0001
Table 3. TAM and FLT3 kinase inhibition profile of example compounds The predicted CNS MPO and BBB (using HBA = 5 and HBD = 3) scores for the compounds of the invention were calculated and the results are shown in Table 4. The scores range from 3.35-4.87 and 2.49-3.48, suggesting that the compounds of the invention exhibit some CNS-penetrance.
Figure imgf000092_0002
Table 4. Predicted CNS MPO and BBB scores for example compounds Pharmacokinetics and brain exposure of Compound 5-02in male Swiss outbred mice No adverse reactions or compound-related side effects were observed in mice following IV administration of Compound 5-02at 3.7 mg/kg. Plasma and brain concentrations of Compound 5-02declined rapidly up to 2 h post-dose, after which concentrations declined with an apparent half-life of ~ 3 h (Table 5). The apparent blood volume of distribution and blood clearance were both high. The apparent in vivo blood clearance (195 mL/min/kg) is higher than the nominal mouse hepatic blood flow (120 mL/min/kg) suggesting that Compound 5-02may be subject to extrahepatic clearance processes.
Figure imgf000093_0001
Table 5. Pharmacokinetic parameters for Compound 5-02in male Swiss outbred mice following IV administration at 3.7 mg/kg Plasma and brain (total) concentration versus time profiles are presented in Figure 1 and the unbound plasma and brain concentration versus time profiles are shown in Figure 2. Compound 5-02was stable in both assay matrices (brain homogenate and plasma) under the conditions of the RED assay, and fraction unbound values (fu) in mouse plasma and brain were 0.018 (± 0.002) and 0.0072 (± 0.0001), respectively. With regards to brain-to-plasma partitioning with, values for Kp,uu were close to unity at all sample times suggesting that Compound 5-02is not subject to efflux at the blood brain barrier. Furthermore, the absence of notable time-dependence in B:P and Kp,uu, suggests that that distributional equilibrium between brain and plasma was achieved soon after dosing. In vivo PET imaging of MERTK using [18F]5-02 C57Bl/6 mice (8-10 weeks, n=8) were administered 0.2% cuprizone via diet for 4 weeks, age matched control mice (n=7) were feed a standard mouse diet. Mice were anesthetized with 4% isoflurane in oxygen in an induction chamber, then maintained on 2% isoflurane via a nose cone for the duration of imaging. A heating pad were used to keep mice warm while under anesthesia, and animals were temperature and heart rate monitored throughout. PET imaging was performed in the prone position (nanoScan PET/CT, Mediso, Hungary). [18F]5-02 was prepared as described above with a specific activity of >104 GBq/ μmol (2.8 Ci/μmol). Radiochemical purity was greater than 99%. For uptake scans, mice were injected via the tail vein with approximately 12−13 MBq (318−363 μCi, decay corrected) of [18F]5-02 in 70 μl of 10% v/v ethanol in 0.9% w/v saline. The acquisition of uptake scans was begun immediately followed by tracer injection. Initially mice were scanned for 90 min (n=2) with data reframed into 20 x 1 min, 5 x 2 min, 6 x 5 min and 3 x 10 min frames, which was subsequently reduced to 60 min (20 x 1 min, 5 x 2 min and 4 x 5 min frames) and then 30 minutes (20 x 1 min and 5 x 2 min frames) due to the rapid kinetics of the tracer. PET dynamic series were reconstructed with the Terratomo 3D ordered subset estimate maximization (OSEM) algorithm (four iterations and six subsets). Voxel size was 0.6mm3 . CT was used attenuation correction as well as correction for randoms and scatter. Each frame of the dynamic series was corrected for radioactive decay and calibrated in SUV. Analysis and interpretation were performed using PMOD (version 4.1, PMOD Technologies, Zurich, Switzerland). Regions of interest were drawn for the whole brain to derive the time-activity curves for each scan performed. The results of this analysis are shown in Figures 3 and 4. Figure 3 shows the brain uptake summed over 10 minutes (standard uptake value) in both a control (left image) and cuprizone challenged (right image, MS model) mouse. As Figure 3 shows, [18F]5-02 is CNS-penetrant. Figure 4 is a time activity curve (whole brain) in a control (black) and cuprizone-exposed (grey) mouse. Rapid uptake and washout are seen with minimal change in tracer concentration in the brain beyond 15 minutes (900 sec). Radiometric protein kinase assay: Reaction Biology A radiometric protein kinase assay (33PanQinaseTM Activity Assay) was used for measuring the kinase activity of the MERTK. MERTK inhibition assay was performed in 96- well ScintiPlatesTM from PerkinElmer (Boston, MA, USA) in a 50 µl reaction volume. The reaction cocktail was pipetted in four steps in the following order: 25 µl of assay buffer (standard buffer/ γ-33P]-ATP), 10 µl of ATP solution (in H2O), 5 µl of test compound (in 10 % DMSO) and 10 µl of enzyme/substrate mixture. The assay contained 70 mM HEPES-NaOH pH 7.5, 3 mM MgCl2, 3 mM MnCl2, 3 µM Na-orthovanadate, 1.2 mM DTT, 50 μg/ml PEG20000, ATP (corresponding to the apparent ATP-Km of the kinase, see Table 6), [γ- 33P]-ATP (approx.9 x 1005 cpm per well), protein kinase (see Table 6), and substrate (see Table 6).
Figure imgf000094_0001
Table 6. The reaction cocktails were incubated at 30 °C for 60 minutes and then the reaction was stopped with 50 µl of 2 % (v/v) H3PO4, plates were aspirated and washed two times with 200 µl 0.9 % (w/v) NaCl. Incorporation of 33Pi was determined with a microplate scintillation counter (Microbeta, Wallac). All assays were performed with a BeckmanCoulter Biomek FX.
Figure imgf000095_0001
Table 7. MERTK kinase inhibition profile of example compounds Pharmacokinetics and brain exposure of Compound 11-02 in male Swiss outbred mice No adverse reactions or compound-related side effects were observed in mice following IV administration of Compound 11-02 at 4.6 mg/kg. Plasma and brain concentrations of Compound 11-02 were detected for the duration of the 7.5 h sampling period, with an apparent half-life of ~ 2.4 h (Table 8). The apparent blood volume of distribution and blood clearance were both high. The apparent in vivo blood clearance (173 mL/min/kg) is higher than the nominal mouse hepatic blood flow (120 mL/min/kg) suggesting that Compound 11-02 may be subject to extrahepatic clearance processes. a
Figure imgf000095_0002
The terminal elimination phase has been estimated on the basis of the last two time points therefore values based on extrapolation to infinity are approximations only. Table 8. Pharmacokinetic parameters for Compound 11-02 in male Swiss outbred mice following IV administration at 4.6 mg/kg Plasma and brain (total) concentration versus time profiles are presented in Figure 5 and the unbound plasma and brain concentration versus time profiles are shown in Figure 6. Compound 11-02 was stable in both assay matrices (brain homogenate and plasma) under the conditions of the RED assay, and fraction unbound values (fu) in mouse plasma and brain were 0.673 (± 0.035) and 0.118 (± 0.028), respectively. With regards to brain-to-plasma partitioning with, values for brain-to-plasma partitioning ratio and Kp,uu showed a clear time dependence, with the highest values being observed from 4 h post-dose being indicative of a substantial delay in the time to reach distributional steady state between plasma and brain. Values for Kp,uu were close to unity once distributional steady state was achieved (i.e. at 4 and 7.5 h post-dose) which is interpreted as indicating that movement of Compound 11-02 across the BBB is a predominantly passive process. *** Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is understood that the invention includes all such variations and modifications which fall within the spirit and scope of the present invention.

Claims

CLAIMS 1. A compound of Formula (I)
Figure imgf000097_0001
Formula (I) wherein Y is -(-C(O)-NH-)- or -(-C(O)-NH-(CH2))-;; n is 0 to 2; dashed lines indicate an optional methylene bridge; R1 is selected from the group consisting of optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; R2 is independently selected from the group consisting of H, optionally substituted C1- 6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of H, OH, optionally substituted C1- C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; R4 is a cyclic group selected from C6-C18aryl, C2-C18heteroaryl, C3-C12cycloalkyl or C1- C12heterocycloalkyl; and is optionally substituted up to four times with R5 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1- C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3-C12cycloalkenyl, optionally substituted C2- C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2- C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3- C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; wherein each R5 is optionally further substituted with R6 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3- C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1- C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2- C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2- C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; and wherein R9 and R10 are independently selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C1-C12heterocycloalkyl, optionally substituted C1-C12alkylamino, optionally substituted C5-C7lactone, optionally substituted C4-C7lactam, and optionally substituted C1-C12haloalkyl; or wherein R9 and R10 when taken together with the atoms to which they are attached form an optionally substituted C3-C12cycloalkyl or an optionally substituted C1- C12heterocycloalkyl; or a pharmaceutically acceptable salt thereof.
2. The compound of claim 1, having a structure of Formula (Ia):
Figure imgf000099_0001
Formula (Ia) wherein Y is -(-C(O)-NH-)-; n is 0 to 2; dashed lines indicate an optional methylene bridge; R1 is selected from the group consisting of optionally substituted C1-6 alkyl or optionally substituted C3-8 cycloalkyl; R2 is independently selected from the group consisting of H, optionally substituted C1- 6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12haloalkyl, optionally substituted C3-8cycloalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C2-C18heteroaryl; R3 is selected from the group consisting of H, OH, optionally substituted C1- C12alkyloxy, NR9R10, optionally substituted C1-C12alkylamino, and halogen; R4 is a cyclic group selected from C6-C18aryl, C2-C18heteroaryl, C3-C12cycloalkyl or C1- C12heterocycloalkyl; and is optionally substituted up to four times with R5 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1- C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3-C12cycloalkenyl, optionally substituted C2- C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1-C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2-C12alkenyloxy, optionally substituted C2- C12alkynyloxy, optionally substituted C2-C10heteroalkyloxy, optionally substituted C3- C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; wherein each R5 is optionally further substituted with R6 which is independently selected from the group consisting of halogen, OH, NO2, CN, SH, NH2, N3, CF3, OCF3, optionally substituted C1-C12alkyl, optionally substituted C1-C12haloalkyl, optionally substituted C2-C12alkenyl, optionally substituted C2-C12alkynyl, optionally substituted C2-C12heteroalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C3- C12cycloalkenyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C12heterocycloalkenyl, optionally substituted C6-C18aryl, optionally substituted C1- C18heteroaryl, optionally substituted C1-C12alkyloxy, optionally substituted C2- C12alkenyloxy, optionally substituted C2-C12alkynyloxy, optionally substituted C2- C10heteroalkyloxy, optionally substituted C3-C12cycloalkyloxy, optionally substituted C3-C12cycloalkenyloxy, optionally substituted C2-C12heterocycloalkyloxy, optionally substituted C2-C12heterocycloalkenyloxy, optionally substituted C6-C18aryloxy, optionally substituted C1-C18heteroaryloxy, optionally substituted C1-C12alkylamino, optionally substituted C1-C12alkylazide, B(OR9)2, CPO(OR9)2, SR9, SO3H, SO2NR9R10, SO2R9, SONR9R10, SOR9, COR9, COOH, COOR9, CONR9R10, NR9COR10, NR9COOR10, NR9SO2R10, NR9CONR9R10, NR9R10, and acyl; and wherein R9 and R10 are independently selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C1-C12heterocycloalkyl, optionally substituted C1-C12alkylamino, optionally substituted C5-C7lactone, optionally substituted C4-C7lactam, and optionally substituted C1-C12haloalkyl; or wherein R9 and R10 when taken together with the atoms to which they are attached form an optionally substituted C3-C12cycloalkyl or an optionally substituted C1- C12heterocycloalkyl; or a pharmaceutically acceptable salt thereof.
3. The compound of claim 1 or claim 2, having a structure of Formula (II):
Figure imgf000101_0001
Formula (II) wherein dashed lines indicate an optional methylene bridge; and R1, R2, R3, and R4 are as defined in claim 1; or a pharmaceutically acceptable salt thereof.
4. The compound of any one of claims 1 to 3, having a structure of Formula (III):
Figure imgf000101_0002
wherein R1 is selected from the group consisting of optionally substituted C1-6alkyl or optionally substituted C3-8cycloalkyl; X is a hetero atom selected from N, O and S; dashed lines indicate optional double bonds; m is 0 to 2; where valency allows, R7 is selected from the group consisting of H, optionally substituted C1-C12alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1- C12haloalkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2- C12heterocycloalkyl, optionally substituted C6-C18aryl, and optionally substituted C1- C18heteroaryl; R2 and R3 are as defined in claim 1; and each R5 is independently as defined in claim 1; or a pharmaceutically acceptable salt thereof.
5. The compound of any one of claims 1 to 4, having a structure of Formula (IV):
Figure imgf000102_0001
Formula (IV) wherein R1 is selected from the group consisting of optionally substituted C1-6alkyl; and each R5 is independently as defined in claim 1; or a pharmaceutically acceptable salt thereof.
6. The compound of any one of claims 1 to 5, wherein each R5 is independently selected from the group consisting of H, CN, halogen, optionally substituted C1-C12alkylsulfonyl, optionally substituted C1-C6alkyl, optionally substituted C1-C12alkyloxy, optionally substituted C1-C12alkylamino, and optionally substituted C1-C12haloalkyl.
7. The compound of any one of claims 1 to 6, wherein R4 has a structure selected from:
Figure imgf000103_0001
Figure imgf000104_0001
8. The compound of any one of claims 1 to 7, wherein R1 is optionally substituted methyl or optionally substituted ethyl.
9. The compound of any one of claims 1 to 8, wherein R3 is OH or halogen.
10. The compound of any one of claims 1 to 9, wherein R2 is H.
11. The compound of any one of claims 1 to 10, which is radio-labelled.
12. The compound of any one of claims 1 to 10, wherein at least one of R5 is radio- labelled.
13. The compound of claim 11 or claim 12, which is radio-labelled with an 18F group.
14. The compound of any one of claims 1 to 10, wherein the compound has the structure:
Figure imgf000105_0001
Figure imgf000106_0001
Figure imgf000107_0001
Figure imgf000108_0001
Figure imgf000109_0001
or a pharmaceutically acceptable salt thereof.
15. The compound of any one of claims 1 to 14, which is CNS penetrant.
16. An imaging composition comprising a radio-labelled compound according to any one of claims 11 to 13.
17. A method of imaging body tissue comprising: a) applying an imaging composition to a subject, wherein said composition comprises a radio-labelled compound according to any one of claims 11 to 13; b) detecting radiation emitted by said composition and forming an image therefrom.
18. The method of claim 17, wherein said body tissue is selected from the group consisting of the brain and spinal cord.
19. The method of claim 17 or claim 18, wherein said imaging is Positron Emission Tomography imaging.
20. A method of identifying, selecting, or diagnosing a disease state in a subject, comprising the steps of: a) applying an imaging composition to a body tissue of the subject, wherein said composition comprises a radio-labelled compound according to any one of claims 11 to 13; b) detecting radiation emitted by said composition and forming an image therefrom, wherein the image indicates a level of microglial subtype present in the body tissue; c) comparing the level with a reference level to determine an increased or decreased level of microglial subtype present in the body tissue, wherein the increased or decreased level of microglial subtype present in the body tissue compared to the reference level is indicative of a disease state or a stage of development of a disease state.
21. The method of claim 20, wherein the reference level of microglial subtype present in the body tissue is the level of expression of microglial subtype present in the body tissue from a normal subject.
22. The method of claim 20 or claim 21, wherein said microglial subtype present in the body tissue is M2 microglia.
23. The method of claim any one of claims 20 to 22, wherein said imaging is Positron Emission Tomography imaging.
24. The method of claim any one of claims 20 to 23, wherein said disease state is a condition associated with MERTK.
25. The method of any one of claims 20 to 24, wherein said disease state is a neuroinflammatory central-nervous system disease.
26. The method of any one of claims 20 to 25, wherein said disease state is multiple sclerosis (MS).
27. The method of any one of claims 20 to 26, further comprising administering to the subject a therapeutically effective amount of an anti-disease state therapeutic.
28. A pharmaceutical composition comprising a pharmaceutically acceptable salt of the compound according to any one of claims 1 to 14, and a pharmaceutically acceptable excipient.
29. A compound of any one of claims 1 to 14 for use in the identification, treatment, prevention, or amelioration of a condition associated with MERTK.
30. The compound of claim 29, wherein the condition is multiple sclerosis.
31. A method of identifying, preventing, treating, or ameliorating a condition associated with MERTK, said method comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of claims 1 to 14.
32. The method of claim 31, wherein the condition is multiple sclerosis.
33. Use of a compound of any one of claims 1 to 14 in the manufacture of a medicament for the identification, treatment, prevention, or amelioration of a condition associated with MERTK.
34. The use of claim 33, wherein the condition is multiple sclerosis.
PCT/AU2023/050497 2022-06-07 2023-06-07 Compounds with mertk activity WO2023235925A1 (en)

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Citations (3)

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Publication number Priority date Publication date Assignee Title
WO2015157115A1 (en) * 2014-04-11 2015-10-15 The University Of North Carolina At Chapel Hill Mertk-specific pyrimidine compounds
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